Therapeutic effect of antimicrobial drugs against experimental infections due toYersinia pestis in mice

Although there are effective antibacterial agents against plague, newer antibacterial agents have been developed which show more potent activity against other bacterial organisms, but have not been tested againstYersinia pestis. A strain ofYersinia pestis was selected (no. 22; National Institute of Infectious Diseases, Tokyo, Japan) that caused a systemic infection in mice.Y. pestis no. 22 was intraperitoneally inoculated into DDY-strain mice, and 13 oral or 6 injectable antibacterial drugs given to the infected mice at varying doses 1 and 24 hours after infection. Levofloxacin, sparfloxacin and ofloxacin were the most effective oral agents against the infection, and prulifloxacin and pazufloxacin were also effective but to a lesser extent. Also, gentamicin and arbekacin were the most potent injectable antibacterial agents againstY. pestis. These results suggest that there are several new drugs, both oral and injectable, which exert excellent in vivo antibacterial activity against a mouse infection model and may be useful for the clinical treatment of plague.     

中国鼠疫菌种资源遗传特征分析

目的 建立《中国甲类传染病菌种资源库》，选择适当的指标，以反映中国鼠疫菌种的遗传背景。方法 使用传统的和分子生物学方法，测定中国鼠疫菌种代表菌株的遗传特征。并对遗传特征的分布及其意义进行分析。结果 利用6项指标，将中国的鼠疫菌划分成15种遗传型，每一型占据一片相对独立的而又自相延续的地理空间。分析遗传型之间的亲缘关系，并根据菌株间的差异，追溯鼠疫耶尔森菌作为一个生物种形成后的进化过程。结论 中国各鼠疫自然疫源地存在着不同特征的鼠疫菌株，但为达到对菌株认同的目标，还需要确定更多的遗传指征。     

聚合酶链反应在鼠疫菌检测中的应用 研究

本文采用聚合酶链反应（ＰＣＲ）技术，通过扩增鼠疫菌９．５ｋｂ质粒ｐｌａ基础上一个４７８ｂｌ片段，用于鼠疫菌检测，被试的４５株鼠疫菌扩增产物经琼脂糖凝胶电泳分析均呈现单一ＤＮＡ带，其分子量大小与预期结果符合。而假结核菌和一株缺６ＭＤ质粒鼠疫菌未见扩增带，洽部实验可在４ｈ内完成。本法具有快速、特异、敏感等优点，可用于不同生态型鼠疫菌的快速诊断和流行病学研究．     

tPA信号肽和鼠疫杆菌保护性抗原V融合基因的构建及免疫效果鉴定

获得含有鼠疫杆菌V抗原编码基因以及tPA信号肽编码序列的重组质粒,并测定其诱导特异性免疫应答的能力。采用PCR扩增鼠疫菌杆菌V基因构建到pVAX1质粒中产生pVAX1/V重组质粒,PCR扩增tPA信号肽编码序列片段并将其插入到pVAX1/V中V基因的上游,构建tPA-pVAX1/V重组质粒;转染COS-7细胞,免疫细胞化学方法鉴定V蛋白的表达;二重组质粒分别加mGM-CSF质粒免疫BALB/c小鼠,观察免疫应答反应;以400个LD50强毒鼠疫杆菌皮下攻击免疫小鼠观察保护效率。结果显示,tPA-pVAX1/V在COS-7细胞中表达了V蛋白;免疫小鼠血清产生了特异性抗体和细胞免疫应答;攻毒保护率达80%。成功构建了分泌型V蛋白的真核表达质粒载体,具有诱导特异性细胞免疫和体液免疫应答的能力,对强毒鼠疫杆菌攻毒有一定的保护效力,为鼠疫杆菌新型疫苗研制奠定了基础。     

口服鼠疫F1-V重组融合基因DNA活菌疫苗的构建及免疫原性研究

目的 构建携带鼠疫耶尔森菌F1-V融合基因的重组减毒沙门菌苗,口服免疫Balb/c小鼠检测其免疫原性,为口服鼠疫活载体DNA疫苗研究打下基础.方法 将F1-V融合基因克隆到真核表达载体asd-pVAX1,进一步依次将重组质粒转化减毒沙门菌X3730、X4550得到重组沙门菌X4550(asd-pVAX1/F1-V),提取重组质粒转染COS-7细胞并做免疫组化和Western-blot检测F1-V融合蛋白在细胞中的表达.以1×109CFU/只的剂量3次口服免疫Balb/c小鼠,ELISA方法检测血清中抗体水平.结果 构建的重组减毒沙门菌转染COS-7细胞后,免疫组化和Western-blot试验证明F1-V融合蛋白在细胞中得到了瞬时表达,ELISA检测到免疫小鼠血清有特异性抗体IgG产生.结论 构建的重组沙门菌能运送DNA疫苗到体内并成功释放质粒刺激机体产生特异性免疫应答,为口服鼠疫活载体DNA疫苗的黏膜免疫研究打下了基础.     

Antibody responses in arthritic and uncomplicatedYersinia enterocolitica infections

Concentrations of antiyersinia antibody isotypes IgG1, IgG2, IgG3, IgG4, IgA and IgM were measured in 33 patients with yersiniosis using a solid-phase radioimmunoassay. Sixteen patients had a complicating reactive arthritis. Throughout the observation period IgG1 and IgM antibodies both constituted approximately one-third of the total antibodies, while IgA accounted for 10%, IgG3 accounted for 1%, and IgG4 antibodies could not be detected. IgG1, IgM, and IgA antibodies (and the total titer) had reached their peak at the beginning of the observation period (ca. day 20 after the onset of symptoms). The levels then gradually decreased; the total titers averaged 40 times the background at the beginning of the observation period and 4 times the background on day 350. IgM antibodies could be detected as late as a year after the infection. The concentration of IgG2 antibodies varied greatly from patient to patient. In most patients it increased until a plateau was reached approximately 2 months after the onset of symptoms. A decline was observed later. Five arthritic but no nonarthritic patients had a pronounced IgG2 response (more than half of the IgG antibodies were IgG2 in one or several samples).     

我国5岁以下儿童小肠结肠炎耶尔森菌感染临床特征及病原学分析

目的 了解我国5岁以下儿童小肠结肠炎耶尔森菌腹泻病例临床与病原学特征,分析其可能的感染来源,为小肠结肠炎耶尔森菌病的防控与诊断提供科学依据。 方法 收集2010—2020年间来自全国10个省市自治区哨点医院儿童腹泻标本、调查及回访问卷;对标本进行小肠结肠炎耶尔森菌的分离鉴定;菌株进行生物分型、血清型鉴定;毒力基因检测以及脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)分型。 结果 2010—2020年共监测11 377例,分离到致病性小肠结肠炎耶尔森菌63株,包括61株O:3血清型、2株O:9血清型菌株,5岁以下腹泻儿童感染率0.55%(63/11 377)。不同性别儿童对致病性小肠结肠炎耶尔森菌的感染率差异无统计学意义,1～5岁感染率高于≤1岁病例(χ²=44.836,P＜0.05),感染患儿中1～5岁发热比例高于≤1岁(χ²=11.508 ,P<0.05),随访病例未发现后遗症。我国儿童感染O:3血清型致病性小肠结肠炎耶尔森菌的PFGE带型存在多样性,优势带型为K6GN11C30021、K6GN11C30012。 结论 我国5岁以下儿童感染致病性小肠结肠炎耶尔森菌的生物血清型以3/O:3为主,偶有4/O:3与2/O:9。根据患儿感染特点与高发季节推测食源为主要感染来源,需进一步调查研究。     

内蒙古自治区不同生态系鼠疫自然疫源地动物鼠疫流行现状及流行风险简析

内蒙古面临着多种类型疫源地鼠疫流行和复燃的风险。蒙古旱獭疫源地虽然暂未在国内发现动物间鼠疫流行,但过去检出过阳性血清,同时蒙古旱獭种群数量持续增加且分布范围不断扩大,毗邻的蒙古国和俄罗斯联邦的蒙古旱獭疫源地动物间鼠疫持续流行;达乌尔黄鼠疫源地动物间鼠疫时隐时现;长爪沙鼠疫源地动物间鼠疫流行猛烈,并频频波及人间。由于蒙古旱獭和达乌尔黄鼠鼠疫耶尔森菌菌株毒力明显高于长爪沙鼠鼠疫耶尔森菌菌株,且在历史上曾给当地带来深重灾难,因此内蒙古自治区鼠疫防控重点应在常规鼠疫监测的基础上,进一步加强蒙古旱獭和达乌尔黄鼠疫源地流行动态的监测,警惕该两个疫源地复燃并波及人间。