目的： 硅是制作神经芯片的良好材料，硅的普通抛光表面使神经细胞黏附极为困难，我们用增加材料表面粗糙度的方法来提高硅的细胞黏附性能。方法： HF溶液对硅片进行表面蚀刻，原子力显微镜测定表面粗糙度。选用胎鼠黑质细胞种植于硅片表面进行培养，72h 后以酪氨酸羟化酶（TH）免疫荧光显色，荧光显微镜及激光共聚焦系统观察细胞的生长状况，并进行扫描电镜观察。结果： 硅材料表面粗糙度的增加可以明显改善细胞的黏附和生长状况，生长于硅片表面的多巴胺能神经元表达TH。结论： 增加表面粗糙度可以提高硅材料对细胞的黏附性能，改善其生物相容性;适于细胞生长的硅片表面平均粗糙度Ra 约为20～50nm，该技术可作为神经芯片材料表面处理的良好方法。

A fast and effective method to enrich large number of neural precursors from the ventricular zone of human fetus by magnetic affinity cell sorting (MACS) is reported. After incubation with phycoerythrin (PE)-conjugated anti CD133 antibodies and anti-PE magnetic beads followed by one cycle of MACS, CD133+ cells were harvested at 85% purity as confirmed by flow-cytometry and immunocytochemistry. In contrast to CD133-cells, these CD133+cells initiated primary and secondary neurospheres in culture, and the progeny of sorted cells could be differentiated into both neurons and glia, indicating that these highly enriched cells are capable of self-renewal and multi-lineage potential.

为了探讨微囊化转酪氨酸羟化酶基因的细胞在帕金森病大鼠模型脑内存活、组织反应及对异常行为的治疗效果。本研究将带有酪氨酸羟化酶基因的载体—质粒pCMV 2TH在体外通过脂质体转染技术转入原代培养的人胚骨骼肌细胞内。将转染后的肌细胞包裹在藻酸盐-多聚赖氨酸-藻酸盐构成的半透膜微囊中，然后将这些微囊化的能表达酪氨酸羟化酶的骨骼肌细胞植入模型大鼠脑的纹状体内。结果表明，微囊化的转基因细胞在脑内可存活至实验结束(16周)，可表达酪氨酸羟化酶。微囊化细胞周围无显著免疫排斥反应及炎症反应。本研究结果表明: 将用于异种移植的转基因细胞微囊化是使基因治疗实用化的有效手段。

用计算机三维重构技术建立了人及大鼠基底神经节的三维数字化模型，并对两者的形态及核团构成进行了比较解剖学研究。在微型计算机上，提取人脑及大鼠脑立体定位图谱中含有基底神经节的连续层面，利用通用的三维建模及动画软件3DStudio MAX，以三维放样的方法，在我国首次建立人及大鼠基底神经节的核团及纤维传导通路的三维数字模型，对人及大鼠的基底神经节进行了比较解剖学研究。并利用重构的三维模型建立虚拟现实文件，在神经解剖学研究中引入浏览器虚拟现实技术,实现了任意拆分、组装及旋转上述核团的功能。本研究显示，无论在形态上还是在毗邻关系上，人与大鼠的基底神经节的差异不大；由于人的直立而导致脑干吻尾轴旋转了一定角度，其内部核团的相互毗邻关系也随之发生变化，从而异于大鼠基底神经节内部核团的毗邻关系；基底神经节各个核团间的投射纤维以尽可能短的路径到达靶核团的相应区域，构成了拓扑投射关系的基础。

目的 观察酪氨酸羟化酶（TH）和胶质细胞系源性神经营养因子（GDNF）基因工程细胞联合移植到纹状体内对帕金森病（PD）大鼠的治疗作用。方法 重组携带TH基因的质粒表达载体pCMVTH及携带GDNF基因的质粒表达载体DCl。neo，分别转染原代培养的成纤维细胞。筛选阳性细胞植入大鼠帕金森病模型纹状体内，动态观察行为学变化，并用高效液相色谱于第4、8、12、16、20周测定多巴胺、3。4-二羟苯乙酸（DOPAc）及高香草酸（HVA）含量的变化。结果 联合移植TH及GDNF转基因细胞致PD大鼠纹状体内可明显减少模型单侧旋转行为，可降至移植前旋转圈数的44.3±5.2%（P<0.01），其效果优于单独移植TH（65.6±6.1%）或GDNF（64.8±5.8%）转基因细胞的PD模型（P<0.05）；显著提高纹状体内多巴胺含量。恢复至移植前水平的21.4±3.47%（P<0.001），其程度高于单独移植TH（12.9=1.4%）或GDNF（14.5±2.2%）转基因细胞的PD模型（P<0.05）。结论 TH及GDNF联合基因治疗具有潜在的临床应用价值。

Objective To induce significant improvement of motor abnormalitie s and striatal dopamine (DA) levels in rat model of Parkinson's disea se (PD), by intrac2 erebral grafting of the genetically modified muscle cells expressing tyro sine hydroxyla se (TH). Me t h o ds Primary myobla st s and myotube s from the rat were prepared by cell culture and a plasmid, pCMVTH, containing TH gene and a promoter of cytomegalovirus (CMV) wa s constructed by DNA recombination technique. The primary muscle cells were transfected with newly constructed pCMVTH DNA vector, by using lipofection. The se genetically modified muscle cells were grafted into the caudate-putamen of 62OHDA2le sionedrats, repre senting PD models. Before and after grafting, the rotational behaviour and the striatal levels of DA and it s metabolitie s were te sted at different po stoperative survival time s. In addition, the immunocy to chemistry for showing TH activity wa s done both in vitro and in vivo. Results The newly constrcuted pla smid, pCMVTH wa s proved to contain TH gene and have correct direction of insertion. The cultured primary myobla st s and myotube s lipofected with pCMVTH were immunocytochemically shown to expre ss TH activity in vitro. After grafting, the se TH2expre ssing muscle cells showed to have a long2term survival cells in vivo and induced a marked decrea se in abnormal locomotion and a increa se in striatal DA levels for PD rat model. Conclusions In experimental gene therapy for PD, the pCMVTH is a useful vector for carrying TH gene. The lipofection is a practical technique for transferring a target gene into eukaryote s and primary cultured muscle cells should be a good vehicle for DNA transfer and intracerebral grafting.

Among the newly discovered spinocerebellar cell groups, theose at lumbar and more caudal levels of the cat's spinal cord were studied with regard to which of the two cerebellar peduncles, the restiform body or the superior cerebellar peduncle, is used by theri axons. Bilateral injections with horseradish peroxidase were made into either of the anterior lobe or the posteriosr cerbellar termination area for spinocerebellar fibers, following unilateral transections of either the superior serebellar peduncle or the resiform body, combined with low contralateral transecions of the lateral and ventral funiculi. Following transection of the superior cerebellar peduncele, labeled neurons were found ipsilateral to the transection in the colun of Clarke and in laminas IV-VI at L3-L7. Contralalterally, labeled neurons were found in the ventromedial nucleus and lamina and in the medial part of lamina VII at L6 and more caudal levels. All these neurons were regarede as semding their axons through the restiform body. Followitn transection of the restiform body, labedled neurons were found in the following areas contralateral to the transection: the dorsolateral nucleus of the L3-K6 segments, the lateral part of lamina VII at L3-L5/6, the medial part of lamina VII in L6 and more caudal segments, and the ventrolatreal nucleus of L4-L5. Ipsilaterally, labeled neurons were found in lamina VIII at L4-L6. All these neurons were regarded as sending their axons through the superior cerebellar peduncle. In addition to new information about the peduncular routes of spinocerebellar neurons, the study has given confirming evidence as to the crossing conditions for different spinocerebellar cell groups. The findings should be useful future studies on the organization of the spinocerebellar system.

The fiber couse of the spinocerebellar tracts in the ventral and lateral funiculi of the cat spinla cord were studied by a new approach making cordotomies at different spinal levels or lesions of the restiform body followed by injectionas of HRP or WGA-HRP into the anterior cerbellar olbe. The retrogradely labeled axons showed characteristic distribution patterns ralated to the level and extent of the lesions. The results show the following. 1) The dorsal spinocerbellar tract (DSCT) originationg ipsilaterally form the thoracic and upper lumbar segments ascends in the dorsolateral fasciculus. It underoes a dorsal shift during its rostral coures. The tract is topially arranged and passes through the restiform body. 2) The ventral spinocerebellar tract (VSCT) arising contralaterally from lower thoractic, lumbar, and more eaudal segments passes via the ventral funiculus and ascends in the ventrolateral fasciculus. This tract is also topically arranged. It maks a lateral and then a dorsal region enetrs via the restiform body. 3) The spnocerebellar fibers oriinationg ipsilaterally from the cervical enlafrgement ascend in the lateralmost part of the lateral funiculus in the area between the dorsolateral and ventrolateral fasciculi, There fibers form two groups, one passing through the dorsolateral and ventrolateral fasciculi. These form two groups, one passing through the restiform body, the other through the superior cerbellar peduncle. 4) The spinocerebellar fibers origination comtralaterally form the central cervical nucaleus pass through the ventral funiculus and ascend in the lateralmost part of the lateral funiculus, mainly inthe ventrolateral fasciculus. Most of the fibers seem to pass through the superior cerebellar peduncle.

The collateral projections of spinocerebellar neurons located in the L 2to Ca 1 spinal segments in the cat were investigated by retrograde fluorescent double labeling technique. Rhodamine labeled latex microspheres (Rm) amd Fast Blue (BF) were used for injections into the cerebellum in 8 cats. Two additonal cats, with injections of Fluoro-Gold (FG) combined with Rm were excluded because lipofuchsin autofluorescence obscured the labeling. Aafter injections with one tracer unilaterally in the paramedian lobule and another tracer bilaterally in the anterior lobe, double labeled neurons were found on the side of the paramedian lobule injection in the anterior lobe, double labeled neurons were fond on the side of the paramedian lobule injection in the column of Clarke at L2-L4, laminae IV-VI at L2-L5 and the dorsolateral nucleus at L2-L6. After bilateral injections of one tracer in lobule VIII Band another in the anterior lobe, double labeled neurons were found bilaterally in the column of Clarke at L2-L4, laminas IV-VI at L2-L5, the medial part of lamina VII at L6-L7 and in certain cell groups at sacro-coccygeal levels. Neurons in the lateral nucleus of L4-L5 were labeled exclusively from. injections in the anterior lobe. The findings indicate that spinocerebellar neurons at lumbar and more caudal levels of the cat spinal cod have different projection patterns in the cerebellum. A certain number of neurons which project to the anteriro lobe have divergent axon collaterals supplying also the postrior vermis and/or the paramedian lobule. Other peurons project to the anterior or to the posterior lobe only.