Cdk13表达对神经元轴突伸长的影响

目的探讨智力障碍相关基因Cdk13对神经轴突伸长的影响。方法通过基因敲除模型、质粒细胞转染等方法,荧光共聚焦显微镜成像技术检测神经元轴突的形态,观察轴突伸长的变化。结果 Cdk12和Cdk13在神经系统中有表达,在敲除Cdk12和Cdk13后轴突长度比对照组减少,两者比较有统计学差异(P0.05),敲除Cdk12和Cdk13基因后Cdk5 mRNA水平降低。结论 Cdk13和Cdk12的表达对神经元轴突的生长至关重要,其失活会引起神经元轴突形态异常,可能是导致智力障碍的病理机制。     

高压氧治疗对慢性脑缺血大鼠认知功能的影响及其作用机制

目的 探讨高压氧（hyperbaric oxygen，HBO）治疗对慢性脑缺血（chronic cerebral ischemia，CCI）大鼠 学习记忆能力的影响及其作用机制。 方法 选取240只雄性SD大鼠随机分为假手术组、CCI组和HBO组，每组80只。采用双侧颈总动脉阻 断法建立CCI模型，HBO组建模12 h后开始进行HBO治疗28 d，压力0.2 MPa，每日1次，每次60 mi n。采用 Morri s水迷宫实验评估7、14、21、28 d大鼠的学习记忆能力，每个时间点20只，检测前均进行3 d训练， 记录各组大鼠的逃避潜伏时间、穿越平台次数。28 d后处死大鼠取海马组织进行HE染色评估神经元 损伤病理变化，RT-PCR法检测Nogo-A mRNA，Western blot法检测Nogo-A蛋白的表达水平。 结果 ①与假手术组比较，CCI 组7、14、21、28 d逃避潜伏时间均延长（均P＜0.05），28 d跨越平台次 数减少（P＜0.05）；与CCI 组比较，HBO组7、14、21、28 d逃避潜伏时间均缩短（均P＜0.05），28 d跨越 平台次数增加（P＜0.05）。②HE染色显示HBO组神经元损伤程度较CCI组减轻；③CCI组Nogo-A mRNA 和Nogo-A蛋白表达水平均较假手术组升高（均P＜0.05），HBO组表达水平均较CCI组下降（P＜0.05）。 结论 HBO治疗可改善慢性脑缺血大鼠认知功能，其机制可能与下调海马组织中Nogo-A的表达水平 有关。     

应用RNA干扰技术抑制神经元E3B1基因表达及对轴突生长的影响

目的:探讨应用RNA干扰(RNAi)技术抑制神经元E3B1基因表达及其对轴突生长的影响.方法:选择针对E381(NCBI:NM-024397)RNAi的3个靶位点Abil1、Abil2、Abil3和1个不针对任何mRNA的RNAi靶位点(阴性对照)以及甘油醛-3-磷酸脱氢酶(GAPDH,阳性对照),用带有neoR选择标志和GFP绿色荧光标志的真核表达载体pGenesil-1构建E381 RNAi质粒,分别转染培养的神经元,在荧光显微镜下观测转染率,经G418筛选得到单一的转染细胞,并用Western blot法检测各转染组神经元E3B1蛋白的表达情况,选出具有最佳抑制效应的siRNA转染神经元,加人轴突生长抑制物,观测轴突生长情况.结果:成功构建了RNAi质粒,各组细胞的转染率相近,均为34%左右.与阴性对照组相比,Abil1、Abil2和Abil3转染组细胞内E3B1 mRNA的表达都受到了不同程度的抑制,其中Abil3转染组细胞的E381 mRNA和蛋白的表达抑制最为显著,加入轴突抑制剂后Abil3转染组神经元轴突仍能继续生长.结论:应用RNA干扰技术能高效特异地抑制神经元E3B1基因的表达,E3B1基因的表达下调可促进神经元轴突的生长.     

轴突分叉点位置计算法适用性的分析

某些中枢神经元通过分叉轴突向两个以上核团投射。轴突分叉点的位置通常由计算轴突主干传导时间予以估测。但是,本工作用该法的4个公式计算细胞内或细胞外记录到的数据未能得到一致和可信的数值。结果提示,计算法的原理是不合理的,并至少对在细胞内或细胞外记录条件下估测轴突分叉点是不适用的。     

匹罗卡品致(癎)大鼠海马中表达生长抑素的中间神经元数目变化及其轴突出芽

Objective To investigate the roles of somatostatin(SS)positive intemeurons in the development and compensation of temporal lobe epilepsy.Methods Piloearpine-induced epilepsy rat model was established.Immunohistochemistry method was used to detect number changes and axonal sprouting of SS positive intemeurons in different domains of the hippocampus at difierent time points.Degeneration of SS positive interneurons and their neurophils were detected by the double immunofluorescence staining with SS and Fluoro-Jade B(FJB)at 7 and 60 days after status epilepticus (SE).Results In the exoerimental rat group,the number of SS positive neurons decreased in each hippocampal domain,and it reached the lowest at 7 days post-SE(There were 11.1±3.3 in hilus,2.8±0.9 in CA1region and 1.8±0.7 in CA1region,t=13.519,9.644 and 8.808,all P<0.01).In chronic phase,the number of SS neurons gradually recovered,and exceeded the control group in CA1 area at 60 days post-SE(12.8±1.5 vs 8.8±1.3,t=-4.506,P<0.01),however,the number of SS neurons in the hilus(25.5±4.6)and CA1 area(4.8±0.8)remained significantly less than normal levels(t value were 4.691 and 3.953.both P<0.01).Increased SS positive fibers were found in the lacunosum-molecular (1m)layer and outer molecular layer of dentate gyrus after 30 days post-SE,and numerous SS positive fibers were seen threnghout the layers of area CA1 at 60 days post-SE.Double immunofluuorescence revealed that a few SS positive interneurons and fibers were also labeled by FJB in area CA1 at 7 days post-SE and in CA domain/hilus at 60 days post-SE.Conclusions SS intemeurons loss plays an important role in the development of temporal lobe epilepsy.The loss is partially caIlsed by the degeneration and death of neurons;SS positive neurophils increase within area CA1 in chronic phase may play a significant role in the generation and compensation of temporal lobe epilepsy.     

弥漫性轴突损伤的磁共振氢谱的变化

弥漫性轴突损伤(Diffuse axonal injury，DAI)普遍存在于各种程度的颅脑损伤中。脑白质中轴突的广泛断裂在重型颅脑损伤病例占50％以上，而在轻度颅脑损伤中仍占30％，检查轴突损伤在了解外伤性脑损害中具有重要的意义。但是如何在活体中检测轴突的病理改变相当困难，虽然明显的病理改变能被传统的影像学(CT或T1、T2加权像MRI)证实，     

壳聚糖神经干细胞复合物修复大鼠完全性脊髓损伤的组织学观察

目的：探讨壳聚糖与神经干细胞复合物修复成年SD大鼠完全性脊髓损伤的可行性。方法：制作大鼠脊髓完全缺损模型，在形成的缺损中植入壳聚糖与神经干细胞复合物(实验组)，或只植入壳聚糖(对照组)，或不加任何材料(空白组)，术后2、4、8周灌注取材，观察结果。结果：壳聚糖与神经干细胞复合物在植入SD大鼠脊髓完全缺损模型后，能桥接缺损两端脊髓；其内的神经干细胞能存活、迁移并分化成多种形态的神经组织细胞，并诱导神经轴突向复合材料内生长。结论：壳聚糖与神经干细胞复合物有可能桥接并修复大鼠脊髓完全缺损性损伤。     

匹罗卡品致(癎)大鼠海马中表达生长抑素的中间神经元数目变化及其轴突出芽

Objective To investigate the roles of somatostatin(SS)positive intemeurons in the development and compensation of temporal lobe epilepsy.Methods Piloearpine-induced epilepsy rat model was established.Immunohistochemistry method was used to detect number changes and axonal sprouting of SS positive intemeurons in different domains of the hippocampus at difierent time points.Degeneration of SS positive interneurons and their neurophils were detected by the double immunofluorescence staining with SS and Fluoro-Jade B(FJB)at 7 and 60 days after status epilepticus (SE).Results In the exoerimental rat group,the number of SS positive neurons decreased in each hippocampal domain,and it reached the lowest at 7 days post-SE(There were 11.1±3.3 in hilus,2.8±0.9 in CA1region and 1.8±0.7 in CA1region,t=13.519,9.644 and 8.808,all P<0.01).In chronic phase,the number of SS neurons gradually recovered,and exceeded the control group in CA1 area at 60 days post-SE(12.8±1.5 vs 8.8±1.3,t=-4.506,P<0.01),however,the number of SS neurons in the hilus(25.5±4.6)and CA1 area(4.8±0.8)remained significantly less than normal levels(t value were 4.691 and 3.953.both P<0.01).Increased SS positive fibers were found in the lacunosum-molecular (1m)layer and outer molecular layer of dentate gyrus after 30 days post-SE,and numerous SS positive fibers were seen threnghout the layers of area CA1 at 60 days post-SE.Double immunofluuorescence revealed that a few SS positive interneurons and fibers were also labeled by FJB in area CA1 at 7 days post-SE and in CA domain/hilus at 60 days post-SE.Conclusions SS intemeurons loss plays an important role in the development of temporal lobe epilepsy.The loss is partially caIlsed by the degeneration and death of neurons;SS positive neurophils increase within area CA1 in chronic phase may play a significant role in the generation and compensation of temporal lobe epilepsy.     

大鼠腓总神经“π”式桥接于胫神经后再生神经的电生理溯源

目的 研究大鼠胫神经原位桥接切断的腓总神经，观察腓总神经再生程度、神经纤维的来源等。方法 将断裂的腓总神经近端和远端分别就近与胫神经施行端侧吻合，存活18个月后，电生理检测再生神经纤维的动作电位传导，取腓总神经远段行光镜及电镜观察神经纤维再生数量及状态。结果 远段腓总神经有明显的神经纤维再生，远段腓总神经通过邻近神经的桥接与近段腓总神经之间有动作电位传导。结论 断裂腓总神经“π”式桥接于胫神经，部分再生神经纤维可能来源于原腓总神经近段，部分来自胫神经。     

荧光染料在视神经脊髓鞘形成期从轴突至少突胶质细胞的扩散

将真蓝(true blue)混悬液注入出生后7、14、28d大鼠眼球内,经过一定时间后,视神经内的轴突和少突胶质细胞被荧光标记。荧光强度在视神经眼球端强于视神经交叉端;出生后14～28d的幼鼠荧光标记明显强于出生后7d的幼鼠;不同年龄组的动物荧光标记普遍在注射荧光染料后的第5d显著增强。本研究表明,荧光染料可被视网膜的节细胞吸收,经轴突输送,然后,横向扩散到少突胶质细胞,扩散的通路可能是朗氏结旁区的轴胶连接。