2型大麻受体的克隆及真核表达体系的构建

目的 克隆人2型大麻受体(hCB2)基因,并构建相应的稳定表达细胞株.方法 利用人T淋巴细胞株HPB-ALL的总RNA,以RT-PCR及酶切、连接等分子生物学方法克隆hCB2基因于质粒载体pFlag-CMV-3内,以HEK293细胞构建稳定表达细胞株,以Western blot方法利用蛋白标记Flag的单克隆抗体(Anti-Flag)和CB2多克隆抗体(Anti-CB2)同时检测质粒的表达.结果 酶切及测序结果表明hCB2基因克隆成功,Western blot检测结果表明Anti-Flag和Anti-CB2都能检测Flag-hCB2蛋白.结论 成功克隆hCB2基因,并获得稳定表达细胞株,同时证实Anti-CB2多克隆抗体有效.     

加强医学本科生读写能力培养的重要性及方法

读写能力是获取信息与表达思维的能力,是反映学生综合素质的重要指标.为培养在信息社会中具备终身学习能力的高素质医学生,教师应该重视对医学本科生读写能力的培养.在教学活动中,通过向学生阐明读写能力的重要性、增加训练量、开设多种形式读写课及第二课堂活动、将读写能力纳入学生能力考核指标等多种行之有效的途径,不断提高学生读写能力,为培养创新性人才作出贡献.     

微波诱导小胶质细胞的促炎症反应

Objective To explore the relationship between microglia] proinflammatory and electro-magnetic radiation and unveil the role of microglia in microwave radiation induced central nervous system in-jury. Methods N9 microglia cells cultured in vitro were exposed to microwave at 90 mW/cm2. Cell flow cy-tometry was used to observe the expression of CD1 lb at different time points after exposure; ELISA was used to detect the concentration of TNF-α in N9 cell culture supernatant; RT-PCR analysis confirmed iNOS mRNA ex-pression in N9 microglia cells; and Nitrate Reductase Method was used to test NO amount in culture super-natant. Results The CD11b positive microglial cells increased significantly at 3 h after microwave exposure (P<0.05), continued to increase until 24 h and peaked at 6 h after exposure. The amount of TNF-α rose dra-matically from 1 h to 24 h after exposure (P<0.01) and peaked at 3 h [(762.1±61.5) pg/ml] after exposure (P< 0.01). The level of NO started to increase at 1 h [(4.48±0.59) μmol/L] and lasted for 24 h after exposure. The expression of iNOS mRNA increased significantly at 1 h(P<0.05), and tripled the original expression at 6 h af-ter exposure, hereafter, it decreased slightly, but all were higher than the control group within 24 h after exposure. Conclusion Microwave radiation could induce the activation of microglia cells. The activated microglia cells could induce microglial proinflammatory by producing large amounts of TNF-α, NO, etc.     

微波诱导小胶质细胞的促炎症反应

Objective To explore the relationship between microglia] proinflammatory and electro-magnetic radiation and unveil the role of microglia in microwave radiation induced central nervous system in-jury. Methods N9 microglia cells cultured in vitro were exposed to microwave at 90 mW/cm2. Cell flow cy-tometry was used to observe the expression of CD1 lb at different time points after exposure; ELISA was used to detect the concentration of TNF-α in N9 cell culture supernatant; RT-PCR analysis confirmed iNOS mRNA ex-pression in N9 microglia cells; and Nitrate Reductase Method was used to test NO amount in culture super-natant. Results The CD11b positive microglial cells increased significantly at 3 h after microwave exposure (P<0.05), continued to increase until 24 h and peaked at 6 h after exposure. The amount of TNF-α rose dra-matically from 1 h to 24 h after exposure (P<0.01) and peaked at 3 h [(762.1±61.5) pg/ml] after exposure (P< 0.01). The level of NO started to increase at 1 h [(4.48±0.59) μmol/L] and lasted for 24 h after exposure. The expression of iNOS mRNA increased significantly at 1 h(P<0.05), and tripled the original expression at 6 h af-ter exposure, hereafter, it decreased slightly, but all were higher than the control group within 24 h after exposure. Conclusion Microwave radiation could induce the activation of microglia cells. The activated microglia cells could induce microglial proinflammatory by producing large amounts of TNF-α, NO, etc.     

电磁辐射后小胶质细胞活化状态和分泌功能的变化

目的:一定量的电磁辐射可引起中枢神经系统神经元的损伤,但其对小胶质细胞的影响尚不清楚。实验观察电磁辐射对小胶质细胞活化状态以及分泌功能的影响,揭示电磁辐射对小胶质细胞及中枢神经损伤的效应关系。方法:实验于2006-08/2007-05在解放军第三军医大学电磁辐射医学防护教育部重点实验室完成。取体外培养的N9小胶质细胞接受X波段脉冲波,平均功率密度为90mW/cm2的电磁波,一次性照射20min,在辐照后0,1,3,6,12,24h等6个时相点观察活化的N9细胞形态学,采用免疫组化的方法观察OX-42的表达情况,用酶联兔疫吸附测定方法检测N9细胞培养上清液中肿瘤坏死因子α的水平,采用硝酸还原酶法检测培养上清液中NO浓度。以未接受电磁波辐照的N9小胶质细胞为假辐照组进行对照。结果:①电磁辐射后3hN9小胶质细胞OX-42表达开始明显增强,并一直持续到辐照后24h,细胞形态由静息状态转变为激活状态;②NO的浓度在辐照后1h开始升高(P<0.05),到辐照后6h达到峰值(P<0.01),12h后趋于恢复,24h后再次明显升高(P<0.05)。③肿瘤坏死因子α水平辐照3h后显著升高(P<0.01),并一直持续到12h,到辐射后24h又再次升高,并达到峰值(P<0.01)。结论:电磁辐射辐照可明显诱导小胶质细胞激活,活化后的小胶质细胞分泌NO、肿瘤坏死因子α等细胞因子的功能增强,分泌大量细胞因子反馈调节引起辐射后期的小胶质细胞激活。     

芦丁对三甲基锡损害学习记忆功能的保护作用与拮抗突触囊泡蛋白表达下调有关

目的探讨芦丁对三甲基锡（Trimethyltin, TMT）损害小鼠学习记忆功能的保护作用及可能机制。方法以6~9周龄雄性
BALB/c小鼠为研究对象,随机分为生理盐水组（对照）、TMT组、TMT+芦丁组、芦丁组,每组各10只;建立TMT（2.25 mg/kg·B.
W.）急性暴露模型,后两组芦丁（10 mg/kg·B.W.）预处理1周,均腹腔注射;TMT给药后24 h,Morris水迷宫测试各组的逃逸潜伏
期,Western检测各组海马和皮层脑区突触囊泡蛋白（Synaptophysin, SYP）的表达情况。结果Morris水迷宫显示TMT给药后
24 h,与TMT组相比,对照组和芦丁组及TMT+芦丁组逃逸潜伏期显著缩短（P<0.05）,与芦丁组及对照组相比,TMT+芦丁组逃
逸潜伏期无统计学意义（P>0.05）;Western显示TMT给药后24 h,与对照组相比,TMT组海马和皮层脑区SYP蛋白表达显著降
低（P<0.05）;与TMT组相比,TMT+芦丁组海马和皮层脑区SYP蛋白表达显著升高（P<0.05）,与芦丁组及对照组相比,TMT+芦
丁组海马和皮层脑区SYP蛋白表达无统计学意义（P>0.05）。结论芦丁预处理对TMT急性暴露损害小鼠学习记忆功能有保护
作用,其保护作用可能与拮抗海马和皮层脑区SYP表达下调有关。
     

雷达部队电磁辐射对作业人员神经行为功能的影响

目的 评价雷达部队作业环境电磁辐射对作战人员情感状态和神经行为的影响并提出相应的防护策略。方法 采用问卷调查进行自觉症状调查。采用神经行为核心测试组合(NCTB)的POMS量表和神经行为功能综合测试仪对辐照人员和对照组进行调查测试。结果 辐照组明显感到视觉疲劳;POMS量表中辐照组疲惫感明显增加;目标追踪实验辐照组总的打点数(χ²=6.808,P<0.05)显著小于对照组。左右手交叉实验总次数辐照组显著小于对照组(χ²=14.820,P<0.01)。结论 电磁辐射对职业人群的心理状态、手部作业能力和作业效率产生明显影响。     

局部解剖学知识在军事训练伤讲解中的应用

军事训练伤是指军事训练直接导致参训人员的组织器官功能阻碍或病理改变,简称训练伤[1]。从训练伤所包括的种类来看,主要有肌肉、骨骼系统的急性、慢性损伤,比较常见的军训伤包括职业性骨骼肌肉疾患或称腰背痛、应力性骨折及肌肉、韧带与关节伤[2]。训练伤内容的讲解需涉及众多损伤部位局部解剖学知识,教学实践表明,将局部解剖学知识与训练伤