Development of aquantitative ELISA detection method for Varicella Zoster Virus(VZV) antigen

目的:建立水痘-带状疱疹病毒(VzV)抗原的双抗体夹心ElISA定量检测方法,用于质控VzV灭活疫苗研发和生产中抗原含量。方法:以VzV中和单抗5f6C8为包被抗体、8H5d1为酶标抗体,构建定量检测VzV抗原的双抗体夹心ElISA方法,并对本方法的特异性、灵敏度、准确性、线性和稳定性等性能进行分析。结果:建立的双抗体夹心定量检测VzV抗原的ElISA方法,线性范围为0.4μg~13μg/Ml,相关系数为r2=0.994,定量限度为0.4μg/Ml;变异系数CV80%。与VzV以外的相关病毒样本没有交叉反应。结论:构建的VzV抗原ElISA定量检测方法的各项性能符合定量检测需要,可用于VzV灭活疫苗的研发和生产过程的抗原含量检测。Objective:To develop a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Varicella Zoster Virus(VZV) antigen.This method was used to determine VZV antigen content at each stage of VZV inactived vaccine developing and manufacturing process.Methods: A double antibody sandwich Q-ELISA was developed to determine concentration of VZV antigen,which was based on the the high-affinity neutralizing monoclonal antibodies 5F6C8 as capture antibodies,and 8H5D1 as HRP-labeled antibody.The performance of reagent were evaluated.Results: The Q-ELISA for VZV antigen content was successfully developed.The reagent had good performance.The quantitation scope was 0.4 μg~13 μg/ml,The coefficient correlation was 0.994,the limit of detection was 0.4 μg /ml,the recovery was between 87.5% and 111.6%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and the reagent was no reaction with other sample except VZV antigen.Conclusion: The Q-ELISA for VZV antigen was developed with good specificity,accuracy and stability.The method can be used to determine VZV antigen content during development and production of VZV inactived vaccine

御寒暖胃膏穴位贴敷对胃癌前病变大鼠胃黏膜的影响

目的:研究御寒暖胃膏贴敷胃经穴对慢性萎缩性胃炎癌前病变大鼠胃黏膜的影响,探讨御寒暖胃膏贴敷胃经穴对慢性萎缩性胃炎癌前病变大鼠胃黏膜损伤修复的作用机制。方法:大鼠随机分为正常组、模型组、御寒暖胃膏贴敷胃经穴组、御寒暖胃膏贴敷对照点组、药物对照组,采用综合干预方法复制慢性萎缩性胃炎癌前病变大鼠模型,肉眼下观察大鼠胃黏膜损伤指数,光镜下观察胃黏膜组织的病理变化,彩色多普勒观察胃黏膜的血流量。结果:与模型组比较,御寒暖胃膏贴敷胃经穴组和药物对照组大鼠胃黏膜损伤指数值均显著降低(P0.05),胃黏膜组织的病理损伤未得到明显修复,胃黏膜血流量未见显著升高(P>0.05)。结论:御寒暖胃膏贴敷胃经穴可以促进慢性萎缩性胃炎癌前病变大鼠胃黏膜损伤的修复,增加胃黏膜的血流量,并且存在经脉-脏腑特异相关性。深圳市科技研发资金项目(JCYJ20130401105615482

The Research Survey of the Degrading-bacteria and Degrading-enzyme of Synthetic Pyrethroid Insecticides

通讯作者：Tel: 0592-2186195; : [email protected][中文文摘]拟除虫菊酯类农药目前已成为我国出口的蔬菜、水果中主要的3类农药残留之一,引起急慢性中毒事件也越来越多,对人类、水生生物和自然环境造成很大危险。而农药生物降解作为去除农药污染的有效手段,逐渐成为环境科学研究的热点。重点综述了拟除虫菊酯类农药降解菌的分离、降解酶的提取、纯化、降解机理、固定化研究,并对以后要解决的问题进行了展望。[英文文摘]Synthetic pyrethroid insecticides have become one of the three major kinds of pesticide residues in the exported vegetabes and fruits in China nowadays. They also caused more and more acute or chronic poisoning affair and do great damage to human beings, aquatic biology and natural environment. As an efficient way to remove pesticides pollution, the biodegradation of pesticides has gradually become the hot issue in the field of environmental sciences. The study on the separation of degrading-bacteria and the extraction, purification, mechanism and immobilization of the degrading-enzyme of pyrethroids is reviewed in this paper. And the problems under resolved in the future work are also prospected.福建省青年科技人才创新项目(No.2007F3094);厦门大学引进人才科研启动费项目(No.0000-X071C3);厦门大学近海海洋环境科学国家重点实验室开放基金(No.MEL0603);国家海洋局近岸海域生态环境重点实验室开放基金(No.200702);国家海洋局第三海洋研究所海洋生物遗传资源重点实验室开放研究基金(No.HY200601

Advance in new determination methods for pesticide residues

第一作者简介：王兆守（1972—），男，博士，讲师。研究方向为有机 污染物的生物降解。电话 0592–2186195；E–mail [email protected]。 联系人：王兆守。[中文文摘]介绍了农药残留检测的步骤与方法。农药残留检测主要分为4个步骤:样品的提取、净化、浓缩和检测。提取方法主要有震荡法、索式提取法、固相微萃取法、超临界流体萃取法、快速溶剂萃取法等。净化方法主要有:液-液分配净化法、柱层析法、磺化法等。浓缩方法主要有:蒸发浓缩、反渗透浓缩、K-D浓缩仪浓缩等。检测方法主要有:气相色谱-质谱联用法、荧光分析法、酶抑制法、免疫分析法、生物传感器检测法、红外光谱法、拉曼光谱法等。评述了这些方法的优点与缺陷,提出了今后的发展方向。[英文文摘]The determination of residual pesticides is mainly divided into four processes:extraction,cleanup,concentration and detection. The methods for extracting samples include shaking extraction,soxhlet’s extraction,solid phase micro-extraction,supercritical fluid extraction，accelerated solvent extraction and so on. The cleanup methods include liquid-liquid partition process,column chromatography,sulfonation and so on. The concentration methods include evaporation and concentration,reverse-osmosis，Kuderna Danish (K-D) concentrator and so on. The detection methods include gas chromatography-mass ectrometry,fluorometry,enzyme inhibition,immunoassay, biosensor and infrared spectrometry,Raman spectrometry,and so on. The advantages and disadvantages of these methods were reviewed and the development tendency in the future was pointed out.福建省青年科技人才创新项目(2007F3094);厦门大学引进人才科研启动费项目(0000-X071C3);近海海洋环境科学国家重点实验室(厦门大学)开放基金(MEL0603);国家海洋局第三海洋研究所海洋生物遗传资源重点实验室开放研究基金(HY200601);国家海洋局近岸海域生态环境重点实验室开放基金(200702

Isolation and identification of a hydrocarbon-degrading bacterium Gordonia sp. S14-10 from the surface water of Atlantic Ocean and analysis on its related characteristics

【目的】本研究的目的是从大西洋表层海水分离筛选新的烷烃降解菌,了解其降解基因及降解特性,为海洋石油污染的生物治理提供材料。【方法】以柴油与原油作为混合碳源从大西洋表层海水样品中富集、并分离筛选出降解能力较强的烷烃降解菌。根据16SrrnA基因和其看家基因SECA1序列确定其系统进化地位。分析了烷烃降解范围、表面活性剂产生能力及其他生理生化特性;利用已报道的兼并引物进行了烷烃羟化酶基因的PCr扩增及系统进化分析。【结果】分离筛选得到1株能够降解C10(C36直链烷烃的菌株S14-10。经16SrrnA基因序列系统发育分析表明它属于戈登氏菌属(gOrdOnIA),与gOrdOnIATErrAET同源性为99.86%,但SECA1序列相似度仅为93.69%,可能为戈登氏属的一个新种。从该菌株中扩增得到了烷烃单加氧酶Alkb和P450基因片段,它们与菌株gOrdOnIASP.Tf6相应的基因序列具有最高相似度,分别为88.76%和99.61%。该菌能够产生糖脂类生物表面活性剂,将发酵液的表面张力降低到31.6Mn/M。【结论】首次从大洋环境中发现了gOrdOnIA属的烷烃降解菌,可能为该属的一个新种。菌株S14-10至少编码两套与烷烃降解相关的单加氧酶系统,对各种直链烷烃降解范围宽,能产表面活性剂,在海洋油溢污染的生物修复中有较好的应用前景。[Objective] The aim of this study is to isolate novel and efficient hydrocarbon-degrading bacteria(HDBs)from pelagic ocean.[Methods] The surface water sample from Atlantic Ocean was enriched in NH medium with crude oil:diesel oil(in a ratio of 1∶1)as the sole carbon source.HDBs were isolated and their degradation ability was tested in MMC medium,and further subjected to genotypic and phenotypic analysis.Polymerase chain reaction with degenerate primers was performed to detect the genes encoding integral-membrane non-heme iron monooxygenase(AlkB)and cytochrome P450 CYP153 family.Meanwhile,the production of biosurfactant was examined by surface tension measurement,then extracted and purified for component characterization.[Results] A hydrocarbon-degrading bacterium named S14-10 was obtained,which can utilize C_ 10 ～C_ 36 as the sole carbon source.Analysis of 16S rRNA sequence showed that it belonged to Gordonia genus showing the highest similarity(99.86%)with type strain of Gordonia terraeT,while secA1 sequence showed a low identity only 93.69%.Furthermore,two genes alkB and CYP153 P450 were obtained with PCR,which had highest similarities 88.76%and 99.61% to that of Gordonia sp.TF6,respectively.In addition,strain S14-10 was found to produce glycolipid biosurfactant,which lowered the surface tension of water to 31.6 mN/m.[Conclusion] Strain S14-10 is possibly a novel species of Gordonia,and the first hydrocarbon-degrading bacterium isolated from open sea.It has potential in bioremediation of oil contaminated environment.国家自然科学基金(30670051);国家自然科技资源共享平台建设项目(2005DKA21209);中国大洋协会项目(DYXM-115-02-2-05);福建省科技项目(2009H0029)---

斑马鱼神经组织特异性增强子及其克隆和应用

本发明公开了一种斑马鱼神经组织特异性增强子及其克隆和应用，涉及一种组织和器官特异性表达基因的增强子捕获技术领域，用于荧光标记斑马鱼胚胎发育过程中的神经细胞。斑马鱼神经组织特异性增强子，其序列为SEQ ID NO.2所示的核苷酸序列。其应用是：①研制神经组织特异表达荧光的转基因鱼，可用于神经器官再生过程研究；②用于驱动任意目的基因在神经组织特异表达，为研究者研究神经系统发育及相关疾病治疗提供有力工具。本发明可用于斑马鱼眼睛发育及损伤再生过程的研究；有助于研究特定基因在神经系统发育及调控方面的作用，为相关疾病治疗提供新材料。</p