AIM:To evaluate the ability of dendritic cells (DCs) loaded with tumor lysate to initiate cell mediated immune responses by stimulating naive T cells, and the efficiency of activated T cells to kill autologous tumor cells in vitro. METHODS: The peripheral blood lymphocytes and monocytes were obtained from the advanced renal cell carcinoma patient by eonglutination method. The immature dendritic cells were generated in the presence of interleukin -4(IL-4) and granulocyte/macrophage colony-stimulating factor (GM-CSF) from monocytes of healthy individuals.These cells were pulsed with tumor lysate or not. Induction of tumor-specific cytotoxic T lymphocytes(CTLs) response by mature dendritic cells (mDCs) was evaluated by the CD95(Fas) expression assay through FCM and the cytotoxic assay a gninst autolognns human tumor cells. RESULTS: Human immature dendritic cells and T cells obtained from healthy donors were stimulated with tumor- pulsed dendritic cells. The immature dendritic cells were applied to the cytotoxicity assay a gainst target autologons tumor cells. The CD95 (Fas) expression, IFN-γ, and TNF -α secreted by the CTLs in tumor lysate-plused DC group were higher than those of other groups. The capacity of the CTLs to kill autolognns tumor cells was significantly different(P<0. 05). Antigen-specific DCs vaccine can induce T cells activation and proliferation, thus we can obtain higher proportion of tumor specific cytotoxic T cells(CTLs), and enhance the CTLs to secret IFN-γ and TNF-α. CONCLUSION: Our results indicate that monocyte-derived human dendritic cells pulsed with tumor lysate could in duce the specific antitumor effect against autologons tumors. This in vitro model offers a new and simple approach to the development of DC + CTL - based immunotherapy.

AIM:To evaluate the ability of dendritic cells (DCs) loaded with tumor lysate to initiate cell-mediated immune responses by stimulating naive T cells, and the efficiency of activated T cells to kill autologous tumor cells in vitro. METHODS: The peripheral blood lymphocytes and monocytes were obtained from the advanced renal cell carcinoma patient by conglutination method. The immature dendritic cells were generated in the presence of interleukin-4(IL-4) and granulocyte/macrophage colony-stimulating factor(GM-CSF) from monocytes of healthy individuals. These cells were pulsed with tumor lysate or not. Induction of tumor-specific cytotoxic T lymphocytes (CTLs) response by mature dendritic cells (mDCs) was evaluated by the CD95(Fas) expression assay through FCM and the cytotoxic assay against autologous human tumor cells. RESULTS: Human immature dendritic cells and T cells obtained from healthy donors were stimulated with tumor-pulsed dendritic cells. The immature dendritic cells were applied to the cytotoxicity assay against target autologous tumor cells. The CD95(Fas) expression, IFN-? and TNF-? secreted by the CTLs in tumor lysate-plused DC group were higher than those of other groups. The capacity of the CTLs to kill autologous tumor cells was significantly different(P

Objective To study the changes of tissue plasminogen activator (t-PA), plasminogen activator inhibitor 1 (PAI-1), and soluble thrombomodulin (sTM) induced by dengue 2 virus (D 2V) infection of vascular endothelial cells. Methods The effects of D 2V infection on the production of t-PA, PAI-1, and sTM of human umbilical cord vein cells were studied. Results D 2V infection significantly induced the secretion of sTM and t-PA but showed no such effects on PAI-1 of human endothelial cells. Antibody against IL-6 inhibited D 2V-induced t-PA production of endothelial cells. A close correlation between serum levels of IL-6 and t-PA was found in dengue hemorrhagic fever (DHF) but not in dengue fever (DF) patients. Conclusion IL-6 can regulate D 2V-induced t-PA production of endothelial cells, suggesting that endothelial cells can be the target for D 2V infection and that D 2V-induced t-PA, TM, and IL-6 production of endothelial cells may contribute to the pathogenic development of dengue haemorrhagic fever/dengue shock syndrome (DHF/DSS).

AIM: The aim of this study is to elucidate the effects of lipopolysaccharide (LPS) on tissue plasminogen activator(tPA) and plasminogen activator inhibitor type-1(PAI-1) expression and secretion in endothelial cells. METHODS: Cultured human umbilical vein endothelial cells (HUVECs) were induced by LPS for different times. Cell viability was then determined by cell counting kit-8. tPA and PAI-1 activities in the media were assayed by fibrin overlay and reverse fibrin autograph, respectively. Cytoplasmic RNA was prepared using the Trizol method and was assayed for PAI-I and tPA mRNA levels by reverse transcript-polymerase chain reaction(RT-PCR). RESULTS: LPS(10 mg/L) did not produce cell toxicity according to LDH determination in culture media. PAI-1 activity in LPS group was high (P0.05). CONCLUSION: LPS (10 mg/L) did not show signs of cell toxicity, but promoted the expression of PAI-1 mRNA and induced an increase activity of PAI-1. However, LPS (10 mg/L) did not change tPA mRNA expression. The time-dependent increase in PAI-1 mRNA expression and activity shifts the local balance toword increased anti-fibrinolytic capacity, which can amplify the extent of acute thrombosis after plaque rupture. This is one of the possible reasons that cause thrombus,blood coagulation and disseminated intravascular coagulation (DIC) during septicemia.

Objective To investigate the risk factors for postoperative liver failure of patients with hepatocellular carcinoma (HCC) and bile duct tumor thrombus through a risk evaluation model.Methods The clinical data of 107 patients with HCC and bile duct tumor thrombus who received hepatic resection at the Eastern Hepatobiliary Surgery Hospital from March 2002 to February 2011 were retrospectively analyzed.All patients were divided into the non-liver failure group (98 patients) and liver failure group (9 patients).Risk factors associated with liver failure were analyzed and a risk evaluation model was established.All data were analyzed using the bivariate regression model,and factors with significance were further analyzed using the multivariate regression model.Results Of the 107 patients,105 received hepatic resection + choledochotomy + thrombectomy and 2 received hepatic resection + extrahepatic bile duct resection + cholangiojejunostomy.The operation time was 2.0-5.5 hours,and the intraoperative blood loss was 200-3500 ml.In the non-liver failure group,5 patients had pleural and peritoneal effusion,3 had biliary bleeding,2 had incisional infection,1 had biliary infection,1 had bile leakage,1 had stress-induced ulcer of upper digestive tract and 1 had thoracic epidural hematoma.The bleeding of the patients with thoracic epidural hematoma was stopped after thoracic spinal decompression,but subsequent paraplegia occurred.In the liver failure group,2 patients died of postoperative acute liver failure,and 7 patients died of postoperative subacute liver failure (death caused by tumor recurrence or medicine was excluded).The results of univariate analysis showed that preoperative total bilirubin,albumin,pre-albumin,albumin/globulin ratio,distribution of tumor thrombus,operative blood loss and ratio of postoperative residual liver volume to the total liver volume were correlated with the postoperative liver failure in patients with HCC and bile duct tumor thrombus (OR =3.017,0.191,0.248,2.681,9.048,4.759,13.714,P ＜ 0.05).The results of multivariate analysis showed that preoperative total bilirubin ＞ 256.5 μmol/L,albumin/globulin ratio ≤ 1.3 and postoperative residual liver volume ＜ 50％ were the independent risk factors of postoperative liver failure (OR =5.537,11.107,172.450,P ＜ 0.05).The risk evaluation model was Z =1.77 × preoperative total bilirubin + 2.408 × preoperative albumin/globulin ratio + 5.150 × ratio of postoperative residual liver volume to the total liver volume-17.288.The risk of postoperative liver failure increased as the increase of Z value.The risk of postoperative liver failure ＞ 50％ when the Z value ＞ 0.Conclusions Preoperative total bilirubin ＞ 256.5μmol/L,albumin/globulin ratio ≤ 1.3 and postoperative residual liver volume ＜ 50％ were the independent risk factors of postoperative liver failure.Risk evaluation model is helpful in screening the risk factors so as to decrease the incidence of postoperative liver failure.

Objective To investigate risk factors of the prognosis of patients with severe fever with thrombocytopenia syndrome (SFTS).Methods From May 2012 to July 2014,17 cases of severe fever with thrombocytopenia syndrome in Renmin Hospital of Wuhan University were treated.Clinical data including history of epidemiology,clinical manifestations,complications,physical examination and laboratory test results on admission and the third day after admission were retrospectively analyzed and compared with the death group and recovery group by application of Spearman correlation analysis.Results Elderly male patients with neuropsychiatric symptoms,or abnormal liver function,or abnormal blood clotting function had higher risk of the poor prognosis.In SFTS patients,AST,ALT was significantly increased,AST 539 U/L (229.73,545.4) U/L (r =0.597,P =0.015) was a risk factor affecting prognosis.Elevated blood ammonia indicated serious liver dysfunction and neurological dysfunction which were manifested as irritability,delirium,and trembling limbs.In SFTS patients,platelets were significantly decreased accompanied with mouth ulcers / bleeding gums,gastrointestinal bleeding.PLT 24.88 × 10 9/L-1 (12.75,35.00) ×10 9/L-1 (r=0.557,P=0.005) or APTT 86.06 s (66.88,114.18) (r=0.798,P=0.001) or D-dimmer 9.79 mg / L (4.09,16.51) mg/L (r =0.597,P =0.015) are risk factors affecting poor prognosis.Conclusions On the third days after admission,AST,WBC,PLT,APTT,Ddimmer are risk factors for prognosis of patients with severe fever with thrombocytopenia syndrome infected by a novel bunyavirus.

AIM To study the effect of Vit E on the MLCK activity and expression in the liver of atherosclerosis model rabbits. METHODS The MLCK activity of rabbit liver was measured by the method of ?- 32P incorporated and its expression was detected by immunofluorescent. RESULTS The model of atherosclerosis was estabilished. After rabbits were fed with cholesterol for four weeks and twelve weeks, the activity of MLCK increased markedly, and there was significantly statistical difference compared with the normal control (P0.05). MLCK expression increased after the rabbits was fed with cholesterol for four weeks, and this increase became more obvious had been the rabbits was fed with cholesterol for twelve weeks. The expression decreased when vitamin E had been added into the cholesterol fed. CONCLUSION The pathology of liver may be associated with the increase of the activity of MLCK. Vit E may reduce MLCK activity and protect hepatocyte from injury.

Objective: To use autophagy inhibitors combined with radiation to treat the oral squamous cell carcinoma CAL-27 and KB cells,and to explore the influence of autophagy in the oral cancer radiation sensitivity and its mechanisms. Methods:The human oral squamous cell carcinoma CAL-27 and KB cells were divided into control group,CQ group,3-MA group,IR group,CQ+IR group,and 3-MA+IR group. The survival rate was detected by MTT method and the autophagy of CAL-27 cells was observed by immunofluorescence method and laser scanning confocal microscope.The expression levels of LC3 and beclin-1 were detected by Western blotting method. The apoptotic rate was determined by flow cytometry with Annexin Ⅴ/PI doulde staining. Results:Compared with IR group,the survival rates in 3-MA + IR and CQ+ IR groups were signifcantly decreased (P < 0.05 ).The autophagy levels of cells in IR group were significantly higher than those in control group, CQ group, 3-MA group,CQ+IR group,and 3-MA+IR group (P <0.05).The expression levels of LC3 and beclin-1 proteins in IR group were significantly higher than those in control group,CQ+ IR group,and 3-MA+ IR group (P < 0.05). The apoptotic rates in IR,3-MA+ IR,and CQ+ IR groups were markedly higher than those in control group. Compared with IR group,the apoptotic rates in CQ+IR and 3-MA+IR groups were significantly increased (P <0.05).Conclusion:Radiotherapy can induce the increase of autophagy level of oral squamous cell carcinoma cells. Inhibiors of autophagy can increase the radio-sensitivity of oral squamous cell carcinoma cells by inhibiting the proliferation and inducing the apoptosis.

Aim To study the effect of melatonin on expression and activity of myosin light chain kinase in the artery wall of atherosclerotic rabbits.Methods The rabbit model of atherosclerosis was induced by a high-cholesterol diet.The blood lipid levels were assayed in the serum of each group.MLCK expression was detected by Western blot and immunohistochemical method.MLCK activity was measured by ?-32P-ATP incorporation into myosin light chain.Results The atherosclerosis model was established successfully.The levels of lipids decreased after MLT treatment.After fed with cholesterol for twelve weeks,the expression and activity of MLCK in the artery of atherosclerotic rabbits increased markedly,whereas there was no obvious difference in expression of MLCK in the artery of atherosclerotic rabbits fed with cholesterol and melatonin for twelve weeks compared with that of control.Conclusions It was suggest that high expression and activity of MLCK in the artery might be closely correlated with the development of atherosclerosis.Melatonin played an important role in inhibiting the development of atherosclerosis by decreasing the expression and activity of MLCK.

Heart failure (HF) has become a major challenge in the treatment of global cardiovascular diseases. Great progress has been made in the drug treatment of HF, however, rehospitalization rate and mortality of patients with HF are still high. Hence, there is an urgent need to explore new treatment strategy and new underlying pathogenic mechanisms. In recent years, some researchers have suggested that regulation of ketone body metabolism may become a potentially promising therapeutic approach for HF. Some studies showed that the oxidative utilization of fatty acids and glucose was decreased in the failing heart, accompanied by the increase of ketone body oxidative metabolism. The enhancement of ketone body metabolism in HF is a compensatory change during HF. The failing heart preferentially uses ketone body oxidation to provide energy, which helps to improve the body's cardiac function. This review will discuss the potential significance of ketone body metabolism in the treatment of HF from three aspects: normal myocardial ketone body metabolism, the change of ketone body metabolism in HF, the effect of ketogenic therapy on HF and its treatment.
Humans ; Heart Failure/metabolism* ; Myocardium/metabolism* ; Ketone Bodies/metabolism* ; Cardiovascular Diseases ; Fatty Acids/metabolism* ; Energy Metabolism