胶质细胞

The pcDNA〓-apoE〓 plasmid was transfected to SK-N-SH Neuroblastoma cell by liposome. It could be screened by G〓. The immunohistochemical assay was shown that neuroblastoma cell transfected by pcDNA〓-apoE〓 expressed the recombinant apoE protein. The intracellular expressed recombinant protein could inhibit the death of neuroblastoma cell induced by beta amyloidal peptides 25-35 fragment.

用pcDNA〓—apoE〓真核表达载体与脂质体共转染神经成纤维胶质瘤细胞，用G〓选择压力筛选，细胞免疫荧光检测表明，pcDNA〓—apoE〓转染的神经成纤维胶质瘤细胞表达了apoE〓重组蛋白，这种内源性的apoE〓重组蛋白对25μM Aβ25—35多肽诱导的神经成纤维胶质瘤细胞的死亡具有拮抗作用。

Methods DCs were prepared from peripherad blood mononuclear induce d with gra-nulocyt e/macrophage colony-stimulating factor and interleukin-4. Apoptosis of glioma cells were induced with γ-radiation. We design these experiment groups including (1) coculture of DCs and apoptotic glioma cells and T cells,(2) coculture of DCs and U937 cells and T cells,(3) coculture of DCs and cultured glioma cell and T cells,(4) coculture of Des and T cell.

用粒－巨噬细胞集落刺激因子加白介素－4(IL-4)从人外周血分化、诱导DCs、γ-射线在体外诱导培养的人脑胶质瘤细胞凋亡，将DCs、T淋巴细胞和凋亡胶质瘤细胞共培养，同时设计不同类型肿瘤细胞（U937及培养胶质瘤细胞）作对照，分离、富集DCs、T淋巴细胞进行免疫应答及肿瘤细胞杀伤试验。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法：1、培养原代的人RPE细胞，经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后，通过AO/PI染色技术确定培养细胞的存活率，描记其生长曲线，第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后，通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后，采用MTT法观察其对细胞的增殖力的作用；通过流式细胞仪观察其对细胞周期的影响；通过扫描电镜观察其对细胞形态的影响；通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用；采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率，评价其对细胞间通讯功能的影响；通过制作RPE细胞损伤模型，观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质，应用等电聚焦电泳和SDS-PAGE双向电泳技术，银染显示分离出的蛋白质斑点，经凝胶图像分析软件对两个样本进行胶图分析，寻找差异蛋白点。

By culturing human brain microvascular endothelial cells and glioma cells separatedly and simutaneously ,and then analyzing their pre- and post-cultune FasL levels, we study the effect of the grouth of glioma cells upon FasL level of human brain microvascular endothelial cells.

我们通过将人脑毛细血管内皮细胞与胶质瘤细胞分别培养及共同培养，对培养前后人脑毛细血管内皮细胞与胶质瘤细胞FasL表达水平的分析，来探讨胶质瘤细胞的生长对人脑毛细血管内皮细胞FasL表达的影响。

Methods At first, retrovirus vectors encoding IFN-γ or IL-4 gene were constructed. They were transfected into PA317 packaging cells by lipofectamine, PA317IFN-γ and PA317IL-4 cells were obtained. C6 glioma cells were infected with replication deficiency retrovius containing IFN-γ or IL-4 gene, cell morphology、cell growth curve and cloning efficiency assay were examined. The intracranial C6 glioma animal model was established in immunocompetent Wistar rats. PA317IFN-γ and PA317IL-4 cells were stereotactically implanted into the tumor areas alone or together. The survival of tumor bearing rats were examined, tumor volumes were measured and immunohistochemical analyses of CD4〓 and CD8〓 T lymphocyte infiltration were performed.

构建和鉴定携带IL-4或IFN-γ基因的逆转录病毒载体，将其导入逆转录病毒包装细胞PA317，通过G418抗性筛选，得到携带目的基因的包装细胞并鉴定；应用携带IL-4或IFN-γ基因的复制缺陷型逆转录病毒感染C6胶质瘤细胞，观察对C6胶质瘤细胞形态、细胞生长曲线和克隆形成率的影响；建立C6胶质瘤大鼠脑内移植动物模型，将携带IL-4和IFN-γ基因的逆转录病毒包装细胞分别或联合注射到脑内荷瘤大鼠的肿瘤组织中，观察其治疗作用，并初步探讨其机制。

In Part 1 of our research, BGSCs were sorted through immunomagnetic beads marking by CD133 and cultured in vitro, and character as a stem cell was identified by stem cell markers (CD133 and Nestin) and differentiated cell markers [ microtubule-associated protein 2(MAP2), glial acidic fibrillary protein and myelin basic protein] , ultrastructure observing with electron microscopeand engrafting to severe combined immunodeficiency mice for tumorigenesis test. The results were as following: Only a small subset of CD133+ glioma cells in glioma cell lines and fresh specimens from various pathologic grade could express stem cell markers CD133 and Nestin, view ultrastructure of a stem cell and be capacity of serial passage in culture. These CD133+ cells possese a marked capacity for multipotent differentiation and could differentiate into tumor cells expressing MAP2,β-TubulinⅢ, GFAP and MBP; When engrafted into SCID mice, they can generate and form tumors that phenotypically resembl the tumor from the patient.

在本课题中，在第一部分实验中采用以CD133为标志的免疫磁珠法从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞并进行体外培养，通过免疫荧光技术检测干细胞标志物CD133、Nestin，诱导分化后检查分化细胞标志物MAP2、GFAP、MBP以及电镜超微结构观察和移植SCID鼠致瘤实验，对其干细胞特性加以鉴定，得到如下结果：不同病理分级的新鲜胶质瘤标本和胶质瘤细胞株中存在一小部分CD133+的胶质瘤细胞，能表达干细胞的标志物CD133和Nestin，符合干细胞的超微结构特点，体外培养能连续传代；具有多向分化潜能：诱导分化后能产生MAP2、β-TubulinⅢ、GFAP、MBP染色阳性的细胞；移植SCID鼠后能形成与亲本肿瘤表型一致的移植瘤。

Neural stem cells have a strong self-renew mechanism and it can transform after a little break. Neural stem cells have a long term survival, which mean that it has more probability of wrong copy than mature cells. These cells are formed glioma stem cells in the end. The genes who adjust neural stem cells can express in glioma stem cells, which hold out glioma stem cells from neural stem cells. There is another presume that glioma stem cells come from differentiated cells. Through the gene break of these cells, they can obtain characteristics of stem cells, then form glioma stem cells.

神经干细胞具有很强的自我更新机制，获得较少突变即有可能恶性转化，而且干细胞存活时间较长，这意味着干细胞比成熟细胞发生细胞复制的错误几率更大，因外界环境的刺激而发生突变的机会更多，最终形成脑胶质瘤干细胞，同时调节神经干细胞增殖和自我更新的基因在脑胶质瘤的脑胶质瘤干细胞中也表达，这也是支持神经干细胞是脑胶质瘤干细胞来源的；也有推测认为它可能起源于已分化的细胞，由这些细胞突变发生去分化得来，并通过基因突变而获得了干细胞自我更新的特性，从而形成脑胶质瘤干细胞。

Glial cell is the important ingredient of central neural system, and it is also the great integrator in the cranial nerve circuit loop.

胶质细胞是中枢神经系统的重要组成部分，也是大脑神经环路活动的重要整合者。

Results Cerebroedema appeared in all animals and they have some behavioral alterations. The meninges tighten, the brain swollen overtly. The adventitia of the brain arteries and the perivascular Virchow－ Robin space dilated irregularly. In some neurons both the nucleus and cytoplasm condensed, chromatin gathered near the nuclear membrane. Some other neurons appeared degeneration or necrosis. There was intracelluar edema of the astrocytes.

结果所有动物均发生脑水肿，表现行为异常，脑膜及脑组织肿胀、充血，小血管壁外膜及Virchow－Robin间隙增宽，内含大量水肿液，毛细血管基底膜肥厚，神经元固缩、轮廓不清晰或肿胀、坏死，胶质细胞增生、肿胀，硬脑膜表面凹凸不平、水肿明显。

A new method and device were first introduced to measure the deformation rate and deformation degree of silastic membrance in the stretch induced cultured cell injury process.

对培养的新生大鼠大脑皮层星型胶质细胞应用本致伤装置采用不同的条件致伤，检测乳酸脱清酶活性和台盼蓝含量，并观察其形态学的改变。

And Pharaoh spoke to Joseph, saying, Your father and your brothers have come to you.

47：5 法老对约瑟说，你父亲和你弟兄们到你这里来了。

Additionally, the approximate flattening of surface strip using lines linking midpoints on perpendicular lines between geodesic curves and the unconditional extreme value method are discussed.

提出了用测地线方程、曲面上两点间短程线来计算膜结构曲面测地线的方法，同时，采用测地线间垂线的中点连线和用无约束极值法进行空间条状曲面近似展开的分析。