Precision of genotyping of Epstein-Barr virus by polymerase chain reaction using three gene loci (EBNA-2, EBNA-3C, and EBER): predominance of type A virus associated with Hodgkin's disease [published erratum appears in Blood 1993 Oct 1;82(7):2268]

To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin's disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31% with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.     

Identification, chromosomal mapping and tissue-specific expression of hREV3 encoding a putative human DNA polymerase zeta

The Saccharomyces cerevisiae REV3 gene encodes the catalytic subunit of a non-essential DNA polymerase zeta, which is required for mutagenesis. The rev3 mutants significantly reduce both spontaneous and DNA damage- induced mutation rates. We have identified human cDNA clones from two different libraries whose deduced amino acid sequences bear remarkable homology to the yeast Rev3, and named this gene hREV3. The hREV3 gene was mapped to chromosome 1p32-33 by fluorescence in situ hybridization. The hREV3 encodes an mRNA of >10 kb, and its expression varies in different tissues and appears to be elevated in some but not all of the tumor cell lines we have examined. In light of recent reports of a putative mouse REV3, these results indicate that mammalian cells may also contain a mutagenic pathway which aids in cell survival at the cost of increased mutation.      

示指双血管蒂岛状皮瓣急诊拇指再造1例

０　引言　拇指外伤后的急诊修复十分重要,以往各种方法各有其优缺点,我们介绍一种新的尝试．１　病例报告　患者男性,４２岁,左拇指机器绞伤２ｈ就诊．查体：全身情况好,左拇指近节中段以远皮肤软组织大部分撕脱,只有背部约１ｃｍ皮肤相连,但已受挤压,远端无充血...     

反相离子对-高效液相色谱法测定河豚毒素

目的建立了反相离子对-高效液相色谱法测定河豚毒素含量的方法。方法选用SHIMADZUODS色谱柱(150×6mm5μm),以0．2％(v／v)醋酸液为流动相,流速为1．2mL／min,检测波长为230nm。结果河豚毒素线性范围20～100μg／mL,r=0．9960(n=5);回收率为93．32％,RSD=5．41％(n=5);日内精密度为6．10％,日间精密度为7．42％(n=5)最低检测限50ng。结论方法准确、快速、简便,可作为迅速确定河豚毒素含量的测定方法。     

高效液相色谱法测定人血清中酒石酸美托洛尔浓度

目的：建立测定人血清中酒石酸美托洛尔浓度的ＨＰＬＣ方法,为临床药理研究提供科学的分析技术。方法：血清样品萃取纯化后溶于流动相中,进样分析。测定使用Ｃ１８色谱柱,甲醇－水相（５０：５０,水相中含０．１６％戊烷磺酸钠和０．０１４％无水乙酸钠）为流动相,以纳多洛尔为内标,用荧光检测Ｅｘ＝２７７ｎｍ,Ｅｍ＝２９９ｎｍ。结果：本方法线性范围为１０～３００ｎｇ／ｍｌ,最低检测浓度为１ｎｇ／ｍｌ。平均回收率为（     

hVEGF165基因转染后骨髓间充质干细胞蛋白分泌功能及成骨活性的检测

目的：血管内皮细胞生长因子（vascular endothelial growth factor，VEGF）在血管再生和骨组织再生过程中起着重要的调节作用。观察hVEGF165基因转染的骨髓间充质干细胞分泌VEGF的功能及转染后细胞的成骨活性。 方法：实验于2004—07／2005—12在江苏省血液研究所国家重点实验室完成。①体外分离、培养大鼠骨髓间充质干细胞，传代培养后以1μg PcDNA3．1-hVEGF165：3μL阳离子脂质体Lipofectamine的比例转染，通过免疫组织化学SP方法检测转染后细胞中外源性VEGF的表达。②体外分离、培养大鼠肾微血管内皮细胞，分别加入hVEGF165基因转染48h及1周后细胞上清、空载体转染及未转染组细胞上清，MTT法观察细胞增殖情况，以了解转染后细胞培养上清中VEGF的生物学活性。③测定在正常条件培养和成骨条件培养下，转染后细胞上清中碱性磷酸酶、骨钙素的水平。 结果：①hVEGF165基因转染的骨髓间充质干细胞能成功分泌VEGF蛋白。②hVEGF165基因转染48h组大鼠肾微血管内皮细胞A值高于空载体转染及未转染组（P〈0．05），但低于hVEGF165基因转染1周组。③成骨条件培养下，基因转染组细胞碱性磷酸酶和骨钙素的分泌量明显高于未转染组（P〈0．05）；而在正常条件培养下，基因转染组细胞碱性磷酸酶和骨钙素的表达分泌量较低。 结论：①采用基因转染技术可将hVEGF165基因转染到骨髓间充质干细胞中并可有效表达具有生物活性的VEGF，且这种生物学活性呈剂量依赖性。②在成骨条件培养下，转染hVEGF165基因后骨髓间充质干细胞的成骨能力增强。