关于几种避孕植物药的药理初筛及成份预试

本文对云南滇西地区民间用于避孕和绝育秘方中的槌果藤,野花椒寄生、花椒寄生、螃蟹树寄生、鸡矢藤及野花椒的醇提取物进行了小鼠最大耐受量测定及小鼠抗生育实验,结果表明,花椒寄生、螃蟹树寄生的醇提取物具有明显抗生育活性,槌果藤也具有一定的抗生育作用。此外,还对槌果藤、野花椒寄生进行了化学成份预试。     

含甲肝病毒基因的重组痘苗病毒在动物体内产生针对甲肝病毒的中和抗体

用DNA重组技术得到的含甲肝病毒基因的重组痘苗病毒,可在家兔体内产生ELISA竞争抑制与中和抗体。基础免疫后,动物体内竞争抑制抗体滴度为10,加强免疫后达到80。由重组病毒产生的抗体中和指数比甲肝病毒产生者略低。     

Construction and packaging of pseudotype retrovirus containing human N—ras cDNA antisense sequence and its biological effects on human hepatoma cells

N-ras is one of the transforming genes in human hepatic cancer cells.It has been found that N-ras was overexpressed at the mRNA and protein level in hepatoma cells.In order to explore the biological roles of N-ras in human hepatic carcinogenesis and the potential application in control of cancer cell growth,a preudotype retrovirus containing antisense sequence of human N-ras was constructed and packaged.A recombinant retrovirus vector containing antisense or sense sequences of N-ras cDNA was constructed by pZIP-NeoSV(X)1.The pseudotype virus was packaged ang rescued by transfection and infection in PA317 and ψ 2 helper cells.It has been demonstrated that the pseudotype retrovirus containing antisense N-ras sequence did inhibit the growth of human PLC/PRF/5 hepatoma cells accompanied with inhibition of p21 expression,while the retrovirus containing sense sequence had none.The pseudotype virus had no effect on human diploid fibroblasts.     

应用纯化的重组Epstein-Barr病毒早期抗原建立检测鼻咽癌病人血清IgA/EA抗体的ELISA方法

利用凝胶和离子交换柱(Mono Q)两次层析,将大肠杆菌表达的EB病毒早期抗原P138片段多肽纯化。以此P138为抗原,增加鼠抗人IgA单克隆抗体以扩大IgA的反应,建立了三步ELISA法。用本法检查了100例鼻咽癌病人和63例正常人血清中抗EB病毒IgA/EA抗体,病人血清的阳性检出率为86％,正常人有3例阳性(4.7％)。此结果表明,三步ELISA法较常用的间接ELISA法(阳性检出率为71％)敏感。     

正常人及肝细胞癌患者肝脏及血浆中丙酮酸激酶同工酶免疫测定

 分别从人肝及鼠肝中高度纯化L型和M_1型丙酮酸激酶(Pyk),制备相应的免抗血清。从抗血清中纯化IgG并水解成F(ab')_2,进而还原成Fab',后者与辣根过氧物酶(HRP)交联成Fab'-HRP,再利用这两种复合物建立了各自的高灵敏夹心ELISA来免疫定量L及M_2型(与鼠M_1-Pyk有交叉免疫)Pyk,结果发现:正常人肝中95％的Pyk为L型,M_2-Pyk仅占5％,而肝细胞癌中L-Pyk降至正常肝的3.5％,M_2-Pyk却增至正常的14.6倍,以致M_2-Pyk占总量的95.5％。免疫组化研究进一步证实定量的结果,正常人肝L-Pyk染色呈强阳性。M_2-Pyk染色较弱,而肝细胞癌恰好相反,L-Pyk染色明显减退,而M_2-Pyk不仅癌组织染色很深,癌周组织也明显深于正常,而且愈近癌巢,染色愈深。测定血浆Pyk,发现正常人L-Pyk占89.9％,M-Pyk(以M_2计)仅占10.1％。肝细胞癌患者血浆L-Pyk略见降低,为正常值的79.6％,p值在0.02～0.05之间,但血浆M型Pyk明显升高至正常的4.59倍,占Pyk总量的39.6％,并证明主要来源于肝癌组织中的M_2,故M_2-Pyk有可能成为肝细胞癌诊断的新指标。     

人黑色素瘤细胞分泌的组织纤溶酶原激活剂(t-PA)的纯化

 本文报道了培养的人黑色素瘤细胞分泌的组织纤溶酶原激活剂(t-PA)的纯化方法。Bowes株人黑色素瘤细胞的分泌产物,经CM-Sephadex C--50层析,赖氨酸-Sepharose 4B,苯甲眯-sepharose 4B亲和层析后,即可得到纯化470倍的蛋白纯品。样品经聚丙烯酰胺凝胶电泳鉴定为均一单带,测得其分子量约为72kD。纯化的t-PA与尿激酶相比较,发现前者有更高亲和纤维蛋白的能力。