人朊蛋白相关蛋白Shadoo的表达、纯化及抗体的制备

目的克隆、表达和纯化人朊蛋白相关Shadoo（SHO）蛋白并制备抗Shadoo的多克隆抗体。方法提取仓鼠各组织的mRNA，分析SHO基因在仓鼠各组织中转录水平的差异;提取人DNA，PCR方法获得人SHO基因，克隆至融合表达载体，在大肠埃希菌中表达人SHO蛋白并纯化;以纯化蛋白为抗原免疫家兔制备抗血清，用Western blot方法分别检测与重组及内源性SHO蛋白的免疫反应性。结果SHO基因在仓鼠各组织中的转录水平差异很大，脑组织转录水平高。在大肠埃希菌中表达了分子质量单位为37 ku的人SHO融合蛋白，制备的SHO蛋白抗体可与重组的SHO蛋白反应，可识别内源性的SHO。结论成功表达纯化人SHO融合蛋白并制备了抗SHO抗体，为研究SHO和正常朊蛋白之间的关系打下初步基础。     

我国脑膜炎奈瑟菌中nalP基因的分布特征

目的 了解我国脑膜炎奈瑟菌（Nm）菌株的nalP基因分布特征,为其功能和作用机制研究提供依据。 方法 采用多位点序列分型方法对187株分离自我国的Nm菌株进行分子分型。利用菌株全基因组框架图获得nalP基因及其两侧序列,对基因序列进行同源性比对及聚类分析。 结果 178株Nm携带nalP基因;Nm中nalP基因与淋病奈瑟菌（Ng）中nalP基因的同源性为95% ~ 96%,但系统进化分析明显分为两支,两侧间隔区同源性＞85%。9株无nalP基因菌株中6株为?nalP2型,新发现3种排布方式。C群患者和健康携带者来源phase on菌株数差异有统计学意义。 结论 Nm中nalP基因可能由Ng获得后各自独立进化,无nalP基因菌株可能是nalP基因丢失,能够表达NalP的C群患者来源菌株可能缺失NalP的补偿机制。      

巴尔通体对19种抗生素体外敏感性分析

目的 检测巴尔通体菌株对10类19种抗生素最低抑菌浓度（MICs）,分析药物敏感性及耐药性,为指导临床用药和耐药监测提供实验和数据参考。方法 采用E试验法,检测巴尔通体属11种、35株菌株对强力霉素、阿奇霉素、利福平等19种抗生素的MICs。将培养的巴尔通体菌制成McFarland（MCF）2.0浊度的菌悬液,均匀涂布于含5%去纤维羊血的胰酶大豆琼脂培养基上,置5% CO2的37℃培养箱培养。培养5~7 d后判读MICs。结果 35株巴尔通体菌在体外对强力霉素、阿奇霉素、红霉素、克拉仙霉素等13种抗生素敏感,MICs 0.016 mg/L;34株对利福平敏感,MICs 0.002 mg/L;对克林霉素、丁胺卡那霉素、万古霉素、多粘菌素和磺胺类5种抗生素不敏感,MICs较高。结论 绝大多数巴尔通体菌对强力霉素等14种抗生素敏感,但也对克林霉素等5种抗生素不敏感,在对巴尔通体病进行治疗时要注意选择敏感药物。     

2015-2016年全国和高发省份伤寒、副伤寒流行病学分析

目的 分析2015-2016年伤寒、副伤寒法定报告病例数,了解全国及高发省份流行病学特点和变化趋势,为防控措施和策略的制定提供依据。方法 采用描述性流行病学方法对网络直报系统中2015-2016年伤寒、副伤寒报告病例数进行分析。结果 2015-2016年全国共报告伤寒、副伤寒病例22 536例,死亡2例,年平均发病率为0.83/10万,较2014年的伤寒、副伤寒发病率下降18.63%。监测点共对14 465份血样标本进行培养,分离出菌株289株,阳性分离率为2.00%,伤寒与副伤寒之比为1.21:1。病例均以农民和民工为主,其次为散居儿童和在校学生,伤寒、副伤寒发病率均以0~4岁组儿童最高（分别为1.70/10万和0.80/10万）。贵州、云南、广西、广东、浙江、湖南和新疆为疫情高发省份,非监测点病例数上升。结论 2015-2016年全国伤寒、副伤寒发病水平处于相对较低的状态,<10岁儿童是防控重点人群,伤寒菌成为优势菌型,应加强非监测点疫情监测。     

A novel linear neutralizing epitope of hepatitis E virus

Hepatitis E virus (HEV) is a serious public health problem that causes acute hepatitis in humans and is primarily transmitted through fecal and oral routes. The major anti-HEV antibody responses are against conformational epitopes located in a.a. 459–606 of HEV pORF2. All reported neutralization epitopes are present on the dimer domain constructed by this peptide. While looking for a neutralizing monoclonal antibody (MAb)-recognized linear epitope, we found a novel neutralizing linear epitope (L2) located in a.a. 423–437 of pORF2. Moreover, epitope L2 is proved non-immunodominant in the HEV-infection process. Using the hepatitis B virus core protein (HBc) as a carrier to display this novel linear epitope, we show herein that this epitope could induce a neutralizing antibody response against HEV in mice and could protect rhesus monkeys from HEV infection. Collectively, our results showed a novel non-immunodominant linear neutralizing epitope of hepatitis E virus, which provided additional insight of HEV vaccine.     

Vaxvec: The first web-based recombinant vaccine vector database and its data analysis

A recombinant vector vaccine uses an attenuated virus, bacterium, or parasite as the carrier to express a heterologous antigen(s). Many recombinant vaccine vectors and related vaccines have been developed and extensively investigated. To compare and better understand recombinant vectors and vaccines, we have generated Vaxvec (http://www.violinet.org/vaxvec), the first web-based database that stores various recombinant vaccine vectors and those experimentally verified vaccines that use these vectors. Vaxvec has now included 59 vaccine vectors that have been used in 196 recombinant vector vaccines against 66 pathogens and cancers. These vectors are classified to 41 viral vectors, 15 bacterial vectors, 1 parasitic vector, and 1 fungal vector. The most commonly used viral vaccine vectors are double-stranded DNA viruses, including herpesviruses, adenoviruses, and poxviruses. For example, Vaxvec includes 63 poxvirus-based recombinant vaccines for over 20 pathogens and cancers. Vaxvec collects 30 recombinant vector influenza vaccines that use 17 recombinant vectors and were experimentally tested in 7 animal models. In addition, over 60 protective antigens used in recombinant vector vaccines are annotated and analyzed. User-friendly web-interfaces are available for querying various data in Vaxvec. To support data exchange, the information of vaccine vectors, vaccines, and related information is stored in the Vaccine Ontology (VO). Vaxvec is a timely and vital source of vaccine vector database and facilitates efficient vaccine vector research and development.     

The tuberculosis vaccine H4:IC31 is safe and induces a persistent polyfunctional CD4 T cell response in South African adults: A randomized controlled trial

BackgroundNew, more effective vaccines to prevent tuberculosis (TB) disease are needed urgently. H4:IC31 is an investigational vaccine that contains a fusion protein of the immunodominant antigens TB10.4 and Ag85B, formulated in IC31^® adjuvant. We assessed the safety and immunogenicity of H4:IC31 in South African adults from a TB endemic setting.MethodsIn this double blind, placebo controlled, phase I trial, Mycobacterium tuberculosis-uninfected, HIV-uninfected, healthy adults with a history of childhood BCG vaccination were randomly allocated to two intramuscular vaccinations with 5, 15, 50 or 150 μg H4 formulated in 500 nmol IC31^®, two months apart. Vaccinees were followed for six months to assess safety; immunogenicity was measured by ELISpot and intracellular cytokine staining assays.ResultsThirty-two participants received H4:IC31 and 8 received placebo. Injection site adverse events were common but mild; mild fatigue was the most common systemic adverse event. Frequencies of adverse events did not differ between dosage groups. Detectable antigen-specific CD4 T cell responses were induced by all doses of H4:IC31, but doses below 50 μg induced the highest frequencies of CD4 T cells, comprised predominantly of IFN-γ⁺TNF-α⁺IL-2⁺ or TNF-α⁺IL-2⁺ cells. These memory responses persisted up to the end of follow up, on study day 182.ConclusionsH4:IC31 demonstrated an acceptable safety profile and was immunogenic in South African adults. In this trial, the 15 μg dose appeared to induce the most optimal immune response.