全血流動下でのフォンビルブランド因子依存性血栓形成における組織因子の機能特性

Von Willebrand factor (VWF) plays an important role in mediating platelet adhesion and aggregation under high shear rate conditions. Such platelet aggregates are strengthened by fibrin-network formation triggered by tissue factor (TF). However, little is known about the role of TF in VWF-dependent thrombus formation under blood flow conditions. We evaluated TF in thrombus formation on immobilized VWF under whole blood flow conditions in an in vitro perfusion chamber system. Surface-immobilized TF amplified intra-thrombus fibrin generation significantly under both low and high shear flow conditions, while TF in sample blood showed no appreciable effects. Furthermore, immobilized TF enhanced VWF-dependent platelet adhesion and aggregation significantly under high shear rates. Neutrophil cathepsin G and elastase increased significantly intra-thrombus fibrin deposition on immobilized VWF–TF complex, suggesting the involvement of leukocyte inflammatory responses in VWF/TF-dependent mural thrombogenesis under these flow conditions. These results reveal a functional link between VWF and TF under whole blood flow conditions, in which surface-immobilized TF and VWF mutually contribute to mural thrombus formation, which is essential for normal hemostasis. By contrast, TF circulating in blood may be involved in systemic hypercoagulability, as seen in sepsis caused by severe microbial infection, in which neutrophil inflammatory responses may be active.博士（医学）・乙第1388号・平成28年11月24日© The Japanese Society of Hematology 2016The original publication is available at www.springerlink.comThe final publication is available at Springer via http://dx.doi.org/10.1007/s12185-016-2086-

Combination of epidermal growth factor and insulin is required for multicellular spheroid formation of rat hepatocytes in primary culture.

We showed that the combination of epidermal growth factor (EGF) and insulin is an essential supplement to Williams' #E medium for the formation of floating multicellular spheroids in primary culture of rat hepatocytes. Isolated hepatocytes assembled to form floating multicellular spheroids within 96 h through transient assembly of monolayer islands within the initial 24 h in dishes coated with liver-derived proteoglycans. However, the assembly of multicellular spheroids was severely suppressed in the absence of either EGF or insulin. The reduction of spheroid assembly was correlated with decreased attachment and subsequent decreased formation of monolayer islands within 24 h. The minimum amounts of EGF and insulin required for the formation of floating spheroids were 1 ng/ml and 0.4 microgram/ml, respectively. These results suggest that the enhancement of hepatocyte attachment provided by the combination of EGF and insulin during the early phase of culture is required for the formation of floating spheroids

A Case of Malignant Gastrointestinal Stromal Tumor of the Transverse Colon: Evaluation of Proliferation Activity

Here, we report a colonic gastrointestinal stromal tumor (GIST) in a middle-aged man above 40. A histologically similar second tumor was detected 2 years after initial surgery. The primary GIST, measuring 5.5 ? 6.0 cm, consisted of spindle tumor cells with a higher number of mitoses and Ki67 labeling index (about 20%) than those of the second tumor, implying a de novo GIST. These markers might be useful in evaluating malignant potential, as well as to differentiate between a de novo and a recurrent tumor in the colonic GIST

Mixed HCV Infection of Genotype 1B and Other Genotypes Influences Non-response during Daclatasvir + Asunaprevir Combination Therapy

Daclatasvir (DCV) + asunaprevir (ASV) combination therapy has become available for patients with hepatitis C virus (HCV) serogroup 1 infection. We studied the efficacy of this therapy by focusing on the factors associated with sustained virological responses (SVR) including resistance-associated variants (RAVs) and mixed infection of different HCV genotypes. We enrolled 951 HCV serogroup 1-positive patients who received this combination therapy at our hospital or affiliated hospitals. The presence of RAVs in non-structural (NS) regions 3 and 5A was analyzed by direct sequencing. HCV genotypes were determined by PCR with genotype-specific primers targeting HCV core and NS5B regions. SVR was achieved in 91.1% of patients. Female sex, age > 70 years, and RAVs were significantly associated with non-SVR (p<0.01 for all). Propensity score-matching results among the patients without RAVs regarding sex, age, and fibrosis revealed that mixed HCV infection determined by HCV NS5B genotyping showed significantly lower SVR rates than 1B-mono infection (p=0.02). Female sex and RAVs were significant factors associated with treatment failure of this combination therapy for patients with HCV serogroup 1 infection. Mixed HCV infection other than 1B-mono infection would be useful for predicting treatment failure

Royal Jelly Increases Hematopoietic Stem Cells in Peripheral Blood: A Double-Blind, Placebo-Controlled, Randomized Trial in Healthy Subjects

Objectives. Royal jelly (RJ), produced by honeybees, influences stem cell functions, such as pluripotency maintenance of mouse embryonic stem cells and prevention of aging-related muscle stem cell functional deterioration. Thus, we hypothesized that RJ administration has various health-promoting effects based on stem cells. However, its effects are unknown in humans. In this study, we have attempted for the first time to clarify whether the administration of RJ in humans affects stem cells. Materials and Methods. This randomized, double-blind, placebo-controlled study was performed on healthy subjects (n = 90) who received protease-treated RJ at a dose of 1200 mg/day or placebo daily for four weeks. Also, the participants with a low number of hematopoietic stem cells (HSCs) in peripheral blood were preferentially selected. HSC counts, endothelial progenitor cell (EPC) counts, blood cell counts in peripheral blood, cytokines in serum, and physical conditions were evaluated. Results and Conclusion. Eligible data from 86 subjects (placebo: 42, RJ: 44) who completed the study were analyzed. There were no significant differences between the two groups regarding the changes in peripheral HSC count (p=0.103), while diastolic blood pressure showed a significant improvement in the RJ group compared to that in the placebo group (p=0.032). The subgroup analysis excluded 14 subjects who complained of cold symptoms at baseline or within five days of the four-week study. The changes in the HSC populations were significantly higher in the RJ group than those in the placebo group (p=0.042). No adverse effects were observed in any of the groups. These results suggest that RJ administration affected the peripheral HSC count and may influence stem cell functions. Further research is needed to reveal the various health-promoting benefits of RJ based on stem cells

Usefulness of Methionine PET for predicting prognosis of patient with adenoc arcinoma in head and neck treated by carbon ion radiotherapy

PURPOSE:C-11 methionine is a tracer for PET that can be used to assess metabolism of amino acid in tumor cells. The purpose of this study is to assess the ability of C-11 methionine PET (MET-PET) to predict prognosis of patient with adenocarcinoma in head and neck treated by carbon ion radiotherapy (CIRT).METHOD AND MATERIALS:We studied consecutive 23 patients with proven adenocarcinoma in head and neck who had been referred to our hospital for CIRT. 15 patients were male and 8 patients were female. The average patient age was 61.9 years (range 40 to 82). Patients were followed for 5.1 to 89.8 months (mean: 31.9 months) after CIRT. We evaluated the lesion semi-quantitatively on the basis of the Tumor-to-Normal-tissue Ratio (TNR). We statistically analyzed the relationship between tumor MET uptake level at pre and post CIRT, recurrence or metastasis rate, and patient prognosis using Kaplan-Meier analysis method.RESULTS:There were significant relationships between the level of tumor activity before CIRT and patients prognosis, recurrence and metastasis rate (p=0.029, 0.043, and 0.014) from Kaplan-Meier analysis. The level of tumor activity after CIRT also significantly related to patients prognosis, recurrence and metastasis rate (p=0.049, 0.043, and 0.014). No statistically significant relationship was observed between tumor uptake change after CIRT and survival rate.CONCLUSIONS:MET-PET is useful to predict patients survival rate, recurrence and metastasis rate in patients with head and neck adenocarcinoma treated by CIRT.89th AAOMS Annual Meeting, Scientific Sessions and Exhibition in conjunction with the Japanese Society of Oral and Maxillofacial Surgeons and the Korean Association of Oral and Maxillofacial Surgeon

Usefulness of 11C-methionine-PET for predicting the efficacy of carbon ion radiation therapy for head and neck mucosal malignant melanoma

AbstractThe aim of this study was to determine whether l-methyl-[11C]-methionine (MET) positron emission tomography (PET) allows the prediction of outcomes in patients with head and neck mucosal malignant melanoma treated with carbon ion radiation therapy (CIRT). This was a retrospective cohort study involving 85 patients who underwent a MET-PET or MET-PET/computed tomography (CT) examination before and after CIRT. MET uptake in the tumour was evaluated semi-quantitatively using the tumour-to-normal tissue ratio (TNR). Local recurrence, metastasis, and outcome predictions were studied in terms of TNR before CIRT (TNRpre), TNR after CIRT (TNRpost), and the TNR change ratio. Kaplan–Meier curves revealed significant differences between patients with higher TNRpre values and those with lower TNRpre values in regard to local recurrence, metastasis, and outcome (log-rank test P < 0.0001 for all three). There were also significant differences in metastasis rates and outcomes between patients with higher and lower TNRpost values (log-rank test P = 0.0105 and P = 0.027, respectively). The Cox proportional hazards model revealed TNRpre to be a factor significantly influencing the risk of local recurrence (hazard ratio (HR) 29.0, P < 0.001), risk of metastasis (HR 2.67, P = 0.024), and the outcome (HR 6.3, P < 0.001). MET-PET or MET-PET/CT is useful for predicting the outcomes of patients with head and neck mucosal malignant melanoma treated with CIRT

C-11 Methionine Positron Emission Tomography and Prediction for Survival Rate in Patients with Head and Neck Adenoid Cystic Carcinoma Treated by Carbonion Radiotherapy

89th AAOMS Annual Meeting, Scientific Sessions and Exhibition in conjunction with the Japanese Society of Oral and Maxillofacial Surgeons and the Korean Association of Oral and Maxillofacial Surgeon