核染质

Cerebral hemorrhage rat models were established via injection of autologous arterial blood in caudate nucleus. Two days after modeling, 5 μL BrdU-labeled human embryonic NSCs suspension was transplanted at four points surrounding hematoma cavity in the rats. After 1 and 2 weeks, rats were sacrificed. Adjacent sections were doubly stained by BrdU/microtubule-associated protein 2 (MAP-2) and BrdU/glial fibrillary acidic protein.

通过注射自体动脉血到尾状核制作大鼠脑出血模型，出血后2 d将标有5'-溴脱氧尿嘧啶的人胚神经干细胞悬液移植到血肿腔周围的4点，1，2周后处死大鼠，相邻脑组织切片行5'-溴脱氧尿嘧啶/微管相关蛋白2和5'-溴脱氧尿嘧啶/胶质纤维酸性蛋白免疫组织化学双染。

METHODS: Cerebral cortex cells of 8-week aborted human fetus were harvested and cultured in vitro to obtain human embryonic NSCs. Cerebral hemorrhage rat models were established via injection of autologous arterial blood in caudate nucleus. Two days after modeling, 5 μL BrdU-labeled human embryonic NSCs suspension was transplanted at four points surrounding hematoma cavity in the rats.

通过注射自体动脉血到尾状核制作大鼠脑出血模型，出血后2 d将标有5'-溴脱氧尿嘧啶的人胚神经干细胞悬液移植到血肿腔周围的4 点，1，2周后处死大鼠，相邻脑组织切片行5'-溴脱氧尿嘧啶／微管相关蛋白2和5'-溴脱氧尿嘧啶／胶质纤维酸性蛋白免疫组织化学双染。

Then we transfected transitorily the recombinant of green fluorescent protein gene and middle molecular weight neurofilament cDNA into wide type N2a (N2a/wt) and N2a/tau40 to observe the effect of tau accumulation on GFP-NFM fusion protein transport in cellular processes in living cells. At last we used an apoptotic inducer, camptothecin (an inhibitor of topoisomerase-1) to treat N2a/wt and N2a/tau40 cell lines, and compared their apoptotic response.

主要结果如下：一、tau蛋白过度表达和聚积对细胞形态的影响：倒置显微镜下观察两种细胞的形态，发现N2a/wt细胞的突起多而长，而N2a/tau40细胞胞体变圆，突起明显缩短；免疫印迹结果显示转染了tau40的细胞内tau的免疫反应约增加14倍，免疫荧光结果显示N2a/tau40细胞胞体内呈现出较强的红色荧光，tau主要分布在核周和突起起始部分的胞质内，而N2a/wt细胞内的荧光很弱。

Methods Amplify the cDNA sequence encoding truncated HCV gene, with a part of carboxyl-terminus deleted, by PCR. Synthesize the mimic epitope at E2 region of HCV and seven T or Th cell epitope genes of NS3-NS5 respectively. Bind HCV core gene with a part of carboxyl-terminus deleted to the synthetic epitope gene by PCR, then clone into eukaryotic expression vector pcDNA3.1 and transiently transfect COS7 cells.

用PCR方法扩增核心区羧基端部分缺失的基因片段；分别合成HCV E2区模拟表位和NS3~NS5 7个T或Th细胞表位基因；PCR方法将羧基端部分缺失的HCV核心区基因与合成的表位基因串联，克隆入真核表达载体pcDNA3.1，并通过脂质体瞬时转染COS7细胞。

Methods The whole-length X gene was directly cloned to pcDNA3.1 vector. The recombinant vector (pcDNA3.1-X) was transfected into HEPG2 cells after selection with hygromycin, steady clones (HEPG2-X cells) were obtained. The expression of RhoC protein was analysed with immunohistochemical stain.

用定向克隆的方法构建X基因的真核表达载体pcDNA3.1-X，脂质体转染HEPG2细胞；潮霉素选择培养稳定表达X基因的HEPG2-X细胞；免疫组化鉴定HEPG2-X细胞RhoC蛋白表达。

METHODS: The total RNA was obtained by TRIzol from an antiricin neutralizing monoclonal antibody hybridoma cell named 4C13 and heavychain and lightchain variable fragments were amplified by overlap PCR and accepted by Genbank of NCBI. VH and VL were linked by3 with a direction of VHlinkerVL and cloned to pCDNA3.1 vector expression system.

用TRIzol提取抗ricin中和性单抗杂交瘤细胞4C13总RNA，扩增其轻、重链可变区基因，登陆GenBank；用Linker3将VH和VL连接，用重叠延伸PCR克隆入真核表达载体pCDNA 3.1，用脂质体转染293T细胞，对瞬时表达产物进行生物学活性检测。

Then the caprine fetal fibroblast cells were transfected with p6.5hLF-EGFP by LipofectamineTM-2000 and selected by G418 for 3 to 4 weeks. The G418 resistant transfectants were identified by PCR and EGFP detection. The results indicated that the transgene was stably integrated into the open region of the chromatin of G418 resistant fibroblast cells.

脂质体介导法转染山羊胎儿成纤维细胞，G418抗性筛选3-4周后，经PCR扩增和报告基因EGFP表达检测，得到稳定整合外源基因的转基因供体细胞系，为制备高效表达人乳铁蛋白的转基因山羊乳腺生物反应器提高可靠的核移植供体细胞。

Results:17 rats were confirmed successful transplantation by MRI,the capacity of the tumor was 88.40±7.62mm3 on 8th day, 986.80±114.46mm3 on 16th day,the volume growth was 8-12 folds;on 8th day following transplantation,the tumor tissue was gray in color with fish homogen state without interior necrosis, the tumor cells were nest-distributed by HE staining,large dense staining of nucleus with manifest heteromorphism,tumor microvessel count increased by immunohistochemistry with buffy yellow staining, VEGF high expressed in the tumor cells with buffy yellow granule shape.

结果：17只大鼠经MRI检查证实均种植成功，肿瘤体积第8天为88.40±7.62mm3，第16天为986.80±114.46mm3，体积增长8-12倍之间；种植第8天，肿瘤组织呈灰白色、鱼肉均质状，内部未见坏死，HE染色肿瘤细胞呈巢状分布，胞核大浓染，异型性明显；免疫组织化学示肿瘤微血管数目较多，呈棕黄色染色，肿瘤细胞VEGF高表达，呈棕黄色细颗粒状。

Methods: Thirty adult male Wistar rats were randomly divided into three groups: the operation group, the transplantation of MSCs group and the injection of saline group. Functional outcome measurements were performed based on the modified neurological severity score at day 1, 7, 14, 21 and 28 after MACO. The survival, migration, expression of Nestin, glial fibriliary acidic protein and neuronspecific enolase of 5bromo2deoxyuridinelabeled MSCs were detected by immunohistochemical double staining.

线栓法建立左侧大鼠MCAO模型，随机分为3组：手术组、MSCs 移植组、生理盐水组。5溴2脱氧尿核苷标记的MSCs移植后，采用改良神经功能损伤评分系统评价大鼠神经功能恢复情况；应用免疫组织化学双染技术检测MSCs的存活、迁移及其巢蛋白、胶质纤维酸性蛋白和神经元特异性烯醇化酶的表达。

Methods:The conservative region of mTOR gene was inserted into pLXIN reversedly, then the vector was packaged in PT67 cells by transfection with lipofectamine and transfected to VSMC. The efficiency of antisense inhibition was verified,and the changes of p70S6k and 4E-BP1 were also determined at the same time. The proliferation activity was determined by flowcytometry and MTT.

采用脂质体介导转染包装细胞后构建mTOR的反义重组逆转录病毒载体(pLXIN-AmTOR)感染VSMC，检测mTOR及其下游底物包括真核细胞启动子4E结合蛋白1(4E-BP1)和核糖体蛋白S6激酶(p70S6k)的表达变化，以及VSMC细胞增殖周期和增殖活性的改变。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐