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Urokinase Receptor Gene Expression Inhibited by siRNA Cocktails in Co-culture of Spermatogenic Cells with Sertoli Cells from 3-Week-Old Rat

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Journal of Reproduction and Contraception 2010年第2期21卷 67-77页

作者： Dong-hui HUANG Hu ZHAO Yu MING Cheng-liang XIONG Family Planning Research Institute Tongji Medical College Huazhong University of Science & Technology Wuhan 430030 China department of Human anatomy Tongji Medical College Huazhong University of Science & Technology Wuhan 430030 China

Objective To study the functions of urokinase receptor （uPAR） during spermatogenesis by adding targeted siRNA cocktails to the co-culture of Sertoli cell with spermatogenic cells. Methods The seminiferous tubule samples from SD rats aged 20-23 d were digested with collagenase for 15-20 rain, then were cut into smaU fragments. Tubular fragments were digested with collagenase again for 5 -10 min, then gently resuspended in F12/ DMEM supplemented with vitamins and hormones. Cell samples were seeded in plate and cultured at 32 ℃ for 2 weeks. After 48 h, siRNA cocktails at a concentration of 15 nmol/L or 30 nmol/L were introduced into this cocultured Sertoli/spermatogenic cells. After then for 48 h, expression of uPAR mRNA was analyzed by real-time RT-PCR. Results Some spermatogenic cells took place morphologic changes and generated an obvious flagellum. The expression levels of uPAR mRNA silencing with siRNA cocktails at a concentration of 15 nmol/L or 30 nmol/L were significantly lower than those in nontransfected group （P〈0.05）; the inhibition rate was 63.5% or 76.7%, respectively. Conclusion The meiosis and spermiogenesis could take place in this culture system, and the siRNA cocktails can effectively inhibit the gene expression of uPAR in co-cultured Sertoli/spermatogenic cells.

关键词： small interfering RNA RNA interference urokinase receptor (uPAR) spermatogenic cell cell co-cultures

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神经生长因子在大鼠心内神经节的表达

解剖学报 2004年第1期35卷 100-102页

作者：刘晓柳李光千赵小明武汉科技大学医学院解剖学教研室武汉430080 华中科技大学同济医学院解剖学系武汉430030

目的　观察成年和中老年大鼠心内神经节神经生长因子 (NGF)阳性神经元的存在及其变化。　方法免疫组织化学法。　结果　成年及中老年大鼠心内神经节均存在NGF阳性神经元 ,但中老年大鼠心内神经节NGF阳性神经元明显减少 ,表达明显降低。... 详细信息

目的　观察成年和中老年大鼠心内神经节神经生长因子 (NGF)阳性神经元的存在及其变化。　方法免疫组织化学法。　结果　成年及中老年大鼠心内神经节均存在NGF阳性神经元 ,但中老年大鼠心内神经节NGF阳性神经元明显减少 ,表达明显降低。　结论　心房后壁心内神经节存在NGF阳性神经元 ,并随年龄增加表达减少 ,提示NGF表达的减少可能与心脏功能的退化相关。

关键词：神经生长因子大鼠心内神经节表达免疫组织化学

雄激素对大鼠心内神经节神经生长因子表达的影响

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解剖学报 2004年第1期35卷 103-105页

作者：刘晓柳李光千殷树仪赵小明范玉华武汉科技大学医学院解剖学教研室武汉430080 华中科技大学同济医学院解剖学系武汉430030

目的　观察神经生长因子 (NGF)在成年雄性大鼠、雌性大鼠、雌性大鼠睾酮处理组、睾丸切除大鼠、睾丸切除后睾酮替代大鼠心房后壁心内神经节的表达及变化。　方法　免疫组织化学染色。　结果　各组大鼠心内神经节均存在NGF阳性神经元 ,... 详细信息

目的　观察神经生长因子 (NGF)在成年雄性大鼠、雌性大鼠、雌性大鼠睾酮处理组、睾丸切除大鼠、睾丸切除后睾酮替代大鼠心房后壁心内神经节的表达及变化。　方法　免疫组织化学染色。　结果　各组大鼠心内神经节均存在NGF阳性神经元 ,但睾丸切除组大鼠心内神经节NGF阳性神经元的数量明显减少 ,表达明显降低。　结论　心内神经节细胞内含NGF ;雄激素可能影响心内神经节细胞的NGF表达。

关键词：雄激素大鼠心内神经节神经生长因子表达免疫组织化学

雌性大鼠心内神经节中雌激素受体及其mRNA的表达

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解剖学报 2003年第4期34卷 372-374页

作者：陈小武祝善乐殷树仪范玉华赵小明华中科技大学同济医学院解剖学系武汉430030

目的　在雌激素受体蛋白及ERmRNA水平提供雌激素对心内神经节中神经元作用的形态学依据。方法　采用免疫组织化学及原位杂交技术。　结果　在心内神经节部分神经元中 ,雌激素受体免疫反应及其mRNA原位杂交反应阳性。雌激素受体免疫反应... 详细信息

目的　在雌激素受体蛋白及ERmRNA水平提供雌激素对心内神经节中神经元作用的形态学依据。方法　采用免疫组织化学及原位杂交技术。　结果　在心内神经节部分神经元中 ,雌激素受体免疫反应及其mRNA原位杂交反应阳性。雌激素受体免疫反应沉淀物呈棕黄色 ,定位于胞核 ,雌激素受体mRNA免疫反应沉淀物呈棕黄色 ,定位于胞浆。　结论　大鼠心内神经节中 ,部分神经元能合成雌激素受体蛋白 ,说明ER阳性神经元可以为雌激素提供结合位点 ,因此 ,这些神经元可能受到雌激素的影响。

关键词：心内神经节雌激素受体雌激素受体mRNA 免疫组织化学大鼠

Inhibition of ethanol-induced toxicity by tanshinone ⅡA in PC12 cells

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Acta Pharmacologica Sinica 2006年第6期27卷 659-664页

作者： Xian-fang MENG~2 Xiao-jing ZOU~3 Bin PENG~2 Jing SHI~2 Xin-min GUAN~2 Chun ZHANG~(4,5)2 department of Neurobiology,tongji medical college,huazhong university of science and technology,wuhan 430030,china 3 department of Anesthesiology,4 department of Nephrology,Union Hospital,tongji medical college,huazhong university of science and technology,wuhan 430022.china department of Neurobiology Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Anesthesiology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Nephrology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To observe the effects of tanshinone IIA(Tan IIA)on the neurotoxicityinduced by ethanol in PC 12 cells and to explore its protective ***:PC 12cell survival was measured by MTT *** formation of reactive oxygenspecies(ROS)and lactate dehydrogenase(LDH)release were detected by 2′,7′-dichlorofluorescin(DCF)fluorescence and calorimetric method,*** of cell apoptosis was monitored by flow *** expression ofp53 was detected by immuno-fluorescence and flow ***:Ethanolsignificantly impaired the survival of PC12 cells as demonstrated by MTT *** also induced significant ROS formation and increased LDH ***-incubation with Tan IIA in the culture medium significantly reversed these *** caused cell apoptosis and the upregulation of p53 *** anti-apoptosis effects of Tan IIA on ethanol-induced toxicity were accompanied by thedownregulation of pro-apoptotic p53 protein ***:Tan IIAcan protect neurons from apoptosis and might serve as a potential therapeuticdrug for neurological disorders induced by ethanol.

关键词： apoptosis ethanol PC12 cells p53 tanshinone ⅡA

Expression of Androgen Receptor in Meningiomas

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Journal of huazhong university of science and technology(medical sciences) 2001年第2期21卷 140-142页

作者：陈坚陈刚 department of Neurosurgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030

In order to investigate the expression of androgen receptor in meningiomas and its relation to tumor proliferative potential, we examined the expression of AR and proliferating cell nuclear antigen by avidine biotin complex immunohistochemistry in 39 cases of meningiomas. Of the 39 cases of meningiomas, 20 showed positive AR immunoreactivity. The AR expression positivity rates were 31 % in benign meningiomas, 58 % in atypical meningiomas, 87.5 % in malignant meningiomas, respectively. In addition to the tumor cells, cells of microvascular endothelial proliferation were frequently AR positive. Malignant meningiomas had a significantly higher percentage of AR positive cells compared with atypical and benign meningiomas . The mean proliferating cell nuclear antigen labeling index was significantly higher in the malignant meningiomas when compared with atypical meningiomas and benign meningiomas . AR positive meningiomas had higher PCNA LI than AR negative meningiomas . The expression of AR in tumor tissues was significantly related with PCNA LI. These data indicated that AR in the meningiomas was correlated with histological grade and AR might participate in the growth of these tumors and tumor angiogenesis. The measurement of AR in these tumors may indirectly represent tumor growth potential.

关键词： meningiomas androgen receptor immunohistochemistry proliferating cell nuclear antigen angiogenesis

Effect of amyloid peptides on serum withdrawal-induced cell differentiation and cell viability

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Cell Research 2004年第6期14卷 467-472页

作者： Yi Peng WANG, Ze Fen WANG, Ying Chun ZHANG, Qing TIAN, Jian Zhi WANG department of Pathophysiology, tongji medical college, huazhong university of science and technology, wuhan, 430030, china department of Pathophyiiology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Abnormal deposition of amyloid-p(Ap) peptides and formation of neuritic plaques are recognized as pathological processes in Alzheimer’s disease (AD) brain. By using amyloid precursor protein (APP) transfected cells, this study aims to investigate the effect of overproduction of Aβ on cell differentiation and cell viability. It was shown that after serum withdrawal, untransfected cell (N2a/Wt) and vector transfected cells (N2a/vector) extended long and branched cell processes, whereas no neurites was induced in wild type APP (N2a/APP695) and Swedish mutant APP (N2a/ APPswe) transfected N2a cells. After differentiation by serum withdrawal, the localization of APP/AP and neurofilament was extended to neurites, whereas those of APP-transfected cells were stillrestricted within the cell body. Levels of both APP and Aβ were significantly higher in N2a/APP695 and N2a/APPswe than in N2a/Wt, as determined by Western blot and Sandwich ELISA, respectively. To further investigate the effect of A0 on the inhibition

关键词： Alzheimer’s disease amyloid β cell differentiation.

Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro

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Asian Journal of Andrology 2003年第2期5卷 113-115页

作者： Ji-Hong LIU, Yang LI, Zheng-Guo CAO, Zhang-Qun YEdepartment of Urology, tongji Hospital, tongji medical college, huazhong university of science and technology,wuhan 430030, china department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 癈. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. Results: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. Conclusion: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

关键词： dibutyryl cyclic AMP forskolin sperm motility

Risk Factors of Learning Disabilities in Chinese Children in wuhan

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Biomedical and Environmental sciences 2003年第4期16卷 392-397页

作者： BIN YAO AND HAN-RONG WUdepartment of Children and Adolescent Health, Maternity and Child Health Care, tongji medical college, huazhong university of science and technology, wuhan 430030, Hubei, china department of Children and Adolescent Health Maternity and Child Health Care Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Chin

Objective To investigate prevalence rate of learning disabilities (LD) in Chinese children, and to explore related risk factors, and to provide theoretical basis for preventing such disabilities. Methods One thousand and one hundred fifty one children were randomly selected in primary schools. According to criteria set by ICD-10, 118 children diagnosed as LD were classified into the study group. Four hundred and ninety one children were classified into the normal control group. Five hundred and forty two children were classified into the excellent control group. The study instruments included PRS (The pupil rating scale revised screening for learning disabilities), Conners' children behavior check-list taken by parents and YG-WR character check-list. Results The prevalence rate of LD in Chinese children was 10.3%. Significant differences were observed between LD and normally learning children, and between the LD group and the excellent group, in terms of scores of Conners' behavior check-list (P china. LD is resulted from a number of factors. Good studying and living environments should be created for LD children.

关键词： Learning disabilities (LD) Behavior problems Character Children Risk factors