自拟方“通痹祛风汤”对大鼠急性痛风性关节炎抗炎作用研究

目的:观察"通痹祛风汤"对急性痛风性关节炎模型大鼠不同时相的影响,探讨其治疗急性痛风性关节炎的作用机制.方法:注射尿酸钠建立大鼠痛风性关节炎模型,大鼠灌胃给药,观察对大鼠关节肿胀率及关节滑膜囊表达IL-1 mRNA和TNF-mRNA(RT-PCR)的影响.结果:"通痹祛风汤"可有效抑制大鼠踝关节肿胀程度;关节滑膜囊TNF-mRNA和IL-1 mRNAr的表达明显减低.结论:"通痹祛风汤"可抑制痛风性关节炎TNF-和IL-1的产生而发挥抗炎作用,明显减轻关节的病理性损伤,有效抑制关节炎的临床进程.     

Carboxypeptidase E and thrombospondin-1 are differently expressed in subcutaneous and visceral fat of obese subjects

The aim of this study was to identify candidate genes for visceral obesity by screening for genes strongly differentially expressed between human subcutaneous and visceral adipose depots. A cDNA microarray with human adipose-derived cDNAs was used as an initial screening to identify genes that are potentially differentially expressed between human subcutaneous and visceral abdominal fat tissues. For the two best candidates, carboxypeptidase E (CPE) and thrombospondin-1 (THBS1) (EST N72406), real-time RT-PCR was performed to confirm their depot specific expression in extremely obese individuals. Both genes appeared to be strongly differentially expressed, having a higher expression in the visceral depot than in the subcutaneous one. For THBS1, the difference in expression between the depots was greater in women than in men. The involvement of CPE and THBS1 in obesity allows us to suggest that the physiological processes controlled by these genes contribute to depot and gender-related differences in the metabolic complications of obesity. Received 7 August 2002; received after revision 19 Septemer 2002; accepted 24 September 2002     

植物病毒RNA提取方法的研究

病毒RNA提取技术是分子生物学中经常应用的重要技术,是对RNA病毒在分子水平进行研究的必要手段.介绍了一种从染病植物组织中提取病毒RNA的方法,利用此方法提取了马铃薯x病毒(PVX)的RNA基因组,并通过RT-PCR得到其外壳蛋白基因,提取的病毒RNA在纯度和产量上都可满足反转录和PCR扩增等基本分子生物学操作.     

人Endostatin基因的克隆及其在Hela细胞中的表达

利用RT-PCR克隆人Endostatin基因,分别构建到双顺反子表达载体pIRES2-EGFP和融合表达载体pEGFP-C1中.两个重组表达载体利用脂质体介导转染真核细胞Hela,48 h后可在荧光显微镜下观察到被转染细胞发出绿色荧光.传代10次后,绿色荧光仍持续表达,说明是稳定转染.以转染细胞的总DNA为模板PCR检测Endostatin基因已整合到细胞染色体上.利用兔抗人Endostatin抗体对已转染细胞进行免疫组化检测,显微镜下观察,转染的细胞显棕红色,表明Endostatin在转染细胞内稳定表达.本试验通过报告基因检测,DNA检测,免疫细胞化学检测三个水平证明Endostatin已整合到细胞的染色体上,为基因治疗以及联合治疗打下基础.     

蝴蝶兰花发育相关基因pPI9的克隆与表达分析

为研究具有复杂而特异花型的蝴蝶兰花发育的分子机制,利用RT-PCR方法从蝴蝶兰花瓣总RNA中分离出834 bp长的cDNA片断.序列分析表明,该cDNA片断与拟南芥的PI基因有60%的同源性,命名为pPI9.它包含一个开放阅读框,具有编码24.5 ku蛋白质的能力.推导的氨基酸序列中,N端包含一个完整的MADS盒,C端有明显的PI基序.半定量PCR结果表明该基因只在植株的生殖器官中表达,而在营养器官中不表达,推测其可能参与花形态建成过程的调节.     

Quantitative detection of ING1 mRNA under different gene regulation based on molecular beacon

As an important tumor suppressor, normal expres- sion of ING1 is associated with the maintenance of cell normal physiological functions. Its low expression of- ten causes aberrant events of cell survive, growth and differentiation[1] which results in loss…     

禽流感病毒三种检测方法的比较研究

对分别应用病毒分离鉴定、反转录-聚合酶链式反应(RT-PCR)、间接免疫荧光(IFA)3种方法对病料组织中的禽流感病毒(AIV) 进行检测与比较研究。结果显示:经鸡胚尿囊腔接种分离到的病毒,通过用禽流感病毒H5、H9亚型单克隆抗体所进行的血凝(HA)和血凝抑制试验(HI),鉴定为AIV H5亚型毒株;应用RT-PCR技术直接对病料组织抽提的RNA样品进行检测,从病料组织能扩增出大小为229bp的AIV通用目的片段和大小为380bp的H5亚型特异性片段;取病料组织的过滤液接种狗肾上皮细胞(MDCK)单层,24h后用H5亚型AIV特异性的单克隆抗体进行间接免疫荧光试验,结果在细胞核、细胞质内观察到特异性的荧光。研究的结果表明,3种技术都可对病料样品中的AIV进行检测,而RT-PCR和IFA两种方法还可直接对病料样品中的病毒进行亚型的鉴定。相对而言,后两者具有更快速、更敏感、操作更简便的特点。     

褐飞虱羧酸酯酶基因cDNA片段的克隆及表达分析

利用反转录多聚酶链式反应(RT-PCR)技术克隆了褐飞虱羧酸酯酶基因编码区的cDNA片段,并进行了序列测定.结果表明,所克隆到的cDNA片段长度为396 bp,经BLAST查找比对发现,该片段所编码的氨基酸序列与来自铜绿蝇、家蝇、沟鼠、黑腹果蝇、线虫和埃及伊蚊的羧酸酯酶的片段存在高度同源性.Northern杂交分析显示,在褐飞虱取食抗性水稻后,羧酸酯酶基因表达水平明显升高.以上结果表明,羧酸酯酶基因的表达受抗性水稻的诱导,该基因在有毒化学物质解毒及增强褐飞虱对抗性水稻的耐受性方面可能起着重要作用.     

褐飞虱细胞色素P450基因cDNA片段的克隆、测序及表达分析

利用反转录多聚酶链式反应(RT-PCR)技术克隆了褐飞虱细胞色素P450基因编码区的cDNA片段,并进行了序列测定.结果表明,所克隆到的cDNA片段长度为237bp,经BLAST查找比对发现,该片段所编码的氨基酸序列与来自烟草天蛾、棉铃虫、埃及伊蚊、家蝇、黑腹果蝇和线虫的CYP6家族的P450的氨基酸序列存在同源性.Northern杂交分析显示,在褐飞虱取食抗性水稻后,P450基因的表达水平明显升高.以上结果表明,P450基因的表达受抗性水稻的诱导,该基因在褐飞虱对抗性水稻的耐受性和解毒方面可能起着重要作用.