Nerve growth factor promotes the repair of corneal nerves in diabetic rats

摘要 目的：探究神经生长因子（NerveGrowthFactor，NGF）对糖尿病大鼠角膜神经的作用，并解释其作用分子机制。 方法：SD大鼠腹腔注射链脲佐菌素（Streptozotocin，STZ）建立Ⅰ型糖尿病大鼠模型，模型建立成功后将正常大鼠和糖尿病大鼠眼球分为四组，正常大鼠左、右眼分别为Ctrl+BSA组和Ctrl+NGF组，糖尿病大鼠左、右眼分别为DM+BSA组和DM+NGF组，并在实验条件下饲养。第6周末检测四组角膜敏感度，然后分别在大鼠左、右眼滴加浓度15ug/mL的牛血清白蛋白（BovineSerumAlbumin，BSA）溶液、NGF溶液，连续2周，每天3次，每次2...Abstract Objective: To explore the effect of nerve growth factor (NGF) on corneal nerves of diabetic rats, and to explain the molecular mechanism. Methods: Type 1 diabetes model rats were induced by intraperitoneal injection of streptozotocin (STZ). Six weeks after the model were successfully established, corneal sensitivity were tested between the control and diabetic rats, then the left or...学位：理学硕士院系专业：医学院_生理学学号：2452014115345

厦门大学教育部双语示范课程《细胞生物学》的建设与发展回顾

双语教学目前在国内各高校的教学中占据越来越重要的地位。双语教学要求使用英文原版教材,从师资配置到教学及考核方法都提出了更高的要求。本文对厦门大学教育部双语示范课程《细胞生物学》的建设与发展过程进行了总结,并对教学过程中存在的问题进行了探讨

早期糖尿病视网膜病变中eEF2K活性变化的实验观察

目的观察早期糖尿病大鼠视网膜中真核生物延伸因子2激酶(e EF2K)的变化,探讨e EF2K活性变化在早期糖尿病视网膜病变(DR)中的意义。方法建立SD大鼠链脲佐菌素(STZ)糖尿病高血糖模型,造模成功后,设立糖尿病大鼠(DM)组和正常对照(NC)组。qRT-PCR检测DM组和NC组4周和8周时大鼠神经视网膜中e EF2K的表达情况。免疫荧光法观察定位神经视网膜中e EF2K激活标志物-磷酸化真核生物延伸因子2(p-eEF2)和Müller细胞活化标志物胶质纤维酸性蛋白(GFAP)表达情况。结果 4周和8周时DM组大鼠神经视网膜中e EF2K的表达水平与NC组相比无明显变化。8周时DM组大鼠神经视网膜各层均出现e EF2K激活表现,即p-eEF2表达增强。神经节细胞层出现细胞凋亡样改变,凋亡样神经节细胞中e EF2的磷酸化水平增强最为明显。神经视网膜中Müller细胞活化,GFAP蛋白表达增强。结论 STZ诱导糖尿病大鼠早期视网膜病变中神经视网膜内e EF2K激活,在凋亡样神经节细胞中e EF2K活性上升最为显著。福建省自然科学基金项目(2016J 01412);;国家自然科学基金青年项目(81700864

p38MAPK induces apoptosis of glioma cell

目的　研究 p38MAPK基因转染大鼠胶质瘤细胞系C6后对其生物学特性的影响 .方法　利用脂质体介导法将p38MAPK基因导入大鼠胶质瘤细胞系 C6中 ,用免疫细胞化学染色检测其在细胞转染前后的表达情况 ,用 HE染色、流式细胞仪等方法研究其对细胞形态、粘着状况和生长周期的影响 .结果　转染 p CMV5 - p38MAPK质粒组 p38MAPK蛋白表达阳性 ,细胞形态发生变化 ,贴壁性降低 ,出现大量凋亡细胞 .结论　转染 p38MAPK基因可诱导胶质瘤细胞凋

福建城市化发展战略博士笔谈

2003年5月,“福建省博士创业促进会”在福州举办了福建省城市发展战略“博士月谈”,与会者主要是省内高校和科研单位的专家、博士。会上,大家用书面发表意见,畅所欲言,崇论宏议。本刊精选部分意见,以飨读者。相信这些意见对福建城市化问题研究的深化将起到积极的推动作用

Regulation of Cell Cycle of Glioma C6 Cells by Regulator of G Protein Signaling 16

目的　探讨G蛋白调节子 16(RGS16)对胶质瘤C6细胞周期的影响。方法　利用脂质体介导法将RGS16基因导入C6细胞中 ,在倒置显微镜下观察细胞形态变化和贴壁生长情况 ;免疫细胞化学法检测转染前后RGS16蛋白的表达情况 ;流式细胞仪检测转染pCMV 5 RGS16和 pCMV 5质粒后每隔 12h后的细胞周期变化。 结果　转染pCMV 5 RGS16质粒 2 4h后 3 0 .0 %细胞贴壁性降低 ,突起收缩 ,细胞变圆 ,72h之后细胞又恢复正常 ;RGS16蛋白的表达呈时相性 ,3 6h时表达率最高 (阳性率为13 .0 % ) ,72h表达终止 ;C6细胞的各期细胞比例变化与RGS16蛋白表达对应 ,在 3 6h时G1期比例从转染前的 70 .5 %降低到60 .2 % ,S期比例从 2 0 .9%增加到 3 4.9% ;在 48h时G1期增加到 76.2 % ,S期减少到 11.4% ;72h各期恢复到正常比例。对照组细胞转染前后形态变化不明显 ,RGS16蛋白表达阴性 ,细胞周期变化不明显。结论　RGS16能促进C6细胞周期的运行 【英文摘要】 Objective To study the effect of regulator of G protein signaling 16(RGS16) on the cell cycle of glioma C6 cell.Methods pCMV5 RGS16 was transfected into C6 cells by lipofectin.The morphological and adhesive changes of the cells were observed under an inverted microscope.Expression of RGS16 was examined by immunocytochemical method both before and after the transfection.Flow Cytometry was adopted to measure the fraction number changes of the cell cycle phase every 12 h.Results 24 hours after the transfec..

Promotion of glioma C6 cells proliferation by overexpressed RGS16

目的　探讨 G蛋白调节子 16 (RGS16 )对大鼠胶质瘤C6细胞的生物学特性的影响 .方法　利用脂质体介导法将RGS16基因导入 C6细胞中 ,在倒置显微镜下观察细胞形态变化和贴壁情况 ;3H- Td R法检测 C6细胞在转染不同梯度p CMV5 - RGS16和 p CMV 5质粒后的增殖情况 ;免疫细胞化学法检测转染前后 RGS16蛋白的表达情况 ;流式细胞仪检测转染 p CMV5 - RGS16和 p CMV5质粒 36 h后细胞周期变化和细胞是否有凋亡发生 .结果　转染 p CMV5 - RGS16质粒 2 4 h后 30 %细胞贴壁性降低 ,突起收缩 ,细胞变圆 ;RGS16蛋白表达阳性 ;3H- Td R法检测显示 C6细胞增殖速度与转染p CMV5 - RGS16的量呈正相关 ;细胞周期结果显示 G1期细胞百分数减少 10 % ,而 S期细胞百分数增多 14 % ;未发现RGS16与凋亡有直接关系 .结论　 RGS16可能促进 C6细胞的增殖 . 【英文摘要】 s: AIM To study the effect of RGS16 on the biological characteristics of glioma C6 cells. METHODS pCMV5 RGS16 was transfected into C6 cells by lipofectin. The morphological and adhesive changes of the cells were observed under an inverted microscope. Proliferation of C6 cells was measured by 3H thymidine ( 3H TdR) assay after gradient transfections of pCMV5 RGS16 and pCMV5. Expression of RGS16 was examined by immunocytochemical method both before and after the transfection. Flow cytometry ...高等学校骨干教师计划资

Inhibition of P38 MAPK reduces tumor conditioned medium-induced angiogenesis in co-cultured human umbilical vein endothelial cells and fibroblasts

Tumor conditioned medium (CM) has been widely used to stimulate endothelial cells to form capillary-like structures in in vitro angiogenesis models. We report herein the effect of HT1080 and A549 CM after they were mixed with microvascular endothelial cells medium-2 (EGM-2) on angiogenesis in human umbilical vein endothelial cells (HUVECs). Both HT1080 and A549 CM decreased HUVEC proliferation, to different extents. While A549 CM significantly increased capillary-like structure formation in a co-culture system, no effect of HT1080 was apparent. Inhibition of p38 mitogen-activated protein kinase (MAPK) blocked both basal and A549 CM induced capillary-like structure formation, but inhibition of extracellular signal-regulated kinases (ERK) and that of c-Jun N-terminal protein kinases (JNK) MAPK had no such effect. Activation of ERK MAPK was inhibited by both CMs, whereas p38 MAPK was inactivated by HT1080 and activated by A549 CM and a control. Neither CM had an effect on JNK MAPK. The results suggest that p38 MAPK played a critical role in capillary-like structure formation in the co-culture, partly via promotion of apoptosis in HUVECs

Prokaryotic expression,purification of chicken Rad51 protein and generation of its antiserum

目的:rAd51蛋白在双链断裂的dnA修复的同源重组中发挥重要作用。因而其过量表达在细胞内的基因操作中被用于在提高基因同源重组的效率。当前在鸡中还未有针对该蛋白的特异性抗体。为此我们对鸡的rAd51进行了体外表达与纯化,并制备了CrAd51抗体,为该蛋白质在鸡细胞中的功能研究及探讨其用于鸡细胞中提高同源重组的研究提供有效工具。方法:对鸡rAd51基因进行全长克隆并构建原核表达载体PET-28A(+),用IPTg诱导该蛋白在rOSSETA菌株中表达,纯化后注射bAlb/C小鼠获得抗体。结果:成功制备鸡rAd51特异多克隆抗体,ElISA显示该抗体血清效价达51 200,该抗体血清可以用于WESTErn等的蛋白检测技术。结论:针对鸡基因产物制备抗体的技术方法有效可行;首次成功获得鸡rAd51蛋白及多克隆抗体,为与该蛋白相关的研究提供了有效工具。Objective:RAD51 protein plays a major role in homologous recombination of DNA double strand break repair.Overexpression of this protein has thus been used to enhance the homologous recombination efficiency in gene manipulations in cells.Currently,there is not yet an antibody available for this chicken protein.We therefore carried out in vitro expression,purification of the chicken Rad51,and tried to prepare the antibody.Methods:Full length of chicken Rad51 coding sequence was cloned and inserted into pET-28a(+) to generate a prokaryotic expression constructs;the expression was induced with IPTG in a Rosseta strain.After the purification,the gene product was procedurally injected into BALB/c mouse to generate antibody.Results:ELISA test showed that the titer of antiserum reached 51200;The antiserum had a good performance in Western blot for both sensitivity and specifity.Conclusion:The protocol adapted in this study works well in generating antiserum against chicken gene product;We have for the first time prepared an antiserum against chicken Rad51 providing a useful tool for studies on related to the chicken Rad51.科技部重大基础研究计划资助(2009CB941601