Differential expression and cellular localization of activin and inhibin mRNA in the rainbow trout ovary and testis

An inhibin cDNA from rainbow trout consisted of 1305 bp, which coded for 352 amino acid residues. The deduced amino acid sequence of mature inhibin was 50 to 60% identical to mammalian sequences. Distribution of inhibin alpha and activin beta A and beta B in different ovarian and testis compartments was studied in rainbow trout by in situ hybridization with complementary RNA probes. In testis tissue, inhibin alpha and activin beta A and beta B were expressed only in the testicular interstitia between the seminal lobules, where Sertoli cells and Leydig cells are distributed. The localizations and intensities of the reactions were constant throughout the maturation of the testis. Within ovarian tissue, the theca cell layers of follicles showed strong reactions of Dig-labeled antisense mRNA probes hybridizing against inhibin alpha and activin beta A and beta B in all samples over the same sampling period. In regressing oocytes, a positive reaction was observed in the granular cell layer of the follicles.     

Estradiol-induced accumulation of vitellogenin mRNA and secretion of vitellogenin in liver cultures of Xenopus

Expiants of male Xenopus liver maintained in a serum-free culture medium respond to stimulation by 2 × 10^?8 M 17β-estradiol with an increasing rate of accumulation of vitellogenin mRNA, as revealed by hybridization of cDNA to the total cytoplasmic RNA extracted from the cultures. A similar response is observed for secretion of ³²PO₄-labeled vitellogenin into the culture medium. The in vitro response is improved in liver tissue of prestimulated animals, and by adaptation of liver expiants to the culture medium prior to hormone treatment, but attains only about 10% of the in vivo response. Since essential features of the in vivo response are maintained in liver expiants, organ culture appears suitable for investigating initial events of estradiol action leading to enhanced synthesis of vitellogenin.     

类风湿关节炎滑膜组织cDNA文库的构建及自身抗原的筛选和鉴定

目的 通过SEREX（serological analysis of recombinant cDNA expression library）技术对类风湿关节炎滑膜组织cDNA文库进行初步筛选,得到新的类风湿关节炎自身抗原.方法 利用SMART技术构建类风湿关节炎患者滑膜组织cDNA文库,并对该文库进行SEREX筛选,获得特异性结合阳性克隆,对阳性克隆同源性分析.然后进行酶联免疫吸附实验（ELISA）验证,分析该阳性克隆对于类风湿关节炎诊断临床敏感度和特异度.结果 构建cDNA文库滴度约为6.89×10^6 pfu/mL,筛选文库获得12个阳性克隆,同源性分析3个阳性克隆插入片段为已知蛋白,分别是：Filaggrin、SERPINE2和p68.进行血清学初步验证,抗-Filaggrin和抗-SERPINE2具有高敏感度和低特异度,而抗-p68具有高敏感度和高特异度.结论 抗-Filaggrin、抗-SERPINE2和抗-p68对于类风湿关节炎的诊断具有不同的敏感度和特异度.     

基因表达谱芯片技术筛选舌癌转移相关基因

目的 应用基因表达谱芯片技术对口腔舌癌细胞系Tca8113及其脑转移细胞株Tb的基因表达谱进行对比分析并筛选差异表达基因,以期从基因水平上探讨舌癌转移机制。方法 以舌癌细胞系Tca8113及其脑转移细胞株Tb作为研究肿瘤转移分子机制的模型,提取mRNA,逆转录为cDNA, 用Ｃｙ3和Ｃｙ5荧光染料标记,制备成ｃＤＮＡ探针,与基因表达谱芯片进行杂交扫描和分析;并应用RT-PCR技术对其中6个基因进行验证。结果 在1 152个基因表达谱的筛选中,37个基因表达有差异（2倍以上）,其中15个基因表达水平上调,22个基因表达水平下调。RT－PCR技术对其中6个基因表达差异的验证结果与基因芯片结果一致。结论 舌癌细胞转移侵袭可能与多个基因相关。