Chromosomal Rearrangement Features of Yersinia pestis Strains from Natural Plague Foci in China

The Yersinia pestis chromosome contains a large variety and number of insert sequences that have resulted in frequent chromosome rearrangement events. To identify the chromosomal rearrangement features of Y. pestis strains from five typical plague foci in China and study spontaneous DNA rearrangements potentially stabilized in certain lineages of Y. pestis genomes, we examined the linking mode of locally collinear blocks (LCBs) in 30 Y. pestis strains by a polymerase chain reaction-based method. Our results suggest most strains have relatively stable chromosomal arrangement patterns, and these rearrangement characteristics also have a very close relationship with the geographical origin. In addition, some LCB linking modes are only present in specific strains. We conclude Y. pestis chromosome rearrangement patterns may reflect the genetic features of specific geographical areas and can be applied to distinguish Y. pestis isolates; furthermore, most of the rearrangement events are stable in certain lineages of Y. pestis genomes.     

蛋白电泳凝胶放置时间对蛋白丰度的影响分析

目的利用高分辨力的双向电泳技术检测蛋白电泳后凝胶中的蛋白含量随时间推移的损失情况。方法用双向电泳蛋白裂解液提取鼠疫EV菌株(无毒疫苗株)全菌蛋白并定量。取800μg EV蛋白,平行进行两块双向电泳凝胶电泳,电泳结束后一块凝胶立刻固定并染色,另一块凝胶4℃放置8h后固定并染色,使用ImageMaster 2DElite软件比较分析两块凝胶蛋白点数量及蛋白丰度。结果相比直接固定染色的凝胶,8h后固定染色的凝胶中蛋白点扩散丢失严重,比电泳后直接固定凝胶少346个蛋白点,且部分相邻蛋白点发生融合,含量在20~350ng的低丰度蛋白65.9%因蛋白扩散丢失。结论蛋白凝胶放置8h后大量低丰度蛋白由于扩散会丢失,并直接影响后续免疫印迹、质谱鉴定等实验结果的可靠性。     

海南地区类鼻疽伯克霍尔德菌PFGE分子分型及同源性分析

目的通过对海南地区类鼻疽伯克霍尔德菌分离株进行脉冲场电泳(PFGE)分子分型分析,探讨海南地区类鼻疽伯克霍尔德菌之间的同源性,了解海南地区类鼻疽流行和分布特征。方法对海南地区2006年7月~2013年4月临床分离的97株类鼻疽伯克霍尔德菌采用PFGE方法进行分子分型,用BioNumerics软件进行聚类分析和同源性比较。结果 97株类鼻疽伯克霍尔德菌分布于海南省18个市县中的14个市县,其中东方市所占比例最高,为17.5%;沿海地区占90.7%(88/97)。感染类型以败血症为主,占61.9%。分离菌的PFGE带型呈现多态性,相似性系数为67.8%。在85%相似度水平以上时,实验菌株分为20个PFGE基因型,其中PFGE12型所占比例最高,占29.8%(29/97),且在本型中有7株菌PFGE条带100%相似,占本型菌的24.1%(7/29)。分离菌中所占比例位于第二和第三的分别是PFGE-9(占18.5%)和PFGE-16型(占14.4%)。结论 PFGE分子分型可较好地对菌株进行同源性分析。海南地区流行的类鼻疽伯克霍尔德菌PFGE型别呈多态性,但存在一些克隆群优势流行。     

Depletion and dysfunction of Vγ2Vδ2 T cells in HIV disease: mechanisms,impacts and therapeutic implications

Infection with human immunodeficiency virus (HIV) disrupts the balance among γδ T cell subsets, with increasing Vδ1+ cells and substantial depletion of circulating Vδ2+ cells. Depletion is an indirect effect of HIV in CD4-negative Vδ2 cells, but is specific for phosphoantigen-responsive subpopulations identified by the Vγ2-Jγ1.2 (also called Vγ9-JγP) T cell receptor rearrangement. The extent of cell loss and recovery is related closely to clinical status, with highest levels of functional Vδ2 cells present in virus controllers (undetectable viremia in the absence of antiretroviral therapy). We review the mechanisms and clinical consequences for Vδ2 cell depletion in HIV disease. We address the question of whether HIV-mediated Vδ2 cell depletion, despite being an indirect effect of infection, is an important part of the immune evasion strategy for this virus. The important roles for Vδ2 cells, as effectors and immune regulators, identify key mechanisms affected by HIV and show the strong relationships between Vδ2 cell loss and immunodeficiency disease. This field is moving toward immune therapies based on targeting Vδ2 cells and we now have clear goals and expectations to guide interventional clinical trials.     

我国食品来源金黄色葡萄球菌种群结构分析

目的了解我国食源性金黄色葡萄球菌(金葡菌)的种群结构。方法通过全基因组测序方法对2006-2020年我国16个省份收集的763株食源性金葡菌进行多位点序列分型、葡萄球菌蛋白A编码基因(spa)和葡萄球菌染色体mec基因盒(SCCmec)分型, 使用BioNumerics 7.5软件创建基于ST类型的最小生成树。国外进口食品分离到的金葡菌31株被纳入基因组系统发育树的构建。结果 763株金葡菌共鉴定出90个ST型和160个spa型别, 其中20种为新ST型别。72个(72/90, 80.0%)ST型属于22个克隆群, 其中主要型别为CC7、CC1、CC5、CC398、CC188、CC59、CC6、CC88、CC15和CC25, 占82.44%(629/763)。其中优势克隆群中ST型和spa型别随着时间的变化呈多态性变化。耐甲氧西林金葡菌(MRSA)的阳性率为7.60%, 共鉴定出7种SCCmec型别, 以ST59-t437-Ⅳa(17.24%, 10/58)、ST239-t030-Ⅲ(12.07%, 7/58)、ST59-t437-Ⅴb(8.62%, 5/58)、ST338-t437-...     

A bivalent outer membrane vesicle-based intranasal vaccine to prevent infection of periodontopathic bacteria

Periodontal disease has become a serious public health problem, not only causing tooth loss, but also inducing chronic disorders of extra-oral organs. The present study assessed an intranasal vaccine strategy to prevent periodontal disease using outer membrane vesicles (OMVs) of two major periodontopathic bacteria, Porphyromonas gingivalis (Pg) and Aggregatibacter actinomycetemcomitans (Aa). We compared the morphology, composition, and immune activity between OMVs of Pg strain ATCC 33277 and Aa strain Y4. Aa OMVs had a smoother surface and stronger lipid A activity compared to Pg OMVs. The in vitro immune activity elicited by Aa OMVs in macrophage-like cells was remarkably stronger than that of Pg OMVs. Intranasal immunization of mice with Aa OMVs alone resulted in robust, humoral immune responses in blood and saliva. Despites the intrinsically low mucosal immunogenicity of Pg OMVs alone, using Aa OMVs as a mucosal adjuvant strongly enhanced Pg-specific immune responses, resulting in both serum IgG and salivary IgA, both of which aggregated Pg and Aa cells. Furthermore, Aa OMVs were found to be a more potent mucosal adjuvant than Poly(I:C) in the context of enhancing the production of Pg-specific IgG (especially IgG2a) and IgA. In addition, in a randomized, blinded study, mice oral challenged with Pg and Aa after intranasal immunization with Pg OMVs and Aa OMVs had significantly decreased numbers of both microorganisms compared to mock-immunized mice. Furthermore, in an intracerebral injection mouse model, there were no serious adverse effects on the brain even after administrating a dose of OMVs as same as that used for intranasal administration. Taken together, the bivalent OMV intranasal vaccine may be effective in preventing colonization of periodontopathic bacteria in the oral cavity and related systemic disorders associated with periodontal diseases.