Language：English   Publisher：Journal of Medical Ultrasonics  

Purpose: Recently, the estimated total atrial conduction time measured using tissue Doppler imaging (PA-TDI duration) has been reported as a more accurate predictor of atrial fibrillation (AF) recurrence after catheter ablation than left atrial volume index (LAVI). The PA-TDI duration is considered to reflect electrical and structural remodeling in the right atrium (RA) and left atrium (LA). We sought to investigate the association between AF recurrence and PA-TDI duration after AF ablation. Methods: We studied 209 patients who underwent radiofrequency ablation for paroxysmal AF and 75 patients who underwent second ablation for AF recurrence. We assessed the duration from the onset of the P wave on the surface electrocardiogram to the atrial electrogram in distal coronary sinus (CS) (PA-CSd duration) indicating electrical remodeling of the atrium, the PA-CS proximal duration (PA-CSp duration) representing electrical remodeling of RA, and the conduction time in CS (proximal to distal) (CSp-CSd duration) reflecting electrical remodeling of LA. We also measured LAVI as a marker of structural remodeling of LA. Results: The PA-TDI duration had a positive correlation with PA-CSd duration. In the patients with AF recurrence, PA-TDI duration, PA-CSd duration, and CSp-CSd duration in the second ablation were significantly longer than those in the first (p < 0.01, respectively), whereas there was no significant difference in LAVI and PA-CSp duration between the first and second ablation sessions. Conclusion: A prolonged PA-TDI duration after AF ablation may indicate advanced electrical remodeling of LA, and may predict AF recurrence after ablation in patients with paroxysmal AF.

DOI： 10.1007/s10396-021-01090-6

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PubMed

Publisher：(株)南山堂  

DOI： 10.15104/j00821.2021134621

Language：English   Publisher：Heart and Vessels  

Platelet functions are thought to contribute to clinical outcomes after heart surgery. This study was conducted to assess the pivotal roles of vascular endothelial growth factor-A (VEGF-A) and microRNA-126 (miR-126) during coronary artery bypass grafting (CABG). Whole blood was collected for platelet isolation from 67 patients who underwent CABG surgery between July 2013 and March 2014. VEGF-A and miR-126 levels in serum, plasma, and platelets were measured at various time points and compared with clinical characteristics. The platelet count was decreased at 3 days after CABG. This dynamic change in platelet count was larger after conventional coronary artery bypass (CCAB) than off-pump coronary artery bypass (OPCAB). VEGF-A in the same number of platelets (IP-VEGF-A) was increased at 3 days after CABG, followed by an increase of VEGF-A in serum (S-VEGF-A) at 7 days after surgery. The miR-126-3p level in serum (S-miR-126-3p) increased rapidly after CABG and then decreased below preoperative levels. The IP-VEGF-A level on day 7 after CABG in patients with peripheral artery disease (PAD), who suffered from endothelial dysfunction, was higher compared with patients without PAD. Conversely, S-miR-126-3p on day 7 after surgery was lower in patients with PAD than in patients without PAD. Low levels of S-miR-126-3p due to endothelial dysfunction may lead to high IP-VEGF-A, which is closely related to complications after CABG.

DOI： 10.1007/s00380-021-01855-6

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PubMed

Language：English   Publisher：Clinical Hemorheology and Microcirculation  

BACKGROUND AND OBJECTIVE: Turbulent blood flow in patients with aortic valve stenosis (AS) results in morphological and functional changes in platelets and coagulation factors. The aim of this study is to determine how shear stress affects platelets and coagulation factors. METHODS: We retrospectively evaluated data from 78 patients who underwent AVR to treat AS between March 2008 and July 2017 at Kagoshima University Hospital. RESULTS: Platelet (PLT) count obviously decreased at three days after AVR, and increased above preoperative levels at the time of discharge. In contrast, platelet distribution width (PDW), mean platelet volume (MPV), and platelet large cell ratio (P-LCR) increased three days after AVR, then decreased to below preoperative levels. No differences were evident between groups with higher (HPPG > 100 mmHg) and lower (LPPG < 100 mmHg) peak pressure gradients (PPG) before AVR, whereas PLT count, PDW, MPV and P-LCR improved more in the HPPG group. Plateletcrit (PCT), which represents the total volume of platelets, increased after AVR due to decreased shear stress. High increasing rate of PCT was associated with lower PLT count, higher PDW and lower fibrinogen. CONCLUSION: Shear stress affects PLT count, PDW, and fibrinogen in patients with AS.

DOI： 10.3233/CH-200928

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PubMed

Language：Japanese   Publisher：一般社団法人 日本血栓止血学会  

DOI： 10.2491/jjsth.32.792

Language：English   Publisher：Journal of Infection and Chemotherapy  

We report an AIDS patient with a high HIV RNA copy number in the plasma who was successfully treated for prolonged Mycobacterium avium bacteremia and other complications. An HIV-infected patient with high fever, anemia, high alkaline phosphatase, cystic lung lesions, hepatitis B virus infection and Kaposi's sarcoma was referred to our hospital. PCR of the blood revealed Mycobacterium avium bacteremia and the time to blood culture positivity was 8 days. The HIV-1 RNA copy number in the plasma was more than ten million copies/ml and the CD4-positive T cell count was 21 cells/μL. Although the high fever resolved five days after therapy for Mycobacterium avium was started, the fever recurred just before starting anti-retroviral therapy (ART) including dolutegravir. The patient experienced repeated but self-limiting bouts of severe inflammation. Mycobacteremia was intermittently detected up to 79 days, suggesting that the recurrent episodes of inflammation were due to the intermittent dissemination of mycobacteria, and that persistent treatment is needed. Five months after the beginning of ART, the HIV-1 RNA copy number in the plasma was still 28,000 copies/ml. An HIV drug-resistance test revealed sensitivity to all anti-retroviral drugs. Eleven months after the initiation of ART, the HIV RNA copy number in the plasma decreased to 45 copies/mL and the CD4-positive T cell count recovered to 205 cells/μL. Our case also suggests that dolutegravir can be effective in cases with prolonged high levels of HIV RNA. Our findings emphasize that prompt diagnosis and persistent therapy for mycobacterial infection are important for successful treatment.

DOI： 10.1016/j.jiac.2019.08.012

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PubMed

Language：English   Publisher：Clinical and Applied Thrombosis/Hemostasis  

Atrial fibrillation (AF) is the most common cardiac arrhythmia in the world and has a high risk of thromboembolism. The most effective approach, catheter ablation, requires evaluation by electrocardiography. The aim of our study was to investigate novel clinical markers that predict restoration of sinus rhythm (SR) after catheter ablation. Seventy-eight consecutive patients with AF underwent catheter ablation and were separated into 2 groups: restored SR and recurrent AF. The levels of 4 blood proteins (serum or plasma) and 3 mature microRNAs (miRNAs) and their primary miRNAs (pri-miRNAs) in serum were measured before and after ablation, and the associations between each parameter were analyzed statistically. Soluble thrombomodulin (s-TM) and plasminogen activator inhibitor-1 (PAI-1) levels increased above baseline after ablation in both the restored SR (s-TM 11.55 [2.92] vs 13.75 [3.38], P <.001; PAI-1 25.74 [15.25] vs 37.79 [19.56], P <.001) and recurrent AF (s-TM 10.28 [2.78] vs 11.67 [3.37], P <.001; PAI-1 26.16 [15.70] vs 40.74 [22.55], P <.001) groups. Levels of C-reactive protein and asymmetric dimethylarginine were not significantly changed. Pri-miR-126 levels significantly decreased after ablation in the recurrent AF group, but the other miRNAs and pri-miRNAs did not. The measurement of s-TM and pri-miR-126 in blood was a useful tool to reflect the condition of AF patients with catheter ablation.

DOI： 10.1177/1076029619851570

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PubMed

Publisher：株式会社医学書院  

DOI： 10.11477/mf.1542201927

Language：English   Publisher：Biochemical and Biophysical Research Communications  

Background: Inhalation of aerosolized Legionella pneumophila, a Gram-negative bacterium, can cause severe pneumonia. During infection, L. pneumophila replicates intracellularly in macrophages. The involvement of host microRNAs (miRNAs) in L. pneumophila infection is not fully understood. Methods: The human macrophage-like cell line U937 was infected with L. pneumophila. The levels of miRNA and messenger RNA (mRNA) were measured using reverse transcriptase polymerase chain reaction. Release of lactate dehydrogenase was used to evaluate cytotoxicity. The expression of RICTOR and related proteins was examined by western blotting of cell lysates. Results: L. pneumophila infection upregulated the expression of miR-218 and the host genes SLIT2 and SLIT3 in U937 cells. The expression of RICTOR, a component of the mechanistic target of rapamycin complex 2 (mTORC2), decreased during L. pneumophila infection. RICTOR protein expression was inhibited by the overexpression of miR-218, whereas knockdown of miR-218 restored the downregulation of RICTOR by L. pneumophila. L. pneumophila infection induced the expression of the proinflammatory cytokines IL-6 and TNF-alpha, which was modulated by knockdown of miR-218 or RICTOR. Conclusions: Our study revealed the involvement of miR-218 in regulating the inflammatory response of macrophages against L. pneumophila infection. These findings suggest potential novel roles for miR-218 and RICTOR as therapeutic targets of L. pneumophila infection.

DOI： 10.1016/j.bbrc.2018.11.093

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PubMed

Language：English   Publisher：Cancer Biomarkers  

BACKGROUND AND AIMS: A striking difference has been observed in structure and functional properties between plasma and platelet von Willebrand factor (VWF). While the existing evidence has revealed a clinical relevance of plasma VWF-Ag in liver regeneration (LR) and different cancers, this study was designed to explore the properties of intra-platelet (IP) and serum VWF-Ag in patients with hepatocellular carcinoma (HCC) undergoing partial hepatectomy. METHODS: A total of 40 patients undergoing partial hepatectomy were prospectively recruited from 3 institutions. VWF-Ag concentrations were evaluated mainly in serum and platelet extracts. Patients were followed-up for postoperative liver dysfunction and HCC recurrence. RESULTS: We observed a post-resection increase in the concentration of VWF-Ag in serum and platelet. Patients with postoperative liver dysfunction had substantially reduced serum and IP VWF-Ag concentrations. After a 2-year follow-up, patients with higher post-resection serum and IP VWF-Ag concentrations were found to develop early HCC recurrence. Likewise, IP VWF-Ag was able to independently predict post-resection early HCC recurrence. CONCLUSION: This multicenter, prospective, pilot study demonstrates a bivalent property of IP VWF in LR and oncological outcome; low preoperative VWF appeared to have a negative association on post-resection liver dysfunction, whereas, patients with higher post-resection VWF-Ag concentrations were found to have early HCC recurrence.

DOI： 10.3233/CBM-190168

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PubMed

Language：English   Publisher：Journal of Oncology  

Purpose. Platelet derived growth factor-BB (PDGF-BB) has emerged as one of the key cytokines in malignant transformation of different cells. PDGF-BB also exhibits a potent mitogenic effect on liver cells; studies have advocated clinical implications of monitoring serum PDGF-BB (sPDGF-BB) in patients with liver disease. We thus investigated the predictive relevance of perioperative sPDGF-BB after curative resection of hepatocellular carcinoma (HCC). Methods. We evaluated perioperative sPDGF-BB in a prospective homogenous cohort of 40 patients diagnosed with HCC. During the first two-year follow-up, patients were evaluated every three months for postresection HCC recurrence. Results. Patients who developed recurrence during two-year follow-up were found to have lower concentration of sPDGF-BB than those without recurrence in both pre- and postoperative settings (P < 0.05 and P < 0.001, resp.). We validated that the reduced postoperative sPDGF-BB (< 2133.29 pg/mL) was associated with an increased incidence of postresection HCC recurrence [area under curve (AUC) > 0.8, 95% confidence interval (CI) = 0.68 - 0.94, P < 0.001]; furthermore, we were able to demonstrate that postoperative sPDGF-BB was an independent predictor of HCC recurrence (hazard ratio = 5.64, 95% CI = 1.56 - 20.30, P < 0.01). Conclusions. These findings provide a new insight into an association between diminished perioperative sPDGF-BB and HCC recurrence. Patients with low perioperative sPDGF-BB progressed early HCC recurrence. Therefore, evaluating perioperative sPDGF-BB may provide useful clinical information to characterize patients with postresection HCC recurrence.

DOI： 10.1155/2019/1925315

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PubMed

Language：English   Publisher：Journal of Cancer  

DOI： 10.7150/jca.30696

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PubMed

Language：English   Publisher：Expert Review of Gastroenterology and Hepatology  

DOI： 10.1080/17474124.2018.1533813

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PubMed

Language：English   Publisher：Journal of Clinical Medicine  

Endothelial cells (ECs) form monolayers and line the interior surfaces of blood vessels in the entire body. In most mammalian systems, the capacity of endothelial cells to divide is limited and endothelial cells are prone to be senescent. Aging of ECs and resultant endothelial dysfunction lead to a variety of vascular diseases such as atherosclerosis, diabetes mellites, hypertension, and ischemic injury. However, the mechanism by which ECs get old and become senescent and the impact of endothelial senescence on the vascular function are not fully understood. Recent research has unveiled the crucial roles of miRNAs, which are small non-coding RNAs, in regulating endothelial cellular functions, including nitric oxide production, vascular inflammation, and anti-thromboformation. In this review, how senescent-related miRNAs are involved in controlling the functions of ECs will be discussed.

DOI： 10.3390/jcm7070170

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PubMed

Language：English   Publisher：International Heart Journal Association  

<p>Endothelial dysfunction is observed in several cardiovascular diseases, where endothelium-dependent vasodilation is impaired by oxidative stress. However, the time course of endothelial function during the perioperative period of a minor-to-moderate surgery, and the effects of atherosclerotic risk factors and employed general anesthetics on recovery of endothelial function, are unknown. Endothelial function of 30 patients was evaluated as the reactive hyperemia index (RHI) of reactive hyperemia peripheral arterial tonometry. RHI was measured on day before surgery (control), immediately after surgery (Day 0), day after surgery (Day 1), and day 4 after surgery (Day 4) in patients with no functional limitations who were scheduled for oral and maxillofacial surgery of around 3 hours. Sevoflurane- or propofol-based anesthesia supplemented with an opioid analgesic remifentanil was employed. The control RHI was 2.26 ± 0.64. The RHI significantly decreased to the lowest level on Day 0 (1.52 ± 0.28), recovered on Day 1 (2.07 ± 0.58), and improved further on Day 4 (2.55 ± 0.83). Multiple linear regression analysis revealed that recovery of the RHI from Day 0 to Day 4 was impaired by diabetes mellitus (<i>P</i> = 0.0313), obesity (BMI ≥ 25; <i>P</i> = 0.0166), hyperuricemia (uric acid ≥ 6.0 mg/dL; <i>P</i> = 0.0416) and sevoflurane-based anesthesia (<i>P</i> = 0.0308). These findings suggest that endothelial function as evaluated by the RHI is severely suppressed on the day of a minor-to-moderate surgery, and that it improves until the 4th postoperative day on average. Recovery of endothelial function is impaired by diabetes mellitus, obesity, hyperuricemia, and sevoflurane-based anesthesia.</p>

DOI： 10.1536/ihj.17-143

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PubMed

Language：English   Publisher：(一社)インターナショナルハートジャーナル刊行会  

小～中等度手術の周術期における血管内皮機能の時間経過と、アテローム動脈硬化症の危険因子および全身麻酔が血管内皮機能の回復に及ぼす影響を調べた。3時間程度の口腔および顎顔面手術を行い、レミフェンタニルを追加したセボフルランまたはプロポフォールベースの麻酔を行った18歳以上の患者を対象とした。血管内皮機能は反応性充血末梢動脈圧力測定の反応性充血指標(RHI)によって手術前日、手術直後、1、4日後に評価した。30例(男性13例、女性17例、平均48.3±18.4歳)のデータ解析を行った。平均RHIは手術前日が2.26±0.64で、手術直後に1.52±0.28と最も低く、術後1日では2.07±0.58と回復し、4日目には更に改善して2.55±0.83となった。重回帰分析により、RHIの術後の改善は糖尿病、肥満、高尿酸血症、セボフルランベースの麻酔によって障害されることが示された。

Language：English   Publisher：Experimental Dermatology  

Psoriasis, a chronic inflammatory skin disease, is closely related to systemic metabolism. An elevated body mass index (BMI) is a risk factor for psoriasis; inflammasomes are activated by adipose tissue macrophages in obese subjects. We hypothesized that hyperlipidaemia is involved in the pathogenesis of psoriasis and examined the role of a high-fat diet (HFD) in the development of psoriasis in imiquimod (IMQ)-treated mice. The body weight and serum level of cholesterol were significantly higher in mice fed an HFD than in a regular diet (RD). HFD mice had higher psoriasis skin scores, and the number of neutrophils infiltrating into the lesional skin was elevated. IL-17A mRNA expression was significantly increased in the skin of IMQ-treated HFD mice; the expression of IL-22, IL-23 and TNF-α mRNA was not enhanced. Caspase-1 and IL-1β were activated in the skin of IMQ-treated HFD mice, and their serum level of IL-17A, TNF-α and IL-1β was significantly upregulated. Our findings strongly suggest that hyperlipidaemia is involved in the development and progression of psoriasis via systemic inflammation and inflammasome activation.

DOI： 10.1111/exd.13484

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PubMed

Language：English   Publisher：Cellular Signalling  

Pulmonary arterial hypertension (PAH) has a major effect on life expectancy with functional degeneracy of the lungs and right heart. Interleukin-13 (IL-13), one of the type 2 cytokines mainly associated with allergic diseases, has recently been reported to be associated with Schistosomiasis-associated PAH which shares pathological features with other forms of PAH, such as idiopathic PAH and connective tissue disease-associated PAH. But a direct pathological role of IL-13 in the development of PAH has not been explored. We examined the effects of recombinant human IL-13 on the function of primary human pulmonary artery endothelial cells (HPAECs) to examine how IL-13 influences exacerbation of PAH. IL-13 increased the expression of Rictor, which is a key molecule of mammalian target of rapamycin complex 2. Treatment of IL-13 induced HPAEC migration via Rictor. Rictor was directly regulated by both miR-424 and 503 (miR-424/503). Therefore, IL-13 increases Rictor level by regulating miR-424/503, causing the increase of HPAEC migration. Since enhancement of HPAEC migration in the lung is thought to be associated with PAH, these data suggest that IL-13 takes some roles in exacerbating PAH.

DOI： 10.1016/j.cellsig.2017.10.019

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PubMed

Language：English   Publisher：Canadian Journal of Gastroenterology and Hepatology  

Liver pathophysiology can, directly and indirectly, impose morphological or biochemical abnormalities of the platelets. Conversely, platelets are also able to regulate the promitogenic and profibrotic signals on liver pathobiology. Platelet contribution to the liver pathophysiology is typically facilitated by the platelet-derived growth factors that are sequestered in different subsets of alpha and dense granules, and the release of these growth factors is synchronized according to the stage and type of liver disease or injury. Thus, platelets harbor clinically relevant information with potential diagnostic and prognostic implications in liver diseases. Hepatocellular carcinoma (HCC) largely influences the platelet kinetics, and a growing body of evidence has recognized its association with HCC occurrence or prognosis. This narrative review summarizes the progress made on implicating platelet as a diagnostic and prognostic tool for HCC; the review also dissects the contradictory results from earlier studies and reflects how combining platelet-based information may enable more reliable test for diagnostic and prognostic evaluation of HCC.

DOI： 10.1155/2018/9142672

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PubMed

Language：Japanese   Publisher：一般社団法人 日本血栓止血学会  

DOI： 10.2491/jjsth.29.545

Language：English   Publisher：Biochemical and Biophysical Research Communications  

In the course of studying crosstalk between inflammation and angiogenesis, high doses of pro-inflammatory factors have been reported to induce apoptosis in cells. Under normal circumstances also the pro-inflammatory cytokines are being released in low doses and are actively involved in cell signaling pathways. We studied the effects of low grade inflammation in growth factor induced angiogenesis using tumor necrosis factor alfa (TNFα) and vascular endothelial growth factor A (VEGF) respectively. We found that low dose of TNFα can inhibit VEGF induced angiogenesis in human umbilical vein endothelial cells (HUVECs). Low dose of TNFα induces mild upregulation and moreover nuclear localization of tumor suppressor protein 53 (P53) which causes decrease in inhibitor of DNA binding-1 (Id1) expression and shuttling to the cytoplasm. In absence of Id1, HUVECs fail to upregulate β3-integrin and cell migration is decreased. Connecting low dose of TNFα induced p53 to β3-integrin through Id1, we present additional link in cross talk between inflammation and angiogenesis.

DOI： 10.1016/j.bbrc.2016.12.096

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PubMed

Language：English   Publisher：Retina  

Purpose: To determine the effects of bevacizumab, ranibizumab, and aflibercept on the permeability and the effects of anti-vascular endothelial growth factor (VEGF) on highly polarized retinal pigment epithelial cells (RPECs) in vitro. Methods: Highly polarized RPECs were cultured in the upper chamber of a Transwell system. Anti-VEGF antibodies were added to the upper chamber, and the concentrations of the drugs in the lower chambers were measured. The permeability rates of the three anti- VEGF drugs through the RPEC layer and the concentration of VEGF in each chamber were determined. Results: The permeability of aflibercept was significantly lower by about 40% than that of bevacizumab through the RPEC layer (P < 0.05). Ranibizumab was significantly more permeable through the RPECs than bevacizumab (180% of bevacizumab, P < 0.05). Although VEGF was almost absent in the upper chamber after exposure to the 3 antibodies, it was decreased more significantly with aflibercept than with bevacizumab in the lower chamber (2.8% vs. 65.8% of control; P < 0.01). Ranibizumab also decreased the VEGF level compared with bevacizumab (31.7% vs. 65.8% of control; P < 0.01). Conclusion: The greater reduction of the amount of VEGF in the lower chamber by aflibercept and ranibizumab than bevacizumab may explain why aflibercept and ranibizumab are more effective than bevacizumab against type 1 choroidal neovascularization.

DOI： 10.1097/IAE.0000000000001117

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PubMed

Language：English   Publisher：Journal of Cancer  

Purpose: Serotonin (5-hydroxytryptamine, 5-HT) is well known for its growth stimulatory effect on several types of carcinoma and tumor cells. Since a large portion of 5-HT is stored and transported by platelets, the aim of this study was to assess the influence of platelet-sequestered 5-HT on post-resection hepatocellular carcinoma (HCC) recurrence. Methods: This pilot study was conducted in a cohort of forty patients diagnosed with HCC undergoing partial hepatectomy. 5-HT levels in serum, plasma and intra-platelet (IP) were monitored preoperatively and four weeks after liver resection. The patients were followed every three months after the surgery. Results: Follow-up was standardized to a fixed length of time. Fifteen patients (37.5%) developed HCC recurrence during 18 months follow-up. Patients with recurrence had significantly reduced serum and IP 5-HT levels at four weeks of liver resection (P = 0.003 and P = 0.014 respectively). Accordingly, in the Cox regression hazard model, serum and IP 5-HT were able to independently predict the recurrence (hazard ratio = 0.1, 95% confidence interval = 0.01 - 0.75 and hazard ratio = 0.1, 95% confidence interval = 0.01 - 0.89 respectively). The optimal cut-off value of 42.77 ng/ml for serum [area under the curve (AUC): 0.78, P = 0.003] and 0.3117 ng per 106 platelets (AUC: 0.733, P = 0.015), on receiver operating characteristic (ROC) curve corresponded to maximum sensitivity and specificity of prediction. The disease free interval was significantly worse in patients with low serum and IP 5-HT (P = 0.001 and P = 0.029 respectively). Conclusion: IP 5-HT monitored during early follow-up, after liver resection may represent a useful marker of early HCC recurrence.

DOI： 10.7150/jca.20971

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PubMed

Language：Japanese   Publisher：Japanese Society of Sonographers  

<p><b>Purpose</b>: Recently, it has been reported that an index of the total atrial conduction time derived by tissue Doppler imaging (PA-TDI duration) has a superior accuracy for prediction of atrial fibrillation (AF) recurrence compared with the left atrial volume index (LAVI) after pulmonary vein isolation (PVI). We performed catheter ablation targeting complex fractionated atrial electrograms (CFAE ablation) on patients with AF and explored the independent predictor of AF recurrence after CFAE ablation.</p><p><b>Subjects and methods</b>: Our study included 160 patients with paroxysmal AF who underwent AF ablation (CFAE ablation or CFAE ablation＋PVI) for the first time. Of these, 107 patients in whom a measurement of PA-TDI duration was possible were divided into no recurrence and recurrence groups. Univariable and multivariable Cox proportional hazards analyses were performed to investigate predictors of AF recurrence after AF ablation. The receiver operator characteristics (ROC) curve was calculated to evaluate the performance of possible independent predictors of AF recurrence after AF ablation was obtained by multivariate analysis. The time-dependent changes of PA-TDI duration and LAVI were observed to investigate the predictors of AF recurrence after AF ablation.</p><p><b>Results</b>: The left atrial dimension (LAD) was significantly larger in the recurrence group than in the no recurrence group (41.1±5.4 mm to 38.1±5.1 mm, p＜0.01). In the multivariate analysis, the left atrial volume (LAV) and LAD were suggested as independent predictors of AF recurrence after AF ablation. However, the ROC curve analyses demonstrated that LAV and LAD had low accuracy of predicting AF recurrence after AF ablation (area under the curve: LAV, 0.609; LAD, 0.659). There were no significant differences in PA-TDI duration or LAVI in both strategies before ablation between the recurrence and the no recurrence groups. Six months after the ablation, the PA-TDI duration in the recurrence group was significantly longer than before ablation with both strategies (CFAE ablation: 142.9±18.5 ms to 155.4±17.6 ms, p＜0.01; CFAE ablation＋PVI: 135.6±20.3 ms to 155.1±21.8 ms, p＜0.01), and PA-TDI duration in the no recurrence group with CFAE ablation was significantly shorter than before ablation (141.8±23.3 ms to 131.1±23.9 ms, p＝0.011). However, there was no significant difference in LAVI between the recurrence and no recurrence groups for both strategies.</p><p><b>Conclusion</b>: The data obtained before ablation could not predict AF recurrence after CFAE ablation for patients with paroxysmal AF. However, longer PA-TDI duration at the 6-month followup may predict AF recurrence after CFAE ablation for paroxysmal AF.</p>

DOI： 10.11272/jss.42.399

Language：Japanese   Publisher：The Japanese Society of Hematology  

<p>"Fibrin is insoluble in blood" is the most important concept to accept the coagulation system. Essentially, blood coagulation is a process to convert blood from soluble to insoluble. Fibrinolysis is another process to convert insoluble protein to soluble protein again. The blood coagulation cascade requires calcium ion and efficiently acts on platelet phospholipid membranes. On the other hand, the fibrinolysis system efficiently acts on fibrin. Unlike other vitamin K-dependent coagulation factors, thrombin can be released from the coagulation site to the systemic circulation and has multiple biological effects regulated through thrombin receptors. Thrombin is captured by thrombomodulin on endothelial cells or anti-thrombin. Low level of coagulation and fibrinolysis idling are always working even under healthy conditions because of shear stress on injured vessels. Measurement of coagulation times, such as PT and APTT, is the basic method used by numerous clinics in coagulation-related examinations. Measurement of coagulation factor activity by one method is based on the concept that excess amounts of coagulation factors circulate in the blood. Under DIC conditions, excess pathological protease activity yields a huge amount of peptides in the blood in addition to coagulation and fibrinolysis markers.</p>

DOI： 10.11406/rinketsu.58.2096

PubMed

Language：English   Publisher：(公財)福田記念医療技術振興財団  

カテーテルアブレーションを施行した心房細動(AF)患者を対象に、術前と術後6ヵ月後の心臓足首血管指数(CAVI)を比較し、CAVIが内皮障害のマーカーとなりうるか検討した。カテーテルアブレーションを施行したAF患者101名(男81名、女20名、平均年齢61.8±8.6歳)を対象とした。6ヵ月後に来院しなかった23名は除いた。78名のうち、洞調律(SR)に回復した患者は33名、AFが再発した患者は45名であった。ベースライン時のCAVIについて、SR回復群とAF再発群との間に有意差はなかった。6ヵ月後のCAVIは両群ともベースライン時と差がなかった。CAVIは年齢と強い相関があり、左室駆出分画と弱い相関がみられた。内皮細胞のバイオマーカーである血清トロンボモデュリン濃度は、SR回復群の方がベースライン時は高値で、両群とも6ヵ月後は上昇していた。ベースライン時の血漿プラスミノーゲンアクチベーターインヒビター1濃度は両群間で差はなく、両群とも6ヵ月後に上昇していた。primary-miR-126とmature miR-126はAF再発群で6ヵ月後に低下し、内皮機能の新しいバイオマーカーとなり得ると考えられた。

Language：English   Publisher：National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee  

Pulsed-field gel electrophoresis (PFGE) is a reliable method for analyzing outbreaks of methicillin-resistant <i>Staphylococcus aureus</i> (MRSA); however, it is time-consuming and technically demanding. A new strain-differentiation method for MRSA, namely phage open reading frame (ORF) typing (POT), is a rapid PCR-based technique, in which the ORFs of lysogenized phage genomes in MRSA are amplified. The aim of this study was to evaluate the utility of the POT method for epidemiological analysis of nosocomial MRSA outbreaks. Forty-four strains from 12 episodes of 3 or more nosocomial MRSA infections in 1 ward within a 4-week period were characterized using PFGE and POT methods. The strains were classified into 16 distinct types using POT and 19 subtypes using PFGE. We defined an outbreak as 3 or more new MRSA infections caused by strains with indistinguishable genetic patterns. The identification of 11 (91.7%) episodes by PFGE, including 4 outbreaks and 7 sporadic events, was consistent with the results of POT analysis. These results suggest that POT is a useful epidemiological tool for evaluating nosocomial MRSA outbreaks.

DOI： 10.7883/yoken.JJID.2015.320

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PubMed

Language：English   Publisher：国立感染症研究所  

三次医療施設におけるMRSA院内アウトブレイクを疫学的に解析するためのphage open-reading frame typing(POT)の有用性を評価した。4週間以内の1病棟のMRSA新規感染症例が3例以上の場合をアウトブレイクと定義し、12エピソード由来の44株について、POT型とPFGEパターンを比較した。44株は16POT型に分類された。最頻度のPOT型は207-117の20株で、PFGEのA型(n=12)、E型(n=6)、I型(n=1)、K1型(n=1)から成っていた。2番目に頻度が高かったのは207-113型の4株で、PFGEのA(n=1)、E1(n=1)、E2(n=1)、I(n=1)から成っていた。4エピソードはPFGEとPOTによりアウトブレイクと特定された。7エピソードはPFGEとPOTのどちらでもアウトブレイクと特定されなかった。PFGEによる、アウトブレイク4と散発例7を含む11エピソード特定率(91.7%)は、POT解析による成績と一致した。POTはMRSA院内アウトブレイクの評価に有用であることが示唆された。

Language：English   Publisher：Obesity  

Objective: Obesity is a chronic inflammatory disease, and adipocytes contribute to obesity-associated inflammation by releasing inflammatory mediators. High mobility group box 1 (HMGB1), a highly conserved DNA-binding protein, mainly localized to cell nuclei, has been recently recognized as an innate pro-inflammatory mediator when released extracellularly. It was hypothesized that HMGB1 is an adipocytokine that acts as an innate pro-inflammatory mediator in white adipose tissue (WAT) of patients with obesity and is associated with insulin resistance. Additionally, it was hypothesized that HMGB1 secretion is regulated by adiponectin. Methods: 3T3-L1 cells were differentiated into mature adipocytes. After tumor necrosis factor-α (TNF-α) stimulation, HMGB1 in culture media was measured. Localizations of HMGB1 in 3T3-L1 adipocytes and human WAT were examined by immunostaining. Results: HMGB1 was secreted from TNF-α-induced 3T3-L1 adipocytes through JNK signaling. HMGB1-activated MAP kinases (ERK1/2, JNK) and suppressed insulin-stimulated Akt phosphorylation in 3T3-L1 adipocytes. The cytoplasm in 3T3-L1 adipocytes and adipocytes of WAT from a patient with obesity was intensely stained with HMGB1. Adiponectin partially inhibited TNF-α-induced HMGB1 secretion from 3T3-L1 adipocytes. Conclusions: These findings suggest that HMGB1 is a pro-inflammatory adipocytokine involved in WAT inflammation and insulin resistance in patients with obesity, which may contribute to the progression of metabolic syndrome, and that adiponectin protects against HMGB1-induced adipose tissue inflammation.

DOI： 10.1002/oby.21549

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PubMed

Language：Japanese   Publisher：(一社)日本臨床検査医学会  

甲状腺刺激ホルモン(TSH)のハーモナイゼーションについて検討した。アーキテクトTSH、エクルーシスTSH、ルミパルスTSH-III、Eテスト「TOSOH」II(TSH)について検討した。NIBSC81/565調製液の濃度に対して、ルミパルスの測定値が最も低く平均94.3%、エクルーシスの測定値も平均97.1%と低値であった。アーキテクトは平均106.1%と高値、Eテスト「TOSOH」は平均118.7%と最も高値であった。臨床検査精度管理調査用試料では、ルミパルス、アーキテクト、Eテスト「TOSOH」の3試薬で比較し、ルミパルスが低値、Eテスト「TOSOH」が高値となった。エクルーシスは4試薬の中で最も高値を示した。各試薬での測定値とAPTMとの差は換算することにより減少した。また、試薬による測定値の変動も換算することにより減少した。

Language：Japanese   Publisher：Japanese Journal of Clinical Chemistry  

Scopus

Language：English   Publisher：PLoS ONE  

Background Liver regeneration (LR) involves an early inductive phase characterized by the proliferation of hepatocytes, and a delayed angiogenic phase distinguished by the expansion of nonparenchymal compartment. The interest in understanding the mechanism of LR has lately shifted from the proliferation and growth of parenchymal cells to vascular remodeling during LR. Angiogenesis accompanied by LR exerts a pivotal role to accomplish the process. Vascular endothelial growth factor (VEGF) has been elucidated as the most dynamic regulator of angiogenesis. From this perspective, platelet derived/Intra-platelet (IP) VEGF-A should be associated with LR. Material and Methods Thirty-seven patients diagnosed with hepatocellular carcinoma and undergoing partial hepatectomy (PH) were enrolled in the study. Serum and IP VEGF-A was monitored preoperatively and at four weeks of PH. Liver volumetry was determined on computer models derived from computed tomography (CT) scan. Results Serum and IP VEGF-A was significantly elevated at four weeks of PH. Preoperative IP VEGF-A was higher in patients with advanced cancer and vascular invasion. Postoperative IP VEGF-A was higher after major liver resection. There was a statistically significant correlation between postoperative IP VEGF-A and the future remnant liver volume. Moreover, the soluble vascular endothelial growth factor receptor-1 (sVEGFR1) was distinctly downregulated suggesting a fine-Tuned angiogenesis at the later phase of LR. Conclusion IP VEGF-A is overexpressed during later phase of LR suggesting its implications in inducing angiogenesis during LR.

DOI： 10.1371/journal.pone.0150446

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PubMed

Language：Japanese   Publisher：The Japanese Society on Thrombosis and Hemostasis  

DOI： 10.2491/jjsth.27.631

Language：Japanese   Publisher：(一社)日本サイトメトリー学会  

症例1は15歳男子で、38℃台の発熱を認め、日中は解熱する熱型で推移した。数日前から眼瞼周囲に紫斑を認め、救急外来で末梢血に異常を認め、救急搬送された。10万を超える白血球の異常増加、リンパ芽球様異常細胞増加、貧血、血小板減少を認めた。細胞形態は好中球と同等の大きさで細胞質は好塩基性に富み、N/C比が大きく、核網微細で大きな核小体を認め一部にブレブを認めた。LDHの高度上昇を認め、Dダイマーが高値であった。CD10陰性、CD15陽性、CD34一部陽性の結果から混合血統白血病(MLL)関連急性リンパ性白血病(ALL)を疑い神経膠抗原2(NG2)抗体を用い追加検査を行った結果、NG2強陽性となり、MLL関連ALL疑いと報告できた。症例2は5歳男児で、鼻出血を繰り返し、左足の紫斑に気付き、2日後から39℃台の発熱を認め、紫斑が増加した。急性白血病が疑われ緊急入院した。リンパ芽球様異常細胞増加、貧血、血小板減少を認めた。骨髄検査でNCC 55.9万/mm3、MgK 12.5/mm3、Blasts 97.0%、LDH高度上昇を認めた。骨髄血のFISH検査ではMLLスプリットシグナルは99.0%であった。NG2抗体を用いたMRD追跡を行った。MLLスプリットシグナル陽性率とNG2の陽性率は同様の変化をしていた。

Language：Japanese   Publisher：Haemophilia  

Introduction: Autoimmune haemophilia-like disease (or haemorrha-philia) due to anti-factor XIII (FXIII; F13 to avoid confusion with FVIII or FXII) antibodies (termed AH13) is a severe bleeding disorder. Although AH13 is thought to be rare, 'the number of its diagnosed patients' has recently increased in Japan. However, its prevalence remains unknown. Aim: To improve understanding of this disease, we examined and diagnosed 32 'new' Japanese patients with AH13. Methods: The presence of antibodies against F13-A subunit and/or F13-B subunit was confirmed by using a dot blot test and enzyme-linked immunosorbent assays. Results: Most of our patients had autoantibodies against the F13-A subunit (88%). A predominance of men (59%) was observed. The mean age and residual F13 activity of our AH13 cohort were 71.7 years and 10.5% of normal, respectively, and 53% of cases were idiopathic. Autoimmune disorders and malignancies were the leading underlying disease (both 16%). Intramuscular and subcutaneous bleeding were the leading symptoms (both 72%). Most of our patients were treated with F13 concentrates (72%) to arrest bleeding and with prednisolone (81%) to eradicate anti-F13 autoantibodies. Cyclophosphamide and rituximab (both 25%) were also administered. The mortality of AH13 was high (22%), and haemorrhage was the major cause of death (71%). Moreover, 13% of our AH13 patients were diagnosed after haemorrhagic death. Conclusion: Physicians/haematologists must raise the awareness of AH13 as a life-threatening disease. This report represents the only experience of a nationwide survey, and may contribute to a diagnosis on potentially overlooked non-Japanese AH13 patients in other countries in the world.

DOI： 10.1111/hae.12677

Scopus

Language：Japanese   Publisher：International Journal of Food Sciences and Nutrition  

To reveal the effect of coffee bean polyphenols (CBPs) on blood vessels, this study aimed to investigate the effect of CBPs on acute postprandial endothelial dysfunction. Thirteen healthy non-diabetic men (mean age, 44.9±1.4 years) consumed a test beverage (active: containing CBPs, placebo: no CBPs) before a 554-kcal test meal containing 14g of protein, 30g of fat and 58g of carbohydrates. Then, a crossover analysis was performed to investigate the time-dependent changes in flow-mediated dilation (FMD) in the brachial artery. In the active group, the postprandial impairment of FMD was significantly improved, the two-hour postprandial nitric oxide metabolite levels were significantly increased and the six-hour postprandial urinary 8-epi-prostaglandin F2α levels were significantly reduced compared to the placebo group. The test meal increased the levels of blood glucose, insulin and triglycerides in both groups with no significant intergroup differences. These findings indicate that CBPs intake ameliorates postprandial endothelial dysfunction in healthy men.

DOI： 10.3109/09637486.2015.1007453

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PubMed

Language：Japanese   Publisher：Biopreservation and Biobanking  

Background: A major concern in both the laboratory-medicine and research communities is the quality of human specimens for analysis. However, there is insufficient scientific evidence regarding optimal conditions for handling and storing routine specimens, especially those in liquid form. Thus, we investigated the stability of clinically relevant samples stored under various conditions. Materials and Methods: Ten clinical laboratories in Japan conducted analyses of the stability of post-clinical (left over after analysis) test samples in relation to temperature and storage duration. We examined serum, whole blood, and urine samples submitted to each laboratory for routine testing. In this study, at least 5 samples for each of 35 tests were analyzed at each laboratory. After completion of routine testing, specimens with sufficient residual volume and values between LL-R/2 (lower limit of reference interval) and UL+R/2 (upper limit) were divided into 300 μL aliquots, where R=UL - LL. Aliquots of serum specimens were stored at either room temperature (23°C), 4°C, -20°C, or -80°C without light exposure. Aliquots of whole blood and urine specimens were stored at either 23°C or 4°C. The storage time was either 1, 3, or 7 days. Average differences between pre- and post-storage test results were evaluated for each laboratory test by two-way ANOVA. F-values for between-day variations were used for judging the statistical significance of storage-related changes in test values, whereas the ratio of between-day SD to between-individual SD (one-fourth of reference interval) was used to indicate the practical significance of the change. Results and Conclusion: Sample denaturation is clearly temperature- and storage-duration dependent for almost all analytes. In general, specimens were most susceptible to denaturation at 23°C, then 4°C, -20°C, and -80°C. This study confirmed the accumulated routine, practice-based, detailed knowledge regarding specimen stability and will help to ensure the reliability of laboratory test results.

DOI： 10.1089/bio.2014.0072

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PubMed

Language：English   Publisher：Journal of Arrhythmia  

Background Complex fractionated atrial electrogram (CFAE)-targeted catheter ablation (CFAE ablation) requires a high rate of atrial fibrillation (AF) termination to provide good outcomes. We determined the optimal settings of CFAE software. Methods In our 430 consecutive patients, AF was terminated in 97 (234/242) and 79% (149/188) of patients with paroxysmal and persistent AF, respectively, by CFAE ablation combined with (31%) or without (69%) pulmonary vein isolation, occasionally with nifekalant infusion. We analyzed 109 consecutive patients who underwent CFAE ablation to determine the optimal settings for comparing subjective versus objective decisions by the CFAE software on CARTO3. We compared three settings: the default setting (0.05-0.15 mV, 50-120 ms) and two modified settings (#1: 0.05-0.30 mV, 40-70 ms, #2: 0.05-0.13 mV, 10-20 ms). We retrospectively analyzed 11,425 points during left atrial mapping before ablation and 10,306 points that were subjectively detected and ablated as CFAE points. An interval confidence level ≥6 denoted a site with CFAE. Results With the default setting, the accuracy, sensitivity, specificity, positive productive value, and negative productive values were 67, 42, 77, 48, and 73%, respectively. With modified setting #1, the values were 78, 55, 87, 74, and 77%, respectively, versus 64, 82, 60, 53, and 91%, respectively, for modified setting #2. Conclusion These data suggest that setting #1 was generally superior to the default setting, whereas setting #2 was optimal for excluding areas not requiring ablation. The optimal CFAE software setting was a voltage of 0.05-0.30 mV and an interval parameter of 40-70 ms.

DOI： 10.1016/j.joa.2014.04.006

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PubMed

Language：English   Publisher：(一社)日本不整脈心電学会  

Complex fractionated atrial electrogram(CFAE)アブレーションを施行した持続性心房細動患者109例(男性87例、女性22例、平均59±10歳)のデータを解析し、CFAEソフトウェアにより主観的および客観的決定を比較するための最適設定について検討した。デフォルト設定(0.05～0.15mV、50～120ms)と2つの修正設定(#1:0.05～0.30mV、40～70ms、#2:0.05～0.13mV、10～20ms)を比較した。アブレーション前の左房マッピング時の11425ポイント、またCFAEポイントとして主観的に検出され焼灼された10300ポイントを後方視的に分析した。デフォルト設定の精度、感度、特異度、陽性的中率、陰性的中率はそれぞれ67%、42%、77%、48%、73%であった。修正設定#1の場合はそれぞれ78%、55%、87%、74%、77%、修正設定#2の場合はそれぞれ64%、82%、60%、53%、91%であった。結論として、修正設定#1は全般的にデフォルト設定より優れており、設定#2はアブレーションを必要としない領域を除外するために最適であった。最適なCFAEソフトウェア設定は0.05～0.30mVの電圧および40～70msの間隔パラメータであった。

Language：Japanese   Publisher：Japan Cytometry Society  

<p>We have recognized the appearance of NG2 antigen in MLL rearrangement-positive ALL. However, it is not clear why NG2 antigen appears in MLL rearrangement-positive ALL. The appearance of NG2 is not distinctive, but it is believed to be useful in MRD detection of MLL rearrangement-positive ALL. Using NG2, we are able to confirm the suspected connection with MLL-related ALL (case 1). And we tried to track the course of MRD in the ALL case (case 2). B cellprecursor acute lymphoblastic leukemia (BCPALL) was initially suspected in case 1. However, positive results for CD79a, CD19 and negative results for CD66c led us to use chondroitin sulfate proteoglycan-type neural-glial antigen 2 antibody (NG2 antibody), with a focus on negative CD10, positive CD15 and partially positive CD34 findings. Positive findings for NG2 confirmed the case 1 as mixed lineage leukemia (MLL)-related ALL. In addition, we confirmed the activity of an idioblast rate by visual observation and the positive rate of the MLL split signal. Tracking of the NG2 positive rate in the course of ALL was effective for the detection of MRD in the case 2.</p>

DOI： 10.18947/cytometryresearch.25.2_21

Language：English   Publisher：PeerJ  

Objective. Allantoin is the primary active compound in yams (Dioscorea spp.). Recently, allantoin has been demonstrated to activate imidazoline 3 (I3) receptors located in pancreatic tissues. Thus, the present study aimed to investigate the role of allantoin in the effect to improve damage induced in pancreatic β-cells by streptozotocin (STZ) via the I3 receptors. Research Design andMethods. The effect of allantoin on STZ-induced apoptosis in pancreatic β-cells was examined using the ApoTox-Glo triplex assay, live/dead cell double staining assay, flow cytometric analysis, and Western blottings. The potential mechanism was investigated using KU14R: an I3 receptor antagonist, and U73122: a phospholipase C (PLC) inhibitor. The effects of allantoin on serum glucose and insulin secretion were measured in STZ-treated rats. Results. Allantoin attenuated apoptosis and cytotoxicity and increased the viability of STZ-induced β-cells in a dose-dependent manner; this effect was suppressed by KU14R and U73112. Allantoin decreased the level of caspase-3 and increased the level of phosphorylated B-cell lymphoma 2 (Bcl-2) expression detected by Western blotting. The improvement in β-cells viability was confirmed using flow cytometry analysis. Daily injection of allantoin for 8 days in STZ-treated rats significantly lowered plasma glucose and increased plasma insulin levels. This action was inhibited by treatment with KU14R. Conclusion. Allantoin ameliorates the damage of β-cells induced by STZ. The blockade by pharmacological inhibitors indicated that allantoin can activate the I3 receptors through a PLC-related pathway to decrease this damage. Therefore, allantoin and related analogs may be effective in the therapy for β-cell damage.

DOI： 10.7717/peerj.1105

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PubMed

Publisher：株式会社医学書院  

DOI： 10.11477/mf.1542200127

Language：Japanese   Publisher：Anticancer Research  

Background: Cluster of differentiation 147 (CD147)/basigin on the malignant tumor cell surface is critical for tumor proliferation, invasiveness, metastasis, and angiogenesis. CD147 expressed on malignant melanoma cells can induce tumor cell invasion by stimulating the production of matrix metalloproteinases (MMPs) by surrounding fibroblasts. Membrane vesicles, microvesicles and exosomes have attracted attention, as vehicles of functional molecules and their association with CD147 has been reported. Cleaved CD147 fragments released from tumor cells were reported to interact with fibroblasts. We investigated the intercellular mechanisms by which CD147 stimulates fibroblasts to induce MMP2 activity. Materials and Methods: CD147 was knockeddown using short hairpin RNA (shRNA). The stimulatory effect of CD147 in cell culture supernatants, microvesicles, and exosomes on the enzymatic activity of MMP2 was examined by gelatin zymography. Results: Supernatants from A375 control cells induced increased enzymatic activity of fibroblasts; such activity was significantly lower in CD147 knock-down cells. Conclusion: Cleaved CD147 plays a pivotal role in stimulating fibroblasts to induce MMP2 activity.

Scopus

Language：English   Publisher：Journal of Cataract and Refractive Surgery  

PURPOSE: To determine the effect of histones on corneal endothelial cells generated during cataract surgery. SETTING: Kagoshima University Hospital, Kagoshima, Japan. DESIGN: Experimental study. METHODS: Standard phacoemulsification was performed on enucleated pig eyes. Histones in the anterior segment of the eye were determined by immunohistochemistry. Cultured human corneal endothelial cells were exposed to histones for 18 hours, and cell viability was determined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitro-phenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt assay. The concentration of interleukin-6 (IL-6) in the culture medium of human corneal endothelial cells was measured using enzyme-linked immunosorbent assay. The effects of signal inhibitors U0126, SB203580, and SP600125 were evaluated. The protective effect of hyaluronan against histones was evaluated in human corneal endothelial cells with and without hyaluronan. RESULTS: Cellular debris containing histones was observed in the anterior chamber of pig eyes after phacoemulsification. Exposure of human corneal endothelial cells to 50 mg/mL of histones or more led to cytotoxic effects. The IL-6 concentration was significantly increased dose dependently after exposure of human corneal endothelial cells to histones (P<.01). The histoneinduced IL-6 production was significantly decreased by extracellular signal-regulated kinases 1/2 and p-38 mitogen-activated protein kinase inhibitors (P<.01). Co-incubation of hyaluronan and histones caused formation of histone aggregates, decreased the cytotoxic effects of the histones, and blocked the increase in IL-6 (P<.01). CONCLUSIONS: Histones were released extracellularly during phacoemulsification and exposure of human corneal endothelial cells to histones increased the IL-6 secretion. The intraoperative use of hyaluronan may decrease the cytotoxic effects of histones released during cataract surgery. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned.

DOI： 10.1016/j.jcrs.2014.07.026

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PubMed

Language：English   Publisher：Journal of Infection and Chemotherapy  

We report a patient with Japanese minor β thalassemia and HIV-1 infection. The patient showed prolonged anemia, which was originally attributed to chronic parvovirus B19 infection. Twelve years later, the patient presented with exacerbation of microcytic anemia following cessation of anti-retroviral therapy; the exacerbation resolved when anti-retroviral therapy was resumed. Sequencing of the β globin gene revealed heterozygosity for a four-nucleotides deletion at codon 41/42 and minor β thalassemia was confirmed. Because HIV-1-infected patients frequently show anemia due to nutritional deficiencies, opportunistic infections, AIDS-related malignancies, drug treatment and a direct effect of HIV-1 on the bone marrow, it is likely to overlook other causes of anemia. Thalassemia should be considered in the differential diagnosis of anemia even in HIV-1 infected patients, when microcytic anemia without iron deficiency is observed. Our case suggested that active HIV infection may have worsened β thalassemia, and early introduction of anti-retroviral therapy is beneficial for the recovery of anemia. © 2014, Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.jiac.2014.01.008

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PubMed

Language：English   Publisher：エルゼビア・ジャパン(株)  

症例は40代前半の男性(2000年3月時点)で、2ヵ月前から重度貧血が続いており、ニューモシスチス肺炎のため入院した。血漿中HIV-1 RNAコピー数とCD4陽性T細胞数からAIDSと診断された。骨髄液のPCRによりパルボウイルスB19感染による重度貧血が疑われた。HIV RNAが減少したため2000年5月に退院した。抗レトロウイルス治療を中断していたが、2012年3月に疲労感のため来院した。4日後に抗レトロウイルス療法を再開し、24日後に増悪は改善した。1年後にヘモグロビンが上昇し、血漿中HIV RNAコピー数の完全阻害が見られた。血漿中鉄濃度と不飽和鉄結合能から患者の小球性貧血は鉄欠乏によるものではないことが示唆された。βグロビン鎖遺伝子のDNA配列決定により、1アリルにおいてコドン41/42で4ヌクレオチドの欠損がある異型接合性が明らかになり、軽症βサラセミアと診断された。

Language：English  

DOI： 10.1038/labinvest.2014.46

PubMed

Language：English   Publisher：(一社)日本動脈硬化学会  

168名の患者(18～89歳)について、手術前日に座位で光電式指尖容積脈波の2次微分波(SDPTG)と心拍数変動(HRV)を測定した。SDPTG指数(b/a、c/a、d/a、e/a)とHRV指数(周波数分析、時間帯分析)を求めた。c/aとアテローム性硬化に基づく状態や危険因子との関連性についても評価した。SDPTG指数c/aは低周波成分(LF)と高周波成分(HF)と正の関連を示した。重回帰分析を行ったところ、LF、HF、心拍数はc/aの独立決定因子であった。c/a値は高血圧、糖尿病、高脂血症の患者で有意に低く、c/aの低値は血清トリグリセリドと総コレステロール濃度と有意に関連した。c/aの低下は末梢血管運動の圧反射反応の低下、心臓の副交感神経活性の低下と関連し、さらに高血圧、糖尿病、高脂血症のようなアテローム性硬化の基礎となる状態と関連すると考えられた。

Language：English  

DOI： 10.1055/s-0034-1371811

PubMed

Language：English   Publisher：Nutrition Research  

Brewed coffee is a widely consumed beverage, and many studies have examined its effects on human health. We investigated the vascular effects of coffee polyphenols (CPPs), hypothesizing that a single ingestion of CPP during glucose loading would improve endothelial function. To test this hypothesis, we conducted a randomized acute clinical intervention study with crossover design and measured reactive hyperemia index (RHI) to assess the acute effects of a 75-g glucose load with CPP in healthy, nondiabetic adult men. Blood glucose and insulin levels were elevated after glucose loading with and without CPP, with no significant differences between treatments. The RHI did not significantly decrease after glucose loading without CPP. With CPP, however, RHI significantly (P < .05) increased over baseline after glucose loading. The difference between treatments was statistically significant (P < .05). No significant changes were observed in an oxidative stress marker after glucose loading with or without CPP. These findings suggest that a single ingestion of CPP improves peripheral endothelial function after glucose loading in healthy subjects. © 2014 Elsevier Inc.

DOI： 10.1016/j.nutres.2013.11.001

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PubMed

Language：English   Publisher：Japan Atherosclerosis Society  

<b><i>Aim</i></b>: The indices of the second derivative of the finger photoplethysmogram(SDPTG) denote stiffness of large arteries, peripheral vascular resistance and vascular aging. However, the association between the autonomic nervous activity and the SDPTG indices has not yet been elucidated.<br>
<b><i>Methods</i></b>: The SDPTG and heart rate variability(HRV) were consecutively measured in the sitting position on the day before surgery in 168 patients 18-89 years of age. The relationships between the SDPTG indices(b/a, c/a, d/a and e/a) and HRV indices(power spectral analysis and time domain analysis parameters) were analyzed. The relationships between c/a and atherosclerosis-based conditions and risk factors for atherosclerosis were also evaluated.<br>
<b><i>Results</i></b>: The SDPTG index b/a was negatively associated and the d/a index was positively associated with the low-frequency(LF)(<i>R</i>=<b>−</b>0.44 and 0.42, respectively) and high-frequency(HF) components(<i>R</i>=<b>−</b>0.31 and 0.35, respectively). The SDPTG index c/a was also positively associated with the LF(<i>R</i>=0.40) and HF(<i>R</i>=0.44) components. A multivariate regression analysis showed that the LF, HF and heart rate were independent determinants of the c/a. Furthermore, the c/a values were significantly lower in the patients with hypertension, diabetes mellitus and hyperlipidemia than in those without these diseases, and a reduced c/a was significantly associated with increased serum triglyceride and total cholesterol concentrations.<br>
<b><i>Conclusions</i></b>: These findings suggest that a decrease in c/a is associated with a reduced baroreflex response of the peripheral vasomotor activity and a decreased cardiac parasympathetic activity. Furthermore, a decrease in c/a was found to be associated with atherosclerosis-based conditions, such as hypertension, diabetes mellitus and hyperlipidemia.

DOI： 10.5551/jat.19877

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PubMed

Language：Japanese   Publisher：The Japanese Society of Internal Medicine  

DOI： 10.2169/naika.102.3174

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PubMed

Language：English  

DOI： 10.1016/j.bbrc.2013.06.117

PubMed

Language：English   Publisher：PLoS ONE  

Introduction:Recent studies have shown that histones, the chief protein component of chromatin, are released into the extracellular space during sepsis, trauma, and ischemia-reperfusion injury, and act as major mediators of the death of an organism. This study was designed to elucidate the cellular and molecular basis of histone-induced lethality and to assess the protective effects of recombinant thrombomodulin (rTM). rTM has been approved for the treatment of disseminated intravascular coagulation (DIC) in Japan, and is currently undergoing a phase III clinical trial in the United States.Methods:Histone H3 levels in plasma of healthy volunteers and patients with sepsis and DIC were measured using enzyme-linked immunosorbent assay. Male C57BL/6 mice were injected intravenously with purified histones, and pathological examinations were performed. The protective effects of rTM against histone toxicity were analyzed both in vitro and in mice.Results:Histone H3 was not detectable in plasma of healthy volunteers, but significant levels were observed in patients with sepsis and DIC. These levels were higher in non-survivors than in survivors. Extracellular histones triggered platelet aggregation, leading to thrombotic occlusion of pulmonary capillaries and subsequent right-sided heart failure in mice. These mice displayed symptoms of DIC, including thrombocytopenia, prolonged prothrombin time, decreased fibrinogen, fibrin deposition in capillaries, and bleeding. Platelet depletion protected mice from histone-induced death in the first 30 minutes, suggesting that vessel occlusion by platelet-rich thrombi might be responsible for death during the early phase. Furthermore, rTM bound to extracellular histones, suppressed histone-induced platelet aggregation, thrombotic occlusion of pulmonary capillaries, and dilatation of the right ventricle, and rescued mice from lethal thromboembolism.Conclusions:Extracellular histones cause massive thromboembolism associated with consumptive coagulopathy, which is diagnostically indistinguishable from DIC. rTM binds to histones and neutralizes the prothrombotic action of histones. This may contribute to the effectiveness of rTM against DIC. © 2013 Nakahara et al.

DOI： 10.1371/journal.pone.0075961

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PubMed

Language：Japanese   Publisher：Atherosclerosis  

To examine the presence of CD34-positive cells and intranuclear factors in acute coronary thrombi, we compared thrombi in patients with type 2 diabetes mellitus (DM, n = 21) and without DM (n = 29). Immunohistochemical staining revealed the constitutive presence of platelets, fibrin, erythrocytes, neutrophils, extracellular high-mobility group box 1 (HMGB-1), and histone H3 in all thrombi. There were significantly more oval or flat CD34-positive cells and significantly larger HMGB-1-positive areas in the thrombi from patients with DM. The flat CD34-positive cells expressed ecto-nucleoside triphosphate diphosphohydrolase (a platelet aggregation inhibitor). The number of CD34-positive cells was negatively correlated with the serum levels of glucose and hemoglobin A1c, whereas the HMGB-1-positive area was positively correlated with the levels of serum glucose. The paucity of CD34-positive cells and the high levels of HMGB-1 expression in acute coronary thrombi from patients with type 2 DM could facilitate thrombus formation. © 2012 Elsevier Ireland Ltd.

DOI： 10.1016/j.atherosclerosis.2012.07.027

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Language：English  

Language：English   Publisher：Journal of Cardiology  

Aims: Esophageal-left atrial (LA) fistula during atrial fibrillation (AF) ablation is a fatal event. We explored the relation of the esophagus-to-ablated point distance and esophageal temperature rise. Methods: Consecutive patients (n=106) underwent complex fractionated atrial electrogram-guided AF ablation using CartoMerge; the pulmonary veins were isolated in 23 patients. Maximum radiofrequency (RF) power near the esophagus was 15. W. Ablated points with esophageal temperature rise (monitored with a probe) to ≥38.0. °C were tagged; if ≥39.0 °C, RF was discontinued. Results: Of 1647 ablated points near the esophagus, 274 were associated with a temperature rise to 38.0-38.9 °C and 241 points to ≥39.0 °C. Distances (mm) from points to esophagus were 5.1. ± 0.6 (no rise), 4.2 ± 3.1 (38.0-38.9 °C), 2.9 ± 2.5 (≥39.0 °C). Altogether, 15.5% of points in the upper LA posterior wall, 41.5% in the middle, and 30.2% in the lower caused rises to ≥38.0 °C; 8.7%, 24.6%, and 11.0% caused rises to ≥39.0 °C. The middle wall was most affected (p<0.01), as shown by multiple logistic regression analysis (both temperatures). Points causing a rise increased significantly as distance decreased (p<0.001). The odds ratio for rise to ≥38.0. °C compared with <4.0 to >5.0 mm distance was 2.28 (p=0.004). The longest distance for ≥38.0 °C rise was 18.5 mm. Conclusion: Distance is an important predictor of esophageal temperature rise. The middle LA posterior wall is most vulnerable. A dose of 15. W is too high for ablation, especially <4.0. mm from the esophagus. Points >20.0. mm away are relatively safe. © 2012 Japanese College of Cardiology.

DOI： 10.1016/j.jjcc.2012.02.009

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PubMed

Language：English   Publisher：(一社)日本心臓病学会  

心房細動(AF)アブレーション中の合併症である食道-左房瘻は致死的事故である。食道と心房アブレーション部位との間の距離と食道温度上昇との関係を検討した。CartoMergeを使用し複合分画化心房電位図ガイド下で106例の患者のAFアブレーションを行った。23例では肺動脈隔離を施行した。食道近位でアブレーションした1647ヶ所の中で274ヶ所では上昇した。食道とアブレーション部位との距離が5.1±0.6mmのときは食道温度は上昇しなかった。距離が食道温度上昇の予測因子で、左房後壁中部アブレーションが最も危険であった。

Publisher：株式会社医学書院  

DOI： 10.11477/mf.1402105993

Language：English   Publisher：Phytotherapy Research  

The Japanese apricot, a commonly consumed food called 'Ume' in Japan, has been used for a traditional Japanese medicine for centuries. MK615, an extract of compounds from 'Ume', has strong antitumorigenic and antiinflammatory effects including the induction of apoptosis and autophagy, and inhibition of cytokine production mediated via the inhibition of MAPKs signaling including ERK-1/2, JNK and p38MAPK. The inhibitor of DNA binding 1 (Id-1), a basic helix-loop-helix (bHLH) transcription factor family, is essential for DNA binding and the transcriptional regulation of various proteins that play important roles in the development, progression and invasion of tumors. In melanoma, Id-1 is constitutively expressed in the late and early stages, suggesting it as a therapeutic target in patients with melanoma. This study reports that MK615 profoundly reduced both the mRNA- and protein expression levels of Id-1 and inhibited cell growth in A375 melanoma cells. MK615 markedly inhibited the phosphorylation of ERK1/2, which is associated with Id-1 protein expression in A375 cells. Id-1-specific RNAi induced the death of A375 cells. Moreover, the expression of Bcl-2 was decreased by both MK615 and Id-1-specific RNAi in A375 cells. The results suggest that MK615 is a potential therapeutic agent for treating malignant melanoma. Copyright © 2011 John Wiley & Sons, Ltd.

DOI： 10.1002/ptr.3645

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PubMed

Language：English  

Language：English   Publisher：(一社)日本血液学会  

健常成人の血小板からの血管内皮増殖因子(VEGF)放出量をin vitroで測定し、血清中のVEGF濃度、血小板数および/またはヘマトクリット値で補正したVEGF濃度との相関を調べた。血小板数とヘマトクリット値の両者で補正した値(補正VEGF)がVEGF放出量と最も強く相関した。次に、再生不良性貧血(AA)の小児患者11名の補正VEGFを測定した。AA患児の補正VEGFは年齢が一致する対照よりも有意に高かった。更に、2年以上治療を要しなかったAA患児の補正VEGFは早期に治療を要したAA患児よりも高かった。以上のデータは、VEGFを増やし、AAの疾患進行を遅らせるための代償機構が働いていることを示唆する。補正VEGFはAAの臨床予後を予測するために有用である。

Language：English  

DOI： 10.1007/s12185-012-1074-1

PubMed

Language：Japanese   Publisher：The Japan Society of Neurotraumatology  

DOI： 10.32187/neurotraumatology.34.2_225

Language：English   Publisher：Graefe's Archive for Clinical and Experimental Ophthalmology  

Purpose/background Although vascular endothelial growth factor (VEGF) is abundant in serum, the intraocular concentration of VEGF in eyes with massive vitreous hemorrhage (VH) is not well-known. The present study was conducted to elucidate the effects of a massive VH on intravitreous VEGF concentration. Methods Vitreous samples were obtained during vitrectomy: 12 samples from eyes with epiretinal membrane without diabetic retinopathy (DR), and nine samples from massive VH with no DR, such as age-related macular degeneration, rhegmatogenous VH, Terson's syndrome and macroaneurysm rupture. Twelve samples were obtained from proliferative DR. VEGF was measured with an enzymelinked immunosorbent assay (ELISA). Samples incubated with or without heparin were also examined for the release of VEGF binding to the vitreous body. The localization of VEGF and type II collagen in the vitreous was evaluated from immunohistochemistry. Results The concentration of VEGF was significantly higher in eyes with proliferative DR (821 ± 949 pg/ml) than in non-DR with massive VH (2.75 ± 7.5 pg/ml, P<0.01, chi-square test) or non-DR with no VH (less than detectable level, P<0.01, chi-square test) There was no statistically significant difference between eyes with massive VH and non-diabetic eyes without VH. Treatment with heparin did not significantly affect the concentration of vitreous VEGF. VEGF was localized mainly in the clot from the results of an immunohistochemical analysis. Conclusions Even with a massive VH, diffusible VEGF does not increase significantly in the liquid phase and is principally present in a clot. VH alone should not be an indication for vitrectomy from the point of view of VEGFrelated pathology. © Springer-Verlag 2011.

DOI： 10.1007/s00417-011-1795-5

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PubMed

Language：English   Publisher：Oncology Reports  

The fucosylated fraction of α-fetoprotein (AFP-L3) is a specific marker for hepatocellular carcinoma (HCC). However, conventional AFP-L3% (c-AFP-L3%) has not always been reliable in cases with low serum α-fetoprotein (AFP) levels. In this study, we evaluated the clinical utility of a newly developed assay, highly sensitive AFP-L3% (hs-AFP-L3%). Subjects included 74 patients with benign liver disease (BLD), including chronic hepatitis and cirrhosis, and 94 with HCC. Serum hs-AFP-L3% was significantly higher than c-AFP-L3% in patients with early-stage HCC (solitary or <20 mm in diameter). Additionally, hs-AFP-L3% was significantly increased in patients with well-differentiated HCC. In patients with serum AFP <20 ng/ml, the sensitivities of c-AFP-L3% and hs-AFP-L3% were 12.5 and 44.6%, respectively, at a cut-off value of 5%. In 59 BLD patients with serum AFP <20 ng/ml, the HCC-positive rate in patients with hs-AFP-L3% =5% was significantly higher compared to those with hs-AFP-L3% <5% during the follow-up period (median, 35 months; range, 5-48 months). Importantly, none of the BLD patients with both serum AFP <20 ng/ml and hs-AFP-L3% <5% developed HCC. These results indicated that hs-AFP-L3% is useful for early detection of HCC in BLD patients, even for those with serum AFP <20 ng/ml. Furthermore, since hs-AFP-L3% increases before HCC is detectable by various advanced imaging modalities, this assay may help identify BLD patients with a higher risk of HCC.

DOI： 10.3892/or.2011.1425

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PubMed

Language：English   Publisher：FEMS Immunology and Medical Microbiology  

Biofilms play a pivotal role in medical device-related infections. However, epidemiological analysis of biofilm formation and genotyping among clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients with orthopaedic infections has rarely been reported. A total of 168 MRSA strains were examined: 23 strains from patients with device-related infection (the device group); 55 from patients with device-non-related infection (the nondevice group); and 90 from asymptomatic nasal carriers (the colonization group). Pulsed-field gel electrophoresis analysis and five genotyping methods including agr typing were performed. Biofilm formation was quantified using a microtitre plate assay. The device group had a significantly higher incidence of agr-2 than the colonization group (78.3% vs. 34.4%, P=0.001). The biofilm index of the agr-2 (0.523 ± 0.572) strains was significantly higher than those of agr-1 (0.260 ± 0.418, P<0.0001) and agr-3 (0.379 ± 0.557, P=0.045). The prevalence of strong biofilm formers in the device group (43.5%) was significantly higher than that in the nondevice group (12.7%, P=0.003) and the colonization group (20.0%, P=0.020). agr-2 MRSA strains may be more likely to cause orthopaedic device infection because of their strong biofilm formation ability. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd.

DOI： 10.1111/j.1574-695X.2011.00821.x

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PubMed

Language：English   Publisher：Laboratory Investigation  

Intraocular neovascularization is the leading cause of severe visual loss and anti-vascular endothelial growth factor (VEGF) therapy is currently performed for choroidal neovascularization (CNV). Despite its potent anti-angiogenic effect, there are concerns about its long-term safety. Non-steroidal anti-inflammatory drugs (NSAIDs) are common therapeutic agents used for treating inflammatory diseases, and their anti-stress effects are attracting attention now. We studied the effects of topical NSAIDs on CNV, focusing on anti-stress proteins. Cultured retinal pigment epithelium (RPE) cells were treated with NSAIDs: bromfenac, indomethacin, or vehicle control. Transcription factor NF-E2-related factor 2 (Nrf2) and its downstream anti-oxidant protein heme oxygenase (HO)-1 were assessed using western blot and immunohistochemistry. As a result, NSAIDs induced translocation of Nrf2 into the nucleus and the robust expression of HO-1 in a dose-and time-dependent manner. Flow cytometric analysis revealed that bromfenac inhibited H 2 O 2-induced apoptosis in cultured RPE cells. Next, we studied the effects of topical bromfenac on laser-induced CNV model in rat. The expressions of Nrf2 and HO-1, infiltrations of ED-1-positive macrophages at CNV lesions and size were analyzed. VEGF in the ocular fluid of these rats was also measured using enzyme-linked immunosorbent assay. Rats administered an inhibitor of HO-1 stannic mesoporphyrin (SnMP) were also studied. The results showed that topical bromfenac led to translocation of Nrf2 and induction of HO-1 in CNV lesions and that the number of infiltrating macrophages at the CNV lesion decreased. The sizes of CNV lesions were significantly smaller in bromfenac-treated rats than control CNV, and the effects were diminished by SnMP. VEGF increased in the ocular fluid after laser treatment and was inhibited by bromfenac and SnMP canceling these effects. NSAIDs inhibit CNV through the novel anti-stress protein HO-1-dependent pathway, indicating its potential therapeutic value for various intraocular angiogenic diseases including CNV. © 2011 USCAP, Inc All rights reserved.

DOI： 10.1038/labinvest.2011.101

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PubMed

Language：Japanese   Publisher：Experimental and Therapeutic Medicine  

Edaravone was originally developed as a potent free radical scavenger and has been widely used to treat cerebral infarction in Japan since 2001. Several free radical scavengers have been developed and some of them have progressed to clinical trials for the treatment of cerebral infarction. One such scavenger, edaravone, has been approved by the regulatory authority in Japan for the treatment of patients with cerebral infarction. Of particular interest is the ability of edaravone to diffuse into the central nervous system in various neurologic diseases. Aside from its hydroxyl radical scavenging effect, edaravone has been found to have beneficial effects on inflammation, matrix metalloproteinases, nitric oxide production and apoptotic cell death. Concordantly, edaravone has been found to have neuroprotective effects in a number of animal models of disease, including stroke, spinal cord injury, traumatic brain injury, neurodegenerative diseases and brain tumors. The proven safety of edaravone following 9 years of use as a free radical scavenger suggests that it may have potential for development into an effective treatment of multiple neurologic conditions in humans.

DOI： 10.3892/etm.2011.281

Scopus

Language：Japanese   Publisher：Proteomics  

We have recently developed a new target plate (BLOTCHIP®) for MALDI-MS. An advantage of this procedure is that it does not require the lowering of protein concentrations in test samples prior to analysis. Accordingly, this new technology enables the detection of peptides present in blood samples, including those that would otherwise be adsorbed to abundant blood proteins and would thus escape detection. Using this technology, we analyzed the peripheral blood of patients with pregnancy-induced hypertension (PIH; the most common serious complication of pregnancy) to test a potential utility of the technology for monitoring of the pathophysiological status. In the present study, we found 23 characteristic peptides for PIH in the blood serum of pregnant women. Offline LC-MALDI MS/MS identified 7 of the 23 peptides as fragments derived from kininogen-1 (three peptides), fibrinogen-α, complement component C4-A/B, α-2-HS-glycoprotein and inter-α-trypsin inhibitor heavy chain H4. 2-D scatter plots with combinations of the peptides found in the present study can be grouped for pregnant women with/without PIH, which would be satisfactory reflected for their status. Additionally, the levels of most of these peptides found were significantly decreased by albumin/IgG depletion prior to BLOTCHIP® analysis in accordance with conventional proteomics procedures. These results indicated that BLOTCHIP® analysis can be applied for discovery study of PIH biomarker candidates. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/pmic.201000753

Scopus

Language：Japanese   Publisher：Experimental and Therapeutic Medicine  

Nosocomial infections caused by microbial opportunistic infections or microbial biofilms may occur during hospitalization and increase patient morbidity, mortality and health care costs. Artificial antibiotic agents were initially used to prevent infection; however, the high prevalence of nosocomial infections has resulted in their excessive use, which has led to microbial resistance to these agents. The increase in microbial resistance to antibiotics and the development of antibiotic agents may be the cause of the production of other microbial resistance. Thus, natural compounds that have no adverse side effects would be a preferred treatment modality. Recently, the monosaccharide 1,5-anhydro-D-fructose (1,5-AF), a natural plant compound derived from starch, has been found to have multifunctional properties, including antioxidant, antiplatelet aggregation by thrombin and anti-inflammatory activities. The results of the present study demonstrate that 1,5-AF suppressed the growth of coagulase negative staphylococci on the hands as well as the growth of Staphylococcus epidermidis, which is a cause of opportunistic infections. Furthermore, 1,5-AF suppressed bio film formation by the methicillin resistant Staphylococcus aureus. In conclusion, 1,5-AF is a natural compound that may be effective in preventing nosocomial infections, without causing adverse side effects.

DOI： 10.3892/etm.2011.245

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PubMed

Language：Japanese   Publisher：The Japanese Society on Thrombosis and Hemostasis  

DOI： 10.2491/jjsth.22.11

Language：Japanese   Publisher：The Japan Society of Neurotraumatology  

DOI： 10.32187/neurotraumatology.33.2_241

Language：Japanese   Publisher：Medical Hypotheses  

Acute stroke, including acute ischemic stroke (AIS) and acute hemorrhagic stroke, (AHS) is a common medical problem with particular relevance to the demographic changes in industrialized societies. In recent years, treatments for AIS have emerged, including thrombolysis with tissue plasminogen activator (t-PA). Although t-PA is the most effective currently available therapy, it is limited by a narrow therapeutic time window and side effects, and only 3% of all AIS patients receive thrombolysis. Edaravone was originally developed as a potent free radical scavenger and, since 2001, has been widely used to treat AIS in Japan. It was shown that edaravone extended the narrow therapeutic time window of t-PA in rats. The therapeutic time window is very important for the treatment of AIS, and early edaravone treatment is more effective. Thus, more AIS patients might be rescued by administering edaravone with t-PA. Meanwhile, edaravone attenuates AHS-induced brain edema, neurologic deficits and oxidative injury in rats. Although edaravone treatment is currently only indicated for AIS, it does offer neuroprotective effects against AHS in rats. Therefore, we hypothesize that early administration of edaravone can rescue AHS patients as well as AIS patients. Taken together, our findings suggest that edaravone should be immediately administered on suspicion of acute stroke, including AIS and AHS. © 2010 Elsevier Ltd.

DOI： 10.1016/j.mehy.2010.07.038

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Language：English   Publisher：American Journal of Pathology  

Stromal cell-derived factor-1 (SDF-1) causes chemotaxis of CXCR4-expressing bone marrow-derived cells. SDF-1 is involved in the pathogenesis of various vascular diseases, including those of the eye. However, the role of SDF-1 in neuronal diseases is not completely understood. Here, we show higher SDF-1 levels in the vitreous humor of patients with retinal detachment (RD) compared with normal patients. SDF-1 correlated positively with the duration as well as the extent of RD. Furthermore, SDF-1 correlated significantly with levels of interleukin-6 and interleukin-8, but not with vascular endothelial growth factor. Western blot analysis results showed significant SDF-1 up-regulation in detached rat retinas compared with normal animals. Immunohistochemistry data showed that SDF-1 was co-localized with the glial cells of the detached retina. SDF-1 blockade with a neutralizing antibody increased photoreceptor cell loss and macrophage accumulation in the subretinal space. The retinal precursor cell line R28 expressed CXCR4. SDF-1 rescued serum starvation-induced apoptosis in R28 cells and enhanced their ability to participate in wound closure in a scratch assay. Our results indicate a surprising, protective role for SDF-1 in RD. This effect may be mediated directly or indirectly through other cell types. Copyright © American Society for Investigative Pathology.

DOI： 10.2353/ajpath.2010.100134

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PubMed

Language：English   Publisher：Biochemical and Biophysical Research Communications  

Proteins are folded properly in the endoplasmic reticulum (ER). Various stress such as hypoxia, ischemia and starvation interfere with the ER function, causing ER stress, which is defined by the accumulation of unfolded protein (UP) in the ER. ER stress is prevented by the UP response (UPR) and ER-associated degradation (ERAD). These signaling pathways are activated by three major ER molecules, ATF6, IRE-1 and PERK. Using HaCaT cells, we investigated ER signaling in human keratinocytes irradiated by environmental doses of ultraviolet B (UVB). The expression of Ero1-Lα, an upstream signaling molecule of ER stress, decreased at 1-4 h after 10 mJ/cm2 irradiation, indicating that the environmental dose of UVB-induced ER stress in HaCaT cells, without growth retardation. Furthermore, expression of intact ATF6 was decreased and it was translocated to the nuclei. The expression of XBP-1, a downstream molecule of IRE-1, which is an ER chaperone whose expression is regulated by XBP-1, and UP ubiquitination were induced by 10 mJ/ cm2 UVB at 4 h. PERK, which regulates apoptosis, was not phosphorylated. Our results demonstrate that UVB irradiation generates UP in HaCaT cells and that the UPR and ERAD systems are activated to protect cells from UVB-induced ER stress. This is the first report to show ER signaling in UVB-irradiated keratinocytes. © 2010 Elsevier Inc.

DOI： 10.1016/j.bbrc.2010.05.128

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PubMed

Language：English   Publisher：Laboratory Investigation  

Granulomatous nephritis can be triggered by diverse factors and results in kidney failure. However, despite accumulating data about granulomatous inflammation, pathogenetic mechanisms in nephritis remain unclear. The DNA-binding highmobility group box-1 protein (HMGB1) initiates and propagates inflammation when released by activated macrophages, and functions as an ‘alarm cytokine’ signaling tissue damage. In this study, we showed elevated HMGB1 expression in renal granulomas in rats with crystal-induced granulomatous nephritis caused by feeding an adenine-rich diet. HMGB1 levels were also raised in urine and serum, as well as in monocyte chemoattractant protein-1 (MCP-1), a mediator of granulomatous inflammation. Injection of HMGB1 worsened renal function and upregulated MCP-1 in rats with crystalinduced granulomatous nephritis. HMGB1 also induced MCP-1 secretion through mitogen-activated protein kinase (MAPK) and phosphoinositide-3-kinase (PI3K) pathways in rat renal tubular epithelial cells in vitro. Hmgb1+/− mice with crystal-induced nephritis displayed reduced MCP-1 expression in the kidneys and in urine and the number of macrophages in the kidneys was significantly decreased. We conclude that HMGB1 is a new mediator involved in crystalinduced nephritis that amplifies granulomatous inflammation in a cycle where MCP-1 attracts activated macrophages, resulting in excessive and sustained HMGB1 release. HMGB1 could be a novel target for inhibiting chronic granulomatous diseases. © 2010 USCAP, Inc. All rights reserved.

DOI： 10.1038/labinvest.2010.64

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PubMed

Language：English   Publisher：Thrombosis Research  

Introduction: Deficiency of Von Willebrand factor (VWF)-cleaving protease (ADAMTS13) causes platelet thrombosis in the microcirculation. Intrarenal coagulation is thought to be associated with the development and progression of diabetic nephropathy. Our aim was to clarify the association between plasma ADAMTS13 antigen (ADAMTS13Ag) levels and diabetic nephropathy. Material and Methods: We measured the plasma levels of VWF antigen (VWFAg) and ADAMTS13Ag, and calculated the VWF/ADAMTS13 ratio in 86 type 2 diabetic patients and 26 healthy volunteers, to investigate the relationship between these levels and renal function. With regard to diabetic macroangiopathy, the relationship between these levels and carotid intima-media thickness (IMT) was also investigated. Results and Conclusions: A significant positive and negative correlation was noted between ADAMTS13Ag and the estimated glomerular filtration rate (eGFR), vWF/ADAMTS13 ratio and eGFR, respectively. The diabetic patients were divided into normoalbuminuria (n = 50), microalbuminuria (n = 8) and overt nephropathy (n = 28) groups. Compared among these three groups and the 26 healthy volunteers, ADAMTS13Ag was significantly lower only in the overt nephropathy group. The mean carotid IMT was measured in 69 patients and was significantly negatively correlated with ADAMTS13Ag and positively correlated with VWF/ADAMTS13 ratio. When these 69 patients were divided into four groups according to eGFR and ADAMTS13 levels (ADAMTS13/eGFR; low/low: n = 12; high/low: n = 7; low/high: n = 25; high/high: n = 25), the mean carotid IMT was increased in patients with a low ADAMTS13Ag in the same eGFR group. The present study suggests that reduced ADAMTS13 might be associated with diabetic nephropathy. © 2010 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.thromres.2010.02.013

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PubMed

Language：English   Publisher：Journal of Immunology  

Vascular endothelial growth factor A (VEGF-A) is a prominent growth factor for both angiogenesis and lymphangiogenesis. Recent studies have shown the importance of VEGF-A in enhancing the growth of lymphatic endothelial cells in lymph nodes (LNs) and the migration of dendritic cells into LNs. VEGF-A is produced in inflamed tissues and/or in draining LNs, where B cells are a possible source of this growth factor. To study the effect of B cell-derived VEGF-A, we created transgenic mice (CD19Cre/hVEGF-Afl) that express human VEGF-A specifically in B cells. We found that the human VEGF-A produced by B cells not only induced lymphangiogenesis in LNs, but also induced the expansion of LNs and the development of high endothelial venules. Contrary to our expectation, we observed a significant decrease in the Ag-specific Ab production postimmunization with OVA and in the proinflammatory cytokine production postinoculation with LPS in these mice. Our findings suggest immunomodulatory effects of VEGF-A: B cell-derived VEGF-A promotes both lymphangiogenesis and angiogenesis within LNs, but then suppresses certain aspects of the ensuing immune responses. Copyright © 2010 by The American Association of Immunologists, Inc.

DOI： 10.4049/jimmunol.0903063

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PubMed

Language：Japanese   Publisher：(一社)日本予防医学会  

介護福祉士の中途退職の要因は腰痛等の健康障害による割合が最多であり、職務継続のためには健康管理が重要であるにも関わらず、基礎教育における1次予防教育または教育評価の報告例はまだない。本研究では、介護福祉学生に対して健康管理状況と介護福祉士になる自信、健康を維持していける自信・不安等の自己効力感について実態を把握し、自己の健康管理能力を高める目的で健康教育を実施し、評価した。評価指標は健康管理への自己効力感と健康不安度の変化等で、教育方法は介入群43名と対照群22名に対してベースラインの評価をした後、対照群には通常の講義演習による基礎教育を行い、介入群には基礎教育に加え集団健康教育ならびに個別健康教育を実施し、評価した。統計解析はStudentのt検定等を用い、統計解析ソフトSPSS12.0Jを用いた。その結果、介入群は対照群と比較して腰痛への不安が軽減(t=3.78、p<0.0001)する他、健康管理に対する自信度が高くなる(t=3.12、p=0.003)、コミュニケーションへの不安が軽減する(p=0.001)等の効果があった。基礎教育において腰痛への不安軽減や健康管理への自己効力感が高まることにより、卒後に介護福祉士としての職務継続の阻害因子となり得る健康不安に対して、予防的に対処できることが示唆された。(著者抄録)

Language：Japanese   Publisher：Experimental and Therapeutic Medicine  

A 77-year-old woman suffered a cardiopulmonary arrest the day after transvenous embolization of dural ateriovenous fistulae. The patient died despite receiving prompt cardiopulmonary resuscitation. Post mortem computed tomography (CT) was performed to determine the cause of death. No lesion was detected on a whole-body plain CT. However, a post mortem contrast-enhanced CT (CECT) performed after the administration of intravenous contrast and cardiac compressions detected pulmonary thromboembolism. Thus, post mortem CECT was useful in determining the cause of sudden death in this case.

DOI： 10.3892/etm_00000079

Scopus

Language：Japanese   Publisher：Vascular Health and Risk Management  

Purpose: Many epidemiological research studies have shown that vital exhaustion and psychosocial factors are associated with the occurrence of cerebrocardiovascular disease (CCVD). Fatigue is thought to induce endothelial dysfunction and may be linked to the occurrence of CCVD; however, no studies have investigated this potential link. We studied to determine the effect of fatigue on endothelial function in healthy subjects with no traditional CCVD risk factors or potential confounding factors to be controlled. Subjects and methods: Peripheral arterial tonometry (PAT) was used to evaluate endothelial function. The influence of the following parameters on endothelial function was analyzed in 74 office workers without traditional CCVD risk factors at health check-ups: endothelial function before and after work, subjective fatigue, lifestyle factors such as sleeping time, and psychosocial factors such as depression and social support. Results: Twenty-five subjects (33.8%) had low endothelial function; reactive hyperemia (RH)-PAT index <1.67, even though no abnormalities were reported in the health check-ups. There was no significant difference in endothelial function before versus after labor. Of note, endothelial function was associated with the individual's level of subjective fatigue (t = 2.98, P = 0.008) and showed a daily fluctuation, sometimes to a pathological degree (<1.67). Conclusion: We showed that, even in healthy people, endothelial function fluctuates diurnally, with an interaction between the individual's cognitive fatigue and the environment, sometimes to a pathological degree. Based on these findings, we suggest that endothelial function is an objective assessment tool of fatigue in healthy individuals. © 2010 Ohno et al, publisher and licensee Dove Medical Press Ltd.

Scopus

Language：English   Publisher：Biochemical and Biophysical Research Communications  

Endocannabinoids including anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are important lipid mediators for immunosuppressive effects and for appropriate homeostasis via their G-protein-coupled cannabinoid (CB) receptors in mammalian organs and tissues, and may be involved in wound healing in some organs. The physiological roles of endocannabinoids in periodontal healing remain unknown. We observed upregulation of the expression of CB1/CB2 receptors localized on fibroblasts and macrophage-like cells in granulation tissue during wound healing in a wound-healing model in rats, as well as an increase in AEA levels in gingival crevicular fluid after periodontal surgery in human patients with periodontitis. In-vitro, the proliferation of human gingival fibroblasts (HGFs) by AEA was significantly attenuated by AM251 and AM630, which are selective antagonists of CB1 and CB2, respectively. CP55940 (CB1/CB2 agonist) induced phosphorylation of the extracellular-regulated kinases (ERK) 1/2, p38 mitogen-activated protein kinase (p38MAPK), and Akt in HGFs. Wound closure by CP55940 in an in-vitro scratch assay was significantly suppressed by inhibitors of MAP kinase kinase (MEK), p38MAPK, and phosphoinositol 3-kinase (PI3-K). These findings suggest that endocannabinoid system may have an important role in periodontal healing. © 2010 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2010.03.080

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PubMed

Language：English  

DOI： 10.2147/vhrm.s8950

PubMed

Language：English   Publisher：International Journal of Immunopathology and Pharmacology  

1,5-anhydrogIucitol (1,5-AG) decreases in diabetic patients and is used as a marker of glycemic control. Type 2 diabetic patients are susceptibile to lipopolysaccharides (LPS), which stimulate macrophages to release large quantities of tumor necrosis factor (TNF)-α and interleukin (IL)-6. This study examines the effects of 1,5-AG on lung inflammation induced by LPS and consequent systemic inflammation to determine whether the decrease of 1,5-AG concentration induces susceptibility to LPS. Before the challenge with LPS (1 mg/kg in vivo and 500 ng/ml in vitro), we pretreated db/db mice and RAW264.7 cells with 1,5-AG at 38.5 mg/kg and 500 μg/mI, respectively. The levels of IL-6, TNF-α, macrophage chemoattractant protein (MCP)-1 and IL-1β in the serum and in the cell supernatants were measured. We also measured macrophage recruitment and the expression of inducible nitric oxide synthase (iNOS) in pulmonary tissues. We found that 1,5-AG attenuated serum cytokine release and protected db/db mice from LPS-induced pulmonary inflammation. In addition, 1,5-AG suppressed cytokine release and iNOS expression by suppressing Akt/NF-κB activity in RAW264.7 cells. These results suggest that 1,5-AG may be a mediator in, as well as marker for diabetes, and 1,5-AG intake may confer tolerance to LPS in patients with type 2 diabetes. © by Biolife, s.a.s.

DOI： 10.1177/039463201002300110

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PubMed

Language：Japanese   Publisher：Experimental and Therapeutic Medicine  

Estimation of the postmortem interval (PMIPMIPMI) is one of the most important tasks in forensic medicine. Numerous methods have been proposed for the determination of the time since death by chemical means. High mobility group box-1 (HMHMGB1), a nonhistone DNADNADNA-binding protein is released by eukaryotic cells upon necrosis. Postmortem serum levels of HMGB1 of 90 male Wistar rats stored at 4, 14 and 24 °C since death were measured by enzyme-linked immunosorbent assay. The serum HMHMGB1 level showed a time-dependent increase up to seven days at 4 °C. At 14 °C, the HMGB1 level peaked at day 3, decreased at day 4, and then plateaued. At 24 °C, the HMHMGB1 level peaked at day 2, decreased at day 3, and then plateaued. Our findings suggest that HMGB1 is related to the PMIPMIPMI in rats.

DOI： 10.3892/etm_00000019

Scopus

Language：English   Publisher：British Journal of Haematology  

Kawasaki syndrome (KS) is an acute febrile vasculitis of childhood. Coronary artery abnormalities (CAA) are a significant problem in KS patients. High dose intravenous immunoglobulin (IVIG) is effective for reducing the occurrence of CAA. Clinical and histopathological findings suggest that vascular endothelial growth factor (VEGF) is involved in CAA. In circulating blood, newly activated platelets are the major source of VEGF, which is released in large amounts in vascular inflammation. The present study analysed 80 KS patients (69 IVIG responders and 11 IVIG non-responders) and evaluated the role of platelet VEGF in KS vasculitis. Serum VEGF and platelet VEGF levels were significantly higher in KS patients than controls (P < 0·001). Platelet VEGF reflected the reactivity of IVIG treatment and was decreased in responders (P < 0·001), but remained increased in non-responders (P = 0·01). Platelet VEGF levels, but not serum VEGF levels, before IVIG were significantly correlated with the maximum CAA z-score (r = 0·524, P = 0·02). Our findings demonstrate that platelet VEGF may reflect the severity of vasculitis related to the pathological development of CAA in KS. Platelet VEGF may be an important feature of KS pathophysiology. © 2009 Blackwell Publishing Ltd.

DOI： 10.1111/j.1365-2141.2009.07922.x

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PubMed

Language：English   Publisher：Biochemical and Biophysical Research Communications  

Aquaporin-4 (AQP4) plays a role in the generation of post-ischemic edema. Pharmacological modulation of AQP4 function may thus provide a novel therapeutic strategy for the treatment of stroke, tumor-associated edema, epilepsy, traumatic brain injury, and other disorders of the central nervous system (CNS) associated with altered brain water balance. Edaravone, a free radical scavenger, is used for the treatment of acute ischemic stroke (AIS) in Japan. In this study, edaravone significantly reduced the infarct area and improved the neurological deficit scores at 24 h after reperfusion in a rat transient focal ischemia model. Furthermore, edaravone markedly reduced AQP4 immunoreactivity and protein levels in the cerebral infarct area. In light of observations that edaravone specifically inhibited AQP4 in a rat transient focal ischemia model, we propose that edaravone might reduce cerebral edema through the inhibition of AQP4 expression following cerebral infarction. © 2009 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2009.09.015

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PubMed

Language：English   Publisher：Biochemical and Biophysical Research Communications  

The Japanese apricot, known as Ume in Japanese, has been a traditional Japanese medicine for centuries, and is a familiar and commonly consumed food. The health benefits of Ume are now being widely recognized and have been strengthened by recent studies showing that MK615, an extract of compounds from Ume, has strong anticancer and anti-inflammatory effects. However, the potential role of MK615 in the periodontal field remains unknown. Here, we found that MK615 significantly reduced the production of pro-inflammatory mediators (tumor necrosis factor-alpha and interleukin-6) induced by Porphyromonas gingivalis lipopolysaccharide (LPS), a major etiological agent in localized chronic periodontitis, in murine macrophage-like RAW264.7 cells. MK615 markedly inhibited the phosphorylation of ERK1/2, p38MAPK, and JNK, which is associated with pro-inflammatory mediator release pathways. Moreover, MK615 completely blocked LPS-triggered NF-κB activation. The present results suggest that MK615 has potential as a therapeutic agent for treating inflammatory diseases such as periodontitis. © 2009 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2009.08.103

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PubMed

Language：English   Publisher：Medical Hypotheses  

Human serum contains thousands of proteolytically derived low-molecular-weight peptide fragments (serum peptidome). The concept of utilizing the serum peptidome for cancer diagnosis has been developed. A pathological serum peptidome appears when the homeostatic balance between proteases and protease inhibitors is disrupted. We hypothesize if analyses of the serum peptidome are of diagnostic value as information on which molecules are disrupted, and the pathological course it will take in unknown pathological conditions and disseminated intravascular coagulation (DIC). We analyzed the serum peptidome in 3 stages (early stage, pre-DIC and DIC stages) in one patient with POEMS (polyneuropathy, organomegaly, endocrinopathy, M protein and skin changes) syndrome, an intractable disease with unknown pathology, using a 1-dimensional gel electrophoresis/matrix-assisted laser desorption/ionization-mass spectrometry (1-DE/MS)-based rapid quantitative approach. A very large number of peptide fragments appeared in the DIC stage, compared to pre-DIC. In addition, we identified fragments of transthyretin (ALGISPFHEHAEVVFTANDSGPR, m/z 2451.18) and α1-antitrypsin (EDPQGDAAQKTDTSHHDQDHPTFN, m/z 2691.02) that significantly increased in the DIC stage, compared to those in the pre-DIC stage. Rapid analyses of the serum peptidome may lead to a diagnostic method that can predict on-going protease activated pathological conditions and help to decide on multilateral strategies including nutritional support and drug therapy. © 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.mehy.2009.04.026

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PubMed

Language：English   Publisher：Biochemical and Biophysical Research Communications  

1,5-anhydro-d-fructose (1,5-AF), a monosaccharide formed from starch and glycogen, exhibits antioxidant and antibacterial activity, and inhibits cytokine release by attenuating NF-κB activation in LPS-stimulated mice. The present study examined whether 1,5-AF inhibits lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) in vitro and in vivo. We found that 1,5-AF significantly blocked the production of NO, and protein and mRNA expression of iNOS, and up-regulated IL-10 production in vitro. We also investigated the effects of 1,5-AF on acute lung inflammation in C57BL/6J mice. We found that protein and mRNA expression of iNOS in lung tissues were inhibited by 1,5-AF pretreatment. In addition, the serum level of IL-10 was upregulated by 1,5-AF. Collectively, the iNOS transcriptional and translational inhibitory effects of 1,5-AF seem to be prolonged and enhanced by the production of IL-10. These results suggest that 1,5-AF could be a useful adjunct in the treatment of acute lung inflammation. © 2009 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2009.06.108

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PubMed

Language：English   Publisher：Journal of Leukocyte Biology  

NPM is a major nucleolar multifunctional protein involved in ribosome biogenesis, centrosome duplication, cell-cycle progression, apoptosis, cell differentiation, and sensing cellular stress. Alarmins are endogenous molecules released from activated cells and/or dying cells, which activate the immune system and cause severe damage to cells and tissue organs. In the present work, stimulation of cells with the alarmin-inducible molecule endotoxin, for 16 h, resulted in NPM release into the culture supernatants of RAW264.7 cells, a murine macrophage cell line. Extracellular NPM was detected in the ascites of the CLP model. NPM was translocated into the cytoplasm from the nucleus in LPS -stimulated RAW264.7 cells; furthermore, NPM was detected in the cytosols of infiltrated macrophages in the CLP model. rNPM induced release of proinflammatory cytokines, TNF-α, IL-6, and MCP-1, from RAW264.7 cells and increased the expression level of ICAM-1 in HUVECs. NPM induced the phosphorylation of MAPKs in RAW264.7 cells. Our data indicate that NPM may have potent biological activities that contribute to systemic inflammation. Further investigations of the role of NPM may lead to new therapies for patients with septic shock or other inflammatory diseases. © Society for Leukocyte Biology.

DOI： 10.1189/jlb.1008644

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PubMed

Language：Japanese  

Language：English   Publisher：Biochemical and Biophysical Research Communications  

High mobility group box-1 (HMGB1), a non-histone DNA-binding protein, is massively released into the extracellular space from neuronal cells after ischemic insult and exacerbates brain tissue damage in rats. Minocycline is a semisynthetic second-generation tetracycline antibiotic which has recently been shown to be a promising neuroprotective agent. In this study, we found that minocycline inhibited HMGB1 release in oxygen-glucose deprivation (OGD)-treated PC12 cells and triggered the activation of p38mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases (ERK1/2). The ERK kinase (MEK)1/2 inhibitor U-0126 and p38MAPK inhibitor SB203580 blocked HMGB1 release in response to OGD. Furthermore, HMGB1 triggered cell death in a dose-dependent fashion. Minocycline significantly rescued HMGB1-induced cell death in a dose-dependent manner. In light of recent observations as well as the good safety profile of minocycline in humans, we propose that minocycline might play a potent neuroprotective role through the inhibition of HMGB1-induced neuronal cell death in cerebral infarction. © 2009 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2009.04.041

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PubMed

Language：English   Publisher：Journal of Pharmacology and Experimental Therapeutics  

Edaravone, a potent free radical scavenger, is clinically used for the treatment of cerebral infarction in Japan. Here, we examined the effects of edaravone on the dynamics of high-mobility group box-1 (HMGB1), which is a key mediator of ischemic-induced brain damage, during a 48-h postischemia/ reperfusion period in rats and in oxygen-glucose-deprived (OGD) PC12 cells. HMGB1 immunoreactivity was observed in both the cytoplasm and the periphery of cells in the cerebral infarction area 2 h after reperfusion. Intravenous administration of 3 and 6 mg/kg edaravone significantly inhibited nuclear translocation and HMGB1 release in the penumbra area and caused a 26.5 ± 10.4 and 43.8 ± 0.5% reduction, respectively, of the total infarct area at 24 h after reperfusion. Moreover, edaravone also decreased plasma HMGB1 levels. In vitro, edaravone dose-dependently (1-10 μM) suppressed OGD- and H2O2-induced HMGB1 release in PC12 cells. Furthermore, edaravone (3-30 μM) blocked HMGB1-triggered apoptosis in PC12 cells. Our findings suggest a novel neuroprotective mechanism for edaravone that abrogates the release of HMGB1. Copyright © 2009 by The American Society for Pharmacology and Experimental Therapeutics.

DOI： 10.1124/jpet.108.149484

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PubMed

Language：Japanese   Publisher：Diabetes and Metabolic Syndrome: Clinical Research and Reviews  

Aim: Although lifestyle changes in university students are highly influential in causing their later obese-related diseases, there is still limited knowledge about obese university students. Metabolic syndrome-related markers and conditions were investigated among obese students in Kagoshima University, Japan. Methods: We evaluated 46 obese students (31 males and 15 females), with high body mass index (BMI) (≥30), aged between 18 and 24 years, and who were recruited at university annual health check-ups in 2006 and 2007. All subjects underwent measurement of waist circumference, laboratory evaluations for the diagnostic criteria (Japan Metabolic Syndrome Criteria Study Group) and levels of uric acid and plasminogen activator inhibitor type-1 (PAI-1), and behavioral assessments using a questionnaire about personal and lifestyle factors. Results: 39.1% of the obese university students fulfilled the criteria for metabolic syndrome. In the metabolic syndrome constituents, higher prevalence was obtained in hypertension and hypertriglyceridemia. Levels of uric acid and PAI-1 were high with statistical significance in metabolic syndrome. A broader expansion of positive correlations between PAI-1 and the metabolic syndrome components as compared with those of uric acid suggested the important roles of PAI-1 in the pathological processes or outcomes of metabolic syndrome. Conclusion: Our findings suggested that strategies to prevent hypertension and/or dyslipidemia are important for the prophylaxis of metabolic syndrome in university students, and that dietary fat lowering has a prophylactic importance in the control of hyperuricemia. Furthermore, a causal relationship between the regularly missed breakfasts and metabolic syndrome was suggested. © 2009 Diabetes India.

DOI： 10.1016/j.dsx.2009.02.009

Scopus

Language：English   Publisher：International Journal of Molecular Medicine  

High mobility group box-1 protein (HMGB1), primarily from the nucleus, is released into the extracellular milieu either passively from necrotic cells or actively through secretion by monocytes/macrophages. Extracellular HMGB1 acts as a potent inflammatory agent by promoting the release of cytokines such as tumor necrosis factor (TNF)-α, has procoagulant activity, and is involved in death due to sepsis. Accordingly, HMGB1 is an appropriate therapeutic target. In this study, we found that an extract of Prunus mume Sieb. et Zucc. (Ume) fruit (Ume extract), an abundant source of triterpenoids, strongly inhibited HMGB1 release from lipopolysaccharide (LPS)-stimulated macrophage-like RAW264.7 cells. The inhibitory effect on HMGB1 release was enhanced by authentic oleanolic acid (OA), a naturally occurring triterpenoid. Similarly, the HMGB1 release inhibitor in Ume extract was found to be OA. Regarding the mechanisms of the inhibition of HMGB1 release, the OA or Ume extract was found to activate the transcription factor Nrf2, which binds to the antioxidative responsive element, and subsequently the heme oxygenase (HO)-1 protein was induced, indicating that the inhibition of HMGB1 release from LPS-stimulated RAW264.7 cells was mediated via the Nrf2/HO-1 system; an essentially antioxidant effect. These results suggested that natural sources of triterpenoids warrant further evaluation as 'rescue' therapeutics for sepsis and other potentially fatal systemic inflammatory disorders.

DOI： 10.3892/ijmm_00000172

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PubMed

Language：English  

DOI： 10.1016/j.ophtha.2008.12.024

PubMed

Language：English   Publisher：Pediatric Infectious Disease Journal  

We examined the serum values of high mobility group box 1 (HMGB1) in 36 patients with Kawasaki syndrome (KS) (29 responders and 7 poor-responders to initial intravenous immunoglobulin treatment). A mean value of HMGB1 of poor-responders was significantly elevated compared with those of responders (P ≤ 0.0042). Among the 6 factors showing significant differences between responders and poor-responders including HMGB1 (admission illness day, white blood cell counts, C-reactive protein, aspartate aminotransferase, lactate dehydrogenase), values of HMGB1 showed the widest area under the receiver operating characteristic curve. In conclusion, an elevated HMGB1 value could be a potential marker for poor-responders. Copyright © 2009 by Lippincott Williams & Wilkins.

DOI： 10.1097/INF.0b013e31818ffe60

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PubMed

Language：English   Publisher：Laboratory Investigation  

High-mobility group box 1 (HMGB1) protein is a multifunctional protein, which is mainly present in the nucleus and is released extracellularly by dying cells and/or activated immune cells. Although extracellular HMGB1 is thought to be a typical danger signal of tissue damage and is implicated in diverse diseases, its relevance to ocular diseases is mostly unknown. To determine whether HMGB1 contributes to the pathogenesis of retinal detachment (RD), which involves photoreceptor degeneration, we investigated the expression and release of HMGB1 both in a retinal cell death induced by excessive oxidative stress in vitro and in a rat model of RD-induced photoreceptor degeneration in vivo. In addition, we assessed the vitreous concentrations of HMGB1 and monocyte chemoattractant protein 1 (MCP-1) in human eyes with RD. We also explored the chemotactic activity of recombinant HMGB1 in a human retinal pigment epithelial (RPE) cell line. The results show that the nuclear HMGB1 in the retinal cell is augmented by death stress and upregulation appears to be required for cell survival, whereas extracellular release of HMGB1 is evident not only in retinal cell death in vitro but also in the rat model of RD in vivo. Furthermore, the vitreous level of HMGB1 is significantly increased and is correlated with that of MCP-1 in human eyes with RD. Recombinant HMGB1 induced RPE cell migration through an extracellular signal-regulated kinase-dependent mechanism in vitro. Our findings suggest that HMGB1 is a crucial nuclear protein and is released as a danger signal of retinal tissue damage. Extracellular HMGB1 might be an important mediator in RD, potentially acting as a chemotactic factor for RPE cell migration that would lead to an ocular pathological wound-healing response. © 2009 USCAP, Inc All rights reserved.

DOI： 10.1038/labinvest.2008.165

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PubMed

Language：English   Publisher：Diabetic Medicine  

Aim: To identify the relationship between vascular endothelial growth factor (VEGF) and diabetic polyneuropathy (DPN). Methods: Two hundred and twenty diabetic patients participated, 113 with DPN and 107 without DPN. All patients were also classified according to the four stages of DPN (no neuropathy: stage 0; asymptomatic neuropathy: stage 1; symptomatic neuropathy: stage 2; disabling neuropathy: stage 3). Serum VEGF concentration was measured using an enzyme-linked immunosorbent assay (ELISA) and levels between the patients with and without DPN and also between the different stages of DPN, were compared. Results: The mean serum VEGF level in all patients was 264.6 ± 218.8 pg/ml. The mean serum VEGF level was higher in patients with DPN (310.1 ± 224.3 pg/ml) than in the patients without DPN (216.5 ± 204.0 pg/ml, P = 0.0014). Serum VEGF was higher in the 'symptomatic' stage (stage 2, 364.8 ± 225.9 pg/ml) in comparison with the 'asymptomatic' (stage 1, 256.7 ± 224.4 pg/ml, P = 0.015) and 'disabling' (stage 3, 180.3 ± 109.4 pg/ml, P = 0.042) stages. The mean serum VEGF level in patients with diabetic retinopathy (261.1 ± 210.6 pg/ml) and in patients with diabetic nephropathy (241.5 ± 185.7 pg/ml) was not increased. Conclusions: The serum VEGF level is increased in patients with DPN, particularly in patients in the neurologically active 'symptomatic' stage. © 2009 Diabetes UK.

DOI： 10.1111/j.1464-5491.2009.02680.x

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PubMed

Language：Japanese   Publisher：Neurocritical Care  

Background: High-mobility group box 1 protein (HMGB1) is a nuclear factor that is a potent proinflammatory mediator, and may trigger increases in other inflammatory cytokines. The inflammatory cytokines in the cerebrospinal fluid (CSF) of patients with subarachnoid hemorrhage (SAH) have been reported previously, but HMGB1 has not. In this study, we measured HMGB1 and the inflammatory cytokines in the CSF of patients with SAH. Methods: CSF samples were collected on days 3, 7, and 14 from the drainage tubes of the postaneurysm clips of 39 patients with SAH. HMGB1, interleukin-6 (IL-6), IL-8, and tumor necrosis factor alpha (TNF-α) were measured in the CSF, and compared between the patients with favorable (good recovery and moderate disability) and unfavorable outcomes (severe disability, vegetative state, and death) at 3 months. Results: In the unfavorable outcome group, HMGB1 (P = 0.017), IL-6 (P = 0.003), IL-8 (P = 0.041), and TNF-α (P = 0.002) were significantly increased. HMGB1 correlated significantly with IL-6, IL-8, and TNF-α (R = 0.672, 0.421, and 0.697, respectively). Conclusions: HMGB1 was increased in the CSF of SAH patients with an unfavorable outcome, as were the other cytokines. These results suggest that HMGB1 and cytokines are related to the brain damage observed after SAH. HMGB1 might play a key role in the inflammatory response in the CNS of SAH patients. © 2009 Humana Press Inc.

DOI： 10.1007/s12028-009-9276-y

Scopus

Language：Japanese   Publisher：The Japanese Society on Thrombosis and Hemostasis  

DOI： 10.2491/jjsth.20.418

Language：English  

DOI： 10.1016/j.bbrc.2008.10.049

PubMed

Language：Japanese   Publisher：International Journal of Molecular Medicine  

High mobility group box 1 (HMGB1) is a nonhistone nuclear protein which is released from the nucleus of activated macrophages into the extracellular space in response to stimuli such as endotoxin or necrosis. The HMGB1 functions as a potent proinflammatory cytokine in the extracellular spaces. Recently, HMGB1 has been implicated in the progression of atherosclerosis. However, the association between HMGB1 and the development of atherosclerosis is poorly understood. Therefore, we examined whether serotonin (5-HT), a key factor involved in the development of atherosclerosis, induced HMGB1 release in human umbilical vein endothelial cells (HUVECs). We found that 5-HT induced the release of HMGB1 but not of ERK1/2 and JNK from HUVECs via the 5-HT receptor (5-HT1B)/p38 mitogen-activated protein kinase (MAPK) signaling pathway. The p38MAPK inhibitor SB203580 and the 5-HT1B antagonist GR55526 markedly inhibited HMGB1 release from 5-HT-stimulated HUVECs. The vascular endothelial growth factor (VEGF) derived from activated macrophages in atherosclerotic lesions also plays an important role in the progression of atherosclerosis. We found that HMGB1 induced VEGF production in macrophage-like RAW264.7 cells. HMGB1 induced the activation of p38MAPK, ERK1/2 and Akt. The PI3-kinase inhibitor LY294002 significantly inhibited VEGF production in HMGB1-stimulated macrophages, while other kinase inhibitors did not. These results suggest that HMGB1 release may contribute as a risk factor in the development and progression of atherosclerosis.

DOI： 10.3892/ijmm_00000066

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Language：English  

PubMed

Language：English   Publisher：Arteriosclerosis, Thrombosis, and Vascular Biology  

Objective - High mobility group box 1 protein (HMGB1) was identified as a mediator of endotoxin lethality. We previously reported that thrombomodulin (TM), an endothelial thrombin-binding protein, bound to HMGB1, thereby protecting mice from lethal endotoxemia. However, the fate of HMGB1 bound to TM remains to be elucidated. Methods and Results - TM enhanced thrombin-mediated cleavage of HMGB1. N-terminal amino acid sequence analysis of the HMGB1 degradation product demonstrated that thrombin cleaved HMGB1 at the Arg10-Gly11 bond. Concomitant with the cleavage of the N-terminal domain of HMGB1, proinflammatory activity of HMGB1 was significantly decreased (P<0.01). HMGB1 degradation products were detected in the serum of endotoxemic mice and in the plasma of septic patients with disseminated intravascular coagulation (DIC), indicating that HMGB1 could be degraded under conditions in which proteases were activated in the systemic circulation. Conclusions - TM not only binds to HMGB1 but also aids the proteolytic cleavage of HMGB1 by thrombin. These findings highlight the novel antiinflammatory role of TM, in which thrombin-TM complexes degrade HMGB1 to a less proinflammatory form. © 2008 American Heart Association, Inc.

DOI： 10.1161/ATVBAHA.107.150631

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PubMed

Language：Japanese   Publisher：(一社)日本医学教育学会  

1)低学年(2年生後期)で受講するPBLテュートリアルの前後で、受動的な学習法(講義を聴く)や能動的な学習法(自分で調べる、発表する、討論する、教える)に対する意識の変化を検討した。2)17年度と18年度の2回、調査を行った。両年度とも、統計的に有意に、能動的な学習法を肯定する意識の変化が観察された。「講義を聴く」については、年度により異なる結果を示した。3)今回のPBLテュートリアルは、学習法に対する意識の変化をもたらした可能性がある。(著者抄録)

Language：English   Publisher：Graefe's Archive for Clinical and Experimental Ophthalmology  

Background: High-mobility group box 1 protein (HMGB1) is recently described as a late mediator of lethal endotoxemia with proinflammatory cytokine-like properties. The purpose of this study was to determine whether HMGB1 is involved in endophthalmitis. Methods: In this retrospective case-control study, vitreous levels of HMGB1 were measured by an enzyme-linked immunosorbent assay in ten eyes with endophthalmitis, and in 12 eyes with idiopathic macular holes which served as controls. Formalin-fixed and paraffin-embedded tissue sections of an enucleated eye with endophthalmitis, and a control eye with recurrent conjunctival malignant melanoma, were analyzed by immunohistochemistry with an anti-HMGB1 antibody. Results: The vitreous HMGB1 level of the patients with endophthalmitis was 13.96±17.17 ng/ml (mean±SD), which was significantly higher than that of the controls (0.236±0.128 ng/ml; P=.0006, Mann-Whitney U test). There were significant correlations between HMGB1 level and disease duration, presenting visual acuity, and final visual acuity. In the eye with endophthalmitis, HMGB1 expression was diffusely observed, particularly in the extranuclear region of the retina and the choroid with infiltrating inflammatory cells. Conclusion: HMGB1 can be released in the vitreous of eyes with endophthalmitis depending on inflammation and tissue damage. Our results suggest that HMGB1 may be a late mediator of endophthalmitis, and related to the progression of endophthalmitis. © Springer-Verlag 2008.

DOI： 10.1007/s00417-008-0827-2

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PubMed

Language：Japanese   Publisher：Brain and Nerve  

Crow-Fukase syndrome is diagnosed based on the presence of chronic sensori-motor polyneuropathy along with other characteristic generalized symptoms denoted by the acronym of POEMS which stands for polyneuropathy, organomegaly, endocrinopathy, M-protein, and skin changes. In this syndrome, the serum levels of the vascular endothelial growth factor (VEGF) are abnormally elevated, and this is a predictive factor for its diagnosis. Although the causes of CFS/POEMS remain unknown, VEGF is evidently correlated with its pathogenesis. Human glioblastoma cells are known to express VEGF. In models of CFS/POEMS, mice that are peritoneally transplanted with human glioblastomas exhibit high serum levels of VEGF, prominent edema with increased circulation volume, and pathological findings in the liver, spleen, and kidney. VEGF that is highly concentrated in platelets may be released in massive amounts due to coagulation in the peripheral tissue and may thus exert its maximal physiological effects and produce the abovementioned diffuse pathological findings. The correlation between polyneuropathy and elevated VEGF remains unclear. However, VEGF may affect the blood-nerve barrier by increased microvascular hyperpermeability, upregulated cytokines such as matrix metalloproteases may induce blood-nerve barrier breakdown and demyelination of the peripheral nerve. Furthermore, microangiopathy due to proliferative endothelial cells and hypercoagulated occlusion also affect axonal damage. Novel strategies that have recently been proposed for the management of this disease include high-dose chemotherapy combined with autologous peripheral blood stem cell transplantation (PBSCT) and molecular-targeted therapy against plasma cells and VEGF. Notably, PBSCT exerts a dramatic effect on polyneuropathy; such an effect has rarely been achieved by the previously described modalities of low-dose melphallan and steroid therapy. PBSCT is observed to induce a rapid and persistent decrease in the serum VEGF levels. In conclusion, VEGF is not only the primary molecule involved in the pathogenesis of CSF, but also an important marker for both the diagnosis and treatment of this disease.

Scopus

Language：English   Publisher：Cardiovascular Pathology  

Background: C-reactive protein (CRP) is widely used as a sensitive biomarker for inflammation. Increasing evidence suggests that CRP plays a role in inflammation. High-mobility group box-1 (HMGB1), a primarily nuclear protein, is passively released into the extracellular milieu by necrotic or damaged cells and is actively secreted by monocytes/macrophages. Extracellular HMGB1 as a potent inflammatory mediator has stimulated immense curiosity in the field of inflammation research. However, the molecular dialogue implicated between CRP and HMGB1 in delayed inflammatory processes remains to be explored. Methods and results: The levels of HMGB1 in culture supernatants were determined by Western blot analysis and enzyme-linked immunosorbent assay in macrophage RAW264.7 cells. Purified CRP induced the release of HMGB1 in a dose- and time-dependent fashion. Immunofluorescence analysis revealed nuclear translocation of HMGB1 in response to CRP. The binding of CRP to the Fcγ receptor in RAW264.7 cells was confirmed by fluorescence-activated cell sorter analysis. Pretreatment of cells with IgG-Fc fragment, but not IgG-Fab fragment, efficiently blocked this binding. CRP triggered the activation of p38MAPK and ERK1/2, but not Jun N-terminal kinase. Moreover, both p38MAPK inhibitor SB203580 and small interfering RNA significantly suppressed the release of HMGB1, but not the MEK1/2 inhibitor U-0126. Conclusion: We demonstrated for the first time that CRP, a prominent risk marker for inflammation including atherosclerosis, could induce the active release of HMGB1 by RAW264.7 cells through Fcγ receptor/p38MAPK signaling pathways, thus implying that CRP plays a crucial role in the induction, amplification, and prolongation of inflammatory processes, including atherosclerotic lesions. © 2008 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.carpath.2007.08.006

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PubMed

Language：English  

DOI： 10.1111/j.1600-0765.2007.00996.x

PubMed

Language：Japanese   Publisher：Japan Society for Medical Education  

1) In problem-based learning (PBL) tutorials during the2nd year, general symptoms (fatigue, weight change, bleeding, fever, pain, and edema) were chosen as subjects.Changes in the students'way of thinking about how to learn were evaluated before and after PBL tutorials.<BR>2) After tutorials students were significantly more likely to believe that their participation in tutorials was a more effective way of learning. The students'views after tutorials on listening to lectures differed between2006and2007.<BR>3) We conclude that PBL tutorials affect medical students'views about learning.

DOI： 10.11307/mededjapan1970.39.267

Language：English   Publisher：Current Eye Research  

Purpose: To identify the roles of stromal-derived factor (SDF-1) and inflammatory cytokines in retinal vein occlusion (RVO). Methods: Samples were collected by vitrectomy and the levels of SDF-1, vascular endothelial growth factor, and inflammatory cytokines (interleukins [IL-1β, IL-6, IL-8, IL-10, IL-12p70]; tumor necrosis factor-α) were measured in 20 eyes with RVO, and 9 eyes with epiretinal membrane served as negative controls. Four eyes with inflammatory diseases were also investigated. Results: SDF-1 levels in active RVO (A-RVO; 4 eyes with iris neovascularization) were significantly higher than those in quiescent RVO (Q-RVO; 16 eyes without iris neovascularization) and the negative controls (p <.01), whereas there were no significant difference between the Q-RVO and the negative controls. There were no significant correlations between the concentrations of SDF-1 and other cytokines. Conclusions: Elevation of intravitreous SDF-1 levels in A-RVO but not Q-RVO suggested a pivotal role of SDF-1 in angiogenic changes during RVO. Copyright © Informa Healthcare USA, Inc.

DOI： 10.1080/02713680701758727

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PubMed

Language：Japanese   Publisher：Hirosaki Medical Journal  

Responses to stimuli in cellar level are diverse and such hierarchical as secretion of stored factors, synthesis of lipid mediators and protein synthesis through genomic transcription. However, how can the cells respond in the case of necrosis? Recently a characteristic intranuclear protein, high-mobility group box 1 protein (HMGB) is released from necrotic cells. The protein is an abundant nuclear protein with a dual function both inside and outside the cells. In physiological state, HMGB1 is present in the nucleus, and binds to DNA, playing a variety of crucial functions, including transcription and keeping the characteristic DNA architecture. However, the protein is released to extracellular space from most of necrotic cells, activated macrophages and dendritic cells. Out of the cells, HMGB1 acts as a signal of tissue damage and can promote inflammation, immune responses, and results tissue regeneration. During sepsis and/or disseminated intravascular coagulation (DIC), however, massive accumulation of HMGB1 in the systemic circulation will cause multiple organ failure (MOF) and subsequent lethal outcome. Thus HMGB1 in the systemic circulation has been considered as a lethal mediator of sepsis, and a promising therapeutic target for sepsis. Recently we identified that thrombomodulin (TM), a natural anticoagulant glycoprotein expressed on the surface of endothelial cells, plays an important role in sequestering HMGB1. TM may prevent HMGB1 from reaching remote organs, thereby restricting the spectrum of HMGB1 action in the site of injury. Here we review recent progress made in defining the physiological and pathological roles of HMGB1 and therapeutic strategies aimed at blocking circulatory HMGB1.

Scopus

Language：English   Publisher：Xenotransplantation  

High mobility group box-1 (HMGB1) protein, primarily from the nucleus, is released into the extracellular milieu either passively by necrotic or damaged cells, or actively by secretion from monocytes/macrophages. Extracellular HMGB1 acts as a potent inflammatory stimulator by promoting cytokine (for example, tumor necrosis factor-α) production, and also has pro-coagulant activity. The signaling pathway initiated by receptor for advanced glycation end-product (RAGE), which is the HMGB1 receptor, also induces complement activation. Recent studies have implicated HMGB1 in acute cardiac allograft rejection, and have identified infiltrating T cells and other damaged cells as its main sources. HMGB1 blockade using the anti-HMGB1 antibody HMGB1 box-A (amino-terminal region) and soluble RAGE rescues mice from acute rejection. We therefore studied the release of HMGB1 in co-cultures of porcine aortic endothelial cells (PAEC) and human leukocytes. Human T cells, but not B cells, monocytes or neutrophils, stimulated significant HMGB1 release in culture with PAEC; this activity required cell-cell contact and was dose-dependent, as determined by Western blotting. The released HMGB1 originated from both cell types, as immunofluorescent microscopy showed that it was present in the cytosol of PAEC in contact with T cells, and had disappeared from the T-cell nuclei. These results demonstrate that direct interactions between PAEC and T cells might be a key factor in triggering HMGB1 release, which suggests that HMGB1 is associated with graft rejection in the early phase. © 2007 Blackwell Munksgaard.

DOI： 10.1111/j.1399-3089.2007.00434.x

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PubMed

Language：English   Publisher：Atherosclerosis  

Vascular endothelial growth factor (VEGF) has been noted in the pathogenesis of atherosclerosis. To examine the usefulness of the serum concentration of VEGF as an index of atherosclerosis, we analyzed the serum VEGF concentrations in 443 adults who underwent a medical checkup. The mean serum VEGF concentration of men (229 ± 147 pg/ml) was significantly higher than that of women (182 ± 112 pg/ml). The platelet count showed a slight correlation with the serum VEGF concentration in both genders (men R = 0.287, women R = 0.296), corresponding with the results of experiments that platelets are the major source of VEGF in circulating peripheral blood. In men, the serum VEGF concentrations correlated with platelet counts, body fat percentages, leukocyte counts, and HDL-cholesterol concentrations (negative correlation). In the multiple regression analysis performed for men's serum VEGF concentrations, the decision coefficient (R2) was maximized (R2 = 0.173) when the leukocyte count, the body fat percentage, and the HDL-cholesterol concentration were taken into account besides the platelet count. Male smokers' serum VEGF concentrations were higher than non-smokers'. Smoking in men significantly affected the sex difference in the serum VEGF concentration, leukocyte count, and HDL-cholesterol concentration. We concluded that the serum VEGF concentration might be closely related to atherosclerosis accelerating factor, especially in men. © 2006 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.atherosclerosis.2006.07.025

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PubMed

Language：English   Publisher：Journal of Biomedical Materials Research - Part B Applied Biomaterials  

We reported earlier that hydroxyapatite (HA) formed on/in agarose gels (HA/agarose) produced by alternate soaking process is a bone-filling material possessing osteoconductive and hemostatic effects. This process could allow us to make bone-like apatite that was formed on/in organic polymer hydrogel matrices. Here, we investigated the mechanism of hemostasis induced by HA/agarose and found that HA/agarose, but not agarose or HA powder, significantly shortened activated partial thromboplastin time (APTT). While HA/agarose did not show significant platelet aggregation, it markedly enhanced adenosine diphosphate (ADP)-induced platelet aggregation. Moreover, Western blot analysis revealed selective adsorption of vitronectin onto HA/agarose. We also observed marked differences between HA powder and HA/agarose in their XRD patterns. The crystallinity of HA powder was much higher compared to that of HA/agarose. Furthermore, 50-100 nm of tube-form aggregations was observed in HA powder on the other hand 100-200 nm of particles was observed in HA/agarose by SEM observation. Thus 100-200 nm of low crystallized particles on the surface structure of HA/agarose may play an important role in hemostasis. Our results demonstrated a crucial role of HA/agarose in the mechanism of hemostasis and suggested a potential role for HA/agarose as a bone-grafting material. © 2006 Wiley Periodicals, Inc.

DOI： 10.1002/jbm.b.30684

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PubMed

Language：English   Publisher：Journal of Thrombosis and Haemostasis  

Background: Sepsis is a life-threatening disorder resulting from systemic inflammatory and coagulatory responses to infection. High-mobility group box 1 protein (HMGB1), an abundant intranuclear protein, was recently identified as a potent lethal mediator of sepsis. However, the precise mechanisms by which HMGB1 exerts its lethal effects in sepsis have yet to be confirmed. We recently reported that plasma HMGB1 levels correlated with disseminated intravascular coagulation (DIC) score, indicating that HMGB1 might play an important role in the pathogenesis of DIC. Objectives: To investigate the mechanisms responsible for the lethal effects of HMGB1, and more specifically, to explore the effects of HMGB1 on the coagulation system. Methods: Rats were exposed to thrombin with or without HMGB1, and a survival analysis, pathologic analyses and blood tests were conducted. The effects of HMGB1 on the coagulation cascade, anticoagulant pathways and surface expression of procoagulant or anticoagulant molecules were examined in vitro. Results: Compared to thrombin alone, combined administration of thrombin and HMGB1 resulted in excessive fibrin deposition in glomeruli, prolonged plasma clotting times, and increased mortality. In vitro, HMGB1 did not affect clotting times, but inhibited the anticoagulant protein C pathway mediated by the thrombin-thrombomodulin complex, and stimulated tissue factor expression on monocytes. Conclusions: These findings demonstrate the procoagulant role of HMGB1 in vivo and in vitro. During sepsis, massive accumulation of HMGB1 in the systemic circulation would promote the development of DIC. © 2007 International Society on Thrombosis and Haemostasis.

DOI： 10.1111/j.1538-7836.2006.02255.x

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PubMed

Language：Japanese   Publisher：(公社)日本人間ドック学会  

目的:従業員の定期健康診断で生活習慣の改善を指示された人を対象に、ステップ台を用いた8週間の運動プログラムを実施し、その前後での運動能力や体重、腹囲、総コレステロール値、血圧などを測定することでベンチステップ運動の有効性を検討する。方法:対象者33名を無作為に2群に振り分け、運動群には運動負荷試験により決定した乳酸閾値に相当する台高と昇降頻度でのベンチステップ運動(踏み台昇降)を行った。運動は10分連続を1単位として1日3単位、週21単位の運動を指示し、試験期間中の食事指導や食事制限は行わなかった。結果:運動群では8週間の試験終了後に運動能力の有意な向上のほか、腹囲や総コレステロール値に有意な改善が認められた。予定されたプログラム(週21単位)の完全実施率は11.8%のみであり、週14単位以上実施した人を合わせても42.2%の実施率であった。しかし週14単位以上実施した人では有意な体重減少が認められ、高血圧の人も全例に血圧の低下が認められた。結論:乳酸閾値でのステップ運動を8週間継続することで腹囲や高コレステロール血症者の総コレステロール値の有意な低下が示された。またこの運動を週14単位以上行うことで体重減少や血圧低下も認められ、生活習慣病を予防・改善するための運動量としては乳酸閾値でのステップ運動を1日2単位、週14単位以上行うことが推奨される。(著者抄録)

Language：Japanese  

Language：English   Publisher：FEBS Letters  

Cepharanthine (CEP), a biscoclaurine alkaloid, has been reported to induce cell death, however, the molecular mechanism of this phenomenon remains unclear. We herein report that CEP induced apoptosis in HuH-7 cells through nuclear fragmentation, DNA ladder formation, cytochrome c release, caspase-3 activation and poly-(ADP-ribose)-polymerase cleavage. CEP triggered the generation of reactive oxygen intermediates, the activation of mitogen activated protein kinase (MAPK) p38, JNK1/2 and p44/42, and the downregulation of protein kinase B/Akt. Antioxidants and SP600125, an inhibitor of JNK1/2, but not inhibitors of p38 MAPK and MEK1/2, significantly prevented cell death, thus implying that reactive oxygen species and JNK1/2 play crucial roles in the CEP-induced apoptosis of HuH-7 cells. © 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.febslet.2005.12.048

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PubMed

Language：English   Publisher：FEBS Letters  

Anandamide (AEA) exhibits anti-inflammatory effects. However, its role in the periodontal field remains unknown. Here, we found that gingival crevicular fluid contained a detectable level of AEA. The cannabinoid receptors CB1 and CB2 were expressed by human gingival fibroblasts (HGFs), and markedly upregulated under pathological conditions. AEA significantly reduced the production of pro-inflammatory mediators (IL-6, IL-8 and MCP-1) induced by Porphyromonas gingivalis LPS in HGFs, and this effect was attenuated by AM251 and SR144528, selective antagonists of CB1 and CB2, respectively. Moreover, AEA completely blocked LPS-triggered NF-κB activation, implying that AEA may regulate hyperinflammatory reactions in periodontitis. © 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.febslet.2005.12.079

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PubMed

Language：Japanese   Publisher：Japan Society of Ningen Dock  

For the prevention or improvement of lifestyle related disease an 8-week exercise program involving a bench step exercise was arranged. Methods: Thirty-three employees who had been recommended to change their lifestyle following a routine health check were enrolled. Before and after the program, their physical performance was evaluated, and their waistline, total cholesterol value and blood pressure was recorded and compared. There were no dietary restrictions. The 33 subjects were randomly divided into two groups: an exercise group and a control group. The exercise group was instructed to execute a bench-stepping exercise over ten consecutive minutes three times a day (i. e. twenty-one times a week) at a bench height and stepping rate that varied depending on each individual's lactate threshold as determined by an exercise stress test. Results: Only 11.8% of the participants completed the step exercise program as planned (21 times/week), but 42.2% executed it 14 or more times a week. These participants showed not only an improved physical performance, but experienced a significant reduction in weight, waistline size and total cholesterol value. Furthermore, the blood pressure of those participants suffering from hypertension was lowered. Conclusion: The step exercise over 8weeks at the lactate intensity threshold leads to a significant reduction in waistline, total cholesterol value and blood pressure. We recommend executing the step exercise at the lactate intensity threshold more than twice a day (i. e.14 or more times/week).

DOI： 10.11320/ningendock2005.21.854

Language：Japanese   Publisher：Conference Proceedings - IEEE International Conference on Systems, Man and Cybernetics  

We used nonlinear methods of Detrended Fluctuation Analysis (DFA) and Recurrence Quantification Analysis (RQA) to analysis of plethysmograms and their peak intervals in subjects with normal and pathological cardiovascular functions. Our results on HRV using DFA showed that scaling exponent indicated quantitatively differentiations among normal and pathologies. The measures of RQA were found to provide an unambiguous and quantitatively characterization of the plethysmogram time series in different cases. Accordingly, the integrations of these two analyses had the ability to identify and quantify normal and pathological cases, implying a possible clinical application. © 2006 IEEE.

DOI： 10.1109/ICSMC.2006.385312

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Language：Japanese  

Language：Japanese  

Language：English   Publisher：Thrombosis Research  

Activated protein C (APC) is an important natural anticoagulant that is proteolytically generated from protein C (PC) by the modulation of thrombin activity in the presence of thrombomodulin on an endothelial surface. Recent studies have demonstrated that, beyond its anticoagulant acitivities, APC had anti-inflammatory and cytoprotective properties. The mechanisms underlying APC's anti-inflammatory effects remain unknown. Our goal was to elucidate and confirm these mechanisms. We first examined the effect of APC on reactive oxygen species (ROS) and inflammatory cytokine production in murine macrophage-like RAW264.7 cells. We further examined the effect of APC on chemically induced lipid peroxidation and advanced glycation end-products (AGE) formation. APC in the range of 10-50 μg/mL could reduce lipopolysaccharide (LPS)-induced ROS generation, nuclear factor κB (NF-κB) activation and resultant proinflammatory cytokine production. Additional cell-free experiments revealed that APC (10-50 μg/mL) had inhibitory effects on lipid peroxidation and AGE formation. These findings suggest that APC, via its intrinsic anti-oxidant properties, may, in settings of oxidant stress, exert important cytoprotective and anti-inflammatory effects that are distinct from its anticoagulant activity as an antioxidant protein. If that is true, APC may contribute to ROS-related chronic disorders including atherosclerosis and diabetes as well as acute shock conditions. © 2004 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.thromres.2004.09.011

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PubMed

Language：English   Publisher：Pathophysiology of Haemostasis and Thrombosis  

Thrombin, a serine protease that plays a pivotal role in blood coagulation, wound healing, and angiogenesis, has also been implicated in the mitogenesis of various cell types. Previously, we showed that thrombin and the thrombin receptor agonist peptide (TRAP-14; SFLLRNPNDKYEPF) for protease-activated receptor 1 (PAR1) induce vascular endothelial growth factor (VEGF) secretion in PC-12 cells. In this study, we show that thrombin and TRAP-14 also stimulate VEGF secretion in the human NB-1 neuroblastoma cells. In these cells, we further show that thrombin-induced VEGF secretion was blocked by cycloheximide and actinomycin D, indicating that de novo protein synthesis is essential for this process. Reduced thrombin-induced VEGF secretion upon treatment with LY294002, calphostin C, or BAPTA, further suggests that the process is dependent on phosphatidyl-inositol-3-kinase, protein kinase C, and calcium. However, the complete loss of thrombin-induced VEGF production upon treatment with argatroban, a derivative of arginine and a potent anticoagulant/antithrombin agent, supports the notion that argatroban serves as a useful therapeutic tool for thrombin-associated pathologic conditions. Here, it appears that argatroban may be effective in controlling disorders linked to thrombin-induced VEGF production in neuronal cells. Copyright © 2005 S. Karger AG.

DOI： 10.1159/000088547

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PubMed

Language：English   Publisher：Clinical Cancer Research  

Islet cell tumors, endocrine neoplasm arising from pancreatic islets of Langerhans, are histologically difficult to diagnose as benign or malignant. Molecular markers are associated with the clinical characteristics that most of insulinoma are usually benign tumors, whereas other islet cell tumors are malignant but have not been identified. In this context, we newly found that an endothelial anticoagulant thrombomodulin was expressed in the normal islet β cells and insulinoma, but not of other islet components or non-insulinoma islet cell tumors. Clinically, all of the subjects (n = 15) of the insulinoma group showed no metastasis together with thrombomodulin expression in the lesions, whereas the other islet cell tumor groups showed a high incidence of metastasis (82%) and a low expression rate of thrombomodulin (6%). To examine the functional role of thrombomodulin, especially regarding the clinical characteristics of islet cell tumors, we tested the effect of exogenous thrombomodulin overexpression on cell adhesiveness and proliferation using MIN6 insulinoma cell line. In cell-based experiments, thrombomodulin overexpression reduced cell proliferation and enhanced Ca2+-independent cell aggregation, possibly through direct interaction with neural cell adhesion molecule. Taken together, these results are suggesting that thrombomodulin may act as antimetastatic molecule of insulinomas. In addition, thrombomodulin is a clinically useful molecular marker not only for identifying β-cell-origin islet cell tumors (i.e., insulinomas) but also for predicting disease prognosis of islet cell tumors.

DOI： 10.1158/1078-0432.CCR-03-0750

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PubMed

DOI： 10.1007/BF02256104

PubMed

Language：Japanese   Publisher：Journal of Biomedical Science  

Plasma nitric oxide (NO) levels in Duchenne muscular dystrophy (DMD) patients were significantly lower than those observed in both healthy controls and in patients with other neuromuscular disorders. The correlation between NO level and ejection fraction was significant (r = -0.384, p = 0.0391) in the DMD group. Disruption of NO systems may contribute to the development of muscular dystrophy and have implications for therapeutic strategies. Copyright © 2004 National Science Council, ROC and S. Karger AG, Basel.

DOI： 10.1159/000077905

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Language：English   Publisher：Shock (Augusta, Ga.)  

Acute hypotension, hypoxemia, cardiac arrhythmias, cardiac arrest, (or a combination of these), and sudden death are well-recognized complications of the cemented hip arthroplasty procedure. Collectively, these are known as the bone cement implantation syndrome (BCIS). The endogenous cannabinoids, anandamide (ANA) and 2-arachidonylglycerol (2-AG), are reported to be strong vasodilators and play a role in the hypotension associated with hemorrhagic and septic shock. In the present study, a potential role for the endogenous cannabinoids in influencing hemodynamic variables in BCIS was investigated. Thirty-five patients (35 hips) entered a prospective, randomized clinical trial. The patients were divided into two groups. Group 1 comprised 16 patients who had the component inserted using a conventional cementing technique, whereas group 2 consisted of 19 patients who had the femoral component inserted without cement. Blood samples were taken at six consecutive time points: before anesthesia, after reaming the femur, 2 min after insertion of stems with or without cement into the femur, and 10 min, 20, and 30 min after stem insertion. In group 1 (with cement), the mean levels of ANA and 2-AG significantly increased after stem insertion. In a comparison of each group after stem insertion, mean ANA and 2-AG levels in group 1 also significantly differed from those in group 2. By contrast, in group 2 (without cement) neither ANA nor 2-AG levels exhibited a significant increase or change at any point in time. In conclusion, we have shown for the first time that endogenous cannabinoids are candidates for lipid mediators of BCIS.

DOI： 10.1097/01.SHK.0000094766.36694.49

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PubMed

DOI： 10.1159/000075053

PubMed

Language：English   Publisher：Journal of Neurochemistry  

Ebselen, a selenium-containing heterocyclic compound, prevents ischemia-induced cell death. However, the molecular mechanism through which ebselen exerts its cytoprotective effect remains to be elucidated. Using sodium nitroprusside (SNP) as a nitric oxide (NO) donor, we show here that ebselen potently inhibits NO-induced apoptosis of differentiated PC12 cells. This was associated with inhibition of NO-induced phosphatidyl Serine exposure, cytochrome c release, and caspase-3 activation by ebselen. Analysis of key apoptotic regulators during NO-induced apoptosis of differentiated PC12 cells showed that ebselen blocks the activation of the apoptosis signaling-regulating kinase 1 (ASK1), and inhibits phosphorylation of p38 mitogen-activated protein kinase (MAPK) and c-jun N-terminal protein kinase (JNK). Moreover, ebselen inhibits NO-induced p53 phosphorylation at Ser15 and c-Jun phosphorylation at Ser63 and Ser73. It appears that inhibition of p38 MAPK and p53 phosphorylation by ebselen occurs via a thiol-redox-dependent mechanism. Interestingly, ebselen also activates p44/42 MAPK, and inhibits the downregulation of the antiapoptotic protein Bcl-2 in SNP-treated PC12 cells. Together, these findings suggest that ebselen protects neuronal cells from NO cytotoxicity by reciprocally regulating the apoptotic and antiapoptotic signaling cascades.

DOI： 10.1046/j.1471-4159.2003.02096.x

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PubMed

Language：English   Publisher：Journal of Thrombosis and Haemostasis  

Background: Vascular endothelial growth factor (VEGF) is an endothelial cell-specific potent mitogen that induces angiogenesis and microvascular hyperpermeability. Recently, it has been reported that megakaryocytes and platelets contain VEGF in their cytoplasm. Objectives: To elucidate and confirm the bioactivity and role of VEGF in platelets (platelet VEGF), which may be closely related to vascular thrombosis and atherosclerosis. Methods: The VEGF localization in mega-karyocytes on bone marrow smears was analyzed by immuno-fluorescence and confocal laser scanning microscopic analysis. The intracellular VEGF expressed in platelets was determined by flow cytometric analysis. Platelet-rich plasma and washed platelets were used to analyze the secretion of VEGF during platelet aggregation by thrombin or gelatinase A (matrix me-talloproteinase-2) stimulation. Immunohistochemical studies for VEGF in the thrombotic region were performed. Results and conclusions: Megakaryocytes and platelets are a very rich source of circulating VEGF. Gelatinase A, which is closely associated with vascular remodeling, enhances the VEGF levels released from platelets. VEGF was clearly detected in the fibrin nets of a thrombus. Taken together, platelet VEGF is bioactive as a direct angiogenic growth factor, and may play a very important role in wound healing and atherosclerosis in con-junction with other platelet cytokines such as platelet-derived growth factor, platelet-derived endothelial cell growth factor, transforming growth factor (TGF)-α, and TGF-β. © 2003 International Society on Thrombosis and Haemostasis.

DOI： 10.1046/j.1538-7836.2003.00475.x

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PubMed

Language：English   Publisher：The Japanese Society of Internal Medicine  

We report a case of a 62-year-old man diagnosed as Crow-Fukase syndrome (POEMS syndrome), in which the serial platelet count and vascular endothelial growth factor (VEGF) concentration were determined before and during the state of disseminated intravascular coagulation (DIC). The serum VEGF concentration was noted to be gradually decreased prior to DIC, after which it abruptly decreased with a corresponding drop in platelet count upon the onset of DIC. The physiological effects of VEGF are viewed as one of the causative factors in DIC and its abrupt and excessive release may have caused the exacerbation of the patient's clinical symptoms.<br>(Internal Medicine 42: 1240-1243, 2003)

DOI： 10.2169/internalmedicine.42.1240

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PubMed

Language：Japanese   Publisher：(株)自然科学社  

Language：English   Publisher：Hepatology  

The endogenous cannabinoid anandamide, a lipid mediator, induces various physiologic events such as vascular relaxation, inhibition of gap-junctions formation, tumor proliferation, neurologic analgesia, and apoptosis. Although increased concentration of anandamide in plasma has been implicated in pathophysiologic states including endotoxin-induced hypotension, the effects of anandamide on hepatocytes still remain unclear. In this study, we present evidence that plasma anandamide concentration is highly increased in severe hepatitis and cirrhosis patients. In addition, concentrations of anandamide within the pathophysiologic range potently induced apoptosis of hepatoma cell line (Hep G2) and primary hepatocytes, suggesting a possible link between increased anandamide level and hepatocyte damage. Anandamide-induced cell death was preceded by G0/G1 cell-cycle arrest, activation of proapoptotic signaling (i.e., p38 MAPK and JNK), and inhibition of antiapoptotic signaling (i.e., PKB/Akt) pathways. Moreover, anandamide increased susceptibility to oxidative stress-induced hepatocyte damage. In this context, methyl-β-cyclodextrin (MCD), a membrane cholesterol depletor, or mevastatin, an HMG-CoA reductase inhibitor, or N-acetyl cysteine, an antioxidant, potently inhibited the anandamide-induced proapoptotic events and cell death, whereas putative cannabinoid receptor antagonists did not exhibit an inhibitory effect on anandamide-induced cell death. Furthermore, binding assay using polymyxin beads revealed that anandamide could interact with cholesterol. In conclusion, our data suggest that cholesterol present in the cell membrane determines the fate of hepatocytes exposed to anandamide, possibly functioning as an anandamide receptor.

DOI： 10.1053/jhep.2003.50459

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PubMed

DOI： 10.11477/mf.1431100337

Language：English   Publisher：Acta Neuropathologica  

We examined the capillaries in muscle biopsy specimens from two patients with Ullrich's disease with collagen VI deficiency by light and electron microscopy. Collagen VI plays an important role in platelet aggregation for binding von Willebrand factor. Using immunohistochemistry, collagen VI was shown to be absent on capillaries from patients with Ullrich's disease, while von Willebrand factor, collagen IV, and vascular endothelial growth factor were normally expressed. Electron microscopy revealed narrow lumens, large nuclei in endothelial cells, and fenestration of a capillary. The number of pinocytotic vesicles per unit endothelial cytoplasm was increased. The cytoplasm of endothelial cells was strongly stained with uranyl acetate and lead citrate. Replication of the capillary basement membrane was observed. On the other hand, easy bleeding and coagulation were not observed in the two patients. These findings suggested that the collagen VI deficiency might have caused the electron microscopic changes of capillaries, while the function of the capillaries is apparently retained.

DOI： 10.1007/s00401-003-0714-1

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PubMed

Language：English   Publisher：Cellular and Molecular Life Sciences  

Thrombin, a serine protease generated by the activation of the blood coagulation cascade following vessel injury, induces vascular endothelial growth factor (VEGF) release. However, the molecular mechanism of thrombin-induced VEGF release is largely unknown. An agonist of protease-activated receptor-1 (PAR1), SFLL-RNPNDKYEPF, mimicked thrombin-induced VEGF release in human vascular smooth muscle (HVSM) cells, as determined by enzyme-linked immunosorbent assay, reverse transcriptase-polymerase chain reaction, and Northern blotting. In contrast, the agonist of PAR3, TFRGAP, did not affect VEGF release or expression. SFLL-RNPNDKYEPF, but not TFRGAP, up-regulated [Ca2+]i. Moreover, the calcium ionophone A23187 was found to trigger VEGF release in HVSM cells. Thrombin-induced VEGF release was blocked by anti-thrombin, heparin, a synthetic thrombin receptor inhibitor E5510, the calcium chelator BAPTA, the protein kinase C inhibitor calphostin C, and the MEK 1/2 inhibitor U0126. Thus, our data show that thrombin caused VEGF release via PAR1 activation in a manner dependent on [Ca2+]i and p44/42 downstream from the receptor activation.

DOI： 10.1007/s00018-003-3140-6

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PubMed

Language：Japanese   Publisher：Thrombosis and Haemostasis  

Anandamide (AEA), an endogenous cannabinoid, is generated by macrophages during shock conditions, and is thought to be a causative mediator of septic shock. Thus, we hypothesized that AEA plays a crucial role in endothelial cell (EC) injury. Here, we demonstrate that AEA induces apoptosis in a time- and dose-dependent manner in human umbilical vein endothelial cells (HUVECs). AEA triggered phosphorylation of c-Jun NH2-terminal kinase (JNK) and p38 mitogen activated protein kinase. AEA also showed a marked increase of interleukin Iβ-converting enzyme (ICE)CED-3 family protease (caspase-3) activity. AEA-induced EC death was inhibited by a selective vanilloid receptor I (VRI) antagonist, capsazepine, and was enhanced by a VRI agonist, capsaicin, indicating that AEA induces apoptosis in ECs via VRI. In conclusion, we propose that AEA may play a crucial role in EC injury under conditions of shock, and that the use of inhibitors of the AEA regulation system may have a therapeutic effect under these conditions.

DOI： 10.1055/s-0037-1613475

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Language：English   Publisher：Arthritis and Rheumatism  

Objective. High mobility group box chromosomal protein 1 (HMGB-1), a nuclear DNA binding protein, was recently rediscovered as a new proinflammatory cytokine. The purpose of this study was to demonstrate HMGB-1 expression in vivo and to identify the role of HMGB-1 in the pathogenesis of rheumatoid arthritis (RA). Methods. HMGB-1 concentrations in synovial fluid (SF) and serum from RA and osteoarthritis (OA) patients were measured by immunoblot analysis. The protein's specific receptor, receptor for advanced glycation end products (RAGE), was examined in SF macrophages (SFMs). We measured levels of proinflammatory cytokines released by SFMs treated with HMGB-1 via enzyme-linked immunosorbent assay and used soluble RAGE (sRAGE) to block the release of tumor necrosis factor α (TNFα). Immunohistochemical analysis and immunofluorescence assay were employed to examine localization of HMGB-1 in RA synovium and its translocation in SFMs after TNFα stimulation. Results. HMGB-1 concentrations were significantly higher in SF of RA patients than in that of OA patients. SFMs expressed RAGE and released TNFα, interleukin-1β (IL-1β), and IL-6 upon stimulation with HMGB-1. HMGB-1 was found in CD68-positive cells of RA synovium, and TNFα stimulation translocated HMGB-1 from the nucleus to the cytosol in SFMs. Blockade by sRAGE inhibited the release of TNFα from SFMs. Conclusion. HMGB-1 was more strongly expressed in SF of RA patients than in that of OA patients, inducing the release of proinflammatory cytokines from SFMs. HMGB-1 plays a pivotal role in the pathogenesis of RA and may be an original target of therapy as a novel cytokine.

DOI： 10.1002/art.10859

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PubMed

Language：English  

DOI： 10.1159/000073088

PubMed

Language：English   Publisher：Acta Neuropathologica  

By electron microscopy, we examined the skeletal muscle from two patients with Ullrich's disease. One patient had a deletion in the collagen VI alpha 2 gene. The muscle biopsy specimens showed no collagen VI immunoreaction, while the expression of collagen IV was increased. Collagen VI is a microfibrillar protein in the extracellular matrix with cell adhesive properties, and collagen IV is a principal component of the basal lamina. Electron microscopy revealed unevenness, extension, and folding of the muscle plasma membrane, and showed thickening and overproduction of the basal lamina. The data show that type VI collagen is certainly one of the important extracellular matrix components maintaining the structural integrity of skeletal muscle, and a defect of the collagen VI protein causes abnormalities of the muscle plasma membrane, dystrophic muscle changes, and upregulation of collagen IV.

DOI： 10.1007/s00401-002-0522-z

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PubMed

Language：English  

DOI： 10.1159/000054727

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PubMed

Language：English   Publisher：The Japanese Society of Internal Medicine  

We encountered a patient with adult T-cell leukemia/lymphoma (ATL) which expressed neural cell adhesion molecule (NCAM). The tumor cells markedly infiltrated the central nervous system (CNS) during the course of the ATL. The patient died 20 months after disease onset, which was considered to be early in the course. During the invasion of the CNS, the surface phenotype of the peripheral blood ATL cells by flow cytometric analysis was CD2+, CD3+, CD4+, CD7-, CD8-, CD16-, NCAM (CD56)+, HLA-DR-. We speculate that the infiltration of ATL cells into the CNS was closely related to the expression of the NCAM in this patient.<br>(Internal Medicine 41: 34-38, 2002)

DOI： 10.2169/internalmedicine.41.34

PubMed

Language：English   Publisher：(一社)日本内科学会  

49歳男.鹿児島県生まれの日本人で健康診断で白血球増多症を指摘され,血清中のヒトT細胞白血病ウイルス抗体が陽性であった.MRIで脳の白質および灰白質にATL細胞のび漫性浸潤を認めた.末梢血中ATL細胞の表現型はCD2+,CD3+,CD4+,CD7-,CD8-,CD16-,NCAM(CD56)+,HLA-DR-であった.発症20ヵ月後に死亡した

Language：Japanese  

PubMed

Language：English  

Language：English   Publisher：Annals of Neurology  

Patients with Ullrich's disease have generalized muscle weakness, multiple contractures of the proximal joints, and hyperextensibility of the distal joints. Recently, we found a deficiency of collagen VI protein in two patients with Ullrich's disease. In this study, we detected a homozygous 26 bp deletion in exon 14 of the collagen VI alpha 2 gene (COL6A2) in one patient. This mutation causes a frameshift and a premature termination codon, and results in a truncated collagen VI alpha 2 chain. Our data suggest that at least some cases of Ullrich's disease result from recessive mutations in COL6A2.

DOI： 10.1002/ana.1120

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PubMed

Language：English   Publisher：ACADEMIC PRESS INC  

Anandamide (ANA) and 2-arachidonoylglycerol (2-AG), two endogenous cannabinoids, can be generated by activated macrophages and platelets, respectively, in the context of endotoxic shock, and are proposed to play a crucial role in the induction of the shock-related hypotension. Taking advantage of our recently discovered function of polymyxin B (PMB) binding to ANA and 2-AG, we developed a new method for measuring ANA and 2-AG by applying PMB-immobilized beads to selectively adsorb them in biological fluids, instead of organic solvent extraction. The eluate from beads can be directly fractionated by reverse-phase high-performance liquid chromatography (HPLC), and the fractionations corresponding to authentic ANA and 2-AG are collected and derivatized with fluorogenic reagent and subsequently quantified by HPLC with fluorometric detection. The calibration graphs of ANA and 2-AG; were linear over a range of 1 to 500 pmol/ml. The limits of detection for ANA and 2-AG were 20 and 50 fmol, respectively. Intraassay precision was 2.24-4.25 and 3.47-5.44%, and interassay was 4.05-6.14 and 4.92-7.28% for ANA and 2-AG, respectively. Using this method, we first determined a 4-fold and 3-fold higher level of ANA and 2-AG, respectively, in the sera of patients with endotoxic shock than in normal serum. This finding should help in elucidating the role of the endogenous cannabinoids in the hypotension of human endotoxic shock. This method is rapid, sensitive, and reliable for simultaneously quantifying ANA and 2-AG in biological fluids, and has potential for clinical usage. (C) 2001 Academic Press.

DOI： 10.1006/abio.2001.5015

Web of Science

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PubMed

Language：English  

DOI： 10.2337/diacare.24.5.891

PubMed

Language：English   Publisher：Neurology  

The authors quantitatively measured levels of matrix metalloproteinases (MMP), tissue inhibitor of metalloproteinases (TIMP), and vascular endothelial growth factor (VEGF) in blood samples of POEMS syndrome. Circulating levels of MMP-1, -2, -3, -9, and TIMP-1 were more increased in patients with POEMS syndrome than in patients with other neurologic disorders or in healthy controls. Serum levels of VEGF and TIMP-1 were strongly correlated with each other. Increased circulating levels of MMP-1, -2, -3, -9, and TIMP-1 may lead to a better understanding the pathogenesis of POEMS syndrome.

DOI： 10.1212/wnl.56.6.807

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PubMed

Publisher：株式会社医学書院  

DOI： 10.11477/mf.1542904637

Language：English  

DOI： 10.2169/internalmedicine.39.1101

PubMed

Language：English   Publisher：Muscle and Nerve  

We report a marked difference in concentration of vascular endothelial growth factor (VEGF) between serum and plasma in patients with Crow-Fukase syndrome (CFS). The serum/plasma VEGF levels in 4 CFS patients were 8,634/152, 5,203/176, 3,724/127, and 868/13 pg/ml, respectively. We also showed that platelets were a major source of this VEGF and that VEGF was released during platelet aggregation by physiological stimulation. It is suggested that in CFS, local VEGF concentration is markedly elevated by aggregation of platelets containing excessive physiological activities of VEGF. Our findings provide important information for developing more effective therapeutic trials. (C) 2000 John Wiley and Sons, Inc.

DOI： 10.1002/1097-4598(200007)23:7<1051::aid-mus7>3.0.co;2-v

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PubMed

Language：English  

DOI： 10.1016/s0022-510x(99)00303-2

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PubMed

Language：Japanese  

PubMed

Language：Japanese   Publisher：Ryōikibetsu shōkōgun shirīzu  

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Language：English   Publisher：The Japanese Society of Internal Medicine  

We report a case of Crow-Fukase (POEMS) syndrome associated with pulmonary hypertension (PH). In this case, the concentration of vascular endothelial growth factor (VEGF) was extremely high in the serum, and the levels of IL-1β, IL-6, TNF-α, and thiamine, which were thought in past reports to be mediators of PH in Crow-Fukase syndrome, were normal. After prednisolone therapy, PH disappeared with a dramatic decrease in serum VEGF. Our results suggest that VEGF is closely correlated with PH in Crow-Fukase syndrome.<br>(Internal Medicine 39: 1101-1104, 2000)

DOI： 10.2169/internalmedicine.39.1101

PubMed

Language：English   Publisher：The Japanese Biochemical Society  

The <i>a</i> subunit of coagulation factor XIII lacks a hydrophobic signal sequence for secretion from cells, while the <i>b</i> subunit has a typical signal sequence. To determine whether the <i>a</i> subunit can be synthesized and released, expression vectors containing the cDNA for either subunit were transfected into baby hamster kidney (BHK) cells. Western blotting analysis and gel filtration chromatography demonstrated that the recombinant <i>a</i> and <i>b</i> subunits (rXIII<i>a</i> and rXIII<i>b</i>) had the same molecular weights and subunit structures (<i>a</i>², <i>b</i>², and <i>a</i>²<i>b</i>²) as the native molecules. rXIII<i>a</i> was enzymatically active when activated by thrombin. Most rXIII<i>b</i> was secreted as measured by ELISA, while most rXIII<i>a</i> was detected in the cytosol by subcellular fractionation. Co-expression with rXIII<i>b</i> in the same cells did not promote the release of rXIII<i>a</i>. Treatment of the cells with brefeldin A, a potent inhibitor of protein transportation, blocked the secretion of rXIII<i>b</i>, although it had no effect on the release of rXIII<i>a</i>. Several drugs and heat stress induced the release of rXIII<i>a</i>, which correlated directly with that of cytoplasmic lactate dehydrogenase. These results suggest that the <i>a</i> subunit is released from cells as a consequence of cell injury, which is independent of the classical secretory pathway.

Language：English   Publisher：(公社)日本生化学会  

凝固因子VIIIaサブユニットは分泌の為の疎水性シグナル配列を欠き,bサブユニットは特有の配列を持つ.aサブユニットが合成・放出されるかを決める為,一方のサブユニットのcDNAを含む表現ベクターをBHK中に移植し,ウェスタンブロット分析とゲル濾過クロマトグラフィーから組換えa・bサブユニット(rVIIIaとrVIIIb)は同じ分子量とサブユニット構造を持つとわかった.rVIIIaはトロンビンで活性化され酵素活性をもつ.rVIIIbはELISAで測定すると分泌され,rVIIIaはsubcellular分割法により細胞ゾル中にみられる.rVIIIbとの共表現ではrVIIIaの放出は促進されず,蛋白輸送阻害剤brefeldin Aで細胞処理するとrVIIIb分泌は抑制されrVIIIaにはこうかがない.幾つかの薬と熱ストレスがrVIIIa放出を誘発し,細胞質のlactase dehydrogenaseの放出と直接関連がある.これよりaサブユニットは細胞傷害の結果放出され古典的分泌経路とは無関係であるといえる

Language：Japanese   Publisher：Journal of Biochemistry  

The a subunit of coagulation factor XIII lacks a Hydrophobic signal sequence for secretion from cells, while the b subunit has a typical signal sequence. To determine whether the α subunit can be synthesized and released, expression vectors containing the cDNA for either subunit were transfected into baby hamster kidney (BHK) cells. Western blotting analysis and gel filtration chromatography demonstrated that the recombinant a and b subunits (rXIIIa and rXIIIb) had the same molecular weights and subunit structures (a2, b2, and a2b2) as the native molecules. rXIIIa was enzymatically active when activated by thrombin. Most rXIIIb was secreted as measured by ELISA, while most rXIIIa was detected in the cytosol by subcellular fractionation. Co-expression with rXIIIb in the same cells did not promote the release of rXIIIa. Treatment of the cells with brefeldin A, a potent inhibitor of protein transportation, blocked the secretion of rXIIIb, although it had no effect on the release of rXIIIa. Several drugs and heat stress induced the release of rXIIIa, which correlated directly with that of cytoplasmic lactate dehydrogenase. These results suggest that the a subunit is released from cells as a consequence of cell injury, which is independent of the classical secretory pathway.

DOI： 10.1093/oxfordjournals.jbchem.a021336

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Language：English   Publisher：The Japanese Biochemical Society  

The <i>a</i> subunit of coagulation factor XIII lacks a hydrophobic signal sequence for secretion from cells, while the <i>b</i> subunit has a typical signal sequence. To determine whether the <i>a</i> subunit can be synthesized and released, expression vectors containing the cDNA for either subunit were transfected into baby hamster kidney (BHK) cells. Western blotting analysis and gel filtration chromatography demonstrated that the recombinant <i>a</i> and <i>b</i> subunits (rXIII<i>a</i> and rXIII<i>b</i>) had the same molecular weights and subunit structures (<i>a</i>², <i>b</i>², and <i>a</i>²<i>b</i>²) as the native molecules. rXIII<i>a</i> was enzymatically active when activated by thrombin. Most rXIII<i>b</i> was secreted as measured by ELISA, while most rXIII<i>a</i> was detected in the cytosol by subcellular fractionation. Co-expression with rXIII<i>b</i> in the same cells did not promote the release of rXIII<i>a</i>. Treatment of the cells with brefeldin A, a potent inhibitor of protein transportation, blocked the secretion of rXIII<i>b</i>, although it had no effect on the release of rXIII<i>a</i>. Several drugs and heat stress induced the release of rXIII<i>a</i>, which correlated directly with that of cytoplasmic lactate dehydrogenase. These results suggest that the <i>a</i> subunit is released from cells as a consequence of cell injury, which is independent of the classical secretory pathway.

Language：English   Publisher：Seminars in Thrombosis and Hemostasis  

Factor XIII consists of two catalytic and two noncatalytic b subunits. The gene for the a subunit is located on chromosome 6, the gene for the b subunit on chromosome 1. Both genes have been characterized. There are several different allelic forms of the a subunit in the normal population and some microheterogeneity for the b subunit. Most patients with congenital factor XIII deficiency lack the a subunit in plasma; few patients appear to have a complete lack of the b subunit. The genes from patients with factor XIII deficiencies were obtained and examined. Based on these analyses a new genetic classification for factor XIII deficiency is proposed: a deficiency of the a subunit (formerly termed type II), a deficiency of the b subunit (formerly known as type I), and a possible combined deficiency of both a and b subunits.

DOI： 10.1055/s-2007-999036

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PubMed

Language：English   Publisher：Journal of Clinical Investigation  

We studied the defect responsible for deficiency of the b subunit for factor XIII in the first known case of this condition. The patient is a compound heterozygote of two genetic defects: deletion of A-4161 at the acceptor splice junction of intron A, resulting in a loss of the obligatory AG splicing sequence; and, replacement of G-11499 by T in exon VIII, resulting in an amino acid substitution of Cys430 by Phe. To determine how the latter mutation impaired b subunit synthesis, recombinant b subunit bearing the mutation was expressed in BHK cells. The mutant as well as wild- type b subunit was synthesized by the cells. However, the apparent molecular weight of the mutant was slightly higher than those of the wild-type and plasma b subunits under nonreducing conditions, probably because of destruction of a disulfide bond. The mutant b subunit was secreted from the cells much less effectively than the wild type and remained susceptible to endoglycosidase H, indicating that it was not transported from the endoplasmic reticulum to the Golgi apparatus where the processing of oligosaccharides occurs. Immunofluorescence study suggested that the mutant protein was retained in the endoplasmic reticulum. These studies demonstrate that a Cys430-Phe mutation does not prevent the de novo synthesis of the b subunit, but alters the conformation of the mutant protein sufficiently to impair its intracellular transport, resulting in its deficiency in this patient.

DOI： 10.1172/JCI117744

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PubMed

Language：Japanese   Publisher：(有)科学評論社  

Publisher：ライフサイエンス出版(株)  

Language：English   Publisher：The Japanese Society of Internal Medicine  

We recently had the opportunity to study a 25-year-old male with both Ehlers-Danlos syndrome (EDS) and von Recklinghausen neurofibromatosis (VRNF). We describe the clinical manifestations of the case and discuss the probable pathomechanism of the combination of the two syndromes, with a review of the literature. As recent literature suggests that both syndromes are linked to chromosome 17, we conclude that their combination is not coincidental, but genetically linked.<br>(Internal Medicine 31 : 671-673, 1992)

DOI： 10.2169/internalmedicine.31.671

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PubMed

Language：Japanese   Publisher：医学図書出版(株)  

Language：Japanese   Publisher：(株)中外医学社  

DOI： 10.1002/hsr2.267

PubMed

Publisher：エルゼビア・ジャパン(株)  

症例は40歳代前半の男性で、HIVに感染しており、高熱、貧血、ALP高値、肺の嚢胞性病変、上胃部痛が持続していたため当院へ紹介され入院となった。当院の検査でMycobacterium avium菌血症、血漿中のHIV-1 RNAコピー数の高値、CD4陽性T細胞数の低値、血清中のB型肝炎ウイルス(HBV)のDNAおよびHBsAg陽性が示された。紹介元の医療施設で行われていた胃の腫瘍に対する生検の所見からはカポジ肉腫が示唆されていた。M.avium菌血症に対し抗抗酸菌治療を開始し、後には抗レトロウイルス療法(ART)を開始するも高熱は収まらなかった。ALP高値から胆管炎を疑いスルバクタム/セフォペラゾン配合剤を投与するも効果はみられなかった。その後も重症の炎症エピソードを繰り返し発症したが、投薬内容を特別に変えなくともそうした炎症エピソードは毎回自然消退していた。M.avium菌血症は当院初診から79日目まで間欠的に検出されており、治療を持続する必要性が示唆された。ART開始から5ヵ月後の時点ではHIV-1のRNAコピー数は依然として高値であったが11ヵ月後には著明に減少し、CD4陽性T細胞数にも回復が認められた。

Publisher：(公社)宮崎県医師会  

血糖測定会に参加した40歳未満の397名を対象として、若年者に潜在する食後高血糖者の割合、背景および関連因子について検討を行った。140mg/dl以上の食後高血糖者は397名中21名(5.3%)で、そのうち19名は過去に尿糖あるいは血糖値異常を指摘されたことがなかった。食後血糖正常群と比較して、食後高血糖群では、男性の割合、食後2時間未満での血糖測定割合が有意に高く、糖尿病の家族歴を持つ割合も高い傾向にあった。多変量解析では、食後2時間未満での血糖測定のみが食後高血糖と独立した関連を有していた。食後高血糖者のほとんどは過去に耐糖能異常を指摘されたことがなかったことから、若年男性の食後2時間未満の血糖測定は、食後高血糖のスクリーニングに寄与する可能性がある。(著者抄録)

Publisher：(一社)日本超音波検査学会  

目的:発作性心房細動のアブレーション治療では、肺静脈隔離術(PVI)が一般的であり、PVI後の再発予測因子として、組織ドプラ法を用いた全心房伝導時間(PA-TDI duration)が左房容積係数(LAVI)より有用であることが報告されている。我々の施設では、発作性心房細動の治療としてComplex fractionated atrial electrogramを指標としたアブレーション(CFAE ablation)を主に施行しているが、本法についての再発予測因子について検討された研究はない。今回我々は、CFAE ablation後の再発予測因子について検討を行った。対象と方法:対象は発作性心房細動で初回アブレーションを施行した160名中、PA-TDI durationの計測が可能であった107名を非再発群と再発群に分けて比較検討した。比例ハザード分析による多変量解析の検討を行い、予測因子となる可能性が示唆された因子については、Receiver Operating Characteristic curve(ROC曲線)による解析を行った。また、PA-TDI duration及びLAVIについては、CFAE ablation単独群とCFAE ablation+PVI群に分別して経過観察を行い心房細動再発との関連について検討した。結果:アブレーション前の心房細動再発予測因子に関して、再発群は非再発群と比較して左房径(LAD)(41.1±5.4mm vs.38.1±5.1mm、p<0.01)が有意に拡大を認めた。比例ハザード分析による多変量解析を行い、左房容量(LAV)、LADが心房細動再発の予測因子となる可能性が示唆されたが、ROC曲線による解析では、ROC曲線下の面積(AUC)が、LAVは0.609、LADは0.651と予測能としては低いという結果であった。また、CFAE ablation単独およびCFAE ablation+PVI両群ともアブレーション前のPA-TDI duration、LAVIは非再発群と再発群に有意差は認めなかったが、6ヵ月後のPA-TDI durationでは、両群とも再発群では、アブレーション前よりも有意に延長を認めた(CFAE ablation単独:142.9±18.5msec vs.155.4±17.6msec、p<0.01。CFAE ablation+PVI:135.6±20.3msec vs.155.1±21.8msec、p<0.01)。非再発群では、CFAE ablation単独例において、アブレーション前より有意に短縮を認めた(141.8±23.3msec vs.131.1±23.9msec、p=0.011)。LAVIでは、CFAE ablation単独及びCFAE ablation+PVI両群とも有意差を認めなかった。結語:アブレーション前の心房細動再発予測因子は、発作性心房細動に対するCFAE ablation後の再発予測因子とはなりえなかった。しかしながら、アブレーション前と比較して6ヵ月後のPA-TDI durationが延長すると、心房細動を再発しやすいことが示唆された。(著者抄録)

PubMed

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PubMed

DOI： 10.3892/etm_00000089

PubMed

DOI： 10.1016/j.bbrc.2009.01.084

PubMed

PubMed

Publisher：(一社)日本内科学会  

62歳男子.Crow-Fukase症候群があり,血管内凝固症候群(DIC)発症前から血小板数と血清中VEGF(血管内皮増殖因子)濃度を経時的に測定した.VEGF濃度はDIC発症に先行して徐々に減少し,発症後は血小板減少とともに急激に低下した

PubMed

Publisher：(一社)日本内科学会  

肺動脈収縮期血圧が62mmHgの肺高血圧を伴った49歳のCrow-Fukase症候群の1例で,血清VEGFが著しく高値であった.プレドニゾロン療法により肺動脈収縮期血圧は31mmHgに低下,血清VEGFも632pg/mlから320pg/mlに低下した

Language：Japanese   Publisher：(一社)日本臨床検査医学会  

マイクロRNA(miRNA)は細胞機能調節分子としての役割がクローズアップされ、さまざまな生理的、病的状態において重要な働きを持つことが証明されている。18〜22塩基の低分子非翻訳一本鎖RNA(small non-coding RNA)は、その役割や組織分布などからカテゴリー化されるようになり、がん関連miRNAはOnco-miRと呼ばれ、多くの腫瘍細胞の増殖能、転移能を増強していることが明らかになった。本稿では、細菌感染症、自然免疫を制御するmiRNAについて、以下の項目で最新の話題を紹介した。1)細菌感染による活性化マクロファージとmiRNA,2)細胞内寄生細菌感染による活性化マクロファージとmiRNA,3)レジオネラ菌感染とmiRNA,4)miR-218のレジオネラ菌感染における意味付け、5)Rictorとマクロファージ、6)細胞外分泌小胞のmiRNA。

Language：Japanese   Publisher：(一社)日本臨床検査医学会  

Language：Japanese   Publisher：(一社)日本臨床化学会  

Language：Japanese   Publisher：日本医療情報学会看護部会  

Language：Japanese   Publisher：(一社)日本血栓止血学会  

Language：Japanese   Publisher：(株)先端医学社  

DOACの副作用としての出血性イベントが少なからず存在する以上、ましてその中には生命を脅かす出血性イベントが含まれるのであるから患者に対する倫理の観点からもDOACを処方する際は、その効能のみならず、出血性イベントを発症した際の対応についてしっかり説明されるべきであろう。予後の改善を目的とした理想的な抗凝固療法の発展のためには、モニタリング法の確立と過度の抗凝固状態あるいは出血性イベント時における中和薬の存在が次世代の課題となるであろう。(著者抄録)

Language：Japanese   Publisher：(一社)日本血栓止血学会  

プロトロンビン時間(PT)の原型は1935年のQuickらの発表に遡る.PT反応に関わる凝固第VII因子(FVII),凝固第X因子(FX),プロトロンビンはCa2+を介してリン脂質膜上のphosphatidylserineに各々の分子のGla残基を介して結合して酵素複合体を形成するが,PTの標準化を困難にする要因の一つとして細胞膜リン脂質成分の組成比の多様性が挙げられる.細胞膜リン脂質成分を反応の場とした組織因子(TF)/FVIIa複合体によるFXの活性化反応,あるいはプロトロンビナーゼ複合体によるトロンビンの生成反応には完全型から不完全型の多様性が存在して,PTはそれらの総和としての結果である.また,Ca2+濃度は至適濃度より高くても低くても酵素複合体の構成を妨げる結果となり凝固時間は延長する.トロンビンは構造上,酵素複合体から遊離できる分子であり反応液中のアンチトロンビンによる不活化の効率もPT反応に影響する.(著者抄録)

Language：Japanese   Publisher：(一社)日本臨床化学会  

Language：Japanese   Publisher：(一社)日本血栓止血学会  

Language：Japanese   Publisher：(株)メディカルレビュー社  

血管(機能)を診ることの意義は、最も直接的に血栓症の発症を予測し、予防することにあると一般的には考えられている。しかしながら、血管の機能破綻は動脈硬化へのプロセスの機序以外にも、"過労死"に代表されるように日常的に存在することを理解すべきである。血管機能の破綻は血栓症の発症に代表されるが、発症の過程には側副血行路をはじめとした進化の賜物とも言える代償機構が働く。しかしながら、過労死に代表されるようなストレスによる血管機能低下に対する代償機構は脆弱であると考えられる。(著者抄録)

Language：Japanese   Publisher：(株)メディカルレビュー社  

Flow mediated dilatation(FMD)のコンセプトを基礎としてreactive hyperemia peripheral arterial tonometory(RH-PAT)が開発された。操作が簡便かつ高感度であり、非駆血側を内部コントロールとして標準化できることから、血管機能に対するいくつかの概念を確認することができた。それは、(1)血管内皮機能は健常人において多様性のあること、(2)血管内皮機能は日内変動を有すること、(3)日常における疲労感は血管内皮機能に影響すること、(4)血管内皮は疲労を感知するセンサーであること、であるRH-PATの開発により血管内皮機能の低下の機序は動脈硬化に至るプロセス以外にも存在することを可視化できたと言える。(著者抄録)

Language：Japanese   Publisher：(一社)日本臨床化学会  

Language：Japanese   Publisher：(株)先端医学社  

核のない細胞におけるアポトーシスの概念が血小板において報告され、そのプロセスはmolecular clockとして報告され血小板の寿命を規定する(default signaling system)。また、赤血球におけるアポトーシスはeryptosisとして報告されている。さらに、血小板は活性化とともに壊死のプロセスをとることが報告され、necrotic plateletsのsubpopulationは高いprocoagulant活性を示す。これらの観察は血栓の質の研究に新たな展開を期待させる。(著者抄録)

Language：Japanese   Publisher：(株)メディカルレビュー社  

血管内皮細胞増殖因子(VEGF-A)は血管新生作用・血管透過性作用を有するangiogenesis、vasculogenesisの鍵分子である。血中のVEGF-Aはそのほとんどが血小板に含まれており止血反応に続く創傷治癒の過程に重要な役割を果たすと考えられる。血小板は骨髄巨核球から産生されることから血小板のVEGF-Aの含量は骨髄機能を反映している可能性を考えている。これまでに血小板VEGF-Aが川崎病の病態と関連していること(Ueno K、et al.Br J Haematol、2010)、また再生不良性貧血において骨髄予備能を反映していること(Kodama、Y、et al.Int J hematol、2012)を報告してきた。血小板の含む増殖因子量の評価は骨髄機能を反映した病態を示す可能性があり、血小板機能における一次止血機能の評価とは別の側面であると考える。(著者抄録)

Language：Japanese   Publisher：(株)医学書院  

Language：Japanese   Publisher：医歯薬出版(株)  

血小板は止血機構における一次凝集反応に加えて免疫系にも深くかかわっている。その関与は循環血中のPAMPs・DAMPsを最前線で感知して自然免疫を活性化するものであり、血小板膜上にはToll様受容体が機能的に発現している。そして血小板と白血球との相互作用における炎症反応の増幅の観察は血小板の研究における大きな進歩であった。一方、核を有さない血小板にもアポトーシス、ネクローシスといったプログラム死のプロセスが存在して組織修復と炎症のバランスを巧みに調節している。これらの観察は血栓症の成因を解明するうえで重要であり、血栓症の治療開発における新しい分子標的を探す手がかりになると考える。(著者抄録)

Language：Japanese   Publisher：(一社)日本臨床化学会  

Publisher：(一社)日本臨床化学会  

Language：Japanese   Publisher：(一社)日本内科学会  

Language：Japanese   Publisher：(一社)呼吸研究  

我々生体内では、DNAやRNA、タンパクなど多くの分子が互いに結合や解離といった特異的な反応を絶えず行うことによって、生命が営まれている。これら生体分子同士の特異的な反応を利用し、特定の分子を検出・同定する実験方法が多く考えられてきた。DNA同士の結合を利用することでDNAを検出するサザンハイブリダイゼーションやDNA-RNAの結合を利用することでRNAを検出するノーザンハイブリダイゼーション、なかでもタンパクの検出においては、優れた特異性と多様性双方をあわせもつ点から免疫反応(抗原抗体反応)を用いた手法が多く開発されている。そのなかの代表的な方法の1つがELISAおよびマルチプレックスである。本稿では、これらの実験法について概説する。(著者抄録)

Language：Japanese   Publisher：(株)日本臨床社  

Language：Japanese   Publisher：(株)メディカルレビュー社  

＜論文のポイント＞[1]凝固・線溶系は健常人においても低次元のレベルで作動している。[2]血管は必ず老化する。[3]ずり応力により血管内皮細胞よりNO(一酸化窒素)が産生され血管は拡張する。[4]血管内皮細胞は疲労感を感知するセンサーと考えることができる。(著者抄録)

Language：Japanese   Publisher：(株)医学出版  

Language：Japanese   Publisher：(株)医学書院  

＜ポイント＞★トロンボプラスチンの概念は1800年代に遡る.★内因系凝固反応を構成する因子は欠損症・異常症の解析により発見されてきた.★APTTはダビガトラン,エドキサバンの治療域における薬効モニタリングにはならない.★APTTは出血傾向の診断法の1つであり,血栓傾向を予知するものではない.(著者抄録)

Language：Japanese   Publisher：(株)医薬ジャーナル社  

高齢化社会が加速するに伴い、QOL(quality of life)を著しく損なう血栓・塞栓症への予防対策は、年を追うごとに重要になってきている。特に心房細動患者の塞栓症発症のリスクは高く、日常の臨床において患者にとって容易かつ安全域の広い経口薬の開発が進められている。数々の国際的大規模臨床試験が着々と進行し、これらの薬剤がワルファリンに替わり血栓・塞栓症予防の主流になってくると思われる。(著者抄録)

Language：Japanese   Publisher：(一社)日本脳神経外傷学会  

Language：Japanese   Publisher：医歯薬出版(株)  

この数年、臨床における血液凝固系の学問領域に炎症とのかかわりにおいて、いくつかの新しい概念が導入されてきた。炎症の環境のなかで、核をもたない血小板が凝固系のイニシエーターである組織因子を翻訳発現できるとする報告や、血小板が単球と複合体を形成することにより単球の炎症活性を増幅する機構が報告されている。そして活性化された好中球によるNETs(neutrophil extracellular traps)の放出は、病原体に対する新しい生体反応と、好中球の壊死でもアポトーシスでもない新しい細胞死の概念(NETosis)をもたらした。(著者抄録)

Language：Japanese   Publisher：(一社)日本血栓止血学会  

Language：Japanese   Publisher：(一社)日本脳神経外傷学会  

Language：Japanese   Publisher：(一社)日本血栓止血学会  

急性炎症とは,生体にとっての非常事態体制である.感染を察知した免疫細胞や,損傷に伴って壊死した細胞は,High mobility group box 1 protein(HMGB1)を放出することによって周囲に非常事態宣言をする.これを受けた周囲の細胞は,炎症,免疫,組織修復のプログラムを立ち上げ,非常事態に対応しようとする.このように,HMGB1は生体防御的役割を担っているが,その一方で,大量に産生されて全身を循環するHMGB1は,全身性炎症反応症候群(SIRS)や播種性血管内凝固症候群(DIC)を引き起こし,致死的に働く.このHMGB1の全身化を防ぐ役割を担っていると考えられるのが,抗凝固タンパク質として知られるトロンボモジュリン(TM)である.TMはトロンビンと結合するとともにHMGB1とも結合し,トロンビン-TM複合体がHMGB1を分解して不活化する.TMは凝固反応や炎症反応を多面的に制御していると考えられる.(著者抄録)

Language：Japanese   Publisher：(株)メディカルレビュー社  

血小板は、止血のみならず炎症との関わりにおいて注目されている。炎症の環境の中で、核を持たない血小板が凝固系のイニシエーターである組織因子(TF)を翻訳発現できるとする報告や、遊走能を持たない血小板が好中球や単球と複合体を形成することにより遊走を可能とし、更には好中球や単球の炎症活性を増幅する機構が報告されている。そして血小板により活性化された好中球によるNETs(neutrophil extracellular traps)の放出と自壊は、好中球の壊死でもアポトーシスでもない新しい細胞死の概念である。(著者抄録)

Language：Japanese   Publisher：(株)中外医学社  

近年,血小板は止血のみならず,炎症との関わりにおいて注目されている.炎症の環境の中で,核をもたない血小板が凝固系のイニシエーターである組織因子を翻訳発現できるとする報告や,遊走能をもたない血小板が好中球や単球と複合体を形成することにより遊走を可能とし,さらには好中球や単球の炎症活性を増幅する機構が報告されている.そして血小板により活性化された好中球によるNETs(neutrophil extracellular traps)の放出と自壊は好中球の壊死でもアポトーシスでもない新しい細胞死の概念である.(著者抄録)

Language：Japanese   Publisher：鳥居薬品(株)  

近年、血小板は止血のみならず、炎症との関わりにおいて注目されている。炎症の環境の中で、核をもたない血小板が凝固系のイニシエーターである組織因子を翻訳発現できるとする報告や、遊走能をもたない血小板が好中球や単球と複合体を形成することにより遊走を可能とし、さらには好中球や単球の炎症活性を増幅する機構が報告されている。そして血小板により活性化された好中球によるNETs(neutrophil extracellular traps)の放出は好中球の壊死でもアポトーシスでもない新しい細胞死の概念である。(著者抄録)

Language：Japanese   Publisher：(株)医学書院  

Language：English   Publisher：弘前大学大学院医学研究科・弘前医学会  

Language：Japanese   Publisher：Molecular Psychiatry  

DOI： 10.1038/sj.mp.4002050

Scopus

Language：Japanese   Publisher：(株)トプコ出版部  

Language：Japanese   Publisher：Medical Hypotheses  

DOI： 10.1016/j.mehy.2006.10.025

Scopus

Language：Japanese   Publisher：(株)永井書店  

Language：Japanese   Publisher：FEBS Letters  

DOI： 10.1016/j.febslet.2006.02.046

Scopus

Language：Japanese   Publisher：FEBS Letters  

DOI： 10.1016/j.febslet.2006.01.067

Scopus

Language：Japanese   Publisher：(株)医薬ジャーナル社  

近年,血管内皮細胞機能検査として,血管の駆血開放後におこる拡張反応flow mediated dilatation(FMD)の測定が知られるようになった.本検査はアセチルコリンによる冠動脈の拡張反応と同じく一酸化窒素(NO)を介する機序とされるが,血管拡張の機序を考察する上で,血管内皮細胞依存/非依存,NO依存/非依存の区別を意識することが大切と思われる.また,FMD測定は内皮細胞依存性かつNO依存性の高い検査法であるが,交感神経活動の影響を含めたデータの再現性に問題が残されている.最近,これらの問題点を考慮したPeripheral Arterial Tonometry(PAT)が注目されつつあるのであわせて紹介する(著者抄録)

Language：Japanese   Publisher：Dermatology  

DOI： 10.1159/000075053

Scopus

Language：Japanese   Publisher：(株)医学書院  

血管内皮増殖因子(VEGF)の持つ生理活性の研究や高VEGF血症を引き起こすヌード・マウスの病理学的検討から,Crow-Fukase症候群における異常に高値のVEGFは,本症候群の種々の特徴的な病態と関連することが次第に明らかになってきた.一方,Crow-Fukase症候群は特徴的な臨床症状がそろう以前の早期の段階では,しばしば診断が困難なことがあり,診断基準の作成が重要である.Crow-Fukase症候群は依然として難治性の疾患であるが,最近,強力な化学療法と自己末梢血幹細胞移植の併用による緩解例の報告が見られるようになり,今後は我が国での検討も必要である

Language：Japanese   Publisher：(株)診断と治療社  

DOI： 10.2169/internalmedicine.41.34

Language：Japanese   Publisher：(株)メディカルレビュー社  

Language：Japanese   Publisher：(一社)日本神経学会  

Language：Japanese   Publisher：(株)中山書店  

Language：Japanese   Publisher：(株)メディカルレビュー社  

VEGF(血管内皮増殖因子)は分泌型の糖タンパク質であり,強力な血管透過性亢進作用をもち,血管新生の全過程との関わりが深い重要な血管新生因子である.低酸素状態でVEGF mRNAが誘導され,内皮細胞の増殖や管腔形成を促進する.また正常の血管新生のみでなく,腫瘍内血管新生,糖尿病網膜症,動脈硬化層,側副血行路,創傷治癒,慢性関節リウマチ等の病態の血管新生にも,VEGFは特に重要な役割を果たすとされている.Crow-Fukase症候群は,VEGFそのものが疾患と一次的に密接に関連している最も代表的な疾患である

Language：Japanese   Publisher：(株)中外医学社  

DOI： 10.1002/ana.1120

DOI： 10.1002/ana.1120

Language：Japanese   Publisher：医歯薬出版(株)  

Language：Japanese   Publisher：(株)日本臨床社  

Language：Japanese   Publisher：(株)医学書院  

VEGFは,血管内皮特異的増殖因子で分泌型の糖蛋白である.強力な血管透過性亢進作用を有し,血管新生の全過程とのかかわりが深い重要な血管新生因子である.低酸素状態でVEGF mRNAが誘導され,内皮細胞の増殖や管腔形成を促進する.また正常の血管新生のみでなく,腫瘍内血管新生,糖尿病性網膜症,動脈硬化層,側副血行路,創傷治癒,慢性関節リウマチ等の病態の血管新生にも,VEGFは特に重要な役割を果たすことが推定されている.抗VEGF抗体やアンチセンスVEGFを用いて病的血管新生を抑え,癌治療などへの試みもなされつつある.又,VEGFの作用を応用して,虚血性疾患への治療にと応用されつつある

Language：Japanese   Publisher：(一社)日本神経治療学会  

DOI： 10.1002/1097-4598(200007)23:7<1051::AID-MUS7>3.0.CO;2-V

DOI： 10.2169/internalmedicine.39.1101

DOI： 10.2169/internalmedicine.39.1101

DOI： 10.1002/1097-4598(200007)23:7<1051::AID-MUS7>3.0.CO;2-V

Language：Japanese   Publisher：(株)メディカルレビュー社  

Language：Japanese   Publisher：(有)科学評論社  

Language：Japanese   Publisher：(一社)日本血栓止血学会  

Publisher：(一社)日本検査血液学会  

凝固検査検体取扱い標準化ワーキンググループでは、院内検査用の凝固検査検体取り扱いのコンセンサスの英文改訂版作成と外注検査用コンセンサスに必要な実験に取り組んだ。院内検査用コンセンサスでは「血漿中残存血小板数が10×10^9/L未満」であれば高速遠心が許容できるかの検証実験を実施した。その結果turnaround time短縮のため、推奨(1,500xg)以上の高速遠心(3,500xg)を実施すると検体血漿中でトロンビン-アンチトロンビン複合体で検出される凝固活性が高まることを明らかにし、凝固検査検体取り扱いとして高速遠心を推奨できないことを追記した。この内容は英語版にも追記した。また外注検査用コンセンサスについては、家子らの報告をもとに長時間搬送MIMIC系での実験を行った。その結果、全血長時間搬送では、4℃静置での変化がより顕著で第VIII因子低下を伴うAPTT延長を認め、血友病と誤診されるレベルであった。凝固因子測定のほとんどを受託施設に頼る本邦の現状を考えると注意が必要である。なお、自施設用コンセンサスの認知度調査も最近報告され、それらを解析すると自施設用コンセンサスの周知には、院内検査室が保持する遠心分離機の複数化や技師会を通じて広いアナウンスの必要性などが改善方法として有用であることを確認した。(著者抄録)

Publisher：(株)南山堂  

Publisher：(株)メディカルレビュー社  

Publisher：(株)メディカルレビュー社  

Publisher：(株)医学書院  

＜文献概要＞血栓傾向を診断する最も代表的なバイオマーカーはDダイマー,フィブリノゲン・フィブリン分解産物(FDP)である.その考え方の基本は,"凝固が起これば線溶が起こる"という二次線溶反応の機序である.凝固反応の結果,フィブリンが生成されると,フィブリンに親和性の高いプラスミノゲンアクチベータとプラスミノゲンがこのフィブリンに結合して,フィブリン上でプラスミノゲンからプラスミンの生成が効率よく起こり,プラスミンはフィブリンを分解してDダイマーが生成される.

Publisher：Journal of Cancer  

DOI： 10.7150/jca.30696

Scopus

PubMed

Publisher：(一社)日本血栓止血学会  

＜ポイント＞・採血は、日本臨床検査標準協議会(Japanese Committee for Clinical Laboratory Standards:JCCLS)の「標準採血法ガイドライン第2版(GP4-A2)」に則って実施されなければならない。その上で結果の正しい解釈が可能である。・凝固系は「溶けない分子」への変換であり、線溶系は「溶ける分子」への再変換というダイナミックな反応系である。・凝固反応はカスケード(増幅)反応であり、カルシウムイオンを必要とする。・凝固系は血小板のリン脂質膜上、線溶系はフィブリンの分子上にて効率よくプロセスされる反応である。・日常臨床における凝固系検査の多くに凝固時間検査が用いられている。(著者抄録)

Publisher：(一社)日本臨床化学会  

Publisher：(一社)日本血栓止血学会  

Publisher：医歯薬出版(株)  

直接経口抗凝固薬(DOAC)の内服症例に対する凝固検査の際は、服薬からの時間・腎機能など全身状態の把握とともに、施設ごとの試薬のDOACに対する感受性を考慮に入れて結果を判断することが必要である。予後の改善を目的とした抗凝固療法を積極的に行うためには、過度の抗凝固状態あるいは残存凝固活性測定法の確立が必須の時代となったといえる。(著者抄録)

Publisher：(一社)日本血液学会-東京事務局  

凝固・線溶データを解釈する上で大切なことは「フィブリンは血液中に溶けない分子である」ことを再認識することであろう。凝固系は可溶性から不溶性への変換であり線溶系は不溶性から可溶性への再変換というダイナミックな反応系である。フィブリンは溶けないからこそ分子としての意味をもち,FDP・Dダイマーは溶けるからこそ分子としての意味をもつ。この大きな概念の中に凝固反応はカスケード(増幅)反応であること,カルシウムイオンを必要とすること,凝固系は血小板のリン脂質膜上,線溶系はフィブリンの分子上にて効率良くプロセスされる反応が含まれる。日常臨床における凝固系検査の多くに凝固時間検査が応用されている。その一つである凝固因子活性測定法(一段法)は生体内の凝固因子は過剰に存在していることがその基本原理である。DICの病態においてはプロテアーゼの活性化により血中に無数のペプチド断片が出現している。(著者抄録)

Publisher：(一社)日本血栓止血学会  

Publisher：(一社)日本臨床検査医学会  

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