4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

急性重度有机磷中毒53例抢救治疗体会

目的总结有机磷中毒的抢救与临床抢救经验,提高抢救成功率。方法53例均给予洗胃、生命体征监测等治疗,5例给予呼吸支持,血液透析。结果经过治疗34例痊愈出院。结论及早洗胃,快速阿托品化,营养支持,预防并发症等是抢救成功的关键。     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

中药静脉注射联合肝炎灵穴位注射治疗慢性肝病62例观察与护理

我院2002年3月～2005年12月采用中药内服和肝炎灵穴位注射治疗慢性肝病62例,疗效较好,现报道如下： 1临床资料一。所有病例均为住院病人，随机分为治疗组和对照组。治疗组：62例，其中，男46例，女16例，年龄10～46岁，平均36岁。乙型肝炎48例，丙型肝炎10例，（乙＋丙）型肝炎4例。对照组：62例，其中男49例，女13例，年龄12～66岁，平均37岁。乙型肝炎51例，丙型肝炎9例，（乙＋丙）型肝炎2例。两组在性别、年龄、分型、肝功能损害等方面比较，差异无统计学意义，具有可比性。[第一段]     

销普钠治疗心力衰竭的观察与护理

心力衰竭是由各种原因引起的心排血量减少,不能满足组织代谢需要的一种综合征,也是常见的发病率高、预后较差的一种循环系统疾病。我院自2005年元月至2006年12月采用硝普钠治疗98例,疗效较好,报告如下。     

TaqMan-MGB探针实时荧光定量PCR检测恙虫病东方体方法的建立

目的建立敏感、特异、定量Real-time PCR方法,用于恙虫病东方体的检测。方法根据恙虫病东方体56 kD外膜蛋白基因序列设计引物和探针,建立实时荧光定量PCR检测方法。结果建立的TaqMan-MGB探针具有良好的特异性,建立的荧光定量PCR标准曲线的循环阈值(Ct)与模板拷贝数呈良好的线性关系(r=0.99),灵敏性评估发现每个20μlPCR反应管中只要有26个拷贝的目的基因即可被检测到,即最低检出浓度为2拷贝/μl,并且具有较好的重复性。结论建立的荧光定量PCR方法具有很高的特异性和敏感性,可用于恙虫病东方体感染的快速检测。     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

首次实验室证实北京平谷地区恙虫病东方体暴发流行

目的了解北京市平谷地区是否存在恙虫病东方体感染。方法采用巢式聚合酶链反应（nPCR）,对平谷地区采集的30份临床标本进行恙虫病东方体热休克蛋白（groEL）基因和56×103蛋白扩增并测序分析。采用间接免疫荧光试验（IFA）对血清标本恙虫病东方体特异抗体检测。结果采用PCR共检测血液标本30份,均阳性（100%）;用IFA检测28份患者血清抗体,25份阳性（89.3%）;长片段PCR扩增,3份标本扩增阳性,阳性结果测序比对所有核苷酸序列相同,与Kawasaki的同源性为96%。结论首次从分子流行病学和血清流行病学的角度证实北京平谷地区存在恙虫病东方体。     

TaqMan MGB 探针实时荧光定量-聚合酶链反应检测人粒细胞无形体Msp2基因方法的建立

目的 建立敏感、特异、实时荧光定量-聚合酶链反应(real-time fluorescence quantitative, FQ-PCR)方法, 用于人粒细胞无形体病的检测。 方法 根据无形体特异外膜蛋白 Msp2基因为靶基因设计引物以及TaqMan MGB探针, 建立FQ-PCR方法,并对湖北省随州和河北省张家口地区的蜱标本进行了检测。 结果 本研究建立的TaqMan MGB探针具有良好的特异性,建立的FQ-PCR标准曲线的循环阈值(Ct)与模板拷贝数呈良好的线性关系(r=0.99),灵敏性评估发现每个20 μl PCR反应管中只要有35个拷贝的目的基因即可被检测到,即最低检出浓度为2拷贝/μl,并且具有较好的重复性。共检测蜱标本426只,其中豪猪血蜱253只,共49组,阳性7份。 结论 本研究建立的FQ-PCR方法具有很高的特异性和敏感性, 可用于人粒细胞无形体感染的快速检测。进一步证实了豪猪血蜱可能是粒细胞无形体的媒介宿主。     

特殊的双硫醒反应2例

1病例介绍 例1：患者，女性，汉族，65岁。于2009年5月29日以“反复胸闷，气喘10年”为主诉住院，诊断为“支气管哮喘并发肺部感染”，治疗于静脉滴注0．9％氯化钠注射液＋注射用头孢哌酮舒巴坦钠4．0g，1日2次静点，5％葡萄糖注射液250ml＋注射用氨茶碱0．25g，1日1次静点，疗效差。于第2天改为0．9％的氯化钠注射液250ml＋注射用氢化可的松100mg，1日2次静点，患者气喘、气憋、胸闷、不适症状明显改善，但于第3天在静点注射用氢化可的松溶液10ml时患者出现面色潮红、出汗、头痛、咽痛不适。