菘蓝中苯丙氨酸解氨酶家族基因的鉴定及分析

目的对菘蓝中苯丙氨酸解氨酶家族的基因进行检定和分析。方法克隆获得菘蓝苯丙氨酸解氨酶基因(Isatisindigotica Fort.phenylalanine ammonia-lyase,LiPAL)全长(CurPAL)序列,并对菘蓝中IiPAL1及IiPAL2进行了生物信息学分析,表达分析及体外酶活测定。通过生物信息学方法,在已经测序菘蓝基因组中预测苯丙氨酸解氨酶(PAL)基因。结果菘蓝IiPAL2的cDNA全长为2115bp,编码705个氨基酸。生物信息学分析表明,IiPAL1与IiPAL2与其它植物中的PAL基因具有较高的同源性。IiPAL1基因与IiPAL2基因均表达在菘蓝分布的茎部、叶子与花中,其中IiPAL1基因在叶中的表达量最高,IiPAL2基因在根部中的表达量最高。结论IiPAL1及IiPAL2对苯丙氨酸具有不同的催化能力,为确定木脂素生源途径的关键酶,可以在此基础上开展次生代谢工程提高菘蓝中药效成分含量。     

日本血吸虫新基因精氨酸酶的扩增及序列分析

目的获取并真核克隆日本血吸虫新基因精氨酸酶全长cDNA.方法对本实验室曾获得精氨酸酶基因利用生物信息学进行分析,发现其5'端尚缺一段序列;根据已知序列设计引物,用5'端单巢式PCR从尾蚴文库中扩增,以获得精氨酸酶基因完整的阅读框(ORF);用生物信息学技术对获得的精氨酸酶编码基因进行结构与功能的分析;并将新基因的完整编码阅读框克隆入真核表达载体pCDNA3.1.结果用5'端单巢式PCR从尾蚴文库中获得了精氨酸酶5'端所缺序列,得到其完整的ORF,其ORF长1 095 bp,编码364个氨基酸.利用生物信息学技术鉴定其为日本血吸虫精氨酸酶的完整cDNA序列.重组质粒经双酶切DCR及测序鉴定证明日本血吸虫精氨酸酶真核表达质粒构建成功.结论成功获得、识别、扩增并克隆日本血吸虫精氨酸酶编码基因的全长cDNA.     

微小隐孢子虫20K亲环蛋白基因克隆及分析

目的 克隆微小隐孢子虫(Cp)南京株(NJ)的20K亲环蛋白(CyP)基因,并对其核苷酸和氨基酸序列进行分析.方法 采用昆明种小鼠建立微小隐孢子虫NJ株感染模型,根据微小隐孢子虫Iowa Ⅱ株20K CyP基因序列设计合成2对引物,应用巢式PCR技术从微小隐孢子虫NJ株基因组DNA中扩增20K CyP基因,并将其克隆到pMD18-T载体上,将阳性克隆重组质粒进行菌落PCR及双酶切鉴定;应用生物信息学方法分析微小隐孢子虫NJ株20K CyP基因与其他虫株核苷酸和氨基酸序列差异,并分析该基因蛋白结构域.结果 巢式PCR扩增得到特异的20K CyP基因,经PCR及双酶切鉴定获得了正确的pMD18-T-20K CyP重组质粒;测序结果显示,微小隐孢子虫NJ株20K CyP基因全长519 bp,编码173个氨基酸,该基因已登录GenBank,登录号为JQ284431;氨基酸序列分析表明,微小隐孢子虫NJ株与Iowa Ⅱ株20K CyP基因编码的氨基酸序列具有100％同源性;对20K Cyp基因进行蛋白结构域分析,显示该基因序列所编码的蛋白具有特异性类亲环蛋白A、B、H的肽脯氨酰顺反异构酶(PPIase)区域,该区域与人的亲环蛋白A、B、H具有相似性.结论 成功克隆微小隐孢子虫NJ株20K CyP基因,该基因具有高度保守性.     

亚洲带绦虫膜联蛋白B3基因生物信息学分析

目的 识别亚洲带绦虫新基因,分析和预测该基因及其编码蛋白的结构和特性,为其生物学功能研究提供依据.方法 利用在线生物信息学网站及工具进行序列分析、预测其编码的膜联蛋白B3的理化特性、抗原表位、翻译后的修饰、功能域、亚细胞定位、拓扑结构、二级结构、三维空间构象、进化树等.结果 该基因全长1176 bp,编码区为70～588 bp,编码173个氨基酸,为全长基因;无跨膜区;具有多个磷酸化位点和B细胞线性表位;蛋白的理化性质稳定,理论分子量为19339.7;没有质体、线粒体定位序列;氨基酸序列高度保守.结论 运用生物信息学方法 从亚洲牛带绦虫成虫cDNA文库中识别出了亚洲牛带绦虫成虫膜联蛋白B3基因,并对其所编码的蛋白质的结构与功能进行预测.     

亚洲带绦虫六钩蚴Ta 18基因克隆及生物信息学分析

目的从亚洲带绦虫(Taenia asiatica)六钩蚴cDNA文库中克隆出亚洲带绦虫六钩蚴Ta 18基因,生物信息学分析预测其编码蛋白的结构和生物学功能。方法利用生物信息网站如美国国家生物技术信息中心(NCBI,http://www.ncbi.nlm.nih.gov/)和瑞士生物信息学研究所的蛋白分析专家系统(ExPASY,http://ca.expasy.org/)中生物信息学分析工具,并结合其他分析软件,分析该基因的结构并预测其编码的蛋白质的结构和功能特征。结果成功克隆出Ta 18基因,全长396 bp,编码131个氨基酸,其序列包含完整开放阅读框,推导出的氨基酸序列与GenBank中牛带绦虫18 kD蛋白的同源性最高,一致性达99%,与猪带绦虫一致性达97%;Ta 18蛋白质的理论分子量为14 810.4 Da,等电点为10.05,蛋白理化性质稳定,分析显示Ta 18蛋白质含有1个信号肽和1个纤连蛋白FnⅢ型结构域。结论成功从亚洲带绦虫六钩蚴中克隆出Ta 18基因并分析预测到所编码蛋白的功能特征。     

环境内毒素相关新基因的克隆测序及生物信息学分析

目的克隆表达环境内毒素相关新基因--人lrg基因(简称Hlrg),并进行生物信息学分析.方法采用RTPCR扩增Hlrg基因,对扩增产物进行序列的测定,利用Internet和GenBank数据库对测序结果正确的Hlrg基因进行生物信息学分析.结果克隆表达了Hlrg基因,测序结果正确;生物信息学分析表明,该序列编码186个氨基酸,GenBank中不含有同源序列.预测相对分子量为21×103,该序列包含有亮氨酸拉链结构,可能具有重要的功能.结论成功扩增、克隆表达了Hlrg基因,对其功能进行了初步预测,为进一步的功能研究打下基础.     

细粒棘球绦虫14-3-3zeta蛋白的生物信息学分析

目的应用生物信息学技术对细粒棘球绦虫(Echinococcus granulosus)14-3-3zeta蛋白的结构和功能进行预测和分析,为进一步的实验研究提供依据。方法利用美国国家生物技术信息中心(NCBI,http://www.ncbi.nlm.nih.gov/)和瑞士生物信息学研究所的蛋白分析专家系统(ExPASY,http://expasy.org/)提供的各种有关基因和蛋白序列、结构信息分析的工具,并结合其它生物信息学分析软件,对该蛋白质的结构和功能进行预测和分析。结果该基因全长为771 bp,编码256个氨基酸,其编码的蛋白相对分子量理论预测值和等电点分别是29.4 kDa和5.04。预测该蛋白无信号肽和跨膜区,二级结构含8个α-螺旋和12个β-折叠股,氨基酸序列中有9个潜在抗原表位。结论初步认识了细粒棘球绦虫14.3-3zeta蛋白的基本特征,为深入研究该蛋白的生物学功能奠定了基础。     

人参的药理作用及临床应用

<正>人参为五加科植物人参的干燥根,始载于我国第一部本草专著《神农本草经》。中医认为人参味甘,有大补元气、生津止渴、调荣养卫的功能。现代化学研究表明,人参中含有13种以上的皂苷,其总皂苷称人参皂苷Rx,皂苷元为人参二醇、人参三醇及齐墩果酸等,皂苷是人参生物活性的物质基础。人参中还含有少量的挥发油、β-榄香烯和人参炔醇,其中含有的氨基酸主要有精氨酸、赖氨酸、甘氨酸等;人参中还含有人参酸、维生素B、维     

日本血吸虫ATP合酶脂结合蛋白样蛋白基因结构与功能分析

目的 利用生物信息学技术对已登录的日本血吸虫(Schistosoma japonicum，Sj)中国大陆株(chinese strain)新基因-ATP合酶脂结合蛋白样蛋白(ATP synthase lipid-binding protein-like protein)基因结构和功能进行分析。方法 使用NCBI，SAPS，TMHMM2．0，PsortⅡ，Protscale和Scanprosite等软件对新基因的结构和功能进行分析。结果 通过Internet在线分析和生物信息学软件分析，对新基因的结构和功能有了进一步的认识，日本血吸虫ATP合酶脂结合蛋白样蛋白基因编码122个氨基酸，分子量为12．7kDa，等电点为9．41，为一跨膜蛋白，含有磷酸化位点和烷基化位点。结论 生物信息学技术有利于日本血吸虫新基因的结构和功能研究。     

日本血吸虫酪蛋白激酶Ⅱβ亚基的序列分析

目的 发现并克隆日本血吸虫新基因。方法 对已获得的日本血吸虫表达序列标签(EST)进行同源性分析。识别血吸虫新基因；根据EST设计引物，用3′RACE从mRNA中扩增获得新基因全长，用生物信息学技术对获得的编码基因进行结构与功能分析，将新基因的完整编码阅读框克隆入原核表达载体pGEX一4T—1。结果 用3′RACE获得一个EST的3′端所缺序列，得到完整编码阅读框，其开放阅读框((ORF)654bp，编码217个氨基酸。利用生物信息学技术确定其为日本血吸虫酪蛋白激酶Ⅱβ亚单位的编码基因。重组质粒经双酶切及测序鉴定，证明日本血吸虫CKⅡβ原核表达质粒构建成功。结论 扩增并成功克隆日本血吸虫酪蛋白激酶Ⅱβ亚单位编码基因的全长cDNA。为进一步的功能研究打下基础。     

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 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.     

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.     

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.     

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.     

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.