随机扩增多态性DNA分型法在铜绿假单胞菌流行病学研究方面的应用

目的摸索适于铜绿假单胞菌随机扩增多态性DNA分型法(RAPD)分型的最佳实验条件,并对我院临床分离到的34株铜绿假单胞菌进行分型研究。方法从随机引物筛选、镁离子浓度、模板量、引物浓度4方面对RAPD反应体系进行优化。结果不同病区分离到的铜绿假单胞菌的指纹图谱互不相同;从烧伤病房分离到的8株高度耐药的铜绿假单胞菌中,其中7株为同一指纹图谱,另外1株为独特的指纹图谱,而外科重症监护病房、肺科病房、干部病房同一病区内分离到的铜绿假单胞菌均为同一指纹图谱。结论铜绿假单胞菌在病区内存在交叉感染的情况,但未出现病区间感染同一流行株。     

白色假丝酵母菌的随机扩增多态性DNA分型研究

目的建立医院感染白色假丝酵母菌的基因多态性DNA分型方法,了解白色假丝酵母菌医院感染的分子流行病学情况,并探讨其基因多态性与真菌药物敏感性检测结果的可能关系。方法收集临床各病区送检的住院患者白色假丝酵母菌感染标本30份和其相应患者资料、抗真菌药物敏感性检测结果,采用经优化的随机扩增多态性(RAPD)DNA方法,对30株白假丝酵母菌进行基因多态性分型,利用Phylip3.67聚类分析软件分别用邻接法和算术平均数的非加权成组配对法,对RAPD扩增条带结果进行聚类分析。结果随机引物OPE-03进行RAPD扩增的指纹图谱清晰,带型稳定,多态性丰富,可以作为分型引物;30株不同来源的菌株可大致分为3种亲缘关系;两种聚类分析结果相同,提示该实验结果可靠。结论白色假丝酵母菌基因多态性分布可能与菌株的来源有关;RAPD多态性可能与其耐药性有关;不同聚类分析方法的RAPD图谱分析结果相同。     

肺炎克雷伯菌随机DNA多态性分型研究

目的 建立肺炎克雷伯菌随机扩增DNA多态性分型技术,以应用于临床菌株流行病学调查。方法 对临床分离的29株肺炎克雷柏菌进行Etest检测,并用酚氯仿法提取其DNA后行RAPD分型。结果 29株临床分离菌有7株ESBL阳性,24株有稳定的RAPD带型,其中ICU分离的5株ESBL阳性株的4株RAPD指纹图谱相同。结论 RAPD基因分型技术不需已知核酸序列,分型率高,分辨力强,简便快捷可为肺炎克雷伯菌株提供分型标志,是分子流行病学研究的有效方法。     

铜绿假单胞菌采用肠杆菌属重复基因间隔共有序列-PCR的分析

目的探讨铜绿假单胞菌(PAE)多重耐药情况,为临床治疗和防治医院感染提供依据。方法从重症监护病房(ICU)>60岁的患者中分离到的14株PAE,进行传统的生化、PCR方法、药敏分型和质粒、肠杆菌重复基因间隔共有序列(ERIC)-PCR多态性分析。结果传统的生化分为一个型,API 20 NE结果为1344575,药敏则呈现出多重耐药,质粒分型有10株得到约23kb的质粒,其余4株则无,质粒的有无与多重耐药无直接关系;运用ERIC-PCR技术进行基因谱系,聚类分析将株菌主要分为3大类,相似系数从0.62~0.88。结论ICU的PAE在老年患者中呈现出散发的状态,ERIC-PCR指纹图谱基因技术分型方法分辨率高,可重复性好,简便快捷,可用于PAE医院感染流行病学监测。     

RAPD技术在唐山市结核分枝杆菌指纹分型研究中的应用

[目的]RAPD DNA指纹分型方法的建立及其在分枝杆菌基因分型中的应用。[方法]收集结核病患者痰标本,分离培养获得分枝杆菌,进行表型鉴定及耐药性检测;提取基因组DNA,在建立RAPD DNA指纹分型方法的基础上对分枝杆菌进行DNA指纹图谱分析。[结果]RAPD指纹图谱分析将102株分枝杆菌分为7种菌型,其中Ⅰ型35株（34.31%）,Ⅱ型28株（27.45%）,Ⅲ型24株（23.53%）,Ⅳ型6株（5.88%）,Ⅴ型3株（2.94%）,Ⅵ型4株（3.92%）,Ⅶ型2株（1.97%）,Ⅰ、Ⅱ、Ⅲ型是主要的3种类型。RAPD扩增特征条带大小为201～1 018 bp。分组统计显示,3种主要的RAPD DNA指纹的分布与患者的居住地、职业、结核菌耐药性有关,而与患者的性别、年龄、结核接触史等无关。[结论]RAPD DNA指纹分型方法是一种简单、快速的分枝杆菌的分型方法;3种主要类型的结核分枝杆菌在唐山地区流行。     

阴沟肠杆菌随机扩增多态性DNA法基因分型

目的建立阴沟肠杆菌(ECL)随机扩增多态性DNA(RAPD)法基因分型图谱.方法以优化的随机引物和反应体系对临床分离的159株ECL进行RAPD法基因分型,并按DNA条带数及片段大小绘制指纹图谱.结果 159株临床分离ECL共得121种RAPD指纹图谱.结论 RAPD技术可将临床分离ECL分型121种.     

医院感染金黄色葡萄球菌的流行与耐药机制

目的对医院感染金黄色葡萄球菌进行基因分型,研究流行菌株的传播及其耐药的分子机制,以控制医院感染的发生。方法对分离的21株金黄色葡萄球菌进行RAPD分型,并进行mecA、gyrA、grlA基因分析,研究其耐药机制。结果21株金黄色葡萄球菌根据RAPD带型可分为3型,其中Ⅰ型12株,是医院感染的主要流行株;在临床分离的17株金黄色葡萄球菌中14株对甲氧西林和喹诺酮类均耐药;在耐药株中除1株(13064)外,其余菌株均检测出mecA基因;14株耐药株中均存在gyrA和(或)grlA突变;gyrA突变点为gyrA 84位TCA→TTA,gylA突变点为grlA80位TCC→TTC。结论RAPD基因分型技术简便快捷,可为医院感染金黄色葡萄球菌准确分型,是分子流行病学研究的有效方法;医院医院感染的金黄色葡萄球菌流行株主要为Ⅰ型,有可能经飞沫传播;17株临床分离株中14株存在多重耐药,并有相应的基因变异。     

随机引物扩增技术对变形菌DNA多态性分型研究

目的建立变形菌随机扩增DNA多态性分型技术,应用于临床菌株流行病学调查. 方法优化随机扩增DNA多态性指纹技术(RAPD)的实验条件,利用RAPD技术成功地检测21株变形菌DNA指纹图谱. 结果 21株变形菌分为18型别,其中16株奇异变形菌分为13个型,5株普通变形菌分为5个型. 结论 RAPD分型技术可简便、快速为变形菌提供分型标志,是分子流行病学研究的有效方法.     

尿源性大肠埃希菌的随机扩增DNA多态性分型及耐药性分析

目的分析尿源性大肠埃希菌随机扩增DNA多态性(RAPD)基因分型同源性及耐药性,以用于医院感染控制和流行病学调查研究。方法收集2009年7-12月从医院分离的60株尿源性大肠埃希菌(UPEC),提取DNA进行RAPD分型,并同时进行耐药性分析及流行病学调查。结果 60株尿源性大肠埃希菌分为16个RAPD型,医院感染患者分离出38株,仅R1、3、7、9、111、6 RAPD分型,而社区获得性感染患者分离出22株,却有R1～8、R10～16 RAPD分型;尿源性大肠埃希菌耐药率普遍较高,ESBLs检出率高达78.9%,门诊分离株抗菌药物敏感性普遍高于住院分离株,除磺胺甲噁唑/甲氧苄啶外差异均无统计学意义。结论尿源性大肠埃希菌耐药率较高,且本院存在RAPD R1型的局部病区流行,RAPD技术能在DNA水平上对尿源性大肠埃希菌分型,其方法快速、简易、特异、可靠,适用于该菌致医院感染的追踪及流行病学研究。     

耐甲氧西林金黄色葡萄球菌分子流行病学研究

目的 研究湖南省4所综合医院分离耐甲氧西林金黄色葡萄球菌(MRSA)分子流行病学特点,为临床预防、临测MRSA的流行,并制定合理的防治方案提供确实可靠的依据.方法 采用质粒图谱分析4所医院MRSA是否存在质粒流行,并分析其型别;采用RAPD分析对住院患者、环境、医务人员分离的MRSA进行同源性分析.结果 对89株MRSA进行质粒图谱分析,有7株未检测到质粒,82株携带有质粒,可分为17型,4所医院MRSA均存在1～2种主要质粒的流行;经RAPD分型89株MRSA可分为A、B、C、D、E 5型,其中A型85株占所有菌株的95.50%,B型1株、C型1株、D型1株、E型1株.结论 质粒图谱分析及RAPD分型方法 可很好用于MRSA分子流行病学分析,医院MRSA临床株与环境株及湖南省4所综合医院之间MRSA 菌株具有同源性,呈暴发流行.     

Calcium intake in the elderly: validation of a dietary questionnaire

Epidemiological studies of calcium and osteoporosis have been hampered by the lack of a suitable tool for assessing calcium intake. This report describes a new frequency and amount questionnaire for measuring present and past calcium intake in the elderly. The validity of the questionnaire was tested against two commonly used standards of dietary assessment, five-day duplicate diets and seven-day weighed dietary inventories. The resulting correlation coefficients were, respectively, r = 0.76 and r = 0.69, while that for repeatability was r = 0.84. Furthermore, the questionnaire categorized subjects into thirds of the distribution of intake with almost no gross misclassification. It is suggested that the present findings may be extended to the majority of normal, healthy elderly subjects, implying wide application for the questionnaire in the assessment of calcium intake in the elderly.     

Self-reported health problems among Swedish miners one year after unemployment

Unemployment is considered to be a public health concern sincedeterioration in the health of the unemployed is often anticipated.However, for some groups, such as miners, unemployment mightimprove health due to a cessation of potentially harmful occupationalexposures. This study evaluates the health of 79 miners in oneSwedish iron-ore mine, and 226 age-matched controls from thegeneral population, during one year after the closure of themine. The participants received a questionnaire regarding medicalhistory and subjective symptoms at the beginning of the studyperiod, and after one year. Statistically significant negativeeffects on self-reported health attributable to unemploymentwere not found, although neuropsychiatric symptoms were morecommon among the unemployed miners. The miners reported a statisticallysignificant improvement in grip force (p=0.031). They had asignificantly higher prevalence of symptoms associated withmining related exposures when compared with the population controls;pain in the upper extremities [relative risk (RR)=2.27, 95%confidence interval (Cl)=1.44–3.59), back pain (RR=1.84;Cl=1.237–2.75), vasospastic disease of the fingers (RR=2.05;Cl=1.18–3.57) and obstructive respiratory symptoms (attacksof dyspnea and wheezing: RR=3.67; Cl=1.167–11.6).     

Excès de cas humains et animaux de tularémie en France au cours de l’hiver 2007–2008 : émergence ou phénomène isolé ?

Context

Tularemia is a zoonosis affecting humans and hares in France. We describe the results of surveillance in both species, in 2007 and 2008.

Methods

Human tularemia cases are mandatorily notifiable in France since 2003. In hares, surveillance relies on volunteer hunter associations in all districts of the country. Data from mandatory reports and volunteer surveillance in 2007/2008 were analyzed and compared with previous results.

Results

In 2007/2008, 144 cases were reported in humans and 117 cases in hares. This was a 100% increase compared to previous years. Human cases differed from those of previous years only by the frequency of contact with breeding animals. Human cases without any documented risk exposure were also more frequent.

Conclusion

An increase of tularemia cases occurred in 2007/2008 in both species. Complementary studies are needed to identify the species reservoir in France to understand the causes of this peak of cases.     

Occupational health hazards in mining: an overview

This review article outlines the physical, chemical, biological, ergonomic and psychosocial occupational health hazards of mining and associated metallurgical processes. Mining remains an important industrial sector in many parts of the world and although substantial progress has been made in the control of occupational health hazards, there remains room for further risk reduction. This applies particularly to traumatic injury hazards, ergonomic hazards and noise. Vigilance is also required to ensure exposures to coal dust and crystalline silica remain effectively controlled.     

Manufacture and use of nanomaterials: current status in the UK and global trends

This paper provides an overview of the production and use of nanomaterials (NMs), particularly in the UK. Currently, relatively few companies in the UK are identifiable as NM manufacturers, the main emphasis being the bulk markets in metals and metal oxides, and some niche markets such as carbon nanotubes and quantum dots. NM manufacturing in the UK does not reflect the global emphasis on fullerenes, nanotubes and fibres. Some assumptions have been made about the types of NM that are likely to be imported into the UK, which currently include fullerenes, modified fullerenes and other carbon-based NMs including nanotubes. Many university departments, spin-offs and private companies have developed processes for the manufacture of NMs but may only be producing small quantities for research and development (R&D) purposes. However, some have the potential to scale up to produce large quantities. The nanotechnology industry in the UK has strong R&D backup from universities and related institutions. This review has covered R&D trends at such institutions, and appropriate information has been added to a searchable database. While several companies are including NMs in their products, only a few (e.g. manufacturers of paints, coatings, cosmetics, catalysts, polymer composites) are using nanoparticles (NPs) in any significant quantities. However, this situation is likely to change rapidly. There is a need to collect more information about exposure to NPs in both manufacturing and user scenarios. As the market grows, and as manufacturers switch from the micro- to the nanoscale, the potential for exposure will increase. More research is required to quantify any risks to workers and consumers.     

深圳公立医院管理体制改革制度设计

深圳公立医院管理体制改革实行政事分开、管办分开,在理事会架构下按法定机构模式组建市医管中心,落实公立医院运营管理自主权.作者从当前公立医院管理体制的弊端入手,介绍了深圳市进行公立医院管理体制改革的基本思路及改革方案设计的主要举措,深入剖析了的改革方案的特点,并对改革效果进行了预测.     

Effects of nitrogen dioxide on red blood cells of rats: Changes in components of red cell membranes during in vivo exposure to NO2

Red cell membranes, prepared from red blood cells of rats exposed to 4, 10, or 20 ppm nitrogen dioxide (NO₂) for 1 to 10 days, were examined for evidence of changes in membrane components. Appreciable changes were not found in contents of phospholipid and cholesterol during exposure to 10 ppm NO₂. By contrast, protein content altered with the time of exposure. Moreover, changes in protein composition were observed by employing sodium dodecyl sulfate — polyacrylamide gel electrophoresis. Twenty-four-hour exposure to NO₂ at the concentration above 10 ppm resulted in a marked increase in the percentage of lysophosphatidylethanolamine (LysoPE) to the total phospholipids. The prolonged exposure to 10 ppm NO₂ gave rise to a further increase in LysoPE, whereas the percentage of phosphatidylethanolamine (PE) showed a gradual decrease. A 1-day exposure to 4.0 ppm NO₂ also caused an increase in sialic acid content and decreases in those of PE and hexose. In addition to contents of these components the percentage of LysoPE increased 5 days after exposure and the elevated values were maintained up to the end of exposure period. These results demonstrate that red blood cells in circulation exhibit different membrane properties in terms of lipid and carbohydrate composition during 10 days of exposure to 4.0 ppm NO₂.     

Investigating clusters in the workplace and beyond

Clusters of disease are common and occur in the workplace and in the general community. They often arouse considerable concern among the population. Investigations have sometimes lead to exciting new knowledge, but in general the investigation of clusters is difficult and often unrewarding, especially for community clusters. In the workplace, investigations are more likely to find associations and even new causes, but still many clusters remain enigmatic. Despite this, there are many reasons for investigating clusters, including allaying community concern and identifying uncontrolled exposures. A structure for investigating clusters in the workplace is suggested.