抗T-2毒素单克隆抗体的制备及鉴定

目的:制备并鉴定抗T-2毒素单克隆抗体杂交瘤细胞株,以期为T-2毒素的快速检测提供有力的抗体工具。方法:采用琥珀酸酐法将T-2毒素活化为T-2HS,T-2HS在DCC与NHS作用下与牛血清白蛋白(BSA)或卵清白蛋白(OVA)偶联合成完全抗原。以T-2HS-BSA为抗原,免疫BALB/c小鼠,用ELISA筛选、建立分泌抗T-2毒素抗体的杂交瘤细胞株。测定单抗免疫球蛋白亚类及单抗效价,用间接竞争法检测单克隆抗体细胞株的特异性。结果:建立了1株稳定分泌抗T-2毒素抗体的杂交瘤细胞株T-1B3,该细胞株所分泌的单隆抗体Ig亚类为IgG1,ELISA检测结果表明抗T-2毒素抗体可以与T-2毒素发生特异性反应,工作浓度为1:6.5×104,IC50为3.6 ng/ml。该抗体与HT-2毒素的交叉反应为65.9%,与黄曲霉毒素无交叉。结论:制备出能分泌具有较高特异性和亲和力的抗T-2毒素单克隆抗体的杂交瘤细胞株。     

抗总黄曲霉毒素单克隆抗体的制备及特性

目的研制能够与黄曲霉毒素B1、B2、G1、G2有较好交叉反应的抗总黄曲霉毒素单克隆抗体。方法利用B细胞杂交瘤技术,建立分泌抗总黄曲霉毒素单克隆抗体杂交瘤细胞株。结果建立了1株稳定分泌抗总黄曲霉毒素抗体的杂交瘤细胞株,命名为2G2。该细胞株所分泌的单克隆抗体Ig亚类为IgG1,参考工作浓度为1∶1.6×106,亲和力常数为8×10-8mol/L。该抗体与黄曲霉毒素B1、B2、G1、G2的交叉反应分别为100.0%、57.5%、104.0%和19.0%,与其他真菌毒素无交叉反应。结论制备出能分泌具有高特异性和亲和力的抗总黄曲霉毒素单克隆抗体的杂交瘤细胞株。     

分泌抗重组葡激酶单克隆抗体杂交瘤细胞株的建立

目的 建立分泌抗重组葡激酶（r-SAK)单克隆抗体杂交瘤细胞株。方法 采用3种方法以r-SAK免疫Balb/c小鼠，取小鼠脾细胞与SP2/O骨髓瘤细胞做细胞融合。结果 经筛选和克隆化后获得10株抗r-SAK单克隆抗体杂交瘤细胞。结论 建立的分泌抗r-SAK单克隆抗体杂交瘤细胞株具有高度特异性，在r-SAK的研制中有很高的应用价值。     

Ⅰ型登革病毒prM抗体的制备及初步鉴定

目的以Ⅰ型登革病毒pr M蛋白为靶抗原,制备相应的多克隆和单克隆抗体,并进行初步鉴定。方法 RTPCR扩增Ⅰ型登革病毒全长pr M基因,转入克隆载体和构建重组表达载体,以原核表达及纯化后的pr M蛋白作为免疫原分别免疫新西兰大白兔和BALB/c小鼠,采集免疫兔血清,制备抗pr M蛋白多克隆抗体;取小鼠脾细胞与骨髓瘤细胞融合,经HAT选择培养、间接ELISA筛选阳性克隆,获得特异性单克隆抗体的杂交瘤细胞株,制备抗pr M蛋白单克隆抗体,应用亲和层析法纯化抗体,Western-blot鉴定抗体特异性,间接ELISA检测抗体效价。结果获得全长pr M的多克隆抗体及1株能稳定分泌抗pr M蛋白的单克隆抗体杂交瘤细胞株,抗体的效价高,特异性好。结论制备了Ⅰ型登革病毒pr M多克隆抗体和1株单克隆抗体,为进一步探讨登革病毒pr M抗体依赖的感染增强作用奠定基础。     

环丙沙星单克隆抗体制备、纯化及鉴定

目的制备抗环丙沙星单克隆抗体.方法用碳二亚胺(EDC)法合成的环丙沙星-牛血清白蛋白完全抗原(CPFX-BSA)免疫BALB/c小鼠,通过杂交瘤技术筛选分泌抗环丙沙星单克隆抗体的杂交瘤细胞株.结果筛选出3株分泌抗环丙沙星单克隆抗体的杂交瘤细胞株1D10、3C6和3G7,其抗体类型分别为IgM、IgG1和IgG1.其中3C6腹水抗体纯度、抗体浓度、亲和常数分别为80.04%、3.96 g/L和1.01×108 L/mol.该单抗与恩诺沙星、诺氟沙星的交叉反应率分别为125.89%、19.50%,与青霉素、卡那霉素和庆大霉素等无交叉反应.结论该单抗可用于动物性食品中CPFX残留ELISA检测方法的建立.     

抗结核分支杆菌相关抗原单克隆抗体的制备及荧光抗体染色方法的建立

目的：制备抗结核分支杆菌的特异单克隆抗体，经纯化标记荧光后，建立直接荧光抗体染色方法用于痰标本的检测。方法：采用常规方法制备单克隆抗体。分泌单克隆抗体阳性杂交瘤细胞株的筛选采用ELISA法，确认采用免疫印迹法。单克隆抗体的纯化采用饱和硫酸铵粗提，Sephadex G-50层析纯化。单抗纯化后采用透析法标记异硫氰酸荧光素，初步应用于临床痰涂片的检测。结果：经筛选获得4株单克隆抗体杂交瘤细胞株。ELISA检测小鼠腹水单克隆抗体效价达到1：6400～1：12800。免疫印迹实验表明4株单抗均为抗结核杆菌38KD蛋白单抗，其中两株产生较强的免疫反应条带。单抗纯化后标记异硫氰酸荧光素，建立直接荧光抗体染色方法，对41份结核病患者的痰标本进行检测，阳性率为90．24％，与抗酸染色法比较，直接荧光抗体染色方法敏感性明显高于抗酸染色法（P〈0．05）。结论：制备抗结核杆菌38KD蛋白的单克隆抗体，并建立直接荧光抗体染色方法，初步应用于临床痰涂片检测中，对于辅助诊断结核病具有一定价值。     

抗黄曲霉毒素M1抗体制备及检测方法建立

目的制备针对黄曲霉毒素M1的单克隆抗体并建立针对黄曲霉毒素M1的间接竞争酶联免疫吸附试验检测方法。方法利用B细胞杂交瘤技术。建立能分泌抗黄曲霉毒素M1单克隆抗体的杂交瘤细胞株，制备抗黄曲霉毒素M1单克隆抗体，建立间接竞争酶联免疫吸附试验检测方法。结果研制出1株能特异性分泌抗黄曲霉毒素M1单克隆抗体的杂交瘤细胞株，命名为2F2。该单克隆抗体的Ig亚类为IgG1，亲和常数为2．8×10^-11mol／L。该抗体与黄曲霉毒素B1、黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2和黄曲霉毒素M2等结构类似物有微弱的交叉反应，具有较高的特异性。在此基础上建立了间接竞争酶联免疫吸附试验检测方法。该方法的最低检出浓度为0．07ng／ml。校正曲线的线性范围为0．02～2ng／ml，线性方程y=-0．4364x＋0．2693（R^2=0．9949）。方法的加标回收率为72．5％～131．3％。结论制备了具有高特异性和亲和力的抗黄曲霉毒素M1单克隆抗体，并建立了快速、灵敏的针对黄曲霉毒素M1的酶联免疫吸附试验检测方法。     

抗腐败西瓦菌单克隆抗体杂交瘤细胞系的建立及初步鉴定

目的 制备和鉴定抗腐败西瓦菌单克隆抗体(mAbs),为建立重要海洋细菌快速ELISA检测试剂盒奠定基础.方法 用灭活腐败西瓦菌免疫小鼠,利用杂交瘤细胞融合技术,制备出抗腐败西瓦菌杂交瘤细胞株,用Giemsa染色对其进行染色体鉴定,并用间接ELISA方法筛选、鉴定其与腐败西瓦菌及其他重要海洋细菌的交叉反应性和效价.结果 共获得4株抗腐败西瓦菌的杂交瘤细胞株(02D6、02H12、03G4、03A7),均符合杂交瘤细胞染色体特点,其培养上清效价为10-1～10-2,具有良好的特异性和免疫反应性.结论 研究制备、筛选出抗腐败西瓦菌的特异性抗体,有助于进一步建立重要海洋细菌快速检测试剂盒.     

H7N9禽流感病毒血凝素线性B细胞抗原表位单克隆抗体的制备和鉴定

目的研制用于H7N9禽流感病毒诊断的血凝素线性B细胞抗原表位单克隆抗体。方法采用H7N9禽流感病毒血凝素特异性线性B细胞抗原表位免疫BALB/c小鼠,通过杂交瘤细胞技术和间接ELISA法筛选、鉴定获得稳定分泌抗H7N9禽流感病毒血凝素单克隆抗体的杂交瘤细胞株;通过SDS-PAGE、间接ELISA和斑点免疫印迹技术鉴定其亚型、效价、纯度等生物学特征。结果共获得2株单克隆抗体(7C8H8和8E9E2),其中7C8H8抗体亚类为IgG2a,腹水抗体效价>1∶512000,且可以特异性结合H7N9禽流感病毒血凝素。8E9E2仅具有较低的H7N9禽流感病毒血凝素蛋白结合力。结论获得1株可分泌高效价、高特异性的抗H7N9禽流感病毒血凝素单克隆抗体的杂交瘤细胞株7C8H8。     

小肠结肠炎耶尔森菌O:9血清型特异性单克隆抗体筛选及其应用

目的利用杂交瘤技术筛选出能分泌小肠结肠炎耶尔森菌O：9血清型特异性单克隆抗体的细胞株,用于该型菌株的鉴定.方法采用ELISA方法筛选克隆株,并用玻片凝集法进行单克隆抗体的特异性鉴定.结果筛选到3株能分泌小肠结肠炎耶尔森菌O：9血清型特异性单克隆抗体的杂交瘤细胞株,分泌的抗体与经鉴定的18株小肠结肠炎耶尔森菌O：9血清型菌株呈阳性反应,而与其他血清型小肠结肠炎耶尔森菌及常见肠道致病菌无交叉凝集反应;应用于小肠结肠炎耶尔森菌野生株分型鉴定时,能准确鉴定出其中的24株O：9血清型菌株,与用日本进口分型血清鉴定、PCR毒力基因检测结果相同.结论筛选获得的细胞株产生的单克隆抗体,用于小肠结肠炎耶尔森菌O：9血清型鉴定,具有很好的型特异性,避免了过去免疫血清制备中繁琐的抗原交叉吸收试验.     

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.