[目的]观察慢病毒介导的CD-TK融合基因转染神经干细胞(NSCs)对C6胶质瘤生长的抑制作用以及疗效.[方法]利用慢病毒介导的大肠杆菌胞嘧啶脱氨酶(CD)和单纯疱疹病毒胸苷激酶(HSV-TK)融合基因转染NSCs,植入C6胶质瘤模型.用流式细胞仪技术检测C6胶质瘤生长凋亡情况.[结果]成功构建CD-TK基因转染NSCs模型.实验组坏死细胞(17.80％)、早期凋亡细胞及晚期凋亡细胞(33.30％)较对照组增加.CDglyTK-NSC+GCV/5-Fc治疗组肿瘤体积为9.36±2.35mm3,小于其他处理组及对照组的肿瘤体积(P＜0.05).[结论]转染NSCs的CD-TK基因对C6胶质瘤细胞有抑制作用,抗肿瘤效果明显,优于单一的自杀基因系统.

Objective To explore the effects of lentivirus-mediated cytosine deaminase(CD)-thymidine kinase(TK) fusion gene transfection on the proliferation of neural stem cells(NSCs) in vitro.Methods Primary NSCs obtained from the subventricular zone(SVZ) of newborn Wistar rats were cultured and identified.After passage and purification in vitro,NSCs were treated with lentiviruses and packaged with CD-TK gene for 3 to 5 d,and then MTT assay was used to detect the proliferation of NSCs.Immunofluorescence technique was used to detect the expressions of glial fibrillary acidic protein(GFAP) and neuronspecific enolase(NSE) in NSCs.Results CD-TK gene transfection showed no influence on the proliferation of the NSCs(P > 0.05).The NSCs was positive for nestin and the naturally differentiated cells were positive for GFAP and NSE by immunofluorescence after incomplete culture.Conclusion CD-TK gene transfection does not affect proliferation of NSCs.

It is well known that puerarin attenuates ischemia–reperfusion injury and promotes function recovery of ischemic region. However, due to its reverse physiochemical properties, puerarin does not easily cross the blood–brain barrier. The aim of the present study is to create puerarin nanoparticles which increase and prolong the puerarin concentration in the brain. Using emulsion solvent evaporation techniques, we designed puerarin-loaded poly(d,l-lactic-co-glycolic acid) nanoparticles. Hydroxypropyl beta cyclodextrin (HP-β-CD) was used to increase the solubility of puerarin and gelatin to enhance viscosity of inner water phase, which improved puerarin entrapment. The drug release kinetics and nanoparticle degradation in phosphate buffered saline (PBS) were analyzed by electronic microscopy and high-performance liquid chromatography. Computerized tomography scans were used to detect the infarction volume and electroencephalogram (EEG) was recorded to estimate the recovery of brain function. The results showed that the combined HP-β-CD and gelatin significantly improved the entrapment efficiency. The infarction volume was significantly decreased on days 3 and 7 after the administration of puerarin nanoparticles compared with that of control and pure puerarin. EEG was also significantly improved. Puerarin nanoparticles are potentially applicable for the brain injury induced by ischemic–reperfusion.

A suicide gene can convert nontoxic prodrugs into toxic products to kill tumor cells. In this study, our aim was to transfect lentivirus-mediated CD/TK fusion gene into Wistar rat’s neural stem cells (NSC) and then implant the NSC into a C6 glioma model to observe a C6 glioma growth inhibition effect. Primary NSC and stable transfection CD/TK fusion gene cell lines were established. To observe the tumor size and rat survival period in different groups, C6 glioma cell apoptosis and cell viability rate were applied to analyze the tumor inhibition effect of the neural stem cells’ transfected CD/TK fusion gene. C6 cell viability showed that CDglyTK-NSC + GCV/5-Fc (group 1) was lower than CDglyTK-NSC (group 2), NSC + GCV/5-Fc (group 3), and control (group 4) from day 2 ( p < 0.05), and the apoptosis rate was higher in group 1 compared with that of other groups (50.6 %, p < 0.05) either in vitro or in vivo (35.47 %, p < 0.05); both cell viability and apoptosis had no significance in the other three groups. In vivo, tumor size in group 1 was 7.76 ± 1.37 mm 3 , which is smaller than the others (group2 27.28 ± 4.11 mm 3 , group3 27.94 ± 2.08 and 28.61 ± 2.97 mm 3 ; p < 0.05). The other groups’ tumor size was not significant ( p > 0.05). Survival time of rats treated with CDglyTK-NSC + GCV/5-Fc (group 1) was significantly longer than that of the other groups ( p < 0.05; group 1 48.86 ± 1.97, group 2 28.67 ± 3.75, group 3 31.5 ± 1.27, group 4 29.3 ± 1.33). We also showed that the transfected C6 cells had a migratory capacity toward gliomas in vivo. Transfected CD/TK fusion gene neural stem cells combined with propyl–guanosine and 5-flucytosine double prodrug significantly inhibit the development of glioma.

A large number of studies show that traditional Chinese medicine is effective in the treatment of glioma,which is characterized by multiple pathways,multiple targets,multi-functions and less side effects.Recently,many researches,both domestic and foreign,focus on the study of active constituents of traditional Chinese medicine,including curcumin,resveratrol,cannabinoids,elemene and Tripterygium wilfordii monomer,which have multiple mechanisms of anti-glioma effects.This paper reviews recent researches on mechanism of active constituents of traditional Chinese medicine against glioma.It will be a reference for the integrated treatment of traditional and western medicine against glioma.

Objective To study the role of heat shock protein 70 (HSP70) in the cerebral vasospasm after subarachnoid hemorrhage (SAH) in rabbits.Methods Fifteen Japanese rabbits were randomly divided into control group (n=3) and experimental groups (n=12),who were redivided into 4 subgroups of 3 animal each according to the time from SAH to sacrifice.The SAH was produced in the rabbits of all the experimental groups by injecting autologous fresh arterial blood into the cisternae magna.The basilar arteries were taken from the animals,who in the experimental group were sacrificed respectively 3,7,14 and 21 days after SAH.The gross and histological changes in the basilar arteries of all the rabbits were observed.The expressions of HSP70 in the basilar arteries of all the rabbits was determined by immunohistochemical technique.Results The gross and histological changes in the basilar arteries after SAH included thickening of the arterial walls,narrowing of the arterial lumina,swelling and deformation of the endothelial cells,wrinkling of the elastic interna and so on.The expression levels of HSP70 in the endothelial cells of the basilar arteries were significantly lower 3,7 and 14 days after SAH than that in the control group (P <0.01).Conclusion It is suggested that the cerebral vasospasm after SAH may be related to the decrease in HSP70 expression level in the cerebrovascular endothelial cells.

目的 探求脑室腹腔分流术在腹腔镜辅助下微创放置分流管腹腔端对肥胖患者的安全性和可行性.方法 回顾性分析了我院2007年至2008年42名肥胖脑积水病人采用腹腔镜辅助下腹腔端置管技术的手术指征和并发症,手术技巧及注意事项.结果 术后并发症主要包括颅内感染1例,脑室端梗阻2例,腹腔端包裹性积液1例.未发生腹腔端异位或腹腔脏器损伤.结论 腹腔镜辅助下分流管腹腔端置管技术安全有效,具有操作简单,对患者损伤轻微,并发症低等特点.

目的 总结经单侧额下入路切除嗅沟大脑膜瘤的手术治疗经验.方法 回顾性分析11例经单侧额下入路手术治疗嗅沟大脑膜瘤.结果 Simpson Ⅰ级切除8例,Ⅱ级切除3例,术后2例出现脑脊液鼻漏,无手术死亡.结论 对于嗅沟大脑膜瘤,术前完善的检查,选择单侧额下入路,并结合显微神经外科技术,可取得的良好疗效.

Objective To investigate the safety and feasibility of ventriculoperitoneal shunt(VPS) by mini-invasive peritoneal catheter placement without laparoscope.Methods The indication,surgical skill and complication with mini-invasive peritoneal catheter placement without laparoscope for VPS were retrospectively analyzed in 85 consecutive patients.All the patients were given a 5 mm skin incision under artificial pneumoperitoneum,and then the shunt catheter was placed into the abdominal cavity without laparoscopy.Results Two patients suffered intracranial infection,2 cerebral ventricular obstruction,1 misplaced ventricular catheter and 1 abdominal pseudocyst,and no bowel injury and distal catheter misplacement happened.Conclusion VPS by minimally invasive peritoneal catheter placement is characterized by minimal invasion,safety,simple operation,saving medical resource and reducing complications,thus being easy to perform in ordinary hospitals.

Objective To discuss the clinical therapeutic effect and value of emergent surgery for intracranial Hunt-Hess grade Ⅳ and Ⅴ aneurysms. Method First we improved patient's condition through decompressive craniotomy, evacuation of hematoma and external ventricular drainage, and then we performed aneurysm surgery. Results A retrospective analysis of 58 patients with intracranial aneurysms were treated with emergent surgery. There were 50 cases of Hunt-Hess grade 1V ,in witch 26 cases had a good outcome, 13 cases had a mild disability, 8 cases had a severe disability and 3 cases died. There were 8 cases of Hunt-Hess grade Ⅴ, in witch 2 cases had a good outcome, 1 case had a mild disability, 2 cases had a severe disability and 3 cases died. Conclusions Emergent surgery is a first choice for hunt-Hess grade Ⅳ and Ⅴ aneurysms and the results were satisfactory.

Objective To investigate surgical indications,techniques and effects for intraeranial arachnoid cyst in children. Methods Clinical data from 82 surgical cases were analyzed retrospectively,and different methods of operation were analyzed comparatively.Four operation-methods were adopted,in which cyst removal were performed in 19,cyst removal plus cisternal opening in 23,cyst-peritoneal shunt in 28,and cyst-peritoneal drainage in 16.Results Upon following up from six months to five years after operation,the chief symptoms got improved in 89.0% of the patients,and CT or MRI scan showed that the size of cvst became smaller in 72 of the patients.Four operation methods were compared,cyst removal was not better than the other three operation methods.Conclusions The treatment of operation methods could make the svmptoms improved and the cyst thrunk of intracranial arachnoid cyst in children.The operation of cyst removal plus cisternal opening or cyst-peritoneal shunt and cyst-peritoneal drainage is preferred.

目的 探讨神经干细胞(NSCs)经枕大池移植到创伤性脑损伤模型大鼠蛛网膜下腔中,观察其向损伤脑组织迁移及治疗效果.方法 体外培养的NSCs取自子鼠皮层,并用5-溴脱氧尿嘧啶(BrdU)标记.采用Feeney\u0027s自由落体脑创伤模型制成大鼠脑损伤模型,伤后24 h将NSCs经枕大池移植到蛛网膜下腔,伤前24 h、伤后24 h及1、2周行动物运动神经功能评分,分别于第1、2周处死取脑,行免疫组织化学染色检测BrdU、微管相关蛋白2(MAP2)、胶质纤维酸性蛋白(GFAP).结果 免疫组织化学染色表明,有BrdU(+)NSCs迁移到脑内损伤灶并分化成MAP2(+)神经元或GFAP(+)胶质细胞.接受NSCs移植组大鼠与损伤组大鼠比较运动功能明显改善.结论 NSCs具有从蛛网膜下腔迁移入脑内损伤组织、分化为神经元及神经胶质细胞并促进或改善神经功能恢复的能力。

颅内蛛网膜囊肿（IAC）是儿童期一种常见的良性病变,病因主要是先天性因素,外伤、炎症等因素较少。现对我院治疗的82例临床资料进行分析。

目的 本实验将神经干细胞经枕大池移植到创伤性脑损伤模型大鼠蛛网膜下腔中,观察其向损伤脑组织迁移、存活及分化能力,探讨神经干细胞更安全、便捷及经济的移植途径.方法体外培养的神经干细胞(NSCs)取自胎鼠皮层,并用5-溴脱氧尿嘧啶(BrdU)标记.采用Feeney\u0027s自由落体脑创伤模型制成大鼠脑损伤模型,伤后24 h将神经干细胞经枕大池移植到蛛网膜下腔,分别于移植后第1、2周处死取脑,行免疫组织化学染色检测BrdU、微管相关蛋白2(MAP2)、胶质纤维酸性蛋白(GFAP).结果免疫组织化学染色表明,BrdU(+)神经干细胞迁移到脑内损伤灶,并分化成MAP2(+)神经元或GFAP(+)胶质细胞.结论神经干细胞具有从蛛网膜下腔迁移入脑内损伤组织、分化为神经元及神经胶质细胞。

目的探讨大鼠脑损伤后,将神经干细胞(neural stem cells,NSCs)经枕大池移植到蛛网膜下腔及立体定向移植到脑内,观察神经功能恢复情况.方法体外培养的NSCs取自胎鼠皮层,采用Feeney自由落体脑创伤模型制成大鼠脑损伤模型,伤后24小时将NSCs经枕大池移植到蛛网膜下腔及经立体定向移植到脑内.伤前24小时、伤后24小时及1、2周行动物运动神经功能评分.结果接受NSCs移植的两组大鼠神经运动功能评分均较损伤对照组明显提高(P＜0.05),两移植组大鼠神经运动功能评分无明显差别.结论经枕大池移植的NSCs具有远距离迁移能力,并能像脑内移植一样明显有助于大鼠神经运动功能的恢复.

背景:基因治疗是国内外对于脑胶质瘤生物学治疗的研究热点。目的:应用已克隆并构建的真核表达载体pCR3-TK,探讨HSV-TK/ACV系统对人脑胶质瘤细胞抑制生长和杀伤作用研究。设计:以细胞为研究对象的实验研究。单位:一所大学医院神经外科、肿瘤内科。对象:实验于2004-01/04在哈尔滨兽医研究所生物技术国家重点实验室完成。所用的真核表达载体pCR3-TK由作者构建,TJ905细胞株由天津神经病学研究所浦佩玉教授惠赠。选择未转染和转染空载体的细胞作为对照组。方法:用阳离子脂质体Lipofectamine将pCR3-Uni及含HSV-TK基因的真核表达载体pCR3-TK转染至人脑胶质瘤细胞株TJ905中,筛选出阳性克隆,阳性细胞克隆给予ACV（50mg/L）,72h后,收集玻片,进行AgNOR染色。主要观察指标:对未转染和转染不同载体的TJ905细胞ACV作用后银染颗粒进行记数。结果:转染HSV-TK基因的细胞,在给予ACV后,细胞增殖活性明显降低,转染pCR3-Uni和pCR3-TK细胞克隆的AgNOR颗粒数分别为14.33和6.67（P＜0.01）。结论:AgNOR计数是一种操作简便、检测细胞增殖活性的方法,为研究HSV-TK/ACV系统的抗肿瘤机制提供帮助。

中央前回深部病灶,手术切除成功的关键在于术前、术中准确的定位和熟练的显微操作技术.在2002年12月至2003年12月间我院在显微镜下切除中央前回深部病灶16例,达到明确病变性质和治疗目的,现报告如下:

Objective To study the effect of the proliferation activity of human glioma cells with HSV\|TK/ACV system.Methods The eukaryotic expressing vectors pCR3\|TK containing the HSV\|TK gene and pCR3\|Uni were transferred into TJ905 cells with cationic liposome, Lipofectamine .After transfection,G418 was used to select the positive clones. The psoitive clones were given ACV(50μg/ml) and collected afer 72 hours. The glass slides were stained with silver.Results The number of silver stained AgNORs in the cell clones (transfected with pCR3\|Uni and pCR3\|TK) was 14.33 and 6.67, respectively .The proliferation activity of glioma transfected with pCR3\|TK was significantly decreased ( P 0.01). Conclusion AgNOR was a simple method of checking the proliferation activity of cells. It provided with an aid on studying the antitumor principle of HSV\|TK/ACV system.