1.Research on the Effect of Silence of Survivin Gene by RNAi on the Proliferation and Apoptosis of Nasopharyngeal Carcinoma CNE1 Cells
Journal of Modern Laboratory Medicine 2014;(6):74-77,80
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.
2.Effects of inhibitory and enhancing polypeptides from HCV core region on cytotoxic T cell functions
Qiaoyu MA ; Yuming WANG ; Fei HAO
Journal of Third Military Medical University 2003;0(19):-
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.
3.A study of the specific cytotoxic T cell of BALB/c mice detected by LDH
Qiaoyu MA ; Yuming WANG ; Fei HAO ;
Journal of Third Military Medical University 1983;0(04):-
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.
4.A preliminary study of effects of hepatitis C virus nucleopolypeptides on specific cytotoxic T cells
Qiaoyu MA ; Yuming WANG ; Fei HAO
Chinese Journal of Infectious Diseases 2001;0(05):-
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.
5.Effects of folic acid deficiency on plasma homocysteine level and arterial lesions in rats
Xuexing MA ; Wangming LIU ; Yuming WANG
Chinese Journal of Geriatrics 2000;0(04):-
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.
6.Research progress of induced pluripotent stem cells and its bottleneck
Chinese Journal of Tissue Engineering Research 2014;(45):7358-7363
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.
7.A retrospective analysis of venous thromboembolism risk and prevention in critically ill patients
Hui ZHANG ; Yu MA ; Yuming WEN ; Li LU
Chinese Journal of Emergency Medicine 2013;22(5):517-520
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.
8.Cell-based Screening and Validation of Human Novel Gene TMEM9B Activated Nuclear Factor Signaling Pathway
Yuming LIU ; Zhi PENG ; Weiwei DENG ; Taiping SHI ; Dalong MA
Progress in Biochemistry and Biophysics 2009;36(11):1423-1428
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.
9.Nutritional Problems and Interventions after Spinal Cord Injury (review)
Yuming WANG ; Ke MA ; Yonggang WU ; Huiming GONG
Chinese Journal of Rehabilitation Theory and Practice 2016;22(1):69-71
Most of the individuals with spinal cord injury (SCI) have nutritional problems in metabolism in energy, glucose, fat and vitamin after injury, which would influence the quality of life. Nutrition intervention program includes personalized nutrition consultation, correcting the imbalance of energy and lipid, supply of creatine and vitamins associated with energy metabolism, monitoring, and health promotion, are helpful to reduce the risk of complications related with metabolism.
10.Criteria and Ethical Issues for Clinical Research of Spinal Cord Injury (review)
Yuming WANG ; Huiming GONG ; Yonggang WU ; Ke MA ; Jianjun LI
Chinese Journal of Rehabilitation Theory and Practice 2016;22(2):156-159
The outcome of spinal cord injury is related to many factors, such as mechanism and severity of injury, and differences of individual. The standard of clinical trial plays a very important role in the reliability of the research. especially the inclusion and exclusion criterias, ethical issues, treatment standardization, informed consent and other issues. The effect of these factors on the clinical trial of spinal cord injury was summarized in this article. The inclusion and exclusion criterias to control the consistency should be developed based on the specific research content. Informed consent for clinical trial is also necessary, especially to the clinical trials with uncertainly benefits and risks, as well as the standardization of the operation procedures and rehabilitation treatment.