nef RNA在人类免疫缺陷病毒早期感染检测中的应用

目的 为进行人类免疫缺陷病毒(HIV)早期感染的检测，预防医院感染。方法 以nef基因引物nef6，nef7和β-肌动蛋白(actin)基因引物BAl，BA4在同一反应体系中对标本进行逆转录聚合酶链反应，用Kodak Digital Science lD软件对条带密度进行分析，并对PCR产物进行Southern印迹杂交，以确定PCR反应所得条带为nef基因扩增产物。结果 所检测的6例HIV—l抗体阴性P24抗原阳性患者标本均扩增出HIV—lnef和β-肌动蛋白的基因产物，通过Kodak Digital Science 1D软件分析得到各标本条带的密度比值，Southern印迹杂交证明PCR反应条带确实为HIV—lnef基因扩增片段。结论 该方法简便、经济，可作为HIV早期感染检测的一种辅助手段，防止医院内HIV的感染。     

乙型肝炎病毒B基因亚型(Ba,Bj)半巢式PCR检测方法的建立

目的 建立一种灵敏特异且简便易行的乙型肝炎病毒(HBV)B基因Ba和Bj亚型分型方法.方法 用DNAStar软件比较分析GenBank中登录的B基因型HBV(HBV/B)和C基因型HBV(HBV/C)的基因序列,分别设计出HBV/B型特异性引物HB及Ba和Bj基因亚型特异性引物BA和BJ.第一轮PCR反应以HBV/B型特异性引物HB作为上游引物,以日本学者Sugauchi等设计的引物HBAS-4V作为下游引物(外引物);第二轮PCR反应同样以HB作为上游引物,亚型特异性引物BA和BJ作为下游引物(内引物),在同一个反应管中进行.根据PCR产物片段的大小判定B基因亚型.随机选取实验室内经型特异性引物PCR法鉴定为HBV/B(56份)及经preS/S区序列测定证实为HBV/C(15份)的HBV慢性感染者血清标本共71份,以研究设计的引物进行半巢式PCR反应,检测Ba和Bj基因亚型,并以载有HBV Bj亚型基因组的质粒作为Bj亚型的阳性对照.然后随机选取15份B型样本的第一轮PCR产物直接测序,利用Blast和DNAStar软件将测序结果与GenBank中登录的Ba及Bj亚型序列进行同源性分析,验证该半巢式PCR法的准确性.结果 56份HBV/B血清标本经该半巢式PCR法检测,均为Ba亚型,随机选取的15份B型PCR产物直接测序,结果均为Ba亚型,与研究中应用的半巢式PCR方法检测结果一致.15份HBV/C的血清标本检测结果均为阴性.结论 建立了HBV Ba和Bj亚型半巢式PCR法.该方法灵敏、特异、简便、易行,可用于临床和流行病学大样本检测.     

HIV流行毒株的DNA macroarray基因分型技术的研究

目的：建立一种特异、相对简便的HIV流行毒株基因分型新技术。方法：从HIV-1抗体阳性标本中提取HIV前病毒DNA。使用HIV-1型共用引物进行PCR扩增。同时设计一条型特异性的探针，然后通过制作DNA macroarray芯片，鉴别样本的HIV亚型。结果：DNAmacroarray分析可以检测我国HIV主要流行株基因型的特异性序列：B、C、CRF01-AE和CRF07-BC。结论：DNA macroarray方法对HIV亚型进行分析是可行的，可以在HIV流行病学调查中应用。     

HIV抗体免疫印迹试验不确定结果的特征及鉴别方法研究

目的 研究HIV抗体确认不确定结果的血清学特征并比较不同实验方法鉴别不确定结果的效果.方法 以解放军艾滋病检测确认实验室2005-2006年确认的42例HIV抗体不确定者为研究对象,对血清样本进行HIV抗体条带免疫印迹检测、HIV病毒载量检测和HIV-1 p24抗原检测,同时进行随访,将随访前后的标本同时检测,比较免疫印迹带型的进展,判断HIV感染的真实状况,以随访的结果为标准,判断不同的方法鉴别不确定结果的效果.结果 (1)42例HIV抗体不确定病例中,共有8种带型:p24(45.2%)、gpl60(30.9%)、gp160±p24(11.9%).(2)对23例进行3个月以上随访,其中22例带型没有变化,判断为HIV抗体阴性;1例的带型在随访后出现显著进展,满足HIV抗体阳性诊断标准;随访证实,22例(95.6%)的不确定是非特异反应,只有1例是早期HIV感染.(3)对23例通过随访确定HIV感染状态的病例,条带免疫印迹检测1例血清学阳转的病例为阳性,22例血清学没有进展的病例中16例为阴性,6例仍为不确定,特异性是72.7%.HIV-1 p24抗原检测23份标本全部为阴性,1例血清学阳转标本漏检.检测7份血浆标本的病毒载量,1例血清学阳转的病例病毒载量为18 000 cp/ml,而6例血清学没有进展的病例检测结果全部低于最低检测限(＜LDL).结论 条带免疫印迹试验能够鉴别大约70.0%的不确定反应,病毒载量检测也是鉴别不确定结果的有效方法;使用这两种方法,可以对HIV抗体不确定进行早期鉴别诊断.     

应用ELISA检测戒毒人员尿液和血液标本HIV-1抗体结果分析

目的 比较应用ELISA（酶联免疫吸附试验）检测尿液和血液标本HIV－1抗体的一致性。方法 对广州市某强制戒毒所的203例曾吸毒人员分别采集尿液和血液标本，应用ELISA（酶联免疫吸附试验）分别采用尿液和血液初筛试验检测尿液和血液标本中的HIV－1抗体。结果 30例中有20例血液标本的HIV－1抗体为阳性，其平行尿液标本中有19例HIV－1抗体为阳性。分析结果表明应用尿液，血液标本检测HIV－1抗体方法的一致性为99．5％。结论 尿液标本HIV－1抗体的ELISA检测结果是可靠的，对采集血液标本有困难的监测人群具有实用价值。     

军团菌D-RAPD基因分型研究

〔目的〕建立军团菌的基因分型方法。〔方法〕采用D—RAPD技术 ,应用 6条简并随机引物对 5株军团菌 (包括 3株嗜肺军团菌 )进行基因分型研究。〔结果〕除引物 4 ,其余 5个随机引物均能对军团菌菌株扩增出特异性的DNA条带 ,其中引物 2的分型效果较好。用引物 2、引物 3两引物 ,引物 2、引物 5两引物 ,引物 3、引物 5两引物 ,分别对 5株军团菌DNA进行扩增 ,结果 3对引物在扩增条带的数量和长度上均能有效区分 5株军团菌。〔结论〕D—RAPD技术快速、简便 ,是在分子水平对致病微生物进行检测、发病机理分析、流行病学研究的理想方法。     

RT-PCR快速检测腮腺炎病毒

目的：建立腮腺炎病毒的核酸快速诊断技术。方法：在腮腺炎病毒核蛋白基因（NP）的保守区域设计一对半引物，采用逆转录聚合酶链（RT-PCR）和半巢式PCR方法扩增腮腺炎病毒特异性核酸片段。结果：该方法能特异性地扩增腮腺炎病毒357bp和232bp的核酸片段，而对麻疹、风疹和流感病毒无交叉反应。检测腮腺炎病毒的灵敏度，1次PCR可检测出10CCID50，2次PCR反应可达0.1CCID50。采用该方法从流行性腮腺炎患含漱液标本中能直接检出腮腺炎病毒核酸条带。结论：该方法能特异、敏感地检测临床标本中腮腺炎病毒，是快速诊断腮腺炎病毒感染的理想方法。     

乙酸镍诱导16HBE细胞恶变过程中的K-ras和P15基因的改变及基因组不稳定性分析

目的：检测乙酸镍对K—ras基因和P15基因的改变及基因组不稳定性的影响，从而进一步探讨镍化合物致癌的分子机制。方法：采用聚合酶链反应一单链构象多态性分析方法探查乙酸镍在诱导16HBE细胞恶变过程中的K—ras基因Exonl和P15基因Exon2存在状况。采用随机扩增多态性技术对乙酸镍在诱导16HBE细胞恶变过程中的基因组不稳定性进行分析。结果：K—ras基因Exonl和P15基因Exon2未发生改变。本实验所选用的7条随机引物均能扩增出清晰、明显的条带，条带数在1-6条之间。7条引物中P4、P7二条引物扩增的片段在实验组和对照组之间无差异。其余5条引物均有差异，对于同一随机引物他们都具有特异的带型。结论：P15基因第2外显子和K—ras基因第1外显子(包括第12、13密码子)可能不是乙酸镍作用的靶部位。在乙酸镍诱发细胞恶性转化过程中，基因组变得逐渐不稳定。     

半巢式RT-PCR法检测呼吸道合胞病毒

目的建立快速、敏感、特异的人呼吸道合胞病毒(Respiratory syncytial virus，RSV)检测方法。方法根据RSVF基因的保守序列设计3条引物，建立半巢式RT-PCR(semi-nested RT-PCR)检测方法。用该方法检测125例患急性下呼吸道感染的婴幼儿的鼻咽吸引物(nasopharyngeal aspirates，NPA)标本，同时对标本进行病毒分离和直接免疫荧光法检测。结果该半巢式RT-PCR灵敏度为0．1TCID50，用该方法在125例标本中共检出RSV阳性标本63例，阳性率50．4％。结论建立快速、敏感、特异的RSV半巢式RT-PCR检测方法，该方法适用于临床早期诊断和流行病学监测。     

1例窗口期HIV感染献血员的追踪与确认

目的对1例HIV RNA阳性献血员进行追踪检测,以确定其HIV病毒的感染情况及感染早期各项指标的变化,为早期诊断和预防提供依据。方法对该献血员初次献血标本及追踪采样的标本,采用两次ELISA筛查及核酸定性、定量检测,并将反应性标本送疾控中心进行确证。结果献血者在高危行为约10d后献血,初次献血标本检测结果为HIV抗体阴性,核酸定性、定量检测均为阳性,随后对该献血员进行了7次追踪随访。在献血第16d后血液标本HIV P24抗原转阳,HIV病毒载量急剧增加;第37d后HIV抗体阳性,确证结果为不确定;第58d后的标本确证为HIV-1抗体阳性。结论该献血员献血时处于HIV感染窗口期。核酸检测能有效降低输血残余风险,对保障血液安全有重要意义。     

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.     

The occurrence of antimicrobial substances in toilet,sink and shower drainpipes of clinical units: A neglected source of antibiotic residues

Antibiotics represent one of the most important drug groups used in the management of bacterial infections in humans and animals. Due to the increasing problem of antibiotic resistance, assurance of the antibacterial effectiveness of these substances has moved into the focus of public health. The reduction in antibiotic residues in wastewater and the environment may play a decisive role in the development of increasing rates of antibiotic resistance. The present study examines the wastewater of 31 patient rooms of various German clinics for possible residues of antibiotics, as well as the wastewater of five private households as a reference.To the best of our knowledge, this study shows for the first time that in hospitals with high antibiotic consumption rates, residues of these drugs can be regularly detected in toilets, sink siphons and shower drains at concentrations ranging from 0.02?μg·L^?1 to a maximum of 79?mg·L^?1. After complete flushing of the wastewater siphons, antibiotics are no longer detectable, but after temporal stagnation, the concentration of the active substances in the water phases of respective siphons increases again, suggesting that antibiotics persist through the washing process in biofilms. This study demonstrates that clinical wastewater systems offer further possibilities for the optimization of antibiotic resistance surveillance.     

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.