霍乱弧菌多重PCR检测方法的建立和运用

[目的]建立一种快速检测霍乱弧菌O1群、O139群、非O1非O139群及是否产毒株的方法。[方法]以霍乱弧菌的外膜蛋白基因（ompW）、编码O1群O抗原基因（O1rfb）和编码O139群O抗原基因（O139 rfb）、霍乱肠毒素A亚单位基因（ctxA）为目的基因,建立一种多重PCR方法对霍乱弧菌进行检测和分型。[结果]根据相关基因携带情况分为6种型别,并分产毒株和非产毒株。以该法对140株霍乱弧菌分型结果与血清学诊断结果相一致,而其它弧菌和肠杆菌等均未检出相关基因。[结论]该方法快速、特异,可应用于霍乱弧菌的检测和分型。     

霍乱弧菌多重PCR检测方法的建立及优化

[目的]建立一种快速检测样品中霍乱弧菌的多重PCR方法,优化霍乱弧菌多重PCR的反应体系和循环参数.[方法]根据霍乱毒素ctxA、toxR、O139群特异性基因、O1群特异性基因序列,设计特异性引物,对引物浓度、PCR缓冲液的浓度、Mg2+浓度、退火温度及其延伸时闻等条件进行优化.[结果]4对引物在不同血清型霍乱弧菌DNA模板中能特异扩增出不同的条带.在同一反应体系中,较低的延伸温度可以提高霍乱弧菌多重PCR检测的灵敏度.[结论]通过对霍乱毒素多重PCR反应体系不同因素的优化,建立了稳定可靠的霍乱弧菌多重PCR检测平台.     

多重PCR快速检测金黄色葡萄球菌四型肠毒素基因的研究

[目的]建立一种快速、高效地检测金黄色葡萄球菌SEA、SEB、SEC和SED基因的多重PCR法并进行优化. [方法]试剂盒提取产肠毒素金黄色葡萄球菌基因组DNA作为模板,根据SEA、SEB、SEC和SED肠毒素基因片段保守区序列设计引物,多重PCR扩增产肠毒素金黄色葡萄球菌基因组中特异靶序列.对反应体系Mg2+浓度、引物浓度和退火温度进行优化,并初步检测方法特异性. [结果]在PCR反应体系中加入设计的4对引物可同时扩增出金黄色葡萄球菌四型肠毒素基因中的特异序列;在总体积为25 μl的反应体系中,最优Mg2+浓度为2.0 mmol/L,引物浓度分别为0.2 μmol/L,退火温度为49℃;以大肠杆菌O157:H7和肠炎沙门菌510041基因组DNA为模板扩增不出特异片段. [结论]成功建立金黄色葡萄球菌四型肠毒素基因多重PCR法,该法能快速、高效地同时检测SEA、SEB、SEC和SED基因,比传统PCR方法省时省力,可以弥补传统PCR法的不足.     

食源性致病菌多重聚合酶链反应快速检测

[目的]建立快速检测食源性致病菌的多重聚合酶链反应(PCR)方法.[方法]以出血性大肠杆菌的编码大肠杆菌不耐热毒素的B亚单位(LTB)基因、福氏志贺菌侵袭性质粒抗原H基因(ipaH)、霍乱弧菌肠毒素细胞外分泌蛋白(EPSM)基因作为目的基因,分别设计一对引物,优化各种工作条件,建立一种多重PCR诊断方法.[结果]应用所建立的多重PCR方法能快速、特异地检测出出血性大肠杆菌194 bp、福氏志贺菌606 bp、霍乱强菌482 bp的目的基因.[结论]该检测方法简单、快速、准确、灵敏度高,可广泛应用于食品卫生检测和临床检验等领域.     

多重聚合酶链反应方法在结核杆菌检测中的应用

[目的]建立结核病诊断及快速检测方法。[方法]将结核杆菌标准菌株、临床病例标本、对照标本采用多重PCR体系进行检测，以研究多重PCR方法对上述菌株检测的敏感性和特异性。[结果]多重PCR体系检测结核杆菌DNA的灵敏性最低为30～210个结核杆菌，对结核病病例和对照标本检测的灵敏度为94．6％，特异度为100％。[结论]多重PCR技术用于结核分支杆菌的鉴定简便、快速、特异，适于在卫生检疫系统和临床实验室应用。     

巢式聚合酶链反应对霍乱弧菌肠毒素快速检测方法的建立

[目的]　建立霍乱弧菌肠毒素快速检测的检验技术。[方法]　运用巢式聚合酶链反应(NESTED-PCR)检测霍乱弧菌肠毒素基因CTX。[结果]　检测霍乱弧菌特异性强,检测下限达4cfu/mL(100cfu/mL级水平)。[结论]　NESTED-PCR检定霍乱弧菌简便、快速、敏感度高且准确性好。     

外膜蛋白（ompw）引物扩增技术在非01群霍乱检测中的应用

[目的]采用霍乱弧菌外膜蛋白合成一对引物，建立一种灵敏、特异的PCR方法检测非01群霍乱弧菌。[方法]在实验室对42株来自外环境和病人中分离出的非01群霍乱弧菌采用PCR技术作了效果观察，并以大肠、痢疾、沙门菌等作为阴性对照其敏感性和特异性进行了验证。[结果]42株非01群霍乱弧菌采用外膜蛋白引物行PCR扩增，阳性率为100％，扩增片段为588bp，阴性对照扩增均为阴性。[结论]霍乱弧菌外膜蛋白引物快速检测非01群霍乱弧菌提供了一种有用的检测手段。     

水产品中霍乱弧菌的监测方法研究

目的:研究水产品中霍乱弧菌的监测方法,及时发现被霍乱弧菌污染的水产品,采取有效防控措施,防范霍乱疫情的发生。方法:用实时荧光PCR法、胶体金法、分离培养法三种方法分别对180份水产品进行霍乱弧菌检测,阳性样品再进行霍乱肠毒素检测。结果:三种方法检出了44份核酸阳性标本,18份霍乱肠毒素阳性标本,分离出8株霍乱弧菌。结论:在进行水产品霍乱监测时,可以先用实时荧光PCR进行初筛,筛出核酸阳性标本,进行霍乱肠毒素检测,及时发现被霍乱产毒株污染的水产品,再对筛出的核酸阳性标本结合胶体金法进行分离培养和分型鉴定,进一步完成霍乱疫情的确证和分析。     

多重PCR快速检测5种重要致病性弧菌

目的建立一种快速检测霍乱弧菌、副溶血性弧菌、创伤弧菌、拟态弧菌和溶藻弧菌5种重要致病性弧菌的多重PCR方法。方法以霍乱弧菌omp W基因、副溶血性弧菌toxR基因、创伤弧菌vvh A基因、拟态弧菌VMH基因和溶藻弧菌gyr B基因为靶基因设计特异性引物,优化建立多重PCR反应体系,系统评价其特异性和检测下限值。结果成功构建致病性弧菌多重PCR检测方法。评价结果显示其特异性为100%,霍乱弧菌、副溶血性弧菌和拟态弧菌的检测下限值为100 CFU/ml,创伤弧菌和溶藻弧菌的检测下限值为1 000 CFU/ml;多重PCR的弧菌检出率明显高于传统分离培养方法(56%vs 17%)。结论该多重PCR方法快速、灵敏、特异性好,可用于海环境和水产品中致病性弧菌的监测。     

非O1非O139群霍乱弧菌多重PCR和SYBR Green实时PCR检测方法的建立

目的建立检测非Ol非0139群霍乱弧菌的多重PCR和SYBRGreen实时PCR方法。方法分别以霍乱弧菌的外膜蛋白基因(ompW)、01和0139群O抗原编码基因(咖)设计引物,建立多重PCR和SYBRGreen实时PCR方法,评价两种方法检测非01非0139群霍乱弧菌的特异性、重复性和一致性及最低检测菌量。结果建立了检测非01非0139群霍乱弧菌的多重PCR和SYBRGreen实时PCR方法,根据特异性条带(多重PCR)和特异性熔解温度(SYBRGreen实时PCR),两种方法均能特异性检测非01非0139群霍乱弧菌,并能鉴别其他弧菌(5种)和肠道杆菌(3种);两种方法的最低检测菌量分别为7×10⁴ cfu／ml(多重PCR)和7×10²cftdml(SYBR Green实时PCR),差异有统计学意义(P<0.05);对实时PCR的重复性检测,批内变异系数(CV)为 0.22％～0.92％,批间cv为0.27％～1.41％;经370株非01非0139群霍乱弧菌的检测,两种方法的一致性均为100％。结论建立的两种方法其特异性、敏感性及重复性均好,可适用于不同条件下非0l非0139群霍乱弧菌的检测和鉴定。     

长春新碱过量引起严重毒副反应1例的护理体会

报道1例霍奇金淋巴瘤患者因误用长春新碱（VCR）10mg一次性静脉推注后治疗护理情况。其出现间断性神志恍惚、眼睑闭合不全、言语不清、口腔黏膜糜烂、全身疼痛、麻痹性肠梗阻、尿潴留、手足麻木等症状,经积极解救,禁食,持续胃肠减压、胃管内注入麻油、开塞露、生理盐水灌肠,合理应用肠外营养,注重疼痛、心理护理,做好口腔、肛周护理,预防感染加重,患者病情得到控制好转出院。     

Correlates of stages of change of smoking among inhabitants of deprived neighbourhoods

BACKGROUND: This study examines the prevalence and correlates of stages of change of smoking, in terms of psychosocial, structural and sociodemographic factors, among inhabitants of deprived neighbourhoods. METHODS: Cross-sectional data were obtained from a survey on health related behaviour. Subjects were 2009 current and former smokers, aged 20-46, living in deprived neighbourhoods in Rotterdam, the second largest city in the Netherlands. Three groups of smokers were formed according to the stages of change-definitions of the Transtheoretical Model: smokers not planning to quit (precontemplators), smokers planning to quit (contemplators/preparators) and former smokers (actors/maintainers). Smokers planning to quit and smokers not planning to quit were compared regarding psychosocial factors (attitude, social norm, self-efficacy), structural factors (neighbourhood problems, material deprivation, financial problems, employment status) and sociodemographic factors (age, gender, marital status, cultural background, educational level). Former smokers were compared with smokers planning to quit regarding structural and sociodemographic factors. Logistic regression was used to assess correlates of stages of change. RESULTS: Smokers planning to quit (prevalence = 19%) reported a more positive attitude, stronger social norms and higher self-efficacy expectations in quitting smoking than smokers not planning to quit (prevalence = 57%). Smokers planning to quit less often were Dutch-born, more often had attended higher vocational schooling or university and more often reported experiencing two or more neighbourhood problems compared to smokers not planning to quit. Former smokers (prevalence = 24%) were older, more often Dutch-born, married, employed and higher educated, compared to smokers planning to quit. Furthermore, former smokers less often reported material deprivation and financial problems than smokers planning to quit. CONCLUSION: Among people living in deprived neighbourhoods, different factors correlate with different stages of change of smoking. Implications for health promotion are discussed.     

Choice of measures of vaccination and estimates of risk of pediatric pertussis

BackgroundVaccination uptake at the individual level can be assessed in a variety of ways, including traditional measures of being up-to-date (UTD), measures of UTD that consider dose timing, like age-appropriate vaccination, and risk reduction from individual doses. This analysis compared methods of operationalizing vaccination uptake and corresponding risk of pertussis infection.MethodsCity-wide case-control study of children in Philadelphia aged 3 months through 6 years, between 2001 and 2013. Multiple logistic regression was used to isolate the independent effects of each measure of vaccination uptake and the corresponding relative odds of pertussis.ResultsBeing UTD on vaccinations was associated with a 52% reduction in risk of pertussis (OR 0.48, 95% CI: 0.34, 0.69). Evaluation of delayed receipt of vaccine versus on-time UTD yielded similar results. There was a decrease in risk of pertussis for each additional dose received with the greatest reduction in pertussis infection observed from the first (OR 0.48, 95% CI: 0.28, 0.83) and second dose (OR 0.17, 95% CI: 0.08, 0.34). Additional doses conferred minimal additional protection in this age group.ConclusionExamining vaccination status by individual doses may offer improved predictive capacity for identifying children at risk for pertussis infection compared to the traditional UTD measure.     

上海市某医院骨科平均住院日影响因素分析

对上海市某医院2003年-2007年骨科出院病人的住院日描述性分析．2003年-2007年骨科的床位利用指数与平均住院日相关性分析．2003年-2007年骨科床位与医护比例分析．2007年骨科前10大病种平均住院目影响因素分别进行单因素相关性分析和多因素逐步回归分析（STATA软件）。通过对骨科10大病种住院日影响因素分析,术前等待天数、手术类型、是否输血分别对10个、9个和8个病种的住院目有影响。输血因素和手术类型是医院不可控、由病人的病情决定的,术前等待天数是管理因素,是最值得医院重视的影响因素。     

两种不同肌瘤剔除术治疗子宫肌瘤的临床疗效比较

目的:比较腹腔镜子宫肌瘤剔除术与腹式子宫肌瘤剔除术的临床疗效。方法:回顾分析63例腹腔镜子宫肌瘤剔除术与61例腹式子宫肌瘤剔除术的临床资料,比较2种手术方式的手术时间、术中出血量、肛门排气时间、使用抗生素时间、术后感染率、体温恢复正常时间、住院时间等情况。结果:两组患者接受子宫肌瘤剔除术顺利完成,腹腔镜组手术时间长于开腹组(P0.05),腹腔镜组术中出血量及剔除肌瘤个数均少于开腹组(P0.05),术后腹腔镜组体温恢复正常时间、肛门排气时间、抗生素使用及住院时间均短于开腹组,两组患者最多随访6个月,腹腔镜组平均康复时间较开腹组短(P0.05),两组随访B超均未发现明显肌瘤结节。结论:腹腔镜子宫肌瘤剔除术损伤小,恢复快,住院时间短,是值得推广的微创手术。     

尘肺病患者145例死因调查

目的分析汉中市尘肺病的发生、发展和死亡原因，以提高尘肺病患者的生存质量。方法对145例尘肺病患者的死因进行回顾性分析，计算其平均发病年龄、死亡年龄及患尘肺病后的存活年限。结果矽肺的平均发病年龄和死亡年龄均早于煤工尘肺和石棉肺，尘肺病的主要死因为患肺部疾患，患尘肺病后的平均存活年限为12a。结论加强职业健康体检工作，对尘肺病人做到早发现早治疗和预防并发症的发生。     

性发育异常发病机制的研究进展

正常的性腺分化可分为3个过程：原始性腺形成、性别决定和卵巢/睾丸发育。在任一环节中,基因表达或调控发生异常均有可能导致性发育异常疾病的发生。性发育异常是指染色体、性腺和解剖性别不典型。随着分子生物学技术的发展,不断地发现新的基因或信号通路参与性腺分化和发育,如SRY、SF1、WT1、Sox9等基因与睾丸发育密切相关,Wnt/Rspo1/B连环蛋白通路、Dax1、Foxl2等基因在卵巢分化中发挥着重要作用,一些非编码RNA和转化生长因子也有重要的调节功能,且睾丸和卵巢发育均为主动过程,即使在出生后因某些基因的改变两者间也可出现横向分化。这些为揭示性分化异常的发病机制提供了可能。     

西林瓶装注射用水的装量检查分析

为了检查西林瓶装注射用水的实际分装量是否符合2005年版《药典》三部要求,按照2005年版《药典》三部中“生物制品分装和冻干规程”,分装规格为0．8ml的西林瓶装注射用水10000瓶,随机在分装过程的前、中、后阶段共抽取100瓶,轧盖、目检。分别用1ml标定注射器及1ml普通无菌注射器抽取每瓶注射用水的实际体积,以t检验法对实际抽取量与标示量的差值进行统计学分析,结果显示其与2005年版《药典》三部要求分装附加量为0．1ml相比有显著性差异（P〈0．001）。     

One-year stability of the Measure of Processes of Care

BACKGROUND: The Measure of Processes of Care (MPOC) is a 56-item self-administered measure designed to examine what parents of a child with a chronic health problem think of the services they and their child receive, and to measure the extent to which these services are family-centred. Reliability and validity of the MPOC were established in prior studies. The aim of the present study was to assess the 1-year stability of the MPOC to justify its use as an evaluative tool. METHODS: Nine paediatric rehabilitation centres in the Netherlands participated in this short longitudinal survey study. Subjects were 205 parents (response rate 74.8%) of children aged 1-18 years who received care in one of the participating paediatric rehabilitation centres. All subjects filled out two MPOCs with a 1-year interval. RESULTS: All correlations between the scale scores of the MPOC at the first and second administration were relatively high and significant (range: 0.443-0.609, all P < 0.001), demonstrating high inter-individual stability. However, all mean scale scores, except for Providing General Information, significantly reduced after 1 year. CONCLUSIONS: The MPOC has a moderate 1-year stability. However, because of its tendency to score lower when repeated after 1 year, its use as an evaluative follow-up instrument to assess the effectiveness of a programme intervention is restricted.     

儿童皮质发育畸形的MRI诊断

目的探讨儿童皮质发育畸形(MCD)的MRI表现特点,为MCD的早期诊断提供依据。 方法选择2010年1月至2012年12月在四川省医学科学院·四川省人民医院放射科经MRI检查确诊为MCD的44例患儿为研究对象。回顾性分析及总结其MRI影像学特征。本研究遵循的程序符合四川省医学科学院·四川省人民医院人体试验委员会制定的伦理学标准,得到该委员会批准,征得受试对象及其监护人知情同意,并与监护人签署临床研究知情同意书。 结果本组44例经MRI确诊为MCD患儿中,巨脑回-无脑回为17例,多小脑回为14例,灰质异位为9例,脑裂畸形为4例。MCD的MRI特点中,巨脑回表现为脑回增宽、皮质增厚及白质变薄;无脑回表现为正常脑沟、脑回消失,皮质增厚及白质变薄;多小脑回表现为脑回增多、细小;灰质异位表现为皮质下及侧脑室周围深部白质区结节状、团块状或带状病灶,信号与灰质信号相同;脑裂畸形表现为从侧脑室到脑表面贯穿大脑半球的裂隙,表面覆盖灰质。 结论MRI是诊断儿童MCD的可靠影像学方法。