心血管疾病基因治疗的基础研究Ⅵ．一种新的体内基因转移方法

将医用缝合线浸泡在质粒ＤＮＡ溶液中，然后缝合在大鼠股四头肌上，可以观察到外源基因在肌肉组织中高效、稳定地表达，其表达量高于磷酸钙共沉淀和ＤＮＡ沉淀块埋植法，较肌肉直接注射法表达量高１０倍左右。     

Sensitive Detection of p53 Gene Mutations in Esophageal Endoscopic Biopsy Specimens by Cell Sorting Combined with Polymerase Chain Reaction Single-strand Conformation Polymorphism Analysis

For the rapid and sensitive detection of p53 gene mutations in esophageal endoscopic biopsy specimens, we combined cell sorting with the polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) analysis. Mutations in exons 5–8 of the p53 gene were investigated by FCR-SSCF analysis using 10³ sorted nuclei obtained from each endoscopic biopsy specimen of 16 patients with esophageal cancer. DNAs extracted from their respective surgical specimens were investigated by a conventional method of PCR-SSCP analysis. Mutations in the biopsy specimens were detected in 6 of the 12 aneuploid tumors but in none of the 4 diploid tumors. After tumor cell enrichment by cell sorting, one mutation in exon 8 became apparent, which could not be detected from the surgical specimen by a conventional method of PCR-SSCP analysis. This method should improve the sensitivity of detecting p53 gene mutations, and provides additional information concerning the DNA ploidy pattern in the tumors.     

Prognostic significance of cytochemical analysis of leukemic M2 blasts

Cytochemical analysis of leukemic blasts from 46 patients with acute myeloblastic M2 leukemia (according to the FAB classification) was performed before and after cytostatic therapy, and compared with findings obtained in 20 age- and sex-matched control subjects. Cytochemical findings for myeloperoxidase (MPO), Sudan black B, acid phosphatase and alpha-naphthyl-acetate esterase (ANAE) were related to the achievement of the first complete remission (CR),i.e. data were compared after the patients had been divided into CR and non-CR groups. The analysis clearly showed that a high proportion of myeloperoxidase- and, to a lesser extent, Sudan black B-positive blasts before treatment may have constituted a significantly unfavourable prognostic factor.     

Preparation and Characterization of a Murine Monoclonal Antibody (MDR3M) Reactive with mdr3 Gene Product

A murine monoclonal antibody (MDR3M) (isotype: IgM) reactive with mdr3 gene product was generated by immunizing mice with mdr3 -specific peptide (H₂N-¹²WRPTSAEGDFELGISSKQKRKKTKTVKMI⁴¹G-COOH) and hybridizing the primed mouse splenic B cells with X63-Ag8,6.5.3 mouse plasmacytoma cells. MDR3M did not cross-react with mdr1 gene product. This monoclonal antibody may be useful for analyzing the role of mdr3 gene product in cells and tissues.     

Point Mutation in the Parkin Gene on Patients with Parkinson''''s Disease

Summary;To investigate the distribution of possible novel mutations from parkin gene in variant sub-set of patients with Parkinson’s disease(PD)in China and explore whether parkin gene plays an im-portant role in the pathogenesis of PD,70 patients were divided into early-onset group and late-onsetgroup; 70 healthy subjects were included as controls.Genomic DNA from 70 normal controls andfrom those of PD patients were extracted from peripheral blood leukocytes by using standard proce-dures.Mutations of parkin gene(exon 1—12)in all the subjects were screened by PCR-single strandconformation polymorphism(SSCP),and further sequencing was performed in tue samples with ab-normal SSCP results,in order to confirm the mutation and its location.A new missense mutationGly284Arg in a patient and 3 abnormal bands in SSCP electrophoresis from samples of another 3 pa-tients were found.All the DNA variants were sourced from the samples of the patients with early-on-set PD.It was concluded that Parkin point mutation a     

正常人及胃癌患者胃蛋白酶原C基因多态性研究

以ＰＧＣ３０１为探讨，对１０例胃癌组织及１１例正常人体组织基因组ＤＮＡ中胃蛋白酶原Ｃ基因的ＥｃｏＲ Ｉ限制性片段长度多态性作了观察分析。发现在正常人体有三种常见等位片段，分别为２０ｋｂ、５．７ｋｂ及３．６ｋｂ；一种稀有片段，３．５ｋｂ。在胃癌患者，未发现与正常人体不同的等位片段。但是，稀有片段及稀有杂交带型的出现频率高于正常组。这一结果对深入探讨胃蛋白酶原Ｃ基因稀有片段及稀有杂交带型对胃癌的诊断价     

致倦库蚊有机磷抗性相关扩增酯酶B基因的多样性

本实验从佛山和成都两地的致倦库蚊群体中筛选出３个酯酶类型不同的有机磷（ＯＰ）抗性品系，ＦＳ－１、ＦＳ－２和ＣＤ－１品系。各品系的酯酶基因Ｂ扩增水平、酯酶活性和ＯＰ抗性水平三者一致。各品系酯酶Ｂ基因限制性酶切片段比较分析表明，我国的ＯＰ抗生库蚊群体中不仅有世界性分布的酯酶Ｂ１和Ｂ２，而且在佛山和成都地区各有一个新的独立扩增的酯酶Ｂ，分别命名为Ｂ６和Ｂ７。     

VEGF和p53在胰腺癌中的表达及意义

目的　探讨胰腺癌组织中血管内皮生长因子 (VEGF)及p5 3蛋白的表达及其临床病理意义。方法　采用SP免疫组织化学方法 ,对 46例胰腺癌组织中VEGF及 p5 3蛋白的表达进行检测。结果　VEGF与 p5 3表达率分别为 65 .2 %和 5 8.7%。p5 3表达与VEGF表达呈明显正相关(P <0 .0 5 )。VEGF表达与胰腺癌淋巴结转移 (P <0 .0 5 )和远处转移 (P <0 .0 1)显著相关 ,而与肿瘤大小 ,病理学分级无关 ,p5 3表达与胰腺癌远处转移 (P <0 .0 5 )显著相关 ,而与肿瘤大小 ,病理学分级及淋巴结转移无关。结论　在胰腺癌中 ,VEGF表达与 p5 3蛋白的表达呈正相关 ,在胰腺癌的转移中起重要作用     

利用cDNA微阵列技术筛选成年期与胚胎期睾丸差异表达基因——钙联接蛋白基因

目的 :用cDNA微阵列技术克隆和筛选成年期与胚胎期睾丸差异表达基因 ,以进行睾丸发育及精子发生的调控研究。　方法 :构建人睾丸cDNA芯片 ,分别用正常成人及胚胎睾丸的mRNA探针进行杂交 ,对成人和胚胎睾丸中差异表达的基因进行高流量的比较。　结果 :发现 1条成人睾丸高表达、胚胎低表达的基因———钙联接蛋白 (CLGN)基因。　结论 :运用cDNA微阵列方法可筛选到成年期与胚胎期睾丸差异表达基因