转基因豆油的检测

目的探讨转基因豆油的检测方法。方法采用PCR扩增法,对通常转基因植物共有的外源基因35s启动子和Nos终止子序列进行检测,确定其是否转基因作物。结论在转基因豆油中成功检测到35s启动子和Nos终止子的基因片段。     

利用聚合酶链式反应方法检测转基因大豆和玉米中的基因修饰物质

采用聚合酶链式反应 (PolymerasechainreactionPCR)来检测转基因大豆 (RR大豆 )和玉米 (Bt 176玉米 )中基因修饰物质 (GeneticallyModifiedOrganismsGMOs)的方法。利用已知GMOs准确含量的转基因大豆和玉米对PCR检测方法进行了探讨 ,检测限可以达到 0 1% ,即 ,可以把食品中含量仅为 0 1%的GMOs检测出来 ,检测为定性检测。检测的步骤包括 :(1)大豆和玉米基因组DNA的提取 ;(2 )对转入的CaMV35s启动子和NOS终止子利用PCR方法进行扩增 ;(3)对食品的house -keeping基因进行PCR扩增 ,以确定我们所检测的食品确实来自于大豆和玉米 ;(4)利用琼脂糖凝胶电泳及XmnI限制性内切酶方法对PCR产物进行描述与确认。RR大豆含有CaMV35s启动子和NOS终止子 ,而Bt 176玉米只含有CaMV35s启动子。由于玉米的淀粉含量较高 ,因此其基因组DNA的提取效果不如大豆 ,最终的电泳图谱也不如大豆的清晰     

PCR定性检测转基因大豆的探讨

目的应用定性PCR方法检测大豆中转基因成分。方法以大豆凝集素（lection）基因为内源参照基因，设计合适引物，对转基因植物中常用的CaMV 35S启动子进行PCR扩增。结果在转基因大豆中扩增出195bp的CaMV 35S启动子片断。结论定性PCR技术是进行转基因食品检测的有效手段。     

PCR法筛选检测转基因食品

目的通过PCR检测花椰菜花叶病毒(camv)35S启动子和胭脂碱合酶(nos)终止子建立筛选食品中有无转基因成分的方法.方法用改良溴化十六烷三甲基铵(CTAB)法制备转基因抗除草剂[大豆RoundUp ReadyTMSoybean(RRS)]和转基因抗虫玉米系列标准品Bt176 Maximaizer的DNA,PCR检测其内参照基因及camv 35 S启动子和nos终止子.结果改良CTAB法制备的DNA用作PCR模板均可扩增出内参照基因,PCR扩增camv 35S启动子可检测出含量为0.5%的RRS和Bt176 Maximaizer,而PCR扩增nos终止子可检测出含1%RRS的食品样品.结论改良CTAB法制备的DNA可用作PCR模板,建立的PCR检测camv 35S启动子和nos终止子方法可用于筛选食品中有无转基因成分.     

SYBR GreenⅠ实时荧光PCR检测食品中转基因成分方法的初步建立

目的：建赢SYBR GreenⅠ实时荧光PCR方法检测食品中转基因成分35S启动子和NOS终止子,并对样品进行检测。方法：以大豆Lectin基因为内源性基因,以转基因食品中常用的花椰菜花叶病毒启动子（CaMV35S）和根癌农杆菌终止子（NOS）基因为目标基因,设计并合成3对特异性引物,优化反应条件,使用SYBR GreenⅠ染料实时荧光PCR检测35S启动子和NOS终止子,并做熔解曲线分析。同时利用该方法对胡萝卜、青豆、玉米、马铃薯等实际样品进行检测。结果：运用该方法检测8份样品,有4份检出35S基因和NOS基因成分。结论：SYBR GreenⅠ实时荧光PCR方法能有效检测出35S和NOS两种转基因成分,简便、快速。     

食品中腰果过敏原成分PCR检测方法建立

目的建立食品中腰果过敏原成分的PCR检测方法。方法采用溴化十六烷三甲基胺(CTAB)法和磁珠法提取的坚果DNA,根据GenBank中腰果3种主要致敏蛋白AnaO1、AnaO2、AnaO3的基因序列,设计9对引物;通过扩增效果和特异性,筛选腰果最佳扩增引物对;采用腰果特异性引物对AnaO3 F3R3和植物通用引物对P lant 159,对腰果、杏仁、开心果等13种坚果成分进行双重PCR扩增,确定方法特异性;通过腰果DNA梯度稀释确定该检测方法灵敏度;通过样品添加试验模拟食品中腰果过敏原成分的检测。结果采用CTAB法提取坚果DNA的A260/A280比值普遍<1.8,说明存在蛋白质污染,采用磁珠法提取DNA的比值>1.8,说明去除蛋白质较干净;9对引物中,引物对AnaO3 F3R3特异性最好,其最佳退火温度为55.4℃;采用该引物与植物通用引物对P lant 159,对腰果、杏仁、开心果等13种坚果DNA进行双重PCR,只有腰果样品能扩增出312 bp的特异性条带;采用该PCR法检测腰果过敏原成分的灵敏度为0.1 pg/μL;可以检出食品中>0.001%的腰果成分。结论建立了一种食品中腰果过敏原成分的双重PCR检...     

发酵乳酸杆菌PCR快速检测方法研究

〔目的〕建立一种特异、灵敏的发酵乳酸杆菌快速检测方法。〔方法〕根据发酵乳酸杆菌纤维二糖tuf基因延伸因子基因tu设计一对PCR引物,进行引物的特异性和灵敏度试验。〔结果〕PCR得到了较好的特异性扩增,其他干扰菌都无扩增,灵敏度达1.3×104cfu/ml。〔结论〕该方法能够实现发酵乳酸杆菌的快速检测,具有灵敏度高、特异性好的特点。     

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。     

结核分枝杆菌的优化PCR检测方法的研究

〔目的〕的建立了检测结核分枝杆菌的高灵敏度和高特异性PCR扩增体系。[方法]以国内结核病人常见病原菌-结核分枝杆菌、牛分枝杆菌特异性抗原Pab编码基因上的419bp段作为扩增靶序列,扩增引物序列为5’-ACC、ACC、GAG、CGG、TTC、CCC、TGA-3’、5’-GAT、CTG、CCG、GTC、GTC、CCA、GGT-3’。〔结果〕变性温度选择69℃-70℃。PCR扩增体系的成份经优化,选择引物浓度为0.30μmol/L,dNTP浓度为150μmol/L,MgC12浓度为3.5mmol/L。〔结果〕该体系对11株标准菌株和12株临床分离菌株提纯的染色体进行扩增,其中卡介苗、结核分枝杆菌、牛分枝杆菌的扩增为阳性,其他分枝杆菌为阴性。〔结论〕以提纯的染色体DNA作模板,该体系可检出5fg结核杆菌染色体DNA,相当于一个结核分枝杆菌。     

PCR法对单核细胞增生李斯特菌食品分离株的鉴定

〔目的〕采用PCR法对常规法分离的八株单核细胞增生李斯特菌 (LM )再鉴定 ,以验证PCR法作为检测LM的有效性及可靠性。〔方法〕八株从食品中分离的LM在BHI培养基纯培养 2 4h后 ,提取基因组DNA ,用PCR扩增hly基因特异性 85 6bp的片段。〔结果〕八株从食品中分离并经传统方法鉴定的LMPCR扩增均出现特异性的条带 ,而其他属的李斯特菌和非李斯特菌未出现特异性的条带。〔结论〕采用hly基因作为PCR扩增的目标基因检测LM具有较好的特异性 ,可用于食品中LM的检测与分离。     

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.