苯并（a）芘代谢产物诱发BALB／3T3细胞程序外DNA合成的研究

为了解苯并（ａ）芘（ＢａＰ）代谢产物对哺乳动物细胞ＤＮＡ的损伤作用，本实验采用单标记和双标记法，分别检测４种苯并（ａ）芘代谢产物（ａｎｔｉ－ＢＰＤＥ，ｓｙｎ－ＢＰＤＥ，３－ＯＨ－ＢＰ和９－ＯＨ－ＢＰ）对ＢＡＬＢ／３Ｔ３细胞ＤＮＡ合成和程序外ＤＮＡ合成（ＵＤＳ）的影响。结果表明：ａｎｔｉ－ＢＰＤＥ、ｓｙｎ－ＢＰＤＥ、Ｅ－ＯＨ－ＢＰ和９－ＯＨ－ＢＰ均在不同程度上使ＢＡＬＢ／３Ｔ３细胞ＤＮＡ合成增加；但     

椰毒假单胞菌酵米面亚种16S rDNA序列测定与分析

本文对３种不同来源样品的４株椰酵假单胞菌的１６ＳｒＤＮＡ的序列进行了测定,以确定其系统发育位置及与其它菌种的遗传进化关系。实验中设计了７条引物,利用聚合酶链式反应（ＰｏｌｙｍａｓｅＣｈａｉｎＲｅ－ａｃｔｉｏｎ,ＰＣＲ）扩增４株细菌的１６ＳｒＤＮＡ,通过扩增产物的克隆测序或直接测序,获得了它们的１６ＳｒＤＮＡ序列,将测得的序列与伯克霍尔德氏菌属中其它菌种的１６ＳｒＤＮＡ序列进行了聚类分析,得到了系统发育进化树状图。树状图及不同菌种之间的同源性比值：椰酵假单胞菌（Ｐ．ｃｏｃｏｖｅｎｅｎａｎｓｓｕｂｓｐ．ｆａｒｉｎｏｆｅｒｍｅｎｔａｎｓ）与唐菖蒲伯克霍尔德氏菌（Ｂｕｒｋｈｏｌｄｅｒｉａｇｌａｄｉｏｌｉ）、椰毒伯克霍尔德氏菌（Ｂｕｒｋｈｏｌｄｅｒｉａｃｏｃｏｖｅｎｅｎａｎｓ）的亲缘关系非常近,与其它菌种的亲缘关系相对较远,椰酵假单胞菌、唐菖蒲伯克霍尔德氏菌和椰毒伯克霍尔德氏菌可归属于一个独立的系统发育系。     

随机引物多聚酶链反应对细菌分型的价值及影响因素探讨

随机引物多聚酶链反应对细菌分型的价值及影响因素探讨高瞻，ＫａｔｈｙＪａｃｋｓｏｎ，ＤａｖｉｄＥ．Ｌｅｓｌｉｅ’踌机引物多聚酶链反应（ＲａｎｄｏｍＡｍｐｌｉｆｉｅｄｐｏｌｙ－ｍｏｒｐｈｉｃＤＮＡＰＣＲ，简称ＲＡＰＤＰＣＲ）是在对所扩增基因一无所知的情况...     

CPB-ST融合基因的构建及表达研究

用限制性核酸内切酶ＥｃｏＲＩ和ＳａｌⅠ双酶切含有大肠杆菌耐热性肠毒素ＳＴ１前体蛋白基因的质粒ｐＸＳＴ１，回收３２５ｂｐ的ＳＴ基因片段，然后，通过Ｔ４ＤＮＡ连接酶将其定向连接于事先经同样的双酶切处理的带有产气荚膜梭菌β－毒素基因（ＣＰＢ）的重组质粒ｐＥＣＢ２中ＣＰＢ基因的下降，转化受体菌ＢＬ２１（ＤＥ３）中，经ＢａｍＨＩ和ＥｃｏＲＩ酶切反应鉴定重组质粒，得到了理想重组质粒ｐＥＣＢ－ＳＴ１。重组菌株Ｂｌ２１（ＤＥ３）（ｐＥＣＢ－ＳＴ１）经ＩＰＴＧ诱导后，其表达产物经ＳＤＳ－ＰＡＧＥ、Ｗｅｓｔｅｒｎ－ｂｌｏｔｉｎｇ及ＥＬＩＳＡ检测，结果表明重组菌株可以表达ＣＰＢ－ＳＴ融合蛋白，而且该融合蛋白无天然β－毒素和ＳＴ１的生物毒性。     

三种利什曼原虫和我国新疆皮肤利什曼原虫kDNA分子…

本研究自新疆克拉玛依地区皮肤利什曼病（ＣＬ）患者的皮肤病变组织内抽提微量的利什曼原虫ｋＤＮＡ，采用引物１３Ａ、１３Ｂ进行ＰＣＲ扩增，获１２０ｂｐ扩增区带（ＣＬ－ＰＣＲ－ＡＰ），并转移到尼龙膜上，分别与地高辛标记的Ｌ．ｔｒｏｐｉｃａ、Ｌ．ｉｎｆａｎｔｕｍ、Ｌ．ｇｅｒｂｉｌｌｉ ｋＤＮＡ探针进行Ｓｏｕｔｈｅｍ印迹杂交。结果：病变组织内利什曼原虫ｋＤＮＡ的ＰＣＲ扩增区带仅与Ｌ．ｔｒｏｐｉｃａ ｋＤＮＡ探     

随机引物多聚酶链反应对革兰氏阴性分型的影响因素研究

随机引物多聚酶链反应对革兰氏阴性分型的影响因素研究高瞻ＫａｔｈｙＪａｃｋｓｏｎＤａｖｉｄＥ．Ｌｅｓｌｉｅ江苏省卫生防疫站（２１０００９南京）随机引物多聚酶链反应（ＲａｎｄｏｍＡｍｐｌｉｆｉｅｄＰｏｌｙｍｏｒｐｈｉｃＤＮＡＰＣＲ，简称ＲＡＰＤＰＣＲ）是...     

前白蛋白cDNA的克隆及真核表达载体的构建

为构建血浆前白蛋白的真核表达载体，采用逆转录ＰＣＲ（ＲＴ－ＰＣＲ）从水囊引产的胎儿肝组织中得到ＰＡｃＤＮＡ。将该序列克隆到ｐＧＥＭ－Ｔ Ｅａｓｙ载体中，再转入真核表达载体ｐＣＤＮＡ３．１（＋）。限制性内切酶消化筛选正向插入重组体。重组体经过序列分析证实，ｐＣＤＮＡ３－ＰＡ中正向插入ＰＡ全长ｃＤＮＡ。     

２０１３－２０１４年杭州地区季节性Ａ （Ｈ３Ｎ２） 流感病毒犘犅１ 基因及其编码蛋白的遗传进化特征研究

摘要：目的　分析２０１３－２０１４年杭州市季节性Ａ （Ｈ３Ｎ２）流感病毒犘犅１ 基因及其编码蛋白的变异情况,
揭示其分子特征与遗传进化趋势。方法　对分离得到的７８株流感病毒犘犅１ 基因进行扩增和测序,采用序
列多重比对、突变位点比较分析和构建进化树等方法进行遗传进化分析。结果　７８ 株流感病毒犘犅１ 基因
的核苷酸序列和编码蛋白序列一致性分别为９７．８９％ ～１００．００％ 和９９．０８％ ～１００．００％,ＰＢ１ Ｆ２ 蛋白短肽
的序列一致性为４４．４４％～１００．００％;ＰＢ１蛋白氨基酸序列与参考株Ａ／Ｔｅｘａｓ／５０／２０１２相比,在十多个位
点均发生了氨基酸突变,其中Ｖ２１２Ｍ、Ｒ２１５Ｋ、Ｅ３３１Ｄ、Ａ３７４Ｓ氨基酸突变频率较高;ＰＢ１ Ｆ２蛋白长度分
析发现了５种不同的ＰＢ１ Ｆ２ 蛋白,其中３ 种为全长蛋白（９０、８７、７９）ａａ,２ 种为截断型ＰＢ１ Ｆ２ 蛋白
（５２,２４）ａａ;构建系统进化树发现７８株Ｈ３Ｎ２毒株犘犅１ 基因及其编码蛋白均在同一个支系上,与同期流
行季的疫苗株进化关系较近,第三个流行高峰（２０１４ 年６～１０ 月） 进化关系相对更集中。结论　２０１３－
２０１４年季节性Ａ （Ｈ３Ｎ２）流感病毒ＰＢ１蛋白氨基酸位点发生了部分重要突变,病毒犘犅１ 基因及其编码蛋
白的遗传进化以及截断型ＰＢ１ Ｆ２蛋白的集中出现,可能促使病毒的复制能力和毒力发生变化,并最终导
致其流行传播趋势的改变。
关键词：Ａ （Ｈ３Ｎ２）流感病毒;ＰＢ１;ＰＢ１ Ｆ２;遗传进化
中图分类号：Ｒ３７３．１＋３　　文献标识码：Ａ　　文章编号：１００９ ６６３９ （２０１７）０３ ０１６１ ０７     

胃腺癌组织中Fas、Bcl-2蛋白表达及其与浸润转移关系的研究

目的：为了解Ｆａｓ、Ｂｃｌ－２蛋白在胃腺癌中的表达及其意义。方法：用免疫组化ＡＢＣ法检测４８例胃腺癌中Ｆａｓ、Ｂｃｌ－２蛋白的表达。结果：中、高分化癌中Ｆａｓ蛋白阳性表达率（７０．６％、７１．４％）显著高于低分化癌（１２．５％）,（Ｐ＜０．０５）,无淋巴结转移组的胃癌Ｆａｓ阳性表达率（５５．６％）高于有淋巴结转移组（２３．８％）,（Ｐ＜０．０５）,浸及粘膜及粘膜下的胃癌Ｆａｓ阳性表达率（７１．４％）高于浸及浆膜及周围组织者（２３．５％）,（Ｐ＜０．０５）。高分化癌中Ｂｃｌ－２的阳性表达率（４２．８％）显著低于低分化癌（８７．５％）,（Ｐ＜０．０５）,Ｂｃｌ－２阳性表达与浸润深度及淋巴结转移无关。结论：Ｆａｓ表达与胃腺癌的分化、浸润转移有关,Ｂｃｌ－２表达仅与胃腺癌分化有关     

我国新型恙虫病立克次体目标基因的PCR／RFLR和序列分析研究

为查明江苏恙虫病立克次体（Ｒｔ）流行株目标基因序列特征，构建Ｒｔｔｓａ５６ｋＤａ群、型特异引物，用ＰＣＲ技术进行检测、分型，将群引物扩增产物进行ＲＦＬＰ分析和热循环自动双向测序法进序列测定。结果所构建的Ｒｔｔｓａ５６ｋＤａ的群、型引物具有Ｒｔ的特异性；江苏地区Ｒｔ流行株型别与日本Ｋａｗａｓａｋｉ型相似；群引物扩增产物经ＲＦＬＰ分析，证明该片段ＤＮＡ无ＨｈａⅠ酶切位点；碱基序列测定结果与Ｋａｗａｓａｋｉ型相应ＤＮＡ片段的碱基序列同源性为９６．８７％，而与其它５株Ｒｔ同源性均少于５２％；该片段的ＨｈａⅠ酶切位点ＧＣＧＣ变异为ＧＴＧＣ结构。结果表明江苏地区Ｒｔ株型属于日本Ｋａｗａｓａｋｉ型，但有遗传差异，是我国新发现的株型     

Enhanced immune protection by a liposome-encapsulated recombinant respiratory syncytial virus (RSV) vaccine using immunogenic lipids from Deinococcus radiodurans

The radiation-resistant bacterium, Deinococcus radiodurans contains a variety of phospho-, glyco- and phosphoglycolipids, the structures of which appear to be largely unique in nature. We show here that such lipids are immunogenic when administered as liposomes intranasally in mice, as evidenced by the induction of serum antibodies which recognize D. radiodurans lipids but not other lipids by thin layer chromatographic immunostaining. By modifying a liposomal vaccine against respiratory syncytial virus (RSV) we find that vaccine efficacy is markedly enhanced by the inclusion of lipids isolated from D. radiodurans. Using dioleoylphosphatidylcholine (DOPC) or D. radiodurans lipids, liposomes were prepared which encapsulated a soluble fragment of the RSV G protein (G(128-188)) fused with a portion of the bacterial thioredoxin (Trx) protein. Mice immunized intranasally with D. radiodurans liposomes showed markedly greater protection against RSV challenge over mice immunized with DOPC liposomes. Enhanced vaccine efficacy was achieved using liposomes prepared from either whole D. radiodurans lipids or from a single isolated phosphoglycolipid previously identified as alpha-galactosylphosphatidylglyceroylalkylamine (lipid 7). Mice immunized and protected against RSV challenge were free of pulmonary eosinophilic infiltration, an undesirable consequence of many RSV vaccines. The results provide further support for liposome-based vaccines for RSV and underline the importance of lipid composition in liposome formulations.     

环氧乙烷灭菌营养琼脂培养皿应用的实验观察

目的：提出了一种培养皿灭菌方法，即营养琼脂培养皿密封后经过环氧乙烷灭菌，解析后保存备用。方法：通过分析残留环氧乙烷的抑菌影响、不同保存方法对细菌培养的影响，应用不同采样方法空气沉降菌采样，对自然菌捕获率进行了比较。结果：营养琼脂培养皿环氧乙烷灭菌后对空气沉降法或对一些表面的采样．收集细菌的效果和培养结果都与普通9cm玻璃培养皿相当。其结果具有可比较性。结论：环氧乙烷灭菌营养琼脂培养皿可以在一些现场采样中使用，尤其在突发事件的紧急处理工作中，减少了采样的准备工作，可争取宝贵的时间。     

Purification and characterization of a novel DNA repair enzyme from the extremely radioresistant bacterium Rubrobacter radiotolerans

Rubrobacter radiotolerans is an extremely radioresistant bacterium. It exhibits higher resistance than the well-known radioresistant bacterium Deinococcus radiodurans, but the molecular mechanisms responsible for the radio-resistance of R. radiotolerans remain unknown. In the present study, we have demonstrated the presence of a novel DNA repair enzyme in R. radiotolerans cells that recognizes radiation-induced DNA damages such as thymine glycol, urea residues, and abasic sites. The enzyme was purified from the crude cell extract by a series of chromatography to an apparent physical homogeneity. The purified enzyme showed a single band with a molecular mass of approximately 40 kDa in SDS-polyacrylamide gel electrophoresis, and was designated as R-endonuclease. R-Endonuclease exhibited repair activity for thymine glycol, urea residues, and abasic sites present in plasmid DNA, but did not act on intact DNA, UV-irradiated DNA and DNA containing reduced abasic sites. The substrate specificity together with the salt and pH optima suggests that R-endonuclease is a functional homolog of endonuclease III of Escherichia coli.     

Restricted replication of the live attenuated influenza A virus vaccine during infection of primary differentiated human nasal epithelial cells

Live Attenuated Influenza Vaccine (LAIV) strains are associated with cold adapted, temperature sensitive and attenuated phenotypes that have been studied in non-human or immortalized cell cultures as well as in animal models. Using a primary, differentiated human nasal epithelial cell (hNEC) culture system we compared the replication kinetics, levels of cell-associated viral proteins and virus particle release during infection with LAIV or the corresponding wild type (WT) influenza viruses. At both 33 °C and 37 °C, seasonal influenza virus and an antigenically matched LAIV replicated to similar titers in MDCK cells but seasonal influenza virus replicated to higher titers than LAIV in hNEC cultures, suggesting a greater restriction of LAIV replication in hNEC cultures. Despite the disparity in infectious virus production, the supernatants from H1N1 and LAIV infected hNEC cultures had equivalent amounts of viral proteins and hemagglutination titers, suggesting the formation of non-infectious virus particles by LAIV in hNEC cultures.     

Outer membrane characteristics of Salmonella enteritidis phage type 4 growing in chickens.

Strains of Salmonella enteritidis belonging to phage type 4 (SE4) were grown in the peritoneal cavities of chickens, and without subculture on laboratory media examined for inducible in vivo phenotypic characteristics. These bacteria expressed three major outer membrane proteins (OMPs) of 33, 35 and 36 kilodaltons (kDa), and iron regulated OMPs of 74, 78 and 81 kDa. Bacteria growing in vivo did not express flagella, or fimbriae with a subunit molecular mass of 14 kDa (14 kDa fimbriae). Two OMPs of 55 and 23 kDa, expressed during culture in nutrient broth, were repressed during growth in chickens. Possession of a 38 MDa ''mouse virulence'' plasmid did not influence the expression of OMPs, flagella or fimbriae. It was concluded that strains of SE4 growing in chicken tissues, use an enterobactin mediated iron uptake system to obtain ferric ions, do not express flagella or 14 kDa fimbriae and appear not to express novel OMPs involved in survival in vivo.     

Functionally Relevant Differences in Plasma Fatty Acid Composition and Expression of Cytotoxic and Inhibitory NK Cell Receptors between Healthy Young and Healthy Elder Adults

(1) Background: In the healthy ageing, NK cell number is not modified; however, their spontaneous cytotoxicity decreases. We postulated that the age-dependent decline in metabolic activities might be responsible for this effect. (2) Methods: The fatty acid profile of 30 healthy young males (23 ± 4 years old, BMI 22.1 ± 1.3) and 30 older males (63 ± 5 years old, BMI 22.9 ± 2.5) donors were evaluated along with the expression of killing (KR) and inhibitory NK receptors (KIR) at basal level and after cultivation with fatty acids for 24 h. (3) Results: Significantly higher levels of oleic (p < 0.01), arachidonic (p < 0.001), lignoceric (p < 0.001), and nervonic acids (p < 0.0001) and significantly lower levels of docosapentaenoic and docosahexaenoic acids (p < 0.01) were found in elders as compared to young adults. At basal levels, significant (p < 0.005) differences in KR and KIR expression were encountered; 12/16 antigens. Treatment of cells with saturated fatty acids or arachidonic acid (AA) significantly enhanced KR expressions (p < 0.001). AA treatment decreased inhibitory KIR expression while docosahexaenoic, and eicosapentaenoic acid increased them. (4) Conclusions: Changes in fatty acids blood levels, and KR and KIR expression in NK cell, are age-dependent. Supplementation of NK cells with eicosapentaenoic or docosahexaenoic acid enhanced inhibitory KIR receptors’ expression which may improve their cell function.     

Characterization,DBPs formation,and mutagenicity of different organic matter fractions in two source waters

Dissolved organic matters (DOM) are critical in the formation of the mutagenic disinfection byproducts (DBPs). In this study, five DOM fractions were isolated and investigated from a contaminated river and a clean reservoir source waters using resin adsorption. The DOM fractions were characterized with excitation–emission matrix, and several typical DBPs formation potentials and the mutagenicity of each DOM fraction were measured. Among these fractions, hydrophobic neutrals (HON), hydrophilic acids (HIA) and hydrophilic bases (HIB) generated the highest carbon-containing DBPs in the river source water, as did HIB and hydrophobic bases (HOB) in reservoir water. Hydrophobic acids (HOA), HON, and HIA were the three most important fractions forming nitrogen-containing DBPs. Following chlorination, the mutagenicity of HON, HIA, HOA and HIB was 1503, 626, 422 and 116 ng 4-NQO/mg DOC in river water, respectively. Only HIA and HOA were mutagenic with 85 and 10 ng 4-NQO/mg DOC in reservoir water, respectively. The soluble microbial products like substances and aromatic proteins contributed significantly to the mutagenicity of river water; whereas the humic acid-like and fulvic acid-like substances were the primary contributors to the mutagenicity of reservoir water.     

Occurrence,origin, and toxicity of disinfection byproducts in chlorinated swimming pools: An overview

Disinfection treatments are critical to conserve the microbiological quality of swimming pool water and to prevent water-borne infections. The formation of disinfection byproducts (DBPs) in swimming pools is an undesirable consequence resulting from reactions of disinfectants (e.g. chlorine) with organic and inorganic matter present in pool water, mainly brought by bathers. A considerable body of occurrence studies has identified several classes of DBPs in swimming pools with more than 100 compounds detected, mainly in chlorinated freshwater pools. Trihalomethanes (THMs), haloacetic acids (HAAs), haloacetaldehydes (HALs) are among the major DBPs in swimming pools. Other DBPs such as haloacetonitriles (HAN), haloamines, nitrosamines, and halobenzoquinones have also been detected. Researchers have been interested in identifying the precursors responsible for the formation of DBPs. In swimming pools, anthropogenic organic loads brought by swimmers increase the complexity of pool water chemistry. When human inputs (e.g. sweat, urine, hair, skin and personal care products) containing very diverse organic compounds are introduced to pools by swimmers, they react with chlorine resulting in the formation of complex mixtures of DBPs. The overwhelming majority of the total organic halide (TOX) content is still unknown in swimming pools. Exposure of swimmers to DBPs can take place through multiple routes, depending on the chemical properties of each DBP. Toxicological studies have shown that swimming pool water can be mutagenic with different potencies reported in different studies. Many DBPs have been shown to be genotoxic and carcinogenic. DBPs were also shown to induce reproductive and neurotoxic adverse effects in animal studies. Epidemiologic studies in humans have shown that exposure to DBPs increases the risk of respiratory adverse effects and bladder cancer. Association between DBPs and other health effects are still inconclusive. Data gathered in the present review (occurrence, toxicity, and toxicological reference values) could be used in conducting chemical risk assessment studies in swimming pools.     

1,25(OH)_2D_3对巨噬细胞吞噬金黄色葡萄球菌过程的影响

目的研究维生素D(VD)对巨噬细胞吞噬金黄色葡萄球菌的吞噬活性及吞菌后巨噬细胞自身凋亡过程的影响。方法巨噬细胞株RAW264.7分为对照组(C),细菌组(B),活性VD+细菌组(VD+B),VD+B组加入终浓度为10-8mol/L活性VD培养24h后,与细菌组一起感染金黄色葡萄球菌1h,用流式细胞仪测定各组巨噬细胞吞噬率、线粒体膜电位、细胞内游离钙离子浓度、活性氧的差异。结果VD+B组巨噬细胞吞噬金黄色葡萄球菌的能力显著高于B组;VD+B组巨噬细胞吞菌后线粒体膜电位显著低于B组;同时,VD+B组的细胞内游离钙离子浓度显著低于B组的;两组间活性氧的生成没有显著性差异。结论维生素D能促进巨噬细胞对金黄色葡萄球菌的吞噬能力,对于吞噬细菌后巨噬细胞自身的凋亡有一定抑制作用。     

河南汉族食管鳞癌与XPD基因多态性关系

目的探讨着色性干皮病基因D(xeroderma pigmentosum group D,XPD)基因多态性与食管鳞癌的关系,为揭示食管癌病因及制定食管癌干预措施提供理论依据。方法采用病例对照研究方法,选取郑州大学附属第一医院食管鳞癌患者235例,按性别与年龄1∶1匹配选取河南省新乡县健康人群235人作为对照,采用聚合酶链反应-限制性片段长度多态性方法,检测XPD 312位点和751位点基因型,采用非条件Logistic回归进行多因素和交互作用分析。结果与野生型GG比较,XPD 312位点GA、AA和(GA+AA)基因型与食管鳞癌易感性无关,分别调整后OR=0.83(95%CI=0.05～13.55),OR=1.12(95%CI=0.06～19.47)和OR=1.35(95%CI=0.73～2.51);与野生型AA比较,XPD 751位点AC、CC和(AC+CC)基因型与食管鳞癌易感性无关,分别调整后OR=1.21(95%CI=0.80～1.83),OR=0.99(95%CI=0.42～2.31)和OR=1.17(95%CI=0.79～1.75);以GA作为对照单体型,GC、AA和AC单体型与食管鳞癌易感性无关,未发现两位点与吸烟或饮酒之间的交互作用。结论 XPD 312和XPD 751位点多态性可能与河南汉族人群食管鳞癌的发生无关,且与吸烟或饮酒不存在交互作用。