Objective To explore the effect of decrease of Survivin expression on the proliferation and apoptosis of the naso-pharyngeal carcinoma cells.Methods To observe the effect of decrease of Survivin expression on nasopharyngeal carcinoma cell morphology using RNAi technology combined with RT-PCR and Western blot.Cell proliferation and apoptosis were de-tected by MTT and TUNEL assay.The expression of PARP,Bcl-2 and Bax was detected by Western blot.Results RT-PCR result showed the survivin mRNA expression in Surviving-siRNA groups was downregulated (about 0.26±0.02)and the inhibition ratio was 43.7% (P<0.05).MTT result showed cell proliferation rates were significantly different between 24,48 and 72 h after transfection.The cell inhibition rates were 21.9%,37.1% and 29.6%,respectively (P<0.05).West-ern blot result showed the Survivin protein expression in Survivin-siRNA2 groups was downregalated and the relative ex-pression level was reduced by 57% (P<0.05).TUNEL assay showed that cell apoptosis rate was increased obviously in surviving-siRNA groups.The expression of PARP (89KD)and Bax wasupregulated (3.9 and 2.4 fold change)and the ex-pression of Bcl-2 was downregulated (0.3 fold change).The phosphorylation of AKT was inhibited when Survivin was down-regulated (about reduced by 5 7%).Conclusion Silencing Survivin gene by siRNA can inhibit the proliferation of Na-sopharyngeal carcinoma cells and induce apoptosis.Survivin may become a potential gene for therapy target of nasopharynge-al carcinoma.

Objective To establish a simple animal model and the cytotoxic T cell(CTL) detecting system for the studies of the effects of CTL on HCV infection. Methods The CTL activity in BLAB/c mice immunized by polypeptides in the core region of HCV was detected with lactic dehydrogenase (LDH) by using SP2/O cells as the target cells. Results CTL activity in BLAB/c mice immunized by polypeptides in the core region of HCV could be detected with LDH. The activity could be enhanced by CPA10(5～23 aa) but inhibited by CPA9(39～74 aa). Conclusion CTL activity in BLAB/c mice can be detected stably by LDH.

BACKGROUND:With the development of the research, induced pluripotent stem cel s are applied to the build of disease model, drug screening, regenerative medicine, and many other research fields, and have made significant achievements, especial y in the study of nervous system diseases.
OBJECTIVE:To summarize the recent development of induced pluripotent stem cel s and to raise problems and prospects based on the latest research in this field.
METHODS:The first author searched the PubMed database for articles about the induced pluripotent stem cel s, including reviews, clinical research and basic research, published from January 2006 to September 2014. The keywords were“iPS, induced pluripotent stem cel”, and final y 60 articles were included in result analysis.
RESULTS AND CONCLUSION:Induced pluripotent stem cel research continues to make breakthrough from its discovery by Yamanaka’s team in 2006 to winning Nobel Prize in 2012. Induced pluripotent stem cel research has broad prospects in the disease model construction, drug screening and regenerative medicine. Currently, problems such as reprogramming methods, cel stability, and clinical transformation stil need to be solved, and further researches are necessary.

Objective To explore the mechanism of CTL dysfunction in HCV infected person, to provide a theoretical basis for further understanding the pathogenesis of hepatitis C and the development of HCV vaccines. Methods HCV nucleopolypeptides were selected and synthesized with the method of solid phase synthesis. BALB/c mice were immunized subcutaneously with HCV nucleopolypeptides, and CTL activity of mice was detected by LDH releasing test. Results CTL of mice could be inhibited by HCV nucleopolypeptides residues 39-74, 67-76, 71-80 and enhanced by HCV nucleopolypeptides residues 5 23,63 72,131 140. Conclusion The function of CTL can be suppressed and intensified by different HCV nucleopolypeptides.

Objective To investigate the pathogenesis of cytotoxic T cell (CTL) dysfunction in patients with HCV infection. Methods CTL detecting system was established. Two polypeptides which could enhance CTL function and two polypeptides which could inhibit CTL function were selected and cross-combined. BALB/c mice were immunized by subcutaneous injection of the combined polypeptides, and the CTL activity in mouse spleen cells was detected by LDH release test. Results CTL activity in BLAB/c mice immunized by polypeptides in the core region of HCV could be enhanced by CPA10 (5-23 aa) and inhibited by CPA9 (39-74 aa). CTL activity in the mice could be enhanced by polypeptides from the HCV core region, CPB2+CPB8, and CPB6+CPB8, respectively. There was no obvious difference between CPB2+CPB7, CPB6+CPB7 and the negative control. Two-factor analysis of variance showed that there was reciprocal action between the inhibitory and enhancing polypeptides from the HCV core region. Conclusion CTL activity in BLAB/c mice can be detected stably by LDH. There is an interactive effect between the inhibitory and enhancing polypeptides from the HCV core region.

Objective To observe effects of the folic acid deficient diet on plasma homocysteine(Hcy) level and lesions of aorta in rats. Methods Twenty male Wistar rats were divided into two groups, folic acid deficent group (FD) and control group (Ctl).The rats were fed with folic acid deficient diet and normal diet for 3 months,respectively. The levels of serum folic acid and plasma Hcy as well as the activities of superoxide dismutase(SOD) and glutathione peroxidase (GPX) in erythrocytes were measured.The histological changes of aorta were also examined by light microscope. Results After 3 months treated with folic acid deficient diet, serum folic acid level 〔(6 08?1 84)?g/L〕 decreased significantly in rats of FD group compared either with pre experimental level 〔(13 32?2 02)?g/L〕 or with control one 〔(12 17?1 67)?g/L〕. Meanwhile, plasma homocycteine level increased significantly in rats of FD group 〔(28 66?6 07)?mol/L〕 compared either with pre experimental level 〔(9 75?1 86)?mol/L〕 or with control one 〔(9 49?1 77)?mol/L〕. The activity of erythrocytic SOD increased but GPX decreased obviously. The morphology lesions were also observed in aortic tissue. Conclusions Folic acid deficient diet induced hyperhomocysteinemia and arterial lesions.The high oxidant stress induced by hyper homocysteinemia may be one of mechanisms of arterial lesion.

Nuclear factor κB (NF-κB) is an important cellular transcription factor. The important role of NF-κB-mediated cell signal transduction pathway in apoptosis is a hot topic at home and abroad. In order to discover new regulators in NF-κB signaling pathway, a high-throughput cell-based screening model based on dual luciferase reporters system was established, a number of genes that can activate NF-κB signal pathway were obtained by screening of 439 novel function genes. Among them, TMEM9B can obviously activate NF-κB signaling pathway. Further experiments showed that TMEM9B activated NF-κB signaling pathway in a dose-dependent pattern. Western blotting and EMSA experiments confirmed that TMEM9B can promote the degradation of IκBα (a cytoplasm inhibitor of NF-κB), and cause NF-κB shift from the cytoplasm to nucleus. At the same time, flow cytometry result demonstrated TMEM9B can induce apoptosis in HEK293T and HeLa cells. In short, a stable and effective screening system for NF-κB has been established, through which TMEM9B was identified to be able to significantly activate NF-κB signal transduction pathway and thus cause cells apoptosis.

Objective To estimate the venous thromboembolism (VTE) risk and prevention in critically ill patients admitted to ICU and discuss the appropriate strategy for prevention.Methods A total of 276 critically ill patients staying longer than 48 hours in ICU were enrolled for a retrospective single-center study.VTE risk assessment,methods for mechanical and pharmacological prophylaxis and demographic data were recorded.Simplified Caprini scores for VTE risk were counted in the first day and 7th day after admission to ICU,and were compared among internal medicine,surgery and trauma subgroups.Relationship between VTE risk and the clinic index was analyzed by Pearson test and Spearman test with SPSS 17.0 software.The prophylaxis strategy applied to patients without low risk of VTE was explored.Results Simplified Caprini scores were (8.71 ± 4.90) and (9.24-± 5.30) on the first day and the 7th day after admission respectively.Simplified Caprini score was significantly related to APACHE Ⅱ score (r =0.397,P =0.027).Meanwhile,simplified Caprini score in surgical and traumatic patients was higher than that in medical ill patients (14.02 ±2.01),(14.5 ± 1.29) vs.(6.55 ±3.98),P ＜0.01.The total rate of early prophylaxis measures used with mechanical prevention (13.43％) and pharmacological prophylaxis (5.22％) was only 18.28％ within 48 hours after admissioin of patients with highest riskof VTE.Even on the 7th day after admission to ICU,the total rate of prophylaxis measure employed with mechanical prevention (11.92％) and pharmacological prophylaxis (11.56％) for VTE was 25.83％.Conclusions Critically ill patients in ICU were subjected to extremely high risk of VTE.The VTE risk related closely to the severity of critically illness existed throughout the whole period of the ICU stay.Constant assessment for VTE risk and bleeding risk should be made with frequent assessment for critically ill patients.

Objective To estimate the clinic features of severe multiple trauma with secondary thrombocytosis as a factor influencing the prognosis.Methods A retrospective single-center study was carried out in 680 patients with severe multiple trauma survived longer than 72 hours in Chongqing Emergency Medical Center from March 2010 through March 2013.The variables including age,gender,ISS (injury severity score),APACHE Ⅱ score,splenectomy and the usages of vasopressors,blood products transfusion,hematopoietic medicines and anticoagulant were analyzed.The prognosis indices including total in-hospital mortality after 72 hours,length of hospital stay and morbidity of thrombo-embolism were explored.The clinic characteristics and prognosis of severe multiple trauma with secondary thrombocytosis (platelet count more than 450 × 109 L-1) were evaluated.T test or rank sum test was used for comparison between measurement data and Chi-square test or Fisher' s exact test was used for comparison between enumeration data.Results Thrombocytosis was identified in 99 (14.56％) patients and it occurred one week after injury with median time of 27 days (ranged from 8 days to 304 days),and maintained for (18.62±4.38) d.The median of platelet count was 584 × 109 L-1 (lowest 478 × 109 L-1,highest 1 072 × 109 L-1) in severe multiple trauma patients with thrombocytosis.The proportions of splenectomy,prolonged use of vasopressors and employment of hematopoietic medicines or anticoagulant were significantly higher in patients with thrombocytosis than those in patients without thrombocytosis (14.14％ vs.7.06％,P=0.03;62.63％ vs.39.07％,P＜0.01; 28.28％ vs.6.71％,P＜0.01; 90.91％ vs.19.45％,P＜ 0.01).The highest D-Dimer level presenting in patients with thrombocytosis during the time of platelet increasing was significantly more common than that in patients of non-thrombocytosis group 7 days after trauma [(11.68 ± 11.90) vs.(5.05 ± 5.11),P =0.004].However,the mortality,length of hospital stay and morbidity of thrombo-embolism were not significantly increased in patients with thrombocytosis compared with patients without thrombocytosis [8.08％ vs.8.78％,P=0.82; 34 d (28.5,54.5) d vs.45 d (23,67) d,P =0.41; 10.10％ vs.10.50％,P =0.91].Conclusion There was a higher rate of secondary thrombocytosis in severe multiple trauma patients.The factors such as splenectomy,vasopressors,hematopoietic medicines and so on might induce the reactive thrombocytosis in trauma patients.Thrombocytosis might increase the incidence of thromboembolism in severe multiple trauma patients without appropriate prophylactic anticoagulation.For the sake of prophylaxis,employment of anti-platelet agent might be the appropriately therapeutic strategy for patients suffering from severe multiple trauma with secondary thrombocytosis accompanying risk factors of arterial thrombo-embolism.

Background and purpose: Peroxiredoxin Ⅱ (PrxⅡ) has the activity of peroxidase. The relevant studies found it played an important role in gastric cancer. This study aimed to investigate the expression of PrxⅡ in human gastric cancer tissues and cells, analyze its relationship with clinicopathological characteristics, and explore the relationship between PrxⅡ and the prognosis and the development of gastric cancer. Methods: The expression of PrxⅡmRNA and protein in gastric cancer tissues and the paired adjacent normal tissues from 45 patients was detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. The same methods were used to detect the expression of PrxⅡ mRNA and protein in GES-1, MGC-803, MKN-45 and MKN-28. Tissue mi-croarray and immunohistochemistry were used to detect the expression of PrxⅡ protein in gastric cancer tissues and the paired adjacent normal tissues from 116 patients. The relationship between the results and clinicopathological char-acteristics was analyzed. The prognosis was analyzed. Results: According to results of RTFQ-PCR and Western blot, we found that PrxⅡ mRNA and protein in gastric cancer tissues were significantly higher than that in adjacent normal tissues (P<0.05). PrxⅡ mRNA and protein in gastric cancer cells were higher than that in normal gastric cells (P<0.01).Immunohistochemistry revealed that the expression of PrxⅡ protein in gastric cancer tissues (76.7％) was also significantly higher (P<0.01) than that in adjacent normal tissues (30.1％). The expression of PrxⅡ protein is significantly related to tumor size, histological differentiation, depth of invasion, TNM stage and lymph node metastasis (P<0.05), but had no significant relationship with the gender, age, tumor location and distant metastasis. Survival in patients with higher PrxⅡ expression significantly shorter than in those with lower expression (P<0.01). PrxⅡ is an independent prognostic factor of gastric cancer (P<0.05). Conclusion: PrxⅡ promotes the development of gastric cancer. It is one of the adverse prognostic factors of gastric cancer and may serve as a new therapeutic target for gastric cancer.