整合素和GFRs介导的信号转导及其相互关系

整合素和GFRs之间可以相互影响和相互作用,并且在信号转导通路上存在多层次的交叉。两者的信号整合的细胞的存活、增殖、和运动等事件中扮演了重要角色。     

RNAi研究进展

基因阻断技术近年来发展迅速。双链RNA介导的、序列特异的转录后基因沉默的过程称为RNAi(RNA interence)。它作为新肖的基因阻断技术,自1998年发现到现在已有很大进展。迄今,在果蝇、线虫、锥虫、小鼠及哺乳动物中相继发现存在RNAi现象。目前许多学者以果蝇、线虫为对象做了RNAi的大量研究人,并相继提出了其作用机制模型。RNAi可能是生物体中存在的一种普遍现象,代表了一古老的细胞反应通路。因此RNAi有望成为今后分析人类基因组功能的有力工具,并可能用于基因的特异治疗。     

线粒体基因组在帕金森病发病机制中的作用

线粒体基因组以及线粒体呼吸链中酶复合体的改变会影响能量的供给,而脑对能量供给的改变非常敏感,甚至因此会引起神经细胞的死亡,导致神经退行性变的发生。由于脑组织不同的区域易感性不同,黑质纹状体部位的神经细胞容易引起氧化应激,导致自由基的升高,mtDNA发生突变,酶复合体功能下降,最终引起神经细胞的死亡,形成帕金森病的临床表现。     

阿拉善盟蒙古族、汉族4顶人类群体遗传学指标的调查

拇指类型、环食指长、指甲类型和足趾长是 4项与手足有关的人类群体遗传学经典指标。这 4项指标目前国内外研究报道较少。指甲类型尚未见研究资料的发表。阿拉善盟位于内蒙古西部 ,总人口 167977人 (1995年资料 ) ,其中蒙古族 4 32 57人 ,其余为汉族、回族。阿拉善蒙古族源于新疆和硕特部与土尔扈特部 ,于清代迁移至内蒙古西部 ,汉族则主要来自于甘肃、宁夏等地。为研究阿拉善盟蒙古族、汉族人的人类学特征 ,我们于1999年 10月赴阿拉善盟进行了蒙古族、汉族 4项指标的调查 ,调查了内蒙古阿拉善盟蒙古族 (男 2 18例 ,女2 2 9例 )、汉族 (男 2…     

EB病毒潜伏膜蛋白1诱导人鼻咽上皮细胞端粒酶的表达

Telomerase activation has been linked to cell immortalization in vitro and tumorigenicity in vivo. In this study, for the first, we reported that Epstein-Barr virus activated the telomerase activity of human nasopharyngeal epithelial cells in the early stage of immortalization as tested by the PCR-ELISA. The telomerase activity in nasopharyngeal epithelial cells was only observed in presenescent cells. It was implicated that Epstein-Barr virus induced the escape of nasopharyngeal epithelial cells from senescence via the activation of telomerase. We further showed that telomerase activation in infected cells was dependent on the protein level of latent membrane protein 1 (LMP1) encoded by Epstein-Barr virus using a Tetracycline regulatory cell line expressing LMP1, pTet-on-LMP1-HNE2. The activity of telomerase in nasopharyngeal cells was decreased when the protein level of LMP1 was blocked by antisense LMP1 plasmid DNA. And the activity of telmerase was also related to the carboxyl terminus of LMP1. It was implicated that the ability of Epstein-Barr virus to suppress senescence is associated with telomerase activation by LMP1.     

HeLa细胞中hR24L基因的表达、DNA损伤修复、G2／M阻滞与电离辐射之间的关系（简报）

The hR24L gene ORF was cloned from the total RNA of HeLa cells by RT-PCR method. There is no mutation of the hR24L gene in HeLa cells. Ion radiation significantly increased the expression of hR24L gene, and the sense hR24L enhanced and accelerated the cell cycle arrest at G2/M phase in HeLa cells. Besides, it should seem that the overexpression of hR24L gene could enhance the repair ability of DNA damage induced by ion radiation, and vice versa.     

α—硫辛酸对6—羟多巴胺诱导的PC12细胞凋亡的影响

PC12 cell line, a clonal cell line derived from a pheochromocytoma of rat adrenal medulla, was used as a model of dopaminergic neuron in vitro to study the effect of alpha-lipoic acid on the 6-OHDA induced apoptosis. The results from MTT method show that 6-OHDA decreased the cell survival rate obviously. Through TUNEL (TdT-mediated dUTP-biotion nick end labeling) and Flow cytometer (FCM) detection, we found that 6-OHDA triggered cell apoptosis and induced necrosis. It was confirmed by the different percentage of cell survival rate and apoptosis concluded from FCM and MTT. alpha-lipoic acid was used as antioxidant to protect the cell from 6-OHDA's injury. The result indicateed that alpha-lipoic acid can partly prevent apoptosis induced by 6-OHDA but fail to prevent necrosis since it can decrease the apoptotic cell from 20.09% to 3.09%, just as increased cell survival rate from 56.8% to 72.6% but can not reach the normal level showed by MTT assay. Biochemical approach showed the cell's antioxidant ability especial for SOD activity and GSH content increased after the treatment of alpha-lipoic acid. The data suggest that alpha-lipoic acid may protect PC12 cells from apoptosis induced by 6-OHDA through the antioxidant path.     

重组人钙调素的制备及对细胞增殖作用影响的研究

The gene coding for human CaM was amplified by PCR in which pUC/hCaM3 cDNA was usd as template. After inserting the hCaM III cDNA into the expression plasmid pBV220, we constructed the hCaM3 cDNA-recombinant expression vector(hCaM3/pBV220). The recombinant plasmid was then transformed into E. coli DH5 alpha. After heat induction, a high level expression of CaM protein was obtained. SDS-PAGE analysis showed that the recombinant E. coli could express a 17 kD protein which accounted for about 20% of the total cellular protein. Western blot analysis showed that anti-CaM monoclonal antibody(McAb) specifically bound to the 17 kD band of expression product. rhCaM was purified by Phenyl-sepharose CL-4B affinity chromatography from recombinant bacterial lysate. 3-4 mg of the purified protein were obtained from 1 liter of bacterial culture. The rhCaM was able to activate NAD kinase to the same extent as the standard human brain CaM (Sigma). K562 cells and SP2/0 cells were seeded in 24-well or 96-well plate and cultured for 48 h with rhCaM and CaM-antagonist trifluoperazine(TFP). Cell proliferation rates was determined by MTT assay. There was a significant positive correlation between the concentrations of rhCaM and the cell proliferation rates. CaM-antagonist TFP had an inhibitory effect on cell proliferation rate. The inhibition could be corrected by the addition of extracellular rhCaM.     

甜橙辣椒红／辣椒玉红素合成酶同源基因的克隆（简报）

The complete sequence of orange homologous capsanthin/capsorubin synthase gene is 3788 bp long with a coding sequence of 1512 bp, which encodes a polypeptide of 503 amino acids. The 5' upstream sequence is 1721 bp long and the 3' downstream sequence is 555 bp long. The amino acid sequence of this gene is 78% and 69% identical to the genes from carrot and pepper, respectively. It is also partially homologous to plant neoxanthin synthase, lycopene beta-cyclase and lycopene epsilon cyclase genes. Isolation of the gene provides a framework for elucidation of the mechanisms involved in inability of citrus to produce capsanthin and capsorubin.     

癌组织中的遗传不稳定性的RAPD分析（简报）

In five kinds of tumors, total 128 specimens were analyzed by RAPD (random amplified polymorphic DNA) PCR with nine 10-base arbitrary primers for detecting instabilities of DNA and chromosome and screening new molecular markers coupled to putative or unknown oncogenes and/or tumor suppressor genes. Bands representing instabilities have been recovered and purified from agarose and cloned into pCAPs vector, and further labeled by DIG as probes for analysis of Southern blot, Northern blot and Sequencing. Results revealed that sample 5 and 3 of the gastric cancers showed the highest genomic changes and the average detectability in five sorts of cancers was up to at least 40% (42.2%-49.4%), and that there were significant differences in the ability of each primer to detect genomic instability, which ranged from 27% to 68%. Despite the highest detectability of genetic instability (68%) in tumor tissues, primer 2 could produce stable profiles of DNA bands in normal tissue genome with good reproducibility. On the contrary, primer 8 was of the lowest one (27%). Band B of single copy found to be allelic losses in gastric and colon cancers according to RFLP analysis was of a novel sequence and registered by Gen-Bank (Accession Number AF151005). Therefore the genetic instabilities often concentrated on some special locuses of chromosome e.g. repetitive sequences etc. and coupled to carcinogenesis. It was impossible or difficult to get great achievements for cancer treatments with the procedure of gene therapy only to one oncogene or one tumor suppressor gene because the extensive DNA variations occurred during the progression of tumor. RAPD assay connected with other techniques was a good tool for the detection of genomic instabilities and direct screening of some new molecular markers related to tumor suppressor genes or oncogenes.