OBJECTIVE:To investigate the regulation effects of oleanolic acid on the mitochondrial nitric oxide synthase(mt-NOS) in human umbilical vein endothelial cells (HUVECs) with oxidative damage. METHODS:HUVECs in exponential phase were divided into normal group,model group and oleanolic acid low,medium and high dose groups(5,20 and 35 μmol/L). After drug acting for 24 h,all groups were given culture solution containing 100 μg/ml ox-LDL to reproduce oxidative damage except normal group. CCK-8 was used to detect cell viability. The mitochondria in cells were extrated,enzyme chemical method was used to detect mtNOS activity and mtNO content,fluorescence microplate method was used to detect fluorescence intensity of reactive oxygen species(ROS),and western blot was used to detect expression of cytochrome C(Cyto-C). RESULTS:Compared with nor-mal group,the cell viability in model group was decreased;mtNOS activity,mtNO content,ROS fluorescence intensity and Cy-to-C protein expression were increased,with significant differences (P<0.05). Compared with model group,the cell viability in oleanolic acid low,medium and high dose groups was increased;mtNOS activity,mtNO content,ROS fluorescence intensity and Cyto-C protein expression were decreased,with significant differences(P<0.05),and they had positive correlation with concentra-tions. CONCLUSIONS:Oleanolic acid can decrease the mtNOS activity of HUVECs,reduce the production of mtNO and Cyto-C, by a mechanism that may be related to the decrease of ROS expression.

Objective To analyze the distribution and influential factors of exam results in medical students' general surgery theory course.Methods 171 medical students were selected as subjects by cluster sampling,and the distribution and influential factors of exam results were analyzed.SPSS 17.0 software was used for statistical analysis,measurement data with (-x) ± s,and normality test with Kolmogorov-Smirnov test.Those quantitative data which do not meet the normal distribution were compared with Mann-Whitney U and Kruskal-Wallis H test.Rank transformation univariate multi-factor variance of LSD (Levene test equal error variance between groups) or Tamhane method (Levene test range error variance between groups) were compared between two groups(3-4) and the influence factors of whether the grade was good was analyzed by single factor and multi factor non conditional Logistic regression model,with the test level of alpha=0.05.Results The distribution of total exam results was normal.77 score was outlier,and the scores of female students were higher than those of male students.Scores of total exam results,multiple-choice questions and essay questions in different classes were significantly different.Data from multivariate logistic regression analysis showed that male students(OR=0.212,95％CI:0.077-0.584) were unfavorable factor for good exam results,while higher scores in noun explanation (OR=12.160,95％CI:1.985-74.495),multiple-choice questions (OR=9.887,95％CI:2.997-32.617),essay questions(OR=18.323,95％CI:6.593-50.928) were favorable factors.Conclusion The cause analysis of score's outlier and sex difference should be strengthened,and the influence of examination items on score should be emphasized.

Aim To research the effects of total flavones of rhododendra(TFR)on transient receptor potential vanilloid receptor 4(TRPV4)in cerebral basilar arteries(CBA)of rats subjected to ischemia/reperfusion(IR)injury.Methods The model of total brain IR was established by four-artery occlusion(4-VO)method in rats.Arterial pressure perfusion and cell membrane potential recording methods were used for surveying the dilatation and hyperpolarization of TFR and ruthenium red(RR,an inhibitor of TRPV4)in the KCl-preconstricted CBA ex vivo in rats subjected to IR.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot were utilized to investigate the TRPV4 mRNA and protein expressions of TFR and RR in cerebrovascular endothelial cells of CBA in vivo in rats subjected to IR.Results 11～2 700 mg·L-1 TFR significantly induced concentration-dependent hyperpolarization and dilatation in the KCl-preconstricted CBA in rats subjected to IR.TFR still produced degenerative hyperpolarization and dilatation by removal of endothelium in CBA,which was remarkably attenuated as compared with endothelium-intact group(P<0.01).After removal of NO and PGI2 vasodilatation,TFR obviously elicited the hyperpolarization and dilatation that were further decreased by RR(an inhibitor of TRPV4)in IR CBAs.TFR pretreatment apparently increased the level of TRPV4 mRNA and protein expressions in IR CBAs.These effects were restrained by RR,an inhibitor of TRPV4.Conclusions TFR could mediate endothelium-dependent and endothelium-independent effects.The endothelium-derived dilatation may be related to the increase of endothelium activity and endothelium-derived hyperpolarizing factor(EDHF)generation and release that have been promoted by TFR,and secondarily activating TRPV4,which results in Ca2+ inflow and subsequent hyperpolarization of vascular smooth muscle cell membrane and vasorelaxation.

OBJECTIVE:To observe the correlation of vitamin D with the incidence of cardiovascular disease and risk factors in elderly patients with type 2 diabetes mellitus. METHODS:94 patients,aged 60 years old above,with type 2 diabetes mellitus were collected retrospectively. Those patients were divided into diabetes group(41 cases)and diabetes complicated with cardiovas-cular disease group(53 cases)according to the condition of disease. 25(OH)D levels and lab indicators of 2 groups were detect-ed. The relationship of cardiovascular lesion with risk factor was analyzed by Multivariate Logistic regression;the correlation be-tween 25(OH)D and risk factors was analyzed by pearson analysis. RESULTS:There was statistical significance in the levels of 25 (OH)D,TC,TG,UA and Ca between diabetes group and diabetes complicated with cardiovascular disease group,with statistical significance(P<0.05). pearson analysis showed that the level of 25(OH)D was negatively related to age,FIB,TC and UA,and positively related to D-dimer and HDL-C,especially correlated with FIB and TC significantly,with statistical significance (P<0.01). Multivariate Logistic regression analysis showed that the deficiency of 25(OH)D was pathogenic factors of type 2 diabetes mellitus. CONCLUSIONS:There is a correlation between the level of 25(OH)D and cardiovascular risk factors as age,FIB,TC, UA,D-dimer,HDL-C. Vitamin D deficiency is the independent risk factors for cardiovascular disease in elderly patients with type 2 diabetes mellitus.

Objective:Prostate cancer frequently metastasizes to the spine. In this study, we investigate the prognostic factors as-sociated with survival in patients with prostate cancer accompanied by spinal metastases at their preliminary diagnosis. Methods:Clin-ical data of 49 patients who were diagnosed with spinal metastasis from prostate cancer between January 2005 and December 2010 were analyzed. Variables including alkaline phosphatase (ALP), previous skeletal-related event, Gleason score, prostate-specific anti-gen (PSA) nadir, and time to castration resistance were obtained. Moreover, the relationship between these variables and overall sur-vival (OS) was analyzed. Survival analysis was performed by using Kaplan-Meier curves. Furthermore, the differences among the OS rates were assessed by using the log rank test. The variables were statistically significant in the univariate analysis (P<0.05) and were included in the multivariate model. Results:The average follow-up time was 64.1 months among the 49 patients. By the end of the follow-up, 41 of these patients were dead;the mean survival was 27 months. The 1-, 3-, and 5-year survival rate was 81.6%, 40.8%, and 20.4%, respectively. Univariate analysis identified that 6 variables were statistically significant prognostic factors of OS:with or without chemotherapy, ALP, previous skeletal-related event, Gleason score, PSA nadir, and time to castration resistance. The multivari-ate analysis showed that the time to castration resistance of ≥19 months and the addition of chemotherapy after disease progression are independent prognostic factors for a high OS. Conclusion:With or without chemotherapy and the time to castration resistance are the independent prognostic factors associated with survival in patients with prostate cancer accompanied by spinal metastases at first diagnosis.

OBJECTIVE:To study the effect and mechanism of physcion 8-O-β-glucopyranoside(PG)on the apoptosis of skin melanoma A375 cells. METHODS:After A375 cells were treated by PG with 0,10,20,50 μg/mL for 24,48,72 h,CCK-8 method was adopted to determine the survival rate of cells. After A375 cells were treated by PG with 0(control),20,50 μg/mL for 48 h,flow cytometry was used to detect the apoptosis rate of cells with membrane protein Ⅴ/propidium iodide (PI) double staining. Immunoblotting was used to detect the protein expressions of Caspase-3 and polyadenyl adenine diphosphate ribose poly-merase (PARP) and protein expressions of cytochrome C inside and outside mitochondria. After A375 cells were treated by PG with 0 (control),5,10 μmol/L for 48 h,enzyme substrate method was used to determine the activities of Caspase-8 and Cas-pase-9. RESULTS:PG can effectively decrease the survival rate of A375 cells. Compared with control,apoptosis rate of cells was obviously increased after treated by PG with 20,50 μg/mL(P<0.01);protein expressions of Caspase-3,PARP in cells and cyto-chrome C in cell matrix were obviously enhanced(P<0.05 or P<0.01);and protein expression of cytochrome C in mitochondria was obviously weakened(P<0.05 or P<0.01). Caspase-9 activity in cells was obviously enhanced after treated by PG with 5,10μmol/L(P<0.05 or P<0.01);and Caspase-8 activity had no obvious changes. CONCLUSIONS:PG can inhibit activity of A375 cells and promote its apoptosis,and its pro-apoptotic effects is achieved by destructing mitochondrial membrane potential and pro-moting cytochrome C outflow.

A wearable wireless health monitoring system for drug addicts in compulsory rehabilitation centers was proposed. The system can continuously monitor multiple physiological parameters of drug addicts in real time, and issue early warning information when abnormal physiological parameters occur, so as to play the role of timely medical practice. In addition, this study proposes a convolutional neural network (CNN)model, which can evaluate the health status of drug addicts based on multiple physiological parameters. Experiments show that the model can be applied to the task of body state recognition in the open physiological parameter data set, and the recognition accuracy can reach up to 100% in a single physiological parameter data set; when the whole physiological data set is used, the recognition accuracy can reach 99.1%. The recognition accuracy exceeds the performance of the traditional pattern recognition method on this task, which verifies the superiority of the model.
Algorithms ; Electrocardiography ; Monitoring, Physiologic ; Neural Networks, Computer ; Wearable Electronic Devices

Objective: To reduce the residual proteins and DNA of host cells in the preparation of H5N1 influenza A virus. Methods: Core 700 was firstly used to remove residual host cell proteins, and then Capto Q was used to remove host cell DNA. Several batches of H5N1 influenza A virus cultured in Madin-Darby canine kidney (MDCK) cells were purified using this method. The efficiency of purification was evaluated using many methods including quantitative real-time PCR, hemagglutination (HA) test and single radial immunodiffusion assay. Moreover, Benzonase nuclease was used for comparison. Results: Without the use of Benzonase nuclease, the overall removal rates of host cell DNA and residual proteins were 99.62% and 98.1%, and the HA antigen recovery rate was 66.96%. Conclusions This study established a purification strategy with good effect for cell-based influenza vaccines. It can efficiently remove host cell DNA and proteins and achieve a high HA recovery rate. The purification result is no worse than that of adding Benzonase nuclease, suggesting the potential of its application in actual vaccine production.