神经生长因子和脑源性神经营养因子在氧化应激诱导细胞凋亡中的作用

目的 探讨神经生长因子（NGF）和脑源性神经营养因子（BDNF）在氧化应激诱导细胞凋亡的作用。方法 脂质体介导入NGF和BDNF基因转染鼠成纤维细胞NIH3T3,使目的基因在细胞中表达,过氧化氢（H2O2）处理转染细胞,吖啶橙荧光染色法和流式细胞检测细胞凋亡。结果 50μmol/LH2O2能诱导NIH3T3细胞凋亡,处理后24h凋亡率为16．7％,而NIH3T3细胞分别经NGF、BDNF和NGF／BDNF转染表达目的基因后,能对抗H2O2诱导的细胞凋亡,其24h凋亡率分别为8．21％、9．51％和6．26％。结论 氧化应激能诱导细胞凋亡,NGF和BDNF能抑制氧化应激诱导的细胞凋亡,两者具有协同作用。     

移植pSVPoMcat微基因修饰雪旺细胞在鼠脊髓内长期存活及其基因表达

目的：观察人Po-5,flanking介导的21．5ku髓鞘碱性蛋白（myelin basic protiein,MBP)微基因（暂命名为pSVPoMcat)修饰雪旺细胞（SC）在鼠脊髓内存活及其基因表达。方法：120只大鼠分为3组,A组为pSVPoMcat微基因修饰SC移植组;B组为高纯化SC移植组;C组为脊髓损伤（SCI）＿对照组,伤后各组分别于2,4,8,12周（每组每次10只）,取移植区脊髓切片,进行S－100蛋白,MBP免疫细胞化学染色及地高辛(DIG)标记的hMBPF2 cDNA探针的原位杂交（ISH）测定。结果：伤后2,4,8,12周,A组S－100,MBP染色细胞和ISH细胞差异无统计学意义（P＜0．05）,其中ISH阳性细胞可见髓鞘形成,B组的S－100染色细胞逐步递减,差异有非常显著性意义（P＜0．01）,未发现MBP染色细胞以及ISH细胞,C组未发现任何阳性细胞,结论：pSVPoMcat微基因修饰SC在鼠损伤脊髓内能长期存活并表达外源基因,且有助于受伤脊髓的髓鞘形成。     

Effect of Microgene pSVPoMcat Implantation to Modify Genetically Schwann Cells on Central Conduction Function after Spinal Cord Injury

We have tested cortical somatasensory evoked potentials (CSEP)after spinal cord injury treated by pSVPoMcat modified Schwann cells implantation and the change of combined behavior score introduced by Gale et al.to explain its effect on central condution after experimental SCI.1 Materials and Methods1.1 SCI model and implantation process Adult SD rats( weighing 200－230g,with no gender regarded,provided by the animal Center of the West China University of medical)were made hemi－transe…     

微基因修饰雪旺氏细胞移植对脊髓损伤中枢传导功能的作用

Objective To approach the effect of microgene pSVPoMcat to modify genetically Schwann cells (SC) on central conduction functionafter spinal cord injury (SCI) . Method Experimental animals were divided into three groups: the group of microgene pSVPoMcat implanted to genetically modify SC (group A), SC implanted group (group B), and the control group (group C). The cortical somatasensory evoked potentials (CESP) and combined behavioral score (CBS) were continuous monitored from operation second week to twelfth week Result The result showed that the peak latency and peak amplitudes of group A, B have a recovery tendency and it was constant with CBS. Conclusion We concluded that the microgene pSVPoMcat to modify genetically Sc may play a promotion role in recovery of central conduction function after SCI.     

pSVPoMcat微基因修饰雪旺细胞脊髓内移植对损伤脊髓细胞的保护作用

Objective To study the protective effects of the intracord transplantation of microgene pSVPoMcat－ genetically－ modified Schwann cells (MSCs)on spinal cord injury (SCI).Method Rats with semi－ division(SD) of the spinal cord was divided into 4 groups.Group S consisted of the rats with SD treated with the transplantation of MSCs, Group B of the rats with SD treated with the transplantation of SCs without genetic modification,Group C of the rats with SD without treatment and Group D was the normal control. 8 hours after operation,the half of the rats of each group were killed and the injured segment of the spinal cord was resected to be examined with atomic absorption spectrophotometry . Another half of the rats of all the groups were examined with neurological function tests to have a combined behavioral score (CBS).Result There was a significant increase of water content and Na＋ and Ca2＋ ions and a decrease of K＋ and Mg 2＋ ions in the injured cord segment of Group C and a statistically significant recovery was observed in Group A. The intracord transplantation of pSVPoMcat genetically modidied SCs improved the neurological outcome of spinal cord injury.Conclusion Our findings indicate that intracord transplantation of pSVPoMcat－ genetically－ modified－ Schwanncells exerts protective effects on the injured segment of the spinal cord through the improvement of the internal ion environment of the spinal cord.     

囊性颅咽管瘤内置管囊小剂量多次化疗的疗效观察

目的探索神经外科综合治疗位于视丘下部附近区域的囊性颅咽管瘤之方法。方法选择16例囊性或以囊性为主的颅咽管瘤,在CT介导立体定向内窥镜下切除部分囊壁或肿瘤实质,排除囊液减压后囊内置入Ommaya管,术后经反复抽吸囊液,并反复注入小剂量博来霉素30～40次。结果16例经治疗后视力、视野障碍明显好转,7例有颅内压增高者术后缓解。CT显示瘤腔缩小,无手术死亡,术后视丘下部功能紊乱反应轻。14例获随访2～3年,临床症状明显改善。结论该疗法安全、简便、有效,是采用微侵袭神经外科手段治疗颅咽管瘤的有效方法,有一定的临床应用价值。     

磷酸钙骨水泥临床应用类型选择及微结构观察

根据磷酸钙骨水泥（CPBCs)的基本性能选择符合临床适用的类型,并观察适用类型CPBC固化体超微结构。制备五种CPBCs粉末,即磷酸八钙－沉淀性羟基磷灰石（OCP－PHA）、磷酸八钙－羟基磷灰石（OCP－HA）、磷酸八钙（OCP）、缺钙性羟基磷灰石（CDHA）、羟基磷灰石（HA).调和液为去离子水和0．25M Na2HPO4/NaH2OPO4等摩尔浓度缓冲液。采用Gillmore双针法测定凝结时间,采用TS－14单颗粒压缩强度自动测量仪－Ⅱ测定压缩强度,按照Ginebra临床应用标准选择适用类型,并用扫描电镜观察适用类型CPBC固化体浸泡仿生液体前后的超微结构。结果表明：5种CPBCs中以OCP－PHA型和CDHA型CPBCs的凝结时间和压缩强度符合Ginebra临床应用标准。SEM观察两种CPBCs固化体为多孔结构,并随浸泡仿生液体中时间的延长而溶解或溶合。OCP－PHA型和CDHA型CPBCs适合于非承重骨的修复,其固化体为多孔结构,利于纤维组织长入及纤维血管化。     

外源性bFGF对大鼠脊髓损伤后低氧诱导因子HIF-1α表达的影响

目的：研究实验性脊髓损伤后低氧诱导因子（HIF-1α）的时序性变化及与创伤性凋亡的关系。探讨其在脊髓损伤中的意义。明确外源性bFGF对HIF-1α表达的影响。方法：以大鼠静压型脊髓损伤模型为研究对象,运用免疫组化方法进行HIF-1α染色,流式细胞术测定凋亡率。结果：脊髓损伤后1天开始,HIF-1α表达明显增加,3-7天达到高峰,14天时HIF-1α阳性染色率下降显著,HIF-1α的时序性变化与凋亡率,变动趋势密切相关;使用外源性HIF-1α后,各时点HIF-1α表达较损伤组都有明显下降。结论：HIF-1α参与了脊髓损伤后缺血缺氧的继发损害过程,它与创伤后凋亡之间有着密切的关系。对其研究的深入可能为找到有效的治疗脊髓损伤的方法提供新的思路。     

大鼠创伤性脑损伤后神经细胞凋亡的实验研究

探讨创伤性脑损伤后神经细胞凋亡机制及其时序空间分布。方法采用末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口末端标记法、琼脂糖凝胶电泳、透射电镜、流式细胞术、凋亡调节蛋白免疫组织化学方法检测大鼠轻、重度脑损伤不同脑域神经细胞凋亡的动态变化。     

人脑胶质瘤低氧诱导因子-1α的基因表达

目的 从人脑胶质瘤中克隆低氧诱导因子－1α（HIF－1α）cDNA,探讨其对人脑胶质瘤发生、发展的作用。方法 从人脑胶质瘤组织中提取总RNA,以逆转发－多聚酶链反应（RT－PCR）方法扩增出全长2481bp的HIF－1α cDNA,经Nhe Ⅰ和BamH Ⅰ双酶切后,将其与表达载体pWAV1连接,转化入E.coli JM109,建立HIF－1α的cDNA克隆;测序后与大鼠HIF－1αcDNA序列进行同源性分析。结果 RT－PCR产物进行琼脂糖凝胶电泳,可见一清晰特异扩增带,长2．5kb,HIF－1α cDNA经筛选、双酶切鉴定后,可见5．3kb的载体片段及2．5kb的插入片段。进行序列分析,与鼠有905的同源性。结论 HIF－1α已成功从人脑胶质瘤中得到克隆和序列分析。