Vero-E6、MDCK、293细胞对SARS冠状病毒敏感性的研究

目的 研究Vero—E6、MDCK、293细胞对SARS冠状病毒的敏感性，为该病的病原学研究、诊断和疫苗制备奠定基础。方法 用Vero—E6、MDCK及293细胞对广东省部分SARS病人的咽拭子标本进行分离培养，并对分离物使用电镜、间接免疫荧光(IFA)及荧光定量聚合酶链反应(FQ—PCR)的方法分别检测细胞和(或)培养上清波中的病毒含量。结果 感染SARS病人的咽拭子标本后，Vero—E6细胞出现病变(CPE)最早，电镜中可观察到Vero—E6细胞中大量的病毒颗粒，IFA试验感染病毒的Vero—E6细胞与SAPS病人的恢复期血清呈强阳性反应，FQ—PCR结果显示Vero—E6感染细胞内的病毒含量10^8拷贝／ml，高于293细胞(10^6拷贝／ml)及：MDCK(104拷贝／ml)。结论 三种细胞对SARS相关病毒的敏感性不同，Vero—E6是SARS相关病毒最敏感的宿主细胞。     

广东省传染性非典型肺炎病例的SARS冠状病毒分离株的分子生物学鉴定

目的 对广东省部分传染性非典型肺炎(SARS)病例的SARS冠状病毒分离株进行鉴定，并初步建立SARS冠状病毒PCR检测方法。方法 采集临床诊断为SARS病例的咽漱液标本，进行多种细胞分离培养，对出现病变的细胞分离物采用冠状病毒特异引物通过逆转录聚合酶链反应(RT-PCR)及巢式聚合酶链反应(Nested—PCR)进行扩增，并对部分扩增片段进行序列测定，应用DNASTAR软件分析比较oN时采用间接免疫荧光(IFA)和ELISA法检测上述患血清中SARS冠状病毒Igc抗体。结果 对21例SARS患的细胞分离物进行PCR扩增，结果均阳性，其中12份患咽漱液标本PCR扩增结果也阳性，序列测定及基因分析表明，广东省SARS病例的病毒分离株为冠状病毒，其基因序列与WHO公布的SARS基因序列一致，氨基酸序列与目前已知的冠状病毒同源性为58％～76％。时对21例患进行血清学检测，其中IFA检测有9例阳性，ELISA检测有12例阳性。结论 从广东省部分SARS病例标本中分离的病毒株是一种新的冠状病毒，PCR检测具有早期、快速的优点。     

Hep-2细胞在严重急性呼吸综合征病原学检测中的应用研究

目的 应用人咽喉上皮癌细胞(Hep-2)对严重急性呼吸综合征(SARS)病例标本进行病原体检测与研究，为该病的预防与控制提供依据。方法 采用Hep-2细胞培养法，对2003年l～2月广东省收集的SARS病例的咽拭子等标本分离致病病原体，并应用电镜形态学、逆转录．多聚酶链反应(RT-PCR)扩增部分基因进行序列分析，并对分离的病原体进行研究。结果 132例SARS病例的3l份咽拭子、100份痰液和l份尸检肺组织标本中有73份标本致单层细胞出现“蚀斑病变”，细胞病变阳性率为55．3％，其中48h阳性率为36．4％。采用巢式PCR(Nested-PCR)对其中25份出现“蚀斑病变”细胞培养物进行检测，能扩增出冠状病毒的特异片段；对中2-18、中2-19等6份Nested-PCR产物进行基因序列分析，结果 显示其核苷酸序列与国内外公布的结果一致，氨基酸序列与已知的人或动物冠状病毒的同源性在58％～76％之间。通过电镜在SARS病例标本(中2-10)病变细胞及l份尸检肺组织标本中观察到衣原体样颗粒和病毒样颗粒。结论 从接种SARS病例标本的Hep-2细胞病变培养物中证实有冠状病毒。     

不同人群血清对人SARS冠状病毒和动物SARS样冠状病毒反应性的比较

目的 探讨不同人群血清中人SARS冠状病毒抗体和动物SARS样冠状病毒抗体所表达的意义及其之间关系。方法 应用人SARS冠状病毒和动物SARS样冠状病毒抗原片进行间接免疫荧光(IFA)定性和半定量试验,检测23份一般人群、25份动物市场从业人员和74份SARS病人的SARS冠状病毒IgG抗体阳性(ELISA法检测)血清样本,并进行比较分析。结果 23份一般人群、25份动物市场从业人员和74份SARS病人的血清样本的IFA阳性率分别为73．91％、68．00％和100．00％,在IFA阳性人群中其血清样本既能对人SARS冠状病毒也能对动物SARS样冠状病毒产生反应的分别占100．00％(17／17)、64．71％(11／17)和94．59％(70／74);70．59％(12／17)一般人群和76．47％(13／17)动物从业人员其血清中的动物SARS样冠状病毒抗体水平高于人SARS冠状病毒;而SARS病人血清中的人SARS冠状病毒与动物SARS样冠状病毒抗体水平以两者相同和前者高于后者为多,分别占56．76％(42／74)和33．78％(25／74)。结论 不同人群血清中的人SARS冠状病毒抗体和动物SARS样冠状病毒抗体两者之间均存在着一定的交叉反应现象,从血清学上提示了人SARS冠状病毒与动物SARS样冠状病毒之间存在着很高的同源性;人SARS冠状病毒抗体和动物SARS样冠状病毒抗体两者各自表达的意义是有所不同的,通过间接免疫荧光半定量试验可有助于区别感染者所染病毒的倾向性。     

广东省各类SARS病例标本实验室检测结果与临床诊断结果比较

目的 比较不同发病时间、有无明确接触史的SARS临床确诊病人和疑似病人实验室检测结果与临床诊断结果，分析实验室检测结果、流行病学资料和临床诊断之间的相互关系，为临床诊断提供更可靠的参考依据。方法 应用荧光定量PCR和巢式PCR技术检测病人咽拭子中SARS病毒特异基因，应用ELISA和间接免疫荧光法检测病人恢复期血清中的SARS病毒IgG抗体。结果 1～5月份，每月临床确诊病例PCR阳性率分别为：100％(2／2)、76．19％(16／21)、73．17％(30／41)、24．56％(14／57)和11．11％(2／18)，IgG抗体阳性率分别为100％(2／2)、78．57％(13／18)、58．33％(32／67)、30．30％(10／35)和9．52％(2／21)，阳性率逐月下降。4～5月份阳性率较低；1～4月份的临床确诊病例中，有明确接触史的病例，PCR和IgG抗体阳性率分别为73．58％(39／53)和74．42％(32／43)，无明确接触史的病例，PCR和IgG抗体阳性率分别为32．14％(27／84)和25．58％(22／86)，有无明确接触史临床确诊病例阳性率差异有显性。4～6月份，疑似病人PCR和IgG抗体的阳性率分别为11．89％(54／454)和7．78％(21／270)，阳性率很低。结论 综合临床症状、流行病学资料和实验室检测结果进行分析，可以得到更准确可靠的诊断结果。     

间接免疫荧光法在检测SARS冠状病毒特异性抗体中的应用

目的 利用间接免疫荧光技术(IFA)建立SARS冠状病毒(SARS—CoV)特异性抗体血清学诊断方法。方法 采用组织培养技术，用SARS病人的咽漱液样本，在Vero—E6细胞中培养，分离SARS冠状病毒；用已制备的SARS病毒抗原片与临床确诊为SARS的病人双相血清和正常人对照血清作用，再用荧光标记羊抗人IgM/IgG抗体与之结合，在荧光显微镜下观察荧光图像。结果 通过建立检测SARS冠状病毒特异性抗体的间接免疫荧光方法，在44份SARS临床病例标本中，检出IgG抗体阳性3l份，与临床诊断符合率为70．45％。31份IgG阳性者的急性期血清中IgM检出率为38．70％，IgM最早产生于发病第3天，发病7d以上的6例的IgM抗体均为阳性，滴度在发病12～15d达到高峰(1：80)；恢复期血清中IgG抗体最早产生于发病第8天，滴度在发病15～20d达到高峰(1：320～1：640)。97份正常人的血清标本中IgG抗体阳性率为3．09％，IgM抗体为阴性。结论 利用此方法，证实被SARS冠状病毒感染后，在机体中产生有特异性的IgM／IgG抗体；本方法检测结果与临床诊断符合率较高，可以用于早期临床诊断SARS病毒感染和辅助临床做出鉴别诊断。     

江苏省传染性非典型肺炎病原学检测研究

目的:建立传染性非典型肺炎(SARS)实验室检测方法,为SARS病例诊断和现场防治提供依据。方法:用逆转录套式PCR、实时荧光定量PCR检测SARS病毒核酸;酶联免疫法检测血清标本SARS病毒IgM抗体;部分PCR检测阳性或SARS病毒IgM抗体阳性病例的标本进行直接电镜观察;简并引物PCR检测冠状病毒属病毒核酸;鉴别诊断IFA检测血清中肺炎支原体等9种常见呼吸道感染病原IgM抗体,PCR检测军团菌以及甲、乙型流感病毒核酸。结果:检测各类临床标本506份。2003年全省报告临床诊断病例7例,5例检测结果阳性;逆转录套式PCR检测各类标本132份,阳性8份;对其中4份PCR扩增产物(108bp)进行了序列分析,与SARS病毒序列100%同源;实时荧光定量PCR检测各类标本30份,阳性3份;酶联免疫法检测血清标本101份,SARS病毒IgM抗体阳性4份;3个诊断病例的各类标本电镜检查发现冠状病毒样颗粒;IFA检测血清标本75份,嗜肺军团菌IgM抗体阳性9份,乙型流感IgM抗体阳性11份,两者同时阳性3份,其他病原体IgM抗体均为阴性。不明发热病例的标本21份简并引物PCR检测冠状病毒,2份咽拭子标本阳性;漱口液、尿液标本各15份PCR检测军团菌、漱口液25份PCR检测甲、乙型流感病毒结果均为阴性。结论:本研究建立的检测方法与临床诊断有比较好的符合性,对于临床诊断和防治工作具有重要的参考价值。PCR检测SARS-CoV特异性较好,而敏感性有待提高。     

荧光定量PCR检测传染性非典型肺炎冠状病毒的分析

目的 探讨荧光定量PCR在检测传染性非典型肺炎(SARS)冠状病毒(SARS—CoV)中的应用。方法 对2003年1～4月广州市各大医院送检的临床确诊或疑似的SARS病例以及密切接触的咽漱液和咽拭子进行荧光定量PCR检测。结果 从57份确诊患标本中检出SARS-CoV阳性38份，检出率为66．7％。14份密切接触标本中检出阳性7份。59份疑似患标本中检出阳性12份。SARS发病10～12d时SARS—CoV检出率最高，达87．5％。结论 荧光定量PCR能早期、快速检出SARS—CoV，临床符合率达66．7％，采样时间在发病1～12d最佳。     

广西野生动物SARS冠状病毒的感染状况及其意义

目的查明广西境内野生动物SARS冠状病毒感染状况。方法对广西境内15种117只野生动物及99只人工养殖的果子狸采用聚合酶链反应和血清学方法检测其病毒核酸。结果15种野生动物中鸟类及爬行类动物血清的SARS冠状病毒抗体阳性率分别为22．2％和20．0％，哺乳类动物SARS冠状病毒抗体阳性率为0，人工养殖的果子狸咽拭子及肛拭子标本未检出SARS冠状病毒核酸。结论野生动物中鸟类及爬行类动物SARS冠状病毒感染率较高，可能是SARS冠状病毒的自然宿主或储存宿主。     

检测SARS冠状病毒的套式RT-PCR方法的建立与初步应用

[目的]建立套式RT-PCR检测SARS冠状病毒的方法。[方法]从广东地区SARS患者尸解肺组织、咽拭子、嗽口水及本实验室分离的SARS冠状病毒（SAPS—CoV）组织培养上清中提取RNA，利用针对SARS冠状病毒多聚酶基因的特异性引物，进行逆转录及套式PCR扩增。扩增出的阳性片段连接入pGEM—T载体中，测序后比较其与其他已知SARS冠状病毒的同源性。[结果]从SARS患者尸解肺组织、咽拭子、嗽口水及SARS冠状病毒组织培养上清中均可扩增出阳性片段，经测序后证实是SARS冠状病毒特有序列。[结论]针对SARS冠状病毒多泵酶基因所建立的套式RT—PCR方法适用于临床标本及组织培养标本中SARS冠状病毒的检测。     

The human drama of genetics: 'hard' and 'soft' media representations of inherited breast cancer

This paper examines media coverage of 'breast cancer genetics', and explores its implications for public understanding. We present a content analysis of coverage in British newspapers and look at a variety of popular forms, including women's magazines, television soap opera and radio drama. Genetic/inherited risk receives a great deal of coverage across a wide range of media formats and outlets. Much of this attention has focused on individuals from 'high risk families' and dilemmas around prophylactic mastectomies. Through examining media coverage, combined with interviews with media personnel and their sources, we show why this story proved so attractive to the media and highlight the different production values which influence coverage. Finally, we introduce preliminary findings from focus group discussions to demonstrate how such 'human interest' framing has engaged audience attention and influenced public understandings. The paper concludes by highlighting the implications for analysing, predicting, and engaging with, media representations of science.     

Worker education in the primary prevention of occupational dermatoses

This paper reports the evaluation of a skin care education programmeconducted on a fine chemicals manufacturing site where over1,000 employees are located. Approximately 60% are involvedin chemical manufacture. Over a 12 month period production staffreceived training in prevention of occupational dermatoses linkedto a site-wide poster initiative. The incidence of new casesof occupational dermatoses fell from 0.055 (70 cases in 1,277employees) to 0.021 (27 cases in 1,277 employees) before andafter the intervention respectively (p<0.0001). After otherfactors such as chemicals handled, observer bias and changesin reporting related to socioeconomic climate were taken intoaccount it is concluded that this study demonstrates the importanceof worker education as a tool for primary prevention of disease.Training materials such as video and poster presentations maybe effectively used in the chemical manufacturing industry asan adjunct to prevention and control of exposure to substanceshazardous to the skin. Such methods may also be used in otherindustries where there are significant risks of dermatoses.     

Regional Variation in Travel-related Illness acquired in Africa,March 1997–May 2011

To understand geographic variation in travel-related illness acquired in distinct African regions, we used the GeoSentinel Surveillance Network database to analyze records for 16,893 ill travelers returning from Africa over a 14-year period. Travelers to northern Africa most commonly reported gastrointestinal illnesses and dog bites. Febrile illnesses were more common in travelers returning from sub-Saharan countries. Eleven travelers died, 9 of malaria; these deaths occurred mainly among male business travelers to sub-Saharan Africa. The profile of illness varied substantially by region: malaria predominated in travelers returning from Central and Western Africa; schistosomiasis, strongyloidiasis, and dengue from Eastern and Western Africa; and loaisis from Central Africa. There were few reports of vaccine-preventable infections, HIV infection, and tuberculosis. Geographic profiling of illness acquired during travel to Africa guides targeted pretravel advice, expedites diagnosis in ill returning travelers, and may influence destination choices in tourism.     

The design of hazard risk assessment matrices for ranking occupational health risks and their application in mining and minerals processing

Two hazard risk assessment matrices for the ranking of occupational health risks are described. The qualitative matrix uses qualitative measures of probability and consequence to determine risk assessment codes for hazard-disease combinations. A walk-through survey of an underground metalliferous mine and concentrator is used to demonstrate how the qualitative matrix can be applied to determine priorities for the control of occupational health hazards. The semi-quantitative matrix uses attributable risk as a quantitative measure of probability and uses qualitative measures of consequence. A practical application of this matrix is the determination of occupational health priorities using existing epidemiological studies. Calculated attributable risks from epidemiological studies of hazard-disease combinations in mining and minerals processing are used as examples. These historic response data do not reflect the risks associated with current exposures. A method using current exposure data, known exposure-response relationships and the semi-quantitative matrix is proposed for more accurate and current risk rankings.     

Psychopathological features of a patient population of targets of workplace bullying

BACKGROUND: A strong association between workplace bullying and subsequent anxiety and depression, indicated by empirical research, suggests that bullying is an aetiological factor for mental health problems. AIMS: To evaluate levels of stress and anxiety-depression disorder developed by targets of workplace bullying together with outcome at 12 months and to characterize this population in terms of psychopathology and sociodemographic features. METHODS: Forty-eight patients (36 women and 12 men) meeting Leymann Inventory of Psychological Terror criteria for bullying were included in a prospective study. Evaluations were performed at first consultation and at 12 months using a standard clinical interview, a visual analogue scale of stress, the Hospital Anxiety and Depression (HAD) scale, the Beech scale of stress in the workplace and a projective test (Picture-Frustration Study). RESULTS: At first consultation, 81% of patients showed high levels of perceived stress at work and 83 and 52% presented with anxiety or depression, respectively. At 12 months, only 19% of working patients expressed a feeling of stress at work. There was a significant change in symptoms of anxiety while there was no change in symptoms of depression. Stress at work and depression influenced significatively capacity to go back to work. At 12-month assessments, workers showed a significantly better score on the HAD scale than non-workers. Over half the targets presented a neuroticism-related predominant personality trait. CONCLUSION: Workplace bullying can have severe mental health repercussions, triggering serious and persistent underlying disorders.     

The rise and fall in incidence of malignant mesothelioma from a British Naval Dockyard, 1979-1999

BACKGROUND: The incidence of malignant mesothelioma in Britain is predicted to rise over the next 15-25 years because of past failure to protect the workforce against inhalation of asbestos. In British Naval dockyards, alternative insulation materials and respiratory protection were introduced from the mid-1960s. Aims This study was carried out to investigate the effects of these control measures on mesothelioma deaths in dockyard workers. METHODS: Cases of mesothelioma of the pleura and peritoneum between 1979 and 1999 in workers from the Devonport Naval Dockyard, south-west England, were sought from coroners' and medico-legal records. RESULTS: Three hundred and one cases were identified, 7% peritoneal. The peak incidence occurred in 1991 with 25 cases per annum (quadratic model fit R(2) = 74.2%, P < 0.001) and we predict that by 2003 the incidence will fall to fewer than five cases per annum. The mean time between first exposure and presentation was 48.5 years [95% confidence interval (CI) = 47.3-49.8], but this was significantly shorter in the more heavily exposed trades, when compared with the less heavily exposed (42 years, 95% CI = 39.0-45.0, versus 49.5 years, 95% CI = 48.2-50.9). Those with higher exposure were also at significantly greater risk of peritoneal disease (P < 0.023, Fisher's exact test). CONCLUSION: The reduction in incidence of mesothelioma is greater than can be accounted for by reduction in numbers of dockyard workers over the last 50 years. Changes in insulation materials and improved industrial hygiene measures introduced into the Devonport Dockyard from the mid-1960s have resulted in an earlier decline in the incidence of malignant mesothelioma than that predicted for the British workforce as a whole.     

Organophosphate toxicity and occupational exposure

The ubiquitous organophosphates present a continuing health hazard in agriculture, public health eradication programmes and as chemical warfare agents. Despite significant progress in understanding the potential mechanisms of toxicity far beyond the commonly accepted mechanism of cholinesterase inhibition in intentional exposures, the precise health effects following occupational exposures are yet to be completely defined. A much greater understanding exists of the clinical features of organophosphate poisoning. These are characterized by a triphasic response involving an initial acute cholinergic phase, an intermediate syndrome (both associated with high mortality) and a disabling but non-lethal delayed polyneuropathy. The delayed polyneuropathy may occur in the absence of the cholinergic or intermediate phases. However, progress is still required in order to improve the quantification and assessment of occupational exposures and the implementation of appropriate preventive measures. Finally, evidence-based guidelines for appropriate or optimal therapeutic interventions following poisoning are required urgently and collaborative work with colleagues in developing countries, where the occurrence of organophosphate exposures is more frequent, may provide the answers.     

Miliaria rubra of the lower limbs in underground miners

This report documents a case series of miliaria rubra of the lower limbs in miners at a deep underground metalliferous mine in tropical arid Australia. During the summer months of February and March 1999, all cases of miliaria rubra of the lower limbs in underground miners seen at the mine's medical centre were clinically examined and administered a questionnaire. Twenty-five patients were seen, an incidence of 56.4 cases per million man-hours. Miliaria rubra was most often located between the ankle and knee (88% of cases). Twenty-four percent had concurrent folliculitis and 20% had concurrent tinea. Thirty-two percent had a personal history of asthma. Walking through ground-water and splashing of the legs was common. Three to 4 weeks of sedentary duties in air conditioning was generally required to achieve resolution of miliaria rubra. The incidence of miliaria rubra of the lower limbs is 38% of the incidence of heat exhaustion at the same mine. The length of disablement is greater, however. Atopics may be at increased risk of miliaria rubra. Control measures are discussed.     

Planning research

The starting point for any research project should be a question.Once this has been defined and the relevant scientific literaturereviewed, a protocol should be drawn up. This will be used notonly as a guide to the conduct of the study and in the preparationof the final report, but also in seeking any financial supportand approvals that are required for the investigation. A protocolis normally arranged in sections covering the background tothe study, the question(s) that it will address, the methodsthat will be used for the collection and analysis of data, thestatistical power of the investigation (where relevant), anyethical considerations, and the financial input that will beneeded. A pilot study is often helpful where aspects of thestudy method are untried or of uncertainvalidity.     

Stress in the Norwegian police service

BACKGROUND: High levels of stress have been attributed to the conditions of police work, but there is little empirical evidence for this. AIMS: To develop a new instrument to measure job stress in the police; to assess the most severe and frequent police stressors; to compare levels of stress according to the demographic and organizational factors; and to study stress in relation to personality traits, work locus of control and coping strategies. METHODS: A comprehensive nationwide questionnaire survey of 3272 Norwegian police at all hierarchical levels, including the Norwegian Police Stress Survey (NPSS), the Job Stress Survey, the Basic Character Inventory, the Work Locus of Control Scale, and the Coping Strategies Scale. RESULTS: Work injuries were appraised as the most stressful but least frequent stressor and job pressure was reported the least severe but most frequent stressor. Females experienced job stressors less frequently, but appraised them as more severe than men did. Older police officers reported more job pressure severity and fewer work injuries. The police in districts where peer support was planned but not implemented, and who worked in districts with more than 50,000 inhabitants, perceived the lack of support more severely than others. The correlations between stress and personality traits, work locus of control, and coping were moderate. CONCLUSIONS: The NPSS captures police-specific stressors that are not adequately measured by global stress instruments. The study of stress in police work should preferably involve a nationwide use of police-specific stress instruments.