柠檬无色藻菌的鉴定方法及体外药敏特点

目的总结临床少见菌柠檬无色藻菌的鉴定过程,认识柠檬无色藻菌的耐药特点,并对该菌感染的临床特征进行分析。方法菌株分离培养后,采用传统的形态、生理生化表型和VITEK-2全自动细菌鉴定仪与16S rRNA序列分析相结合的方法来鉴定菌株。结果多次传代后,综合菌株形态、生理生化表型、VITEK-2全自动细菌鉴定仪以及16S rRNA序列分析的结果,确定该分离株为柠檬无色藻菌;药物敏感定量试验显示红霉素、四环素、复方新诺明、三代头孢菌素及喹诺酮类药物均敏感,唯有万古霉素和替考拉宁耐药。结论柠檬无色藻菌引起的血流感染比较罕见;16S rRNA基因序列分析方法可以较好地鉴定常规方法难以鉴定的不典型及罕见菌株。对万古霉素及替考拉宁耐药可以作为无色藻菌与其他阳性球菌鉴别的重要依据。     

1株少见贪铜菌的分离鉴定及耐药性分析

目的对从桶装饮用纯净水中分离出的疑似铜绿假单胞菌菌株进行后续分析。方法分离株依次进行生化鉴定、16S rRNA基因测序与药敏试验。结果该分离株生化鉴定为少见贪铜菌;16S rRNA基因的BLAST序列比对结果与少见贪铜菌LMG 3413T的16S rRNA基因序列同源性为99%;药敏试验显示分离株对阿米卡星、庆大霉素、氨曲南、美罗培南、氨苄西林、磷霉素和妥布霉素耐药,对头孢菌类、内酰胺酶抑制剂、喹诺酮类等抗生素敏感。结论少见贪铜菌是比较罕见的菌血症致病菌之一,免疫力低下患者感染较多。此次研究结果可作为少见贪铜菌相关疾病检测、鉴别、诊断和治疗的依据。     

1株从败血症患者血液分离的念珠状链杆菌的生物学特征

目的采用形态学方法结合分子生物学方法分析1株临床败血症患者血液分离的念珠状链杆菌(Streptobacillus moniliformis),为临床罕见致病性病原菌的鉴定提供参考。方法采用革兰染色、DL-96E,VITEK2 Compact自动细菌鉴定仪、MALDI-TOF MS质谱仪结合16S rRNA基因序列进行细菌鉴定,同时采用E-test法开展药敏试验。结果传统方法和MALDI-TOF MS无法鉴定出该菌,16S rRNA鉴定出该菌为念珠状链杆菌,药敏试验结果显示该菌对青霉素、头孢噻肟、亚胺培南、四环素和环丙沙星敏感,对氨基糖苷类(妥布霉素)、阿米卡星、庆大霉素和磺胺甲噁唑/甲氧苄啶耐药。结论 16S rRNA分子生物学方法结合形态观察和生化反应可以很好地鉴定出念珠状链杆菌,青霉素、头孢噻肟、亚胺培南、四环素和环丙沙星可用于该菌临床治疗。     

基于细菌16S rRNA基因扩增临床不常见病原菌的价值

目的探讨16S rRNA基因扩增与测序在临床不常见病原菌鉴定中的价值，指导临床相关感染的诊治。方法选择临床微生物实验室常规方法难以准确鉴定、无法鉴定或有特殊表型的细菌12株，采用聚合酶链反应（PCR）扩增其16S rRNA基因，测序后进行BLAST比对鉴定菌种，并分析相关感染的临床特点。结果12株菌经PCR扩增均得到阳性条带（约1 500 bp），均鉴定到种（相似度≥99%），分别为产单核细胞李斯特菌、马耳他布鲁杆菌各2株，死亡梭杆菌、空间罗氏菌、鼻疽奴卡菌、解糖葡萄球菌、放射性根瘤菌、二路普雷沃菌、解甘露醇罗尔斯顿菌及阴道阿托波菌各1株。16S rRNA基因扩增灵敏度高，大肠埃希菌ATCC 25922的最低检测限为1.5×101 CFU/mL。12例患者临床资料显示上述菌可引起临床多部位、多类型感染，选用针对性抗菌药物治疗后11例好转，1例死亡。结论16S rRNA基因测序方法可快速、准确鉴定少见菌、厌氧菌及难培养细菌，为临床不同类型感染的病原学诊断和治疗提供实验室依据。     

罕见非嗜肺军团菌的鉴定

目的对非嗜肺军团菌进行分离培养和分子分型鉴定。方法依据《公共场所集中空调通风系统卫生规范》(2006),对广州市海珠区公共场所集中空调冷却塔水进行军团菌检测。用常规分离培养法检测军团菌,进一步运用生化和血清学凝集试验、实时荧光PCR技术及16S rRNA基因、mip基因测序对菌株进行种属鉴定。结果在BCYE平板上生长,在L-半胱氨酸缺失的BCYE平板及血平板中均不生长的菌株,通过实时荧光PCR技术及16S rRNA基因、mip基因测序鉴定为菲氏军团菌4株和釜山军团菌1株,这是本区首次从集中空调冷却塔水中检出的非嗜肺军团菌。结论运用常规分离培养方法和分子生物技术相结合检测出菲氏军团菌和釜山军团菌(国内罕见)等多种军团菌种,同时提示海珠区公共场所集中空调冷却塔水存在非嗜肺军团菌污染;16S rRNA基因、mip基因核酸测序鉴定有助于发现军团菌新种。     

玉树地震受伤患者伤口感染梭状芽胞杆菌培养鉴定分析

目的了解玉树地震受伤患者伤口感染梭状芽胞杆菌情况,为临床诊治及突发公共卫生灾害细菌的感染救治提供参考依据。方法 3例伤口感染的地震受伤患者入院时立即采集伤口分泌物进行细菌厌氧和需氧培养,并采用ATB自动微生物分析仪进行细菌鉴定;鉴定出的梭状芽胞杆菌菌株经PCR扩增细菌16S rRNA基因,DNA测序分析,进一步确定细菌种类。结果 3份标本经自动微生物分析仪鉴定出1株产气荚膜梭菌、4株疑似梭状芽胞杆菌;PCR扩增显示5株菌均出现目的大小条带;测序证实5株菌有1株为产气荚膜梭菌,另4株为需氧芽胞杆菌属细菌。结论自动微生物分析仪鉴定和细菌16S rRNA基因测序分析联合应用可快速、准确为临床提供诊治和预防控制信息。     

16S rRNA基因序列分析在非典型菌株鉴定中的应用

目的：建立16SrRNA基因序列分析鉴定细菌的方法，评价其对常规方法不能鉴定菌株的鉴定效果。方法：选择细菌的16SrRNA基因为靶序列，在两端保守区设计引物，对三株生化、形态不典型菌株提取模板进行PCR扩增，PCR产物纯化后进行克隆，测序。结果：三株细菌的16SrRNA基因序列与Genbank比较，YC1序列与大肠杆菌相似性〉99％，YC2序列与肺炎克雷伯菌相似性〉99％，YC3序列与缓症链球菌相似性〉98％。结论：16SrRNA基因序列分析的方法可以较好地鉴定常规方法难以鉴定的不典型菌株。     

云南省首例人感染猪链球菌病的病原学分析

目的 对猪链球菌菌株进行鉴定,了解其病原学特征.方法 对菌株进行了菌落形态、生化鉴定、血清凝集、PCR 检测、16S rRNA 细菌种属鉴定、PFGE 检测和MLST 分型.结果 菌落形态、生化鉴定、血清凝集、16S rRNA 细菌种属鉴定,证明此菌株为猪链球菌2 型;经PCR 检测16SrRNA 和毒力基因均为阳性;PFGE 检测结果表明此株菌同3 株参考菌株相似性很低;MLST 分型属于高致病性ST1 型.结论 此次分离到菌株的为云南省首例人感染的猪链球菌2 型,是强毒力株.     

16S rRNA、16S-23S rRNA基因测序分析检测主要血流感染病原菌比较

目的比较细菌16S rRNA、16S-23S rRNA 基因测序分析在血流感染病原菌检测中的作用。方法提取临床上血流感染常见的金黄色葡萄菌、表皮葡萄球菌、大肠埃希菌、粪肠球菌、肺炎链球菌、铜绿假单胞菌、阴沟肠杆菌、鲍曼不动杆菌、洛菲不动杆菌、肺炎克雷伯杆菌、化脓性链球菌、奇异变形杆菌、潘尼变形杆菌、屎肠球菌、粘质沙雷菌、宋内志贺菌、产气肠杆菌、小肠结肠炎耶尔森菌、腐生葡萄球菌基因组 DNA,运用 16S rRNA、16S-23S rRNA 基因进行 PCR 扩增。扩增产物经测序后在美国国家生物技术中心(NCBI)上进行比对分析,确定菌种。结果在所分析的 19 种临床血流感染常见细菌中,16S rRNA 基因测序分析可将除粘质沙雷菌外的细菌鉴定到种的水平,但无法完全区分近缘种属;16S-23SrRNA 成功鉴定 17 种细菌,除大肠埃希菌、宋内志贺菌外所有细菌均成功鉴定到单一种的水平。结论 16S-23S rRNA 基因可作为血流感染细菌检测较好的分子靶标。     

呼吸道感染患者分离奈瑟菌属菌种的生物学与16S rRNA鉴定

目的 了解引起呼吸道感染的常见致病性奈瑟菌属菌种以及从慢性呼吸道感染患者分离的致病性奈瑟菌属的鉴定方法.方法 采集249例不同年龄慢性呼吸道感染患者的痰标本,分别进行革兰染色镜检和接种血琼脂培养基分离培养;分离的奈瑟菌样革兰阴性双球菌,分别进行常规细菌学方法和染色体16S rRNA基因检测鉴定以及药物敏感(K-B法)试验.结果 在184例患者的痰标本内分离出227株.占73.9％,呈明显优势生长甚至惟一生长的革兰阴性双球菌,常规细菌学方法鉴定分别为干燥奈瑟菌、黏液奈瑟菌、微黄奈瑟菌、金黄奈瑟菌、多糖奈瑟菌、嗜乳糖奈瑟菌、脱氨奈瑟菌及灰色奈瑟菌;各菌种通过16S rRNA基因PCR检测和序列测定,发现与常规细菌学方法鉴定的符合率＞64.0％;来自不同患者的各种奈瑟菌对临床常用抗菌药物的敏感性相似,存在许多耐药或多药耐药菌株.结论 人体上呼吸道正常菌群奈瑟菌属的许多菌种,是引起呼吸道感染患者慢性继发性呼吸道感染的常见致病菌,这些菌种的生物学特性及其广泛的耐药性,使其成为影响慢性呼吸道感染的病原学诊断与治疗效果的重要因素,常规细菌学与16S rRNA检测相结合,能够提高呼吸道条件致病性奈瑟菌鉴定的准确率.     

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.