胶质的

Consequently, the properties of asphaltene film will change, but the properties of resin films have been rarely affected.

胶质在此浓度范围没有象沥青质这种自缔合现象，胶质膜的表面浓度几乎不受这些因素的影响。

Northern blot revealed that DAT is expressed weaklt but specificlly in the brain tissue, and in situ hybridization DAT is restrictedly expressed in the adult brain tissues including substantia nigra and striatum, suggesting an important function in the dopaminergic system.

利用cRNA探针的原位杂交组织化学研究，发现DRP在成年大鼠脑组织神经元和胶质细胞均有表达，以胶质细胞广泛表达是其重要特征；而DAT在成年脑组织仅限于少部分核团的有限表达，包括黑质和纹状体。

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

We introduced improved primary mixed glial culture and different-attachment method to isolate and purify the OPCs, the cells were proliferated in serum-free medium, flow cytometry and immunohischemistry methods were employed to estimate the purity of cultured OPCs. Their abilities of differentiation and expression of trophic factors were identified by RT-PCR and immunostaining. Several methods including TUNEL and MTT were adopted to estimate the protective effects of conditioned culture medium from oligodendrocyte lineage cells on the primary cultured cerebellar granule neurons. Intravitreal transplant of OPCs, combined with retrograde fluorescent labeling the superior colliculus and intraorbital optic nerve transection, were used to investigate the protective effects of OPCs on the axotomized RGCs in vivo. Intravitreal transplantof OPCs or NSCs on the newborn rats, and retinal transplant of OPCs on the young rats were performed, to observe the myelin formation in the retina at different stages after cellular transplantation. Optic nerve transection was carried out on some rats with myelinated retinae, to study the influence of myelination on the injuried RGCs.

为此，本研究采用改良的胶质细胞混合培养与差速贴壁方法获得大鼠OPCs，使用无血清培养基进行扩增、培养，用免疫组织化学和流式细胞技术对培养细胞的纯度进行鉴定，对少突胶质系细胞表达部分营养因子的情况进行检测；采用TUNEL、MTT等方法对少突胶质系细胞条件培养基对原代培养小脑颗粒神经元的保护作用进行检测；将OPCs移植入成年SD大鼠玻璃体内，利用上丘逆行荧光标记技术，观察眼内移植的OPCs对眶内视神经切断时的视网膜神经节细胞的保护作用及其持续时间；将OPCs或NSCs移植入新生和幼年SD大鼠玻璃体或视网膜内，观察不同时期视网膜内髓鞘形成与分布特点，分析髓鞘的超微结构，并观察眼内髓鞘形成对损伤神经节细胞的保护作用。

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。

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。

The expression of PCNA mirrors the general rule of cell multiplication and differentiation of glioma.

PCNA的表达反应了脑胶质瘤细胞增殖与分化的一般规律， PTEN表达的降低在脑胶质瘤的发生、发展过程中可能起着非常重要的作用，PCNA和PTEN的联合检测有利于更准确地判断胶质瘤生物行为和评估患者预后。

The expression of PCNA mirrors the general rule of cell multiplication and differentiation of glioma. The decrease of the expression of PTEN may have very important effect on development and growth of glioma.

PCNA的表达反应了脑胶质瘤细胞增殖与分化的一般规律， PTEN表达的降低在脑胶质瘤的发生、发展过程中可能起着非常重要的作用，PCNA和PTEN的联合检测有利于更准确地判断胶质瘤生物行为和评估患者预后。

TCM may modulate the glial cells through the following mechanisms: inhibiting expression of iNOS, thereby minishing the toxicity of NO and its oxygen free radicals; inhibiting overactivation of astrocytes and overexpression of GFAP in acute cerebral ischemia and cerebral hemorrhage; promoting glial cells' secretion of NGF which can be caused by ischemic injury; inhibiting the nervous system's immune inflammatory response; and so on.

中药调节神经胶质细胞活性的机制可能包括：（1）抑制iNOS表达，从而抑制NO及其产生的氧自由基的毒性；（2 抑制脑缺血和脑出血急性期星形胶质细胞的过度活化和GFAP的过度表达；(3)上调缺血损伤导致的神经胶质细胞分泌NGF；(4)抑制神经系统的免疫炎症反应等各种途径。

The results show that though the strain is growing slower in the nitrogen-free medium than in the nitrogen-containing medium, the content of capsular polysaccharides produced by the strain in the nitrogen-free medium is higher than those in the nitrogen-containing medium, the content of capsular polysaccharides produced by the strain in the nitrogen-free medium is higher than those in the nitrogen-containing medium. The capsular polysaccharides produced by the strain in the culture which contains different mineral powders will reach the highest content on the third or the fourth day in its growing period. The strain's capability of releasing potassium from k-feldspar and biotite in the nitrogen-containing medium is higher than that in the nitrogen-free medium because in the nitrogen-free medium, the strain and its production of glucoprotein are less than those in the nitrogen-containing medium.

结果表明，尽管该菌在无氮培养条件下的菌体数量远小于有氮培养条件，但无氮培养条件下所提取的细菌多糖多于在有氮条件下培养所提取的细菌多糖；胶质芽孢杆菌在以添加钾长石粉或黑云母粉制作的无氮培养基中生长可形成大量多糖，采用丙酮法进行细菌培养液多糖提取，发现细菌培养的第3天所提多糖含量最高；胶质芽孢杆菌在以添加钾长石粉或黑云母粉制作的有氮培养基中生长亦可形成较多的多糖，且在细菌培养的第4天所提多糖含量最高；胶质芽孢杆菌在有氮条件下对含钾矿物的释钾率高于在无氮条件下的释钾率，这可能与该菌在有氮条件下生长更快、可产生较多菌体细胞有关。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失