原位PCR和免疫组化检测垂体腺癌P16基因

目的研究人垂体腺瘤P16基因的改变,同时探索原位PCR技术的适宜条件和结果确认,以及用于基因缺失检测的可行性。方法原位PCR和免疫组化技术。结界绝大部分间质细胞为P16蛋白免疫组化阴性,而一部分垂体肿瘤细胞呈P16阳性;针对P16基因的原位PCR信号则可见于垂体肿瘤细胞和间质细胞等各种细胞,即P16组化阴性的细胞同样表明有P16基因存在。结论原位PCR可以是P16基因研究的一种有效手段,提示垂体腺癌的P16基因改变形式可能主要是表达过度而不是基因缺失。     

上海地区汉人瘦素受体基因G1n223Arg变异的研究

目的探讨瘦素受体基因Gln223Arg变异与肥胖的关系。方法359例无亲缘关系的上海地区汉人及66例白种人运用PCR-RFLP方法（MspI限制性内切酶）检测瘦素受体基因Gin223Arg变异的基因型。结果中国人中GG、GA及AA基因型频率分别为0.789,0.200及0.011;G及A等位基因频率分别为0.889及0.111,与本研究组中白种人的基因型及等位基因频率分布有显著差异（P=2.98×10-16,3.46×10-18）。在中国人正常体重组与超重组间该基因变异的基因型及等位基因频率分布无显著差异（P=0.369,0.344）。结论瘦素受体基因Gln223Arg变异,在中国人肥胖的发病中可能无重要作用。     

阳离子脂质体介导细胞因子基因对小鼠肝癌抑制的作用

目的观察阳离子脂质体介导细胞因子ＴＮＦ－ａ及ＩＬ－２基因对小鼠肝癌的抑制作用。方法构建含细胞因子ＴＮＦ－ａ及ＩＬ－２的质粒载体,用阳离子脂质体ＬｉｐｏｆeｃｔＡＭＩＮＥ或ＤＯＳＰＥＲ介导的含细胞因子ＴＮＦ－ａ及ＩＬ－２的质粒ＤＮＡ分别体外转染小鼠肝癌细胞株ＭＭ４５Ｔ．Ｌｉ,并在荷瘤小鼠体内分别直接注射上述阳离子脂质体－ＤＮＡ复合物,观察瘤体大小及小鼠生存期。结果两种阳离子脂质体介导的细胞因子转染后都具有生物学活性,转染效率为１０％左右,瘤内注射后均能延长荷瘤小鼠的生存期。ＤＯＳＰＥＲ介导细胞因子的治疗效果较佳。结论ＬｉｐｏｆｅｃｔＡＭＩＮＥ或ＤＯＳＰＥＲ介导细胞因子ＴＮＦ－ａ及ＩＬ－２基因均具有抑制小鼠肝癌生长、延长荷瘤小鼠生存期的作用,ＤＯＳＰＥＲ的效果较好。     

Variability of the immunoglobulin light chain in the Siberian sturgeon, Acipenser baeri

All sturgeon VL segments isolated in this study belong to a single family, VLI, which can be divided into two subfamilies. Of the 79 cDNA clones isolated, 76 belong to the larger subfamily, VLIa, and only 3 clones constitute the smaller subfamily, VLIb. To evaluate variability, the Shannon entropy was estimated for each individual amino acid position, and to facilitate comparisons of variability between species the mean entropy of the CDR regions was calculated. In such a comparison, the sturgeon was found to have CDR1 and CDR3 variability approaching those found in mouse and clawed frog, but showed very low variability for CDR2. Amino acid position 50 does however display variability in the range of mouse and clawed frog. It is further confirmed that the sturgeon has numerous J segments, but that the junctional diversity does not contribute greatly to the diversity of the light chain. Comparisons of cDNA clones and a genomic VL segment indicate that the VL undergoes changes, particularly in the CDR regions, in a manner that can be explained by somatic hypermutation and/or gene conversion.     

肺鳞状细胞癌p53 过度表达与肿瘤生物学行为及预后的关系

目的：探讨肺鳞状细胞癌组织ｐ５３ 蛋白的过度表达与肿瘤生物学行为及预后的关系。方法：应用单克隆抗 ｐ５３ 抗体对石蜡包埋的肺癌组织标本进行免疫组织化学染色。结果：６８ 例获随访的肺鳞状细胞癌病例中有 ４４ 例 ｐ５３ 表达阳性,总阳性率为 ６４７％ ,且其阳性率随肿瘤病理分级、临床分期的增高而增加（ Ｐ＜ ００５）;与术后５ 年存活情况有关（ Ｐ＜ ００５）。结论：提示ｐ５３ 基因的异常表达在肺鳞状细胞癌的发展过程中起重要作用,对肺鳞状细胞癌病人的预后判定有一定意义。     

Mutation of P_(53) tumor suppressor gene in colorectal cancer and its relationship with histologic grade and Dukes'''' stage

目的：探讨大肠癌组织类型、临床分期与P53基因突变的相关关系。方法：采用PCR—SSCP方法对30例大肠癌P53基因突变状况进行了检测。结果：发现6例（20％）P53基因突变,其中,1例在第4外显子,5例在第5外显子,第2,3外显子未检出突变。在不同组织分化程度中,P53基因突变率分别为高分化腺癌和中分化腺癌均为1／8（12．5％）,低分化腺癌为3/10（30．0％）,未分化腺癌为1／4（25．0％）。在临床分期中,Dukes’B期为1／10（10．0％）,C期为2／12（16．7％）,D组为3／8（37．5％）。结论：P53基因突变与Dukes’分期有关,并且P53基因突变很少发生第2,3外显子。     

Experimental study on the antitumor effect of mouse B7-1 gene

ＴｗｏｓｉｇｎａｌｔｈｅｏｒｙｆｏｒＴｃｅｌａｃｔｉｖａｔｉｏｎｐｒｏｖｉｄｅｄａｎｅｗａｐｐｒｏａｃｈｆｏｒｔｕｍｏｒｇｅｎｅｔｈｅｒａｐｙ［１－２］．Ｅｘｐｒｅｓｓｉｏｎｏｆｃｏｓｔｉｍｕｌａｔｏｒｙｍｏｌｅｃｕｌｅｇｅｎｅｉｎｔｒａｎｓｆｅｃｔ...     

The construction ofSchistosoma Japonicum vaccine BCG-Sj26GST and its identification

Summary The expression of foreign gene,Schistosoma Japonicum 26 ku antigen (Sj26GST),in Bacillus Calmette-Guerin (BCG),Mycobacterium CM. smegmatis) andEscherichia coli (E. coli) were studied. The cDNA fragment encoding Sj26GST was amplified by PCR using plasmid pGEX, which could express Sj26GST inE. coli as template. The Sj26GST cDNA was cloned into the downstream of humanM. tuberculosis heat shock protein (hsp) 70 promoter with correct reading frame, and then the DNA fragment containing hsp70 promoter and Sj26GST gene were subcloned together intoE. coli-Mycobacteria shuttle plasmid pBCG-2000 to construct the expression shuttle plasmid pBCG-Sj26. The recombinant BCG andM. smegmatis mc²155, which were electroplated with pBCG-Sj26, could express Sj26GST and the recombinantSchistosoma Japonicum vaccine BCG-Sj26GST was made. The recombinant Sj26GST (rSj26GST) were soluble and could be observed on SDS-PAGE at molecular weight of 26 ku. The content of rSj26GST accounted for 15% and 10% of total bacterial protein in BCG andM. smegmatis respectively. The results of Western blot showed the combination of rSj26GST with antibody of GST. This project was supported by Primary Foundation of China (No. 94-Y-19), Chinese National Nature Science Foundation (No. 339480022) and Chinese Health Foundation (No. 94-1-131).