泛素连接酶E4B通过U-box基序可将经泛素活化酶(Ubiquitin-activating enzyme,E1)和泛素结合酶(Ubiq-uitin-conjugating enzyme,E2)传递的泛素(Ubiquitin,UB)标记至底物蛋白质.探究3个蛋白质:金属蛋白酶M20家族蛋白质2(Peptidase M20 domain-containing protein 2,PM20D2)、多腺苷酸结合蛋白质1(Polyadenylate-binding pro-tein 1,PABPC1)、细胞色素C氧化酶I翻译激活剂(Translational activator of cytochrome C oxidase I,TACO1)是否可被E4B介导泛素化.从HEK293细胞中提取总RNA,反转录为cDNA,以其为模板调取底物基因并构建重组质粒,利用大肠杆菌表达系统表达并纯化泛素化过程所需的各个相关蛋白质,通过蛋白质体外泛素化实验,对3个蛋白质进行泛素化验证.结果表明3个蛋白质均可以在蛋白质水平被E4B泛素化,证明3个蛋白质都是E4B的底物,为进一步在细胞中研究其泛素化的机理奠定基础.

Objective We evaluated the molecular epidemiology of Clostridium difficile strains in patients with inflammatory bowel disease in our hospital so as to provide evidence for effective surveillance of C. difficile infection. Methods From June 2014 to June 2015, a total of 222 stool samples were collected from inpatients with inflammatory bowel disease and sent to clinical microbiological lab to test C. difficile by VIDAS fluorescence enzyme immunoassay and culture. We analyzed the clonal relatedness of bacterial strains by multilocus sequence typing (MLST), antimicrobial resistance pattern by agar dilution method and prevalence of toxin genes by conventional PCR. Simultaneously, we also examined the environment of the ward where the patients stayed. Results The C. difficile was identified in 30 (13.5%) of the 222 patients. The incidence of C. difficile infection (CDI) was 15.7%(22/140) in patients with Crohn’s disease and 9.8% (8/82) in patients with ulcerative colitis. Four strains of C. difficile were isolated from the ward environment. All the 22 clinical strains from stool samples were typed into 14 STs by MLST analysis. The most common type was ST54 (5 strains). Most (72.7%) of the 22 strains belonged to toxin A+B+. However, no isolates contained binary toxin genes. Of the C. difficile strains evaluated, 63.6% displayed resistance to clindamycin and eight strains were resistant to moxifloxacin. All the strains were susceptible to the other six drugs, i.e., chloramphenicol, tetracycline, ampicillin, metronidazole, vancomycin and meropenem. Conclusions Strains with toxin A+B+ were the most common C. difficile isolates. ST54 was the most predominant STs in patients with inflammatory bowel disease in our hospital. It’s necessary to focus on the surveillance of C. difficile infection in patients with inflammatory bowel disease.

Objects To investigate the correlation of the single nucleotide polymorphism (SNP) of clopidogrel related gene CYP2C19, ABCB1, PON1 to the occurrence of clopidogrel resistance (CR) and TEG among patients after percutaneous coronary intervention. Methods A total of 299 patients after PCI were enrolled from April 2015 to December 2015.It genotyped the CYP2C19(rs4244285, rs4986893)ABCB1(rs1045642) and PON1(rs662) gene, measured clopidogrel response by TEG. Accordingly, all the enrolled 299 patients were then divided into CR group (n=17) + non-CR (NCR) group (n=282) or CLR group (n=54) + non-CR (NCR) group (n=245) by TEG(%). All the patients were divided into EM、IM and PM group by CYP2C19 genotype. The age of patient in CR (71.1±11.1) years old is higher than NCR (65.02±10.51) years old (t=2.559, P<0.05). Results CYP2C19 PM was associated with decreased of TEG(ADP) (Z=-2.065, P=0.039), while it was not related to the age of patient(Z=0.405, P>0.05). There was no significant difference between CR(χ2=0.175, P=0.916) CLR(χ2=1.589, P=0.452)and the level of TEG(ADP) (Z=-0.030, P=0.976) in PON1(rs662) polymorphism. There was no significant difference between CR(χ2=1.722, P=0.423) CLR(χ2=0.176, P=0.916)and the level of TEG(ADP) (Z=-0.331, P=0.741) in ABCB1(rs1045642) polymorphism. Conclusions CYP2C19 PM is associated with decreased of TEG(ADP). It is considered that no correlation exists between ABCB1(rs1045642) and PON1(rs662) polymorphism and clopidogrel resistance in patients with coronary heart diseases. The loss of function of ABCB1(rs1045642) and PON1(rs662) is not associated with decreased of TEG(ADP) in CYP2C19 PM patients.(Chin J Lab Med, 2016, 39: 678-684) Key words: Ticlopidine; Aromatase; Aryldialkylphosphatase; P-Glycoproteins; Polymorphism, single nucleotide; Platelet aggregation

金黄色葡萄球菌是临床重要的致病菌,引起的感染可以累及全身各脏器,严重威胁人类健康.ESAT-6分泌系统广泛存在于革兰阳性菌.在金黄色葡萄球菌中,经典的ESAT-6通路由esxA、esxB及其他9个分子组成的基因簇构成,该系统分泌的ESAT-6/WXG100家族蛋白对金黄色葡萄球菌的毒力和免疫防御功能起到重要作用.该文就金黄色葡萄球菌ESAT-6分泌系统的研究进展进行综述.

近年来，研究发现人类免疫缺陷病毒（Human immunodeficiency virus ，HIV）感染人数增多并呈现低龄化的趋势。HIV感染人体后，主要攻击、破坏CD4+T淋巴细胞，经过一段时间（一般是6~8年）的潜伏期，人体细胞免疫功能下降而导致机体发生一系列机会性感染或肿瘤，最终人体丧失对各种疾病的抵抗能力而导致死亡[1]，同时，大量研究发现HIV感染可造成精子活力、精子活率、精子密度、精子总数降低和畸形率增加[2]；并且HIV可通过父婴垂直传播。HIV病毒也可感染人体各期生殖细胞，其中，病毒颗粒是通过结合在精子细胞膜上和（或）存在于精子胞浆中，使病毒RNA整合到宿主精子基因组内。因而，受精时携带HIV 基因的人类精子会进入正常卵母细胞。在胚胎发育过程中， HIV 基因随宿主细胞分裂复制和表达，从而使子代患病[3]。因此，临床上对精液捐献者进行HIV检测意义重大，它可大大减少供精人工授精及体外受精后代患AIDS的风险，从而在一定程度上可降低HIV的传播。

Cilia serve as an important regulator of various signaling pathways,as well as the Hedgehog(Hh) signaling pathway.Recent studies have revealed that there is a close relationship between Hh signaling pathway and intraflagellar transport of cilia,and further evidences from genetics and cytobiology have also proved the relationship between primary cilia and Hh signaling pathway.All of the core components of Hh signaling pathway are found on the primary cilia,and intraflagellar transport proteins are required for the transmission of information between the signaling molecules that mediate the activity of Hh signaling pathway.Flagella of sperm share the same structure with cilia,and ciliopathies can also lead to the defects of spermatic flagella.Ciliopathies and other diseases caused by the defects of structure and function of cilia could affect Hh signaling.Regulatory molecules such as Foxj1 contribute to the construction of cilia,which brings new ideas in the field of infertility and contraception.The research progress of regulatory mechanism of Hedgehog signal in cilia is introduced in this paper.

目的探讨精液冷冻管承载工具对人类精液活动力及复苏率的影响,优化精子冷冻保存的方案.方法取本库20例健康自愿供精的精液,同一例供精者的精液分成两份:实验组冷冻管用棉花包裹,对照组不做任何处理.通过相差显微镜镜检,利用计算机辅助精液分析(CASA)系统分析两组冷冻前后人类活动精子的运动参数,并作相关的统计学分析及比较.结果实验组冷冻复苏率为(66.71±6.51)%,对照组冷冻复苏率为(59.67±7.74)%(平均提高了7.04%)(P<0.01).结论冷冻管包裹棉花的冷冻前处理对精子有一定的保护作用,因而冷冻管承载工具的专门设计对精液冷冻有实质性意义.