视网膜特异性表达人血管内皮生长因子 基因载体的构建

目的构建在视网膜组织特异性表达的人血管内皮生长因子（VEGF）₁₆₅基因。方法用聚合酶链反应(PCR)方法从BLAB/C鼠全基因组扩增能在视网膜组织特异性表达的rho启动子，经限制性内切酶纯化后克隆于质粒pcDNA^3.1+-VEGF₁₆₅中，建立重组质粒pcDNA^3.1+-rho-VEGF₁₆₅，通过限制性内切酶酶切分析及PCR鉴定筛选出正确重组质粒pcDNA^3.1+-rho-VEGF₁₆₅,由jetPEI介导转染人视网膜色素上皮细胞和人脐静脉内皮细胞，并通过免疫组织化学染色以及绘制细胞生长曲线检测在人视网膜色素上皮细胞和人脐静脉内皮细胞中VEGF蛋白的表达。结果在人视网膜色素上皮细胞中，重组质粒pcDNA^3.1+-rho-VEGF₁₆₅比质粒pcDNA^3.1+-rho-VEGF₁₆₅的VEGF蛋白表达强，在人脐静脉内皮细胞，两者的表达量无明显差别。结论pcDNA^3.1+-rho-VEGF₁₆₅载体的构建为进一步研究VEGF在视网膜新生血管形成中的致病机理提供基础材料，并为进一步建立视网膜特异性表达VEGF转基因鼠模型建立了基础。(中华眼底病杂志,2005,21:106-109)     

恶性和非恶性肿瘤患者正常组织细胞DNA含量参数的对比研究

目的 :探讨恶性和非恶性肿瘤患者正常组织细胞生物学特性的区别。　方法 :用流式细胞术对 2 37例恶性肿瘤患者和 148例非恶性肿瘤患者正常组织细胞的DNA含量进行了检测 ,并对二者的DNA指数 (DI)、DNA倍体类型、细胞程序性死亡水平 (Apo)和增殖活性 (SPF)做了对比分析。　结果 :2 37例恶性肿瘤患者正常组织的DI值为1.0 8± 0 .2 3,DNA异倍体检出率为 18.99% ;148例非恶性肿瘤患者正常组织的DI值为 1.0 0± 0 ,DNA倍体类型均为二倍体。恶性肿瘤患者正常组织的Apo和SPF均显著高于非恶性肿瘤患者正常组织 (P <0 .0 5 )。即使剔除DNA异倍体病例 ,其Apo和SPF仍显著高于非恶性肿瘤患者的正常组织 (P <0 .0 5和P <0 .0 1)。　结论 :恶性肿瘤患者癌旁远处组织DNA二倍体细胞的Apo和SPF显著高于非恶性肿瘤正常组织 ,不能真正代表正常组织细胞的生物学特性     

生物医学测量及控制技术新进展

介绍生物医学测量及控制技术领域中的一些研究进展。     

图像分析DNA含量对判断膀胱移行细胞癌分级和预后的意义

采用IBAS图像分析系统检测37例膀胱移行细胞癌病人的肿瘤细胞DNA含量，其中35例随访18个月至7年（平均5l个月），结果显示DNA含量和AN出现率增加与肿瘤的恶性程度有关（P＜0.01和P＜0.05）；肿瘤复发亦与非整倍体细胞（AN）出现率密切相关，AN出现率小于10％无复发，＜30％5例复发，＞30％9例复发。研究资料表明，图像分析技术测定细胞核的DNA含量对判断膀胱移行细胞癌病人的分级和预后有重要意义。     

巢湖水有机提取物致鱼红细胞DNA损伤的初步观察

目的　研究巢湖水有机提取物致突变性。方法　单细胞凝胶电泳技术测定鱼红细胞DNA损伤效应。结果　巢湖源水引起慧星细胞的百分率最高 (5 7.2 5 % ) ,滤前水最低 (2 7.6 3% ) ,出厂水经过二次加氯后慧星细胞的百分率有所上升 (4 4 .0 0 % )。结论　巢湖源水具有潜在致突变性 ,经混凝、活性炭吸附及沉淀处理后其DNA损伤作用有所下降 ,但氯化消毒可增加水中有机提取物的DNA损伤作用。     

Brain and kidney of progressive multifocal leukoencephalopathy patients contain identical rearrangements of the JC virus promoter/enhancer

The kidneys of six progressive multifocal leukoencephalopathy (PML) patients were examined by PCR amplification for the presence of JC virus. Amplification of three different areas of the viral genome from multiple samples of each kidney revealed three that were positive for the virus. The use of a PCR-based typing assay on all tissue samples, and cloned sequences from the viral coding region from each positive kidney showed that the same viral genome was present in the kidney as in the brain of the patient. Regulatory region clones all had the archetypal promoter/enhancer structure. However, when PCR fragments from the regulatory region were digested with a restriction enzyme which cuts in region D, the region most often deleted in PML-type promoters, a low level of undigested DNA remained. This DNA refractory to digestion had a rearranged sequence identical to that of the unique rearranged promoter in the brain of each patient. © 1994 Wiley-Liss, Inc.     

16株假单胞菌DNA的G＋C百分含量测定

采用改良的热变性温度法，将DNA升温及恒温处理与紫外吸收值的测定分两步进行，检测3株标准菌株和16株假单胞菌DNA的G Cmol% 含量。结果表明，14株假单胞菌与其相应标准菌种的DNAG＋Cmol%含量基本一致，与其生物学性状亦相符；另2株DNA的G＋Cmol%含量与其生物学性状不符，其分类学位置尚待探讨。     

Cerivastatin inhibits fetal calf serunvinduced DNA synthesis in cultured rat mesangial cells

SUMMARY: Inhibition of mevalonate synthesis by several statins has been shown to suppress DNA synthesis in glomerular mesangial cells. In the present study, we investigated the effect of a new statin, cerivastatin, on fetal calf serum (FCS)-induced DNA synthesis of cultured rat mesangial cells. Cultured rat mesangial cells were stimulated by 10% FCS in the presence or absence of cerivastatin and mevalonate. 5-bromo-2-deoxyuridine (BrdU) incorporation was used to assess DNA synthesis. the present study showed that 10% FCS caused marked stimulation of DNA synthesis in the mesangial cells. Cerivastatin inhibited FCS-stimulated BrdU incorporation in a dose-dependent manner. IC₅₀ was approximately 1 umol/L. Exogenous mevalonate, farnesyl pyrophosphate and geranylgeranyl pyrophosphate significantly prevented the inhibitory effect of cerivastatin on DNA replication. It appears that cerivastatin, by inhibiting the synthesis of mevalonate, may suppress DNA synthesis in the mesangial cells.     

Andrology: Application of different in-situ hybridization detection methods for human sperm analysis

The detection of some types of aneuploidy in human spermatozoacan be based on the use of the fluorescence in-situ hybridizationtechnique (FISH). One of the crucial steps for FISH is to achievea proper decondensation and denaturation of the DNA in the specimen,so as to obtain efficient hybridization results. However, afterDNA decondensation the morphology of sperm heads is partly distortedand the majority of the tails is lost. This situation leadsto problems in the distinction between disomic and diploid spermatozoa,as well as between abnormal spermatozoa and somatic cells. Double-and triple-target FISH can partly solve this discriminationproblem. To improve these procedures we adapted the steps ofdecondensation and visualization of the single sperm cells.Firstly, DNA decondensation with 25 mM dithiothreitol in 1 MTris at pH 9.5 resulted in sperm cells with intact morphologyof both the head and the tail, and allowed efficient single-,double- and triple-target ISH to be performed. Secondly, weapplied a novel detection method, based on enzyme immunocyto-chemicalreactions, with coloured precipitation products. Thirdly, thisISH procedure was combined with Diff-Quik staining and bright-fieldmicroscopy. This absorption method has the advantage of a permanentsignal, and the adapted cytoplasmic staining of the sperm plasmamembrane allows the visualization of the outline of the singlespermatozoon. Using this approach, therefore, it is possibleto discriminate between disomic, diploid and abnormal spermatozoa,somatic cells and spermatozoa that overlap, because the morphologyof the cells is not distorted and the tails of the spermatozoaare intact and properly visualized.     

Higher-order organization and compartmentalization of satellite DNA PIM357 in species of the coleopteran genus Pimelia

The PIM357 satellite DNA family is present in 26 Pimelia taxa (Tenebrionidae, Coleoptera) with endemic congeneric species from the Canary Islands showing higher interrepeat variability than continental ones. In this paper, we compare the repetitive DNA sequences of a Canarian species that has distinct subfamilies of repeat units, P. radula ascendens, with another without such subfamilies, P. sparsa sparsa. The chromosomal localization of the repeat units and the comparison of the variability of randomly cloned monomers to the one estimated by comparing repeat units from dimers and trimers suggest the absence of satellite subfamilies in P. sparsa sparsa. Hence, the repeat units of this species seem to be uniformly and randomly distributed throughout all chromosomes out of one chromosomal pair. On the contrary, P. radula ascendens shows four divergent subfamilies of repeat units supported by several diagnostic nucleotide substitutions. These subfamilies seem to form four distinct repeat units: monomer subfamily 1, monomer subfamily 4 and two higher-order units (dimer linking subfamily 1 and 4, and dimer linking subfamily 2 and 3). Moreover, monomers of subfamily 1 are present in three chromosomal pairs only. We discuss the effect of different potential factors acting in the concerted evolution and the genomic organization of stDNA sequences in these taxa. This revised version was published online in July 2006 with corrections to the Cover Date.