Inactivation of Dipeptidyl Peptidase IV Attenuates the Virulence of <Emphasis Type="Italic">Streptococcus suis</Emphasis> Serotype 2 that Causes Streptococcal Toxic Shock Syndrome

Di-peptidyl peptidase IV (DPP IV), originally recognized as CD26 in eukaryotic cells, is distributed widely in microbial pathogens, including Streptococcus suis (S. suis), an emerging zoonotic agent. However, the role of DPP IV in S. suis virulence remains unclear. Here, we identified a dpp IV homologue from highly invasive isolate of S. suis 2 (SS2) causing streptococcal toxic shock syndrome (STSS). Enzymatic assays reproduced its enzymatic activity of dpp IV protein product as a functional DPP IV, and ELISA analysis demonstrated that SS2 DPP IV can interact with human fibronectin. An isogenic SS2 mutant of dpp IV, Δdpp IV, was obtained by homologous recombination. Experimental animal infection suggested that an inactivation of dpp IV attenuates greatly its high virulence of Chinese virulent strains of SS2. Functional complementation can restore this defect in SS2 pathogenicity. To our knowledge, it may confirm, for the first time, that DPP IV contributes to SS2 virulence. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Authors Junchao Ge, Youjun Feng and Hongfeng Ji contributed equally.     

外源糖原调控猪链球菌基因表达的转录组分析

【背景】碳水化合物的利用与猪链球菌在宿主体内的定殖和致病性密切相关。感染期间,宿主细胞释放的糖原可能是猪链球菌重要的碳源。【目的】从转录组学角度解析猪链球菌全基因转录水平对外源糖原诱导的响应,特别是毒力基因。【方法】将猪链球菌2型强毒株分别用糖原和葡萄糖进行液体培养,通过高通量转录组测序,比较分析糖原对猪链球菌代谢通路和毒力基因差异表达的影响,并通过体外试验和攻毒试验进行验证。【结果】猪链球菌在糖原培养基中生长良好。转录组数据显示,糖原培养条件下的猪链球菌共有908个基因差异表达,基因组占比46.07%,其中501个基因上调表达,407个基因下调表达。富集分析结果表明,糖原影响了猪链球菌广泛的基础代谢过程,但糖酵解途径保持稳定。30个毒力基因的表达水平发生变化,重要的毒力因子SLY、ApuA、ArcABC等的基因转录水平大幅度升高(倍数>20)。糖原培养后的猪链球菌的溶血活性、黏附和侵入能力显著上升,对受试动物的毒力增强,证实猪链球菌能够响应糖原诱导,糖原能调控猪链球菌的致病性。【结论】外源糖原的利用显著影响了猪链球菌的基因表达谱,这种对碳源的响应是细菌对不断变化的生存环境的适应...     

In vitro transcriptome analysis of porcine choroid plexus epithelial cells in response to Streptococcus suis: release of pro-inflammatory cytokines and chemokines

The Gram-positive zoonotic bacterium Streptococcus suis (S. suis) is responsible for a wide range of diseases including meningitis in pigs and humans. The blood-cerebrospinal fluid (CSF) barrier is constituted by the epithelial cells of the choroid plexus, which execute barrier function also after bacteria have entered the central nervous system (CNS). We show that the bacterial capsule, a major virulence factor, strongly attenuates adhesion of S. suis to the apical side of porcine choroid plexus epithelial cells (PCPEC). Oligonucleotide microarray analysis and quantitative PCR surprisingly demonstrated that adherent wild-type and capsule-deficient S. suis influenced expression of a pronounced similar pattern of genes in PCPEC. Investigation of purified capsular material provided no evidence for a significant role of the capsule. Enriched among the regulated genes were those involved in “inflammatory response”, “defense response” and “cytokine activity”. These comprised several cytokines and chemokines including the interleukins 6 and 8, which could be detected on protein level. We show that after infection with S. suis the choroid plexus contributes to the immune response by actively producing cytokines and chemokines. Other virulence factors than the bacterial capsule may be relevant in inducing a strong inflammatory response in the CNS during S. suis meningitis.     

猪链球菌分子流行病学研究方法

猪链球菌（Streptococcus suis）是猪重要的细菌性病原,它可以导致猪的脑膜炎、败血症和关节炎等症状,给养猪业带来严重经济损失;同时该菌还可感染人,是一种人畜共患病原菌。应用分子流行病学方法,阐明猪链球菌病的流行病学特征,明确其毒力分型、时空分布、传播途径、传染源,确定传播的遗传决定因素等,将有助于猪链球菌病的防控。目前常用的分子流行病学方法主要有多位点序列分型、脉冲场凝胶电泳、全基因组测序和基于PCR的方法等。本文介绍了上述方法的原理以及在猪链球菌流行病学中的应用,并分析这几种方法的优缺点,从而为更好地揭示猪链球菌流行病学特征、制定猪链球菌病的防控策略提供参考。     

布鲁氏菌入侵相关基因IalB缺失株的构建及生物学特性分析

入侵相关基因(invasion-associated locus B, ialB)的同源基因在布鲁氏菌所属的根瘤菌目中是广泛保守的,但其在布鲁氏菌中的功能研究几乎为空白。根据有限的报道资料,猜测ialB的功能可能与布鲁氏菌入侵细胞以及适应胞内环境胁迫有关。【目的】探究ialB在布鲁氏菌中的生物学功能,揭示其在布鲁氏菌黏附和入侵细胞以及胞内存活中的作用。【方法】以猪种布鲁氏菌S2株为亲本,运用同源重组的方法构建布鲁氏菌ialB缺失株ΔialB,并通过表达质粒转化的方法构建其回补株CΔialB,比较3种菌株的生长特性、对体外应激的敏感性;通过扫描电镜观察ialB缺失对布鲁氏菌形态的影响,通过实时荧光定量PCR检测3种菌株极性延长相关基因的表达;通过免疫荧光和平板计数的方法分析ialB缺失对布鲁氏菌黏附、入侵RAW264.7细胞以及胞内存活的影响。【结果】成功构建了菌株ΔialB和CΔialB;ΔialB与布鲁氏菌S2株相比,生长受限,活力降低,在酸应激、高渗应激、低渗应激、多黏菌素B应激条件下存活率降低,在氧化应激条件下存活率上升;而且,ΔialB的菌体形态发生改变,菌体变短,直径增加,极...     

Epigallocatechin gallate inhibits Streptococcus pneumoniae virulence by simultaneously targeting pneumolysin and sortase A

Streptococcus pneumoniae (pneumococcus), the causative agent of several human diseases, possesses numerous virulence factors associated with pneumococcal infection and pathogenesis. Pneumolysin (PLY), an important virulence factor, is a member of the cholesterol‐dependent cytolysin family and has cytolytic activity. Sortase A (SrtA), another crucial pneumococcal virulence determinate, contributes greatly to the anchoring of many virulence‐associated surface proteins to the cell wall. In this study, epigallocatechin gallate (EGCG), a natural compound with little known antipneumococcal activity, was shown to directly inhibit PLY‐mediated haemolysis and cytolysis by blocking the oligomerization of PLY and simultaneously reduce the peptidase activity of SrtA. The biofilm formation, production of neuraminidase A (NanA, the pneumococcal surface protein anchored by SrtA), and bacterial adhesion to human epithelial cells (Hep2) were inhibited effectively when S. pneumoniae D39 was cocultured with EGCG. The results from molecular dynamics simulations and mutational analysis confirmed the interaction of EGCG with PLY and SrtA, and EGCG binds to Glu277, Tyr358, and Arg359 in PLY and Thr169, Lys171, and Phe239 in SrtA. In vivo studies further demonstrated that EGCG protected mice against S. pneumoniae pneumonia. Our results imply that EGCG is an effective inhibitor of both PLY and SrtA and that an antivirulence strategy that directly targets PLY and SrtA using EGCG is a promising therapeutic option for S. pneumoniae pneumonia.     

猪链球菌2型硫氧还蛋白还原酶介导细菌应激与致病作用

【背景】硫氧还蛋白还原酶（thioredoxin reductase,TRR）是硫氧还蛋白系统关键组成部分,对病原菌应对体内外氧化应激、调节细菌稳态和介导致病过程具有重要作用。【目的】探究硫氧还蛋白还原酶TRR在人畜共患猪链球菌2型感染过程中参与的生物学效应。【方法】同源重组法构建猪链球菌2型硫氧还蛋白还原酶trr基因缺失株（Δtrr）及回补株（cΔtrr）,通过细菌染色、点板计数、体外细胞和动物感染模型等试验比较分析trr基因对细菌形态、抗应激反应及致病过程的影响。【结果】缺失trr对猪链球菌2型形态与生长特性的影响不大,但可增强细菌抗热应激、氧化应激和酸应激能力,缺失株对上皮细胞黏附力下降,侵袭进入脑血管内皮细胞作用显著降低,易于被吞噬细胞吞噬清除,对小鼠模型致病效应显著减弱。【结论】猪链球菌2型TRR因子参与细菌应激反应,介导细菌黏附、侵袭等致病过程,是猪链球菌2型新的潜在毒力因子。     

中国青藓科植物分类学修订(Ⅱ):燕尾藓属

基于燕尾藓属(Bryhnia Kaurin)被处理为狭义青藓属(Brachythecium Schimp.s.str.)的异名,燕尾藓属已不复存在。笔者对原分布于中国的5种燕尾藓属植物进行了相应的分类学修订,结果如下:短尖燕尾藓(B.hultenii E.B.Bartram)、燕尾藓[B.novae-angliae(Sull.Lesq.)Grout]和毛尖燕尾藓(B.trichomitria DixonThér.)被移入狭义青藓属并处理为短尖青藓[Brachythecium hultenii(E.B.Bartram)Min LiY.F.Wang]、燕尾青藓[B.novae-angliae(Sull.Lesq.)A.Jaeger]和东亚青藓[B.trichomitrium(DixonThér.)Huttunen,Ignatov,Min LiY.F.Wang];短枝燕尾藓(B.brachycladula Cardot)被移入鼠尾藓属(Myuroclada Besch.),并处理为羽枝鼠尾藓[M.longiramea(Müll.Hal.)Min Li,Y.F.Wang,IgnatovHuttunen]的异名;密枝燕尾藓[B.serricuspis(Müll Hal.)Y.F.WangR.L.Hu]移入拟异叶藓属(Pseudokindbergia Min Li,Y.F.Wang,IgnatovB.C.Tan),并处理为拟异叶藓[P.dumosa(Mitt.)Min Li,Y.F.Wang,IgnatovB.C.Tan]的异名。     

一株分离自流浪猫的猪链球菌9型的分子生物学鉴定北大核心CSCD

【目的】猪链球菌是一种能感染人和猪的人畜共患病病原,并且还可零星感染多种哺乳动物。本试验旨在调查流浪猫携带猪链球菌的情况。【方法】在流浪猫身上分离猪链球菌,经血清凝集实验和PCR检测,鉴定其血清型;经多序列位点分型分析,鉴定其ST型;将所分离的细菌与Gen Bank上已公布的猪链球菌构建16S rRNA的系统发育树,分析该菌株与其他猪链球菌的亲缘关系;药敏纸片法分析其耐药性;小鼠攻毒试验分析其毒力。【结果】本试验在流浪猫身上分离到一株猪链球菌,命名为m70,其血清型为9型。多序列位点分型显示,m70株属于一个新的ST型。与Gen Bank上已公布的猪链球菌16S rRNA进行系统发育树分析,结果显示m70属于一个单独的分支。m70与临床菌株的耐药情况相似,对四环素耐药,对红霉素中介耐药,对氨苄西林敏感。小鼠攻毒试验显示,感染10~8 CFU剂量m70的小鼠,死亡率达到60%–80%（3/5–4/5）,3次攻毒试验的平均LD（50）为5.1×107 CFU;而本实验室保存的猪链球菌强毒株HA9801感染小鼠的平均LD（50）为3.9×107 CFU,两者之间没有显著差异（P〈0.05）。【结论】从流浪猫身上分离得到的猪链球菌m70属于优势血清型,且毒力较强,提示一些流行血清型的猪链球菌强毒株具有从流浪猫传染人的潜在风险。     

水玉霉(Pilobolus)属的种的重新划分

我们在过去的工作中承认水玉霉(Pilobolus)属的9个种(郑、胡,见戴,1979)。近年来我们重新分离得到了这些分类群并对它们进行了再研究。研究结果表明,尽管它们是彼此可以互相区分的分类群,但是,包括我们过去的概念在内,目前被普遍接受的用于这个属的分类的种概念太小.为了与整个毛霉目的其它属的分类系统相一致,我们把这9个分类群重新划分为由9个变种组成的5个种:晶澈水玉霉原变种[Pilobolus crystallinu (Wigg.) Tode var. crystallinus],晶澈水玉霉透孢变种新组合[P. crystallinus var.hyalosporus (Boedijn) Hu &Zheng, comb. nov.],晶澈水玉霉克莱因变种新组合[P. crystallinus var.kleinii (van Tieghem) Zheng &G.-q. Chen, comb. nov.],豆状水玉霉原变种(P. lentiger Corda var. lentiger),豆状水玉霉小型变种新组合[P. lentiger var. minutus (Speg.) Zheng &G.-q. Chen, comb. nov.],长型水玉霉(P. longipes van Tieghem),厚壁水玉霉(P. Oedipus Mont.),露水玉霉原变种[P. roridus (Bolt.) Pers. var. roridus],露水玉霉突囊变种新组合[P. roridus var. umbonatus (Butler) Hu &Zheng, comb. nov.]。水玉霉属先后报道过的种或种下分类群名称共计50个左右,其中一些异名往往被不同的作者归到不同的正名下面。为了解决它们的正确归属问题,我们对全部原始描述作了细致的文献考证然后决定其位置。对那些找不到原始描述或从原始描述中得不出结论的则作为可疑名称处理。可疑名称共计12个:Mucor obliquus Scop., M. urceolatus Dicks.;Pilobolus urceolatus Purt., P. pestis-bovinae Hallier(=P. hallierii Rivolta), P. nanus van Tieghem, P. intermedius, (Coem.) P. A. Karsten(=P. Oedipus Mont. var,intermedius Coem.), P. pullus Massee, P.proliferens McVickar, P. ramosus McVickar, P. simplex McVickar, P. lentiger forma leinii Reyn. &Laysa, P. lentiger forma minutus Reyn. &Laysa.     

Disporum sinovietnamicum sp. nov. (Colchicaceae) from southwestern Guangxi,China

Disporum sinovietnamicum R. C. Hu & Y. Feng Huang sp. nov. is described and from southwestern Guangxi, China. It is morphologically closest to D. jinfoshanense X. Z. Li, D. M. Zhang & D. Y. Hong, but differs by having stems that are 35–90 cm tall, narrowly lanceolate and thinly leathery leaves, and ovate and glabrous tepals.     

Update on the praying mantises (Insecta: Mantodea) of South-East Vietnam

A report of Mantodea species collected in South-East Vietnam (provinces Dong Nai, Binh Phuoc and Lam Dong) is presented. New for Vietnam are the subfamily Phyllothelyinae, the tribe Anaxarchini of the subfamily Hymenopodinae and the species Leptomantella lactea (Saussure, 1870), Tropidomantis gressitti Tinkham, 1937, Amantis bolivari Giglio-Tos, 1915, Anaxarcha graminea Stål, 1877, Creobroter nebulosa Zheng, 1988, Phyllothelys breve (Wang, 1993), Ceratocrania macra Westwood, 1889, Ceratomantis saussuri Wood-Mason, 1876 and Theopompa tosta Stål, 1877, as well as five additional species whose precise identifications are less certain. Eighteen species are new for the South-East region and seven are new for the provinces sampled. The fauna of Vietnam now contains 69 species of Mantodea, 40 of which occur in the South-East region. Morphological, taxonomical and identification details for selected species are discussed and illustrated.     

云南蚊科三新种 （双翅目：蚊科）

记述了采自云南蚊科3新种,即：蓝带蚊属伪费蚊亚属一新种瞿氏蓝带蚊Uranotaenia (Pseudoficalbia) qui Dong, Dong et Zhou, sp. Nov.,采自西双版纳优乐山竹穴积水中,与新糊蓝带蚊Ur. Novobscura Barraud和二斑蓝带蚊Ur. Bimaculata Leicester近缘;领蚊属一新种类孟连领蚊Heizmannia (Heizmannia) menglianeroides Dong, Dong et Zhou, sp. Nov.,采自云南省景洪县森林竹筒积水中,与孟连领蚊Hz. Menglianensis Lu et Gong, 1986为近缘;库蚊属泰蚊亚属一新种陈氏库蚊Culex (Thaiomyia) cheniDong, Wang et Lu, sp. Nov.,采自云南西北部海拔1795 m的腾冲县竹筒积水,与海南库蚊Cu. (Th.) hainaensis Chen, 1977近缘。讨论了各新种与近缘种的鉴别特征。新种分别存放在云南省疟疾防治研究所和贵阳医学院微生物教研室。     

The SspA subtilisin-like protease of Streptococcus suis triggers a pro-inflammatory response in macrophages through a non-proteolytic mechanism

Background  

Streptococcus suis is a major swine pathogen worldwide that causes meningitis, septicemia, arthritis, and endocarditis. Using animal models, a surface-associated subtilisin-like protease (SspA) has recently been shown to be an important virulence factor for S. suis. In this study, we hypothesized that the S. suis SspA subtilisin-like protease may modulate cytokine secretion by macrophages thus contributing to the pathogenic process of meningitis.     

Improvement of arachidonic acid and eicosapentaenoic acid production by increasing the copy number of the genes encoding fatty acid desaturase and elongase into <Emphasis Type="Italic">Pichia pastoris</Emphasis>

Genes encoding Δ6 desaturase, Δ6 fatty acid elongase, and Δ5 desaturase from the alga, Phaeodactylum tricornutum, were co-expressed in Pichia pastoris to produce arachidonic acid (ARA; 20:4 Δ^{5, 8, 11, 14}) and eicosapentaenoic acid (EPA; 20:5 Δ^{5, 8, 11, 14, 17}). A panel of Pichia clones carrying progressively increasing copies of the heterologous gene expression cassette was created using an in vitro multimerization approach. ARA and EPA accumulated up to 0.3 and 0.1% of total fatty acids, respectively, in the recombinant P. pastoris carrying with double copies of these three heterologous genes, as compared to 0.1 and 0.05%, respectively, in the recombinant P. pastoris with single copy. Yun-Tao Li and Mao-Teng Li contributed equally to this work.     

RETRACTED: High-level expression and purification of recombinant huGM-CSF (9–127)/IL-6 (29–184) fusion protein in Escherichia coil

This article has been retracted at the request of the Editor-in-Chief and authors.The article was published previously in substantially identical form in another journal prior to appearing in Protein Expression and Purification. The authors deeply regret this duplication and wish to withdraw the article from publication. The article was published previously in Sheng Wu Gong Cheng Xue Bao [Refolding and purification of the huGM-CSF(9-127)-IL-6(29-184) fusion protein, Sun QM, Liu HY, Dai CB, Ma YB, Sun MS, Xu WM. Sheng Wu Gong Cheng Xue Bao (Chinese Journal of Biotechnology) 2002 May;18(3):291-294]. Chinese. PMID: 12192859.     

Characterisation of the laccase-encoding gene abr2 of the dihydroxynaphthalene-like melanin gene cluster of Aspergillus fumigatus

Aspergillus fumigatus is an important pathogen of the immunocompromised host. Previously, it was shown that the polyketide synthase encoded by the pksP (alb1) gene represents a virulence determinant. pksP is part of a gene cluster involved in dihydroxynaphthalene (DHN)-like melanin biosynthesis. Because a putative laccase-encoding gene (abr2) is also part of the cluster and a laccase was found to represent a virulence factor in Cryptococcus neoformans, here, the Abr2 laccase was characterised. Deletion of the abr2 gene changed the gray-green conidial pigment to a brown color and the ornamentation of conidia was reduced compared with wild-type conidia. In contrast to the white pksP mutant, the susceptibility of the Δabr2 mutant against reactive oxygen species (ROS) was not increased, suggesting that the intermediate of DHN-like melanin produced up to the step catalysed by Abr2 already possesses ROS scavenging activity. In an intranasal mouse infection model, the Δabr2 mutant strain showed no reduction in virulence compared with the wild type. In the Δabr2 mutant, overall laccase activity was reduced only during sporulation, but not during vegetative growth. An abr2p-lacZ gene fusion was expressed during sporulation, but not during vegetative growth confirming the pattern of laccase activity due to Abr2.     

Gac two-component system in Pseudomonas syringae pv. tabaci is required for virulence but not for hypersensitive reaction

Pseudomonas syringae pv. tabaci 6605 causes wildfire disease on host tobacco plants. To investigate the regulatory mechanism of the expression of virulence, Gac two-component system-defective mutants, ΔgacA and ΔgacS, and a double mutant, ΔgacAΔgacS, were generated. These mutants produced smaller amounts of N-acyl homoserine lactones required for quorum sensing, had lost swarming motility, and had reduced expression of virulence-related hrp genes and the algT gene required for exopolysaccharide production. The ability of the mutants to cause disease symptoms in their host tobacco plant was remarkably reduced, while they retained the ability to induce hypersensitive reaction (HR) in the nonhost plants. These results indicated that the Gac two-component system of P. syringae pv. tabaci 6605 is indispensable for virulence on the host plant, but not for HR induction in the nonhost plants. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. The nucleotide sequence data reported in this paper have been submitted to the DDBJ/GenBank/EMBL databank with the accession numbers AB266103, AB266104, AB266105, AB266106, AB266107, AB266108.     

甘南铁线莲, 甘肃毛茛科一新种

该文描述了自甘肃南部发现的毛茛科铁线莲属的一新种,甘南铁线莲。