人抵抗素基因cDNA的克隆

目的克隆人抵抗素基因(hRETN)cDNA,为进一步研究RETN的结构和功能提供实验基础.方法应用RT-PCR方法从中国人网膜脂肪垫总RNA中扩增出RETN 基因cDNA,克隆入载体pMD18-T中,形成重组载体pMD18-T/hRETN.通过蓝白斑筛选出阳性克隆,限制性内切酶酶切鉴定后对其进行测序.结果从脂肪组织总RNA中扩增得到363 bp片段hRETN基因,其cDNA序列与Genbank hRETN基因序列基本相同.结论成功地克隆中国人hRETN cDNA.     

山医群体中国地鼠近交E家系RAPD分析

目的　分析山医群体中国地鼠E家系的遗传纯度。方法　应用经过筛选的 3 1条随机引物对中国地鼠E家系 12只个体基因组DNA进行RAPD扩增 ,计算近交个体间的相似系数。结果　所有样品的相似系数0 943 1到 0 9978之间变动 ,平均相似系数为 0 9749,聚类分析得到了这些个体的同源树资料。结论　山医群体E家系有较高的遗传纯度     

宫颈癌细胞中DAPK1基因CpG岛甲基化及其表达

目的 :探讨宫颈癌细胞中凋亡相关蛋白激酶 1(DAPK1 )基因CpG岛甲基化及其表达与宫颈癌的相关性 .方法 :应用甲基化特异性PCR和SABC免疫组化方法 ,检测 32(鳞癌 1 8,腺癌 1 4 )例宫颈癌组织DAPK1基因CpG岛甲基化修饰及蛋白表达 .结果 :宫颈癌中DAPK1基因甲基化扩增阳性率明显增高 5 6 .3% ,与正常宫颈比较有显著性差异 (P <0 .0 5 ) ,鳞癌与腺癌甲基化扩增阳性率无显著差异 (P >0 .0 5 ) ;宫颈癌中DAPK1蛋白表达阳性率降低 1 5 .6 3% ,与正常宫颈比较有显著性差异 (P <0 .0 5 ) ,鳞癌和 7例与腺癌无显著性差异 (P >0 .0 5 ) .结论 :DAPK1基因CpG岛异常甲基化修饰能抑制DAPK1的转录 ,使DAPK1蛋白不表达 ,丧失抑癌作用 ,是促进正常宫颈上皮细胞癌变的一个重要因素     

硒抑制镍诱导的人肺成纤维细胞的DNA损伤

目的　探讨三氧化二镍(Ni2O3)对人肺成纤维细胞(human lung fibroblasts,HLF)的DNA损伤作用以及Na2SeO3的保护作用。方法　应用单细胞凝胶电泳(single cell gel electrophoresis,SCGE)检测DNA损伤。结果　Ni2O3 处理HLF细胞4 h,SCGE检测出DNA损伤程度高于对照组,差异有显著性(P<0.01)。而Ni2O3 Na2SeO3 处理组DNA损伤程度低于Ni2O3 组,差异有显著性(P<0.01)。结论　硒可抑制镍诱导的人肺成纤维细胞的DNA损伤。     

Standardized molecular typing of Candida albicans strains

Summary. A method is presented for the standardization of Candida albicans DNA fingerprinting, which is based on Southern hybridization of Eco RI-digested chromosomal DNA with the moderately repetitive DNA element CARE-2 and the subsequent rehybridization of the blots with a molecular size marker also included in each DNA sample. This method resulted in extremely precise alignment of all strain-specific CARE-2 hybridization patterns, even when analysed on different gels, and will enhance the accuracy of genetic relationship determinations in epidemiological studies including large numbers of strains.
Zusammenfassung. Zur Standardisierung des DNA-Fingerprinting von Candida albicans wurde eine Methode entwickelt, die auf der Southern Hybridisierung Eco RI-gespaltener chromosomaler DNA mit dem mittelrepetitiven DNA-Element CARE-2 und der darauffolgenden Rehybridisierung der Blots mit einem auch in den Proben enthaltenen molekularen Größenmarker beruht. Dies resultierte in einer äußerst präzisen Größen-bestimmung der hybridisierenden Fragmente, so daß alle stammspezifischen CARE-2-Hybridisierungsmuster exakt verglichen werden konnten, auch wenn die Isolate auf verschiedenen Gelen analysiert wurden. Die Methode erhöht die Genauigkeit der Bestimmung genetischer Verwandtschaftsbeziehungen in epidemiologischen Untersuchungen, in denen eine große Anzahl von Stämmen analysiert wird.     

Genetic analysis of the yeast NUD1 endo-exonuclease: a role in the repair of DNA double-strand breaks

The deoxyribonucleases (DNases) have been shown genetically to be important in the vital processes of DNA repair and recombination. The NUD1 gene, which codes for an endo-exonuclease of Saccharomyces cerevisiae, was analyzed for its role in the DNA double-strand break (DSB) repair processes. While the nud1 strain is only slightly sensitive to ionizing radiation, expression of the HO-endonuclease to introduce a DSB at the MAT locus in that strain results in cell death. Cell survival is inversely proportional to the duration of HO-endonuclease expression. Analysis of the surviving colonies from the nud1 strain indicated that many of the survivors are sterile and that the proportion of these sterile survivors increases with the time of HO-endonuclease expression. On the other hand, the surviving colonies from the isogenic NUD1 strain are mating-proficient. Interestingly, double mutants of nud1 rad52 are more resistant to ionizing irradiation than the rad52 strain and have a cell-survival fraction of 32% for rad52-1 nud1 and 9% for rad52::URA3 nud1 following prolonged HO-endonuclease expression, indicating that nud1 has a suppressor effect on the DSB-induced lethality in rad52. Polymerase chain reaction analysis showed that many of the nud1 survivors contained small alterations within the MAT locus, suggesting that the survivors arose through the process of non-homologous end-joining. These results suggest that the endo-exonuclease acts at a DSB to promote DNA repair via the homologous recombination pathway. Received: 20 July / 20 September 1998     

乙肝核酸疫苗在BALB/c小鼠体内的生物分布情况

目的：了解DNA疫苗在BALB／c小鼠体内的生物分布情况，建立DNA疫苗体内生物分布情况研究方法。方法：乙肝核酸疫苗(HBV DNA)经^32P标记、分离、纯化、鉴定后，胫前肌注射给药，结合三氯乙酸(TCA)沉淀，研究肌注乙肝核酸疫苗后BALB／c小鼠体内的生物分布情况。结果：质粒DNA疫苗除注射局部肌肉分布较多外，其他一些重要组织也有分布，其中腺体组织(肾上腺、胰腺、胸腺)中的浓度最高，其次为排泄物(尿、肠内粪)，脑组织中浓度最低。各组织中可沉淀放射性按血浆浓度时间曲线下面积(AUC)从高到低排列依次为胸腺、肾上腺、膀胱、生殖腺、脂肪、肠内粪、胰腺、颌下腺、脾、小肠、眼球、肝、肠内容、肾、甲状腺、肺、淋巴结、对侧肌肉、心脏和脑。结论：^32P标记DNA疫苗后各组织β计数结合三氦乙酸沉淀法研究核酸疫苗生物分布情况，方法灵敏、可靠，易于分析。     

DNA flow cytometry in stage IB and II cervical carcinoma

In a retrospective study the prognostic significance of nuclear DNA content was investigated, as measured by flow cytometry, of the tumor specimens from 212 women with nonpretreated FIGO stage IB and II cervical cancer. One-hundred and thirty cases (62%) were found to be diploid, whereas 82 (38%) were aneuploid. Univariate analysis of the follow-up data showed an increased relative risk (RR) for recurrence free survival (RFS) for stage II tumors (RR = 1.87, 95% CI: 1.13–3.10, P = 0.015) and for age (RR = 1.52, 95% CI: 0.66–3.52 and RR = 2.35, 95% CI: 1.19–4.65, P = 0.032). Ploidy showed a relative risk of 1.33 (95% CI: 0.83–2.13, NS). In addition, univariate analysis of overall survival (OS) revealed similar results. For the subgroup of patients with primary surgery ( n = 151), positive pelvic nodes (RR = 5.38, 95% CI: 2.70–10.71, P = 0.0001) and parametrial extension (RR = 2.53, 95% CI: 1.24–5.17, P = 0.011) were significant factors for OS after univariate analysis, the estimated effects on RFS were slightly smaller. Multivariate analysis of RFS for the whole study population showed age, histologic grade and stage with a slightly increased risk, but no effect was significant. Ploidy with an RR of 0.97 (95% CI: 0.58–1.62) seems to have no influence on prognosis. For the subgroup with primary surgery, ploidy again failed statistical significance with an RR of 1.20 (95% CI: 0.58–2.49). Our results suggest that abnormalities of the nuclear DNA content in this homogeneous group of patients are associated with clinical and morphological prognosticators, however, ploidy is not an independent prognostic factor for RFS, or for the whole study population or for the subgroup with primary surgery.     

Evolutionary aspects of “chloroplast-like” trnN and trnH expression in higher-plant mitochondria

Two identical “chloroplast-like” tRNA^Asn genes, trnN1 and trnN2, have been identified in the potato (Solanum tuberosum) mitochondrial genome. The flanking sequences of trnN1 are unrelated to the corresponding authentic potato chloroplast regions, whilst those of trnN2 are very similar to the chloroplast sequences. The trnN1 copy is present in the mitochondrial genome of various plants whereas the second copy, trnN2, is absent from all the other plant genomes studied so far. Interestingly, both trnN copies are expressed in potato mitochondria. Sequences flanking the chloroplast-like tRNA^His gene (trnH), present as a single copy in the potato mitochondrial DNA, are unrelated to the corresponding chloroplast sequences, whereas chloroplast-derived sequences have been maintained in the vicinity of the maize chloroplast-like mitochondrial trnH gene. However, both the potato and the maize trnH are expressed in mitochondria. Received: 10 April / 1 August 1997     

Study on the Relationship between Cytogenetics and Phenotypic Effect in Turner＇s Syndrome

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