微流控芯片和比色法双模同时检测副溶血性弧菌和鼠伤寒沙门氏菌

由病原微生物引起的食源性疾病日益引起全球关注,迫切需要开发同时快速检测多种食源性致病菌的方法.本研究建立了一种基于磁性适配体捕获探针的微流控芯片-比色双模式传感器,实现了副溶血性弧菌(V.P)和鼠伤寒沙门氏菌(S.T)的快速定性和同时定量分析.此探针包括磁珠及其表面修饰的含ATP、待测物适配体及重复G碱基核苷酸片段的D...     

生物标志物用于污染物的快速检测及毒性评价

生物标志物是指示污染物危害效应的生物信号。文中概括性讲述了生物标志物的定义、分类、特点及优越性,介绍了生物标志物常用的检测方法,归纳了生物标志物在快速检测各类环境污染物含量和评价其毒性方面的应用。还讨论了它的缺陷和不足,展望了生物标志物的发展前景。     

新型微流控芯片上鼠伤寒沙门氏菌的免疫捕获与原位荧光检测

食源性致病菌仍然严重影响人类健康,发展中国家情况更为严重,以沙门氏菌所引起的死亡为例,发展中国家的死亡率比发达国家高24%。而现有的细菌检测方法存在成本高、操作复杂或检测灵敏度不理想等缺陷,20世纪90年代发展起来的微流控芯片分析技术,以其独特的优势正逐步成为细胞/细菌的高效分析测试和研究平台。重庆大学化学化工学院徐溢教授课题组将微流控芯片分析技术、MEMS加工技术和纳米探针结合,开展细菌分离、检测、细菌浸染及其群     

污染水产去甲基化表观遗传毒性EGFP评价方法

pEGFP-C3质粒经过体外人工甲基化处理后,被转染进入HepG2细胞以构建重组细胞株．以5-AZA为阳性去甲基化毒物与重组细胞共培养,通过亚硫酸氢钠测序法定量检测EGFP基因启动子区甲基化状态,通过实时定量PCR检测EGFP基因表达,借助流式细胞术和荧光摄片定量检测共培养细胞的绿色荧光强度,在DNA甲基化、EGFP基因mRNA表达、GFP蛋白等多个层次研究5-AZA染毒处理与其去甲基化能力和荧光表达改变的响应关系．对天津污染水产的去甲基化能力进行了实际样品测试．结果表明,5-AZA与重组细胞的DNA甲基化、基因表达、蛋白产物变化之间存在显著关联,具有较低的检出浓度和良好的重复性．天津污染海域的水产去甲基化能力较强．本文初步建立了一种污染物去甲基化表观遗传毒性评价方法．     

食品药品检测实验室质量管理标准化评价方法

分析和总结食品药品检测实验室质量管理标准化的评价指标.以实验室质量管理和检测质量保证为研究对象,采用清单式评价和可回溯性管理等质量控制方法,将实验室管理的全过程进行分解,确保评价指标的全面性.并以某食品检测实验室为例,对实验室的质量管理进行定量分析.实验结果表明该检测项目的总评分为1.227,满足管理指标设定要求.对实...     

LDI学生成绩评价方法的构建——以精细化学品化学课程为例

根据我院化工专业精细化学品化学课程改革的实际,围绕以评促学、以评助教,构建了LDI学生课程成绩评价方法。该方法以教师和学生为评价主体,从3个环节（理论学习、课内实验、创新实践）、4个维度（课前自学、课堂学习、课内实验与创新实践、期末考试）入手,将评价内容分为多个项目来引导学生学习、实践,并全面考核学生的知识与能力。该法不仅能提高学生学习的主动性,还能潜移默化地培养学生的创新意识和团队精神。     

高职院校有机化学课程过程性评价方法探究

针对高职院校目前有机化学课程中过程性评价方法存在的问题,初步探索了由单元知识结构图、创新型作业、期中考试和日常考勤四部分组成的过程性评价方法。实践证明,该评价方案在客观地反映学生平时学习情况的同时,能很好地激励学生自主学习,提升学习效果,促进师生共同进步。     

一种基于纳米金化学发光快速检测三聚氰胺的方法

本发明公开了一种基于纳米金化学发光快速检测三聚氰胺的方法,包括以下步骤:(1)移取纳米金溶液和含有三聚氰胺的溶液于一离心管中,充分混匀,作用5 min使得纳米金达到最佳的团聚态;(2)移取纳米金溶液和三聚氰胺溶液的混合溶液于化学发光池中,采用静态注射的方式注入鲁米诺–过氧化氢化学发光试剂,通过IFFL–D流动注射化学发光分析仪测定并记录其化学发光强度,根据测定结果进行判定。本发明检出限为8.6×10–14 g/m L,达到目前固相萃取–化学发光分析法的灵敏度,且较之简单方便;分析测试时间缩短为10 min以内;整个操作实验条件较温和,有利于自动化的分析操作。     

布绒玩具中致敏性芳香物质的快速筛查方法研究

建立了基于热解吸-气相色谱-质谱联用(TD-GC-MS)测定布绒玩具中38种致敏性芳香剂的快速筛查方法。样品无需前处理,采用直接热解吸进样,经气相色谱质谱测定其在一定条件下的挥发量,外标法定量。各物质线性范围在0.005~10μg(相关系数均大于0.99),定量限(LOQ,S/N=10)在0.05~0.5 mg/kg之间。另外,本研究建立了挥发量与总量之间的相关性方程,实现了通过挥发量对残留量的合理预测。将本方法应用于实际玩具样品的检测,结果满意。     

标准差标准化方法修正的PBBs多效应3D-QSAR模型及其在环境友好性分子修饰中的应用

构建了标准差标准化方法修正的兼具多溴联苯(PBBs)分子红外振动强度、 生物富集性和毒性3种效应的CoMFA模型, 分析了PBBs分子力场对其综合值的影响, 确定取代位点, 并进行兼具易红外光谱检出、 低生物富集性和毒性特征的PBB分子修饰(以PBB-153为例). 研究结果表明, 构建的CoMFA模型对PBBs分子的红外振动强度、 生物富集性和毒性3种效应综合值具有较好的预测和拟合能力, 且具有较好的稳定性, 静电场和立体场的贡献率分别为59.9%和40.1%. 根据模型三维等势图选择正电性高于Br原子的5种取代基团对目标分子PBB-153进行单、 双取代, 筛选出6个3种效应综合值上升的PBB-153衍生物. PBBs衍生物分子单效应计算或预测结果验证表明所构建的兼具PBBs分子红外振动强度、 生物富集性和毒性3种效应综合值的CoMFA模型可以有效应用于PBBs分子的修饰. 设计的PBB-153衍生物分子具有较好的稳定性, 同时阻燃性与目标分子相当, 环境持久性及迁移性方面优于目标分子. 2D-QSAR模型表明, PBBs分子的偶极矩、 最负电荷及邻位Br原子数对其红外振动强度、 生物富集性和毒性单效应和综合值影响趋势一致.     

In-Silico Screening of High Production Volume Chemicals for Mutagenicity using the mcase QSAR Expert System

Computational screening is suggested as a way to set priorities for further testing of high production volume (HPV) chemicals for mutagenicity and other toxic endpoints. Results are presented for batch screening of 2484 HPV chemicals to predict their mutagenicity in Salmonella typhimurium (Ames test). The chemicals were tested against 15 databases for Salmonella strains TA100, TA1535, TA1537, TA97 and TA98, both with metabolic activation (using rat liver and hamster liver S9 mix test) and without metabolic activation. Of the 2484 chemicals, 1868 are predicted to be completely nonmutagenic in all of the 15 data modules and 39 chemicals were found to contain structural fragments outside the knowledge of the expert system and therefore suggested for further evaluation. The remaining 616 chemicals were found to contain different biophores (structural alerts) believed to be linked to mutagenicity. The chemicals were ranked in descending order according to their predicted mutagenic potential and the first 100 chemicals with highest mutagenicity scores are presented. The screening result offers hope that rapid and inexpensive computational methods can aid in prioritizing the testing of HPV chemicals, save time and animals and help to avoid needless expense.     

Immunoassay for Highly Water-Soluble Nitenpyram: Evaluating the Analytical Performance of an Easy-to-Use Screening Method for Agricultural Samples

Abstract

Nitenpyram, a neonicotinoid insecticide, is highly soluble in water (>5.9?×?10⁵?mg/L). Specifically examining its physicochemical properties, we have established a simple and rapid residue analytical method for nitenpyram using an enzyme-linked immunosorbent assay (ELISA) which requires no organic solvents. The I₅₀ value found with this ELISA method using a selective monoclonal antibody toward nitenpyram was 4.8?ng/mL. A hand-shaking method using water was applied for ELISA analysis to extract nitenpyram from fruiting vegetable samples of four kinds. Matrix effects caused by interfering substances coexisting in aqueous sample extracts were reduced by dilution with water. No problems were found in the analytical values obtained using ELISA. From analyses of nitenpyram-incurred fruiting vegetables with the proposed ELISA and the reference HPLC protocols, high mutual correlation was found, which suggests that the measurement accuracy of the proposed ELISA method is comparable to the reference HPLC procedure.     

Methodology for Bioassay-Directed Fractionation Studies of Air Particulate Material and Other Complex Environmental Matrices

Abstract

A normal phase HPLC methodology using a semi-preparative polyaminocyano column in conjunction with a selection of short-term genotoxicity assays has been developed for bioassay-directed fractionation studies of complex environmental mixtures. To illustrate the effectiveness of this methodology, an organic extract prepared from respirable air particulate samples collected in Hamilton, Canada was separated into a non-polar aromatic fraction and a polar aromatic fraction using a combination of alumina and Sephadex LH20 chromatography. These fractions were evaluated for their genotoxic potential using the Salmonella/microsome (Ames) assay with six different strains of Salmonella.

The non-polar aromatic fraction was analyzed by normal phase HPLC and the eluent was collected in one-minute subfractions; these subtractions were bioassayed in three different Salmonella strains (YG1021 -S9, YG1024 -S9 and YG1029 +S9) to afford three different mutation profiles of this sample. Some subfractions which exhibited high mutagenic responses were subjected to further chemical analyses using GC/MS in order to identify those compounds responsible for the genotoxic responses. The nitroarene compounds 2-nitrofluoranthene, 1-nitropyrene and 2-nitropyrene and higher molecular weight polycyclic aromatic hydrocarbons such as benzo[a]pyrene and indeno[l,2,3-cd]pyrene were identified and quantified in some of the biologically active subfractions. The normal phase gradient conditions afforded very reproducible retention times for a series of polycyclic aromatic standards with a broad range of compound polarities. In addition, polycyclic aromatic hydrocarbons (PAH) were observed to elute from the normal phase HPLC column in a series of peaks; successive peaks contained PAH of increasing molecular weight while any individual peak was shown to contain PAH of the same molecular weight.     

Protective Effects of Cinnamaldehyde on the Inflammatory Response,Oxidative Stress,and Apoptosis in Liver of Salmonella typhimurium-Challenged Mice

Salmonella typhimurium infection is associated with gastrointestinal disorder and cellular injury in the liver of both humans and animals. Cinnamaldehyde, the main component of essential oil from cinnamon, has been reported to have anti-inflammatory, anti-oxidative, and anti-apoptotic effects. However, it remains unknown whether cinnamaldehyde can alleviate Salmonella typhimurium infection-induced liver injury in mice. In the present study, we found that cinnamaldehyde attenuated Salmonella typhimurium-induced body weight loss, the increase of organ (liver and spleen) indexes, hepatocyte apoptosis, and the mortality rate in mice. Further study showed that cinnamaldehyde significantly alleviated Salmonella typhimurium-induced liver injury as shown by activities of alanine transaminase, aspartate transaminase, and myeloperoxidase, as well as malondialdehyde. The increased mRNA level of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, and IFN-γ) and chemokines (CCL2 and CCL3) induced by Salmonella typhimurium were significantly abolished by cinnamaldehyde supplementation. These alterations were associated with a regulatory effect of cinnamaldehyde on TLR2, TLR4, and MyD88. 16S rDNA sequence analysis showed that Salmonella typhimurium infection led to upregulation of the abundances of genera Akkermansia, Bacteroides, Alistipes, Muribaculum, and Prevotellaceae UCG-001, and downregulation of the abundances of genera Lactobacillus, Enterorhabdus, and Eggerthellaceae (unclassified). These alterations were reversed by cinnamaldehyde supplementation. In conclusion, cinnamaldehyde attenuated the inflammatory response, oxidative stress, and apoptosis in the liver of Salmonella typhimurium-infected mice. Supplementation of cinnamaldehyde might be a preventive strategy to alleviate liver injury caused by Salmonella typhimurium infection in humans and animals.     

2,8-二氨基-10-羟基-5-甲基-5,10-二氢磷杂吖嗪-10-氧化物的合成及其诱变性研究

在升温条件下,二苯胺和三氯化磷反应,产物经水解、氧化得次膦酸:10-羟基-5,10-二氢磷杂吖嗪-10-氧化物(产率45%).经酰化后,与甲醇钠作用,生成相应的次膦酸甲酯.次膦酸甲酯经NaH处理后,在120℃下发生甲基迁移,形成5-甲基次膦酸(产率62%).用45倍摩尔量的硝化剂将5-甲基次膦酸硝化,得到双硝基产物(产率59%).在5%Pd/C催化下,双硝基产物又被氢气还原.考察催化剂用量对该还原反应的影响,并在最佳催化剂用量时得到2,8-二氨基-10-羟基-5-甲基-5,10-二氢磷杂吖嗪-10-氧化物(产率70%).用NMR,IR和质谱确定了所合成的5个中间体结构.对合成的氨基化合物进行Salmonella/mam-malianmicrosomeassay测试,结果表明,该氨基化合物表现为非诱变性.     

Fast Method for Synthesis of Ylidenemalononitriles

Ylidenemalononitrile can be prepared from aromatic aldehydes and malononitrile by grinding at room temperature, catalyzed by potassium carbonate under solvent‐free conditions.     

A Sensitive Gas Chromatographic Method for the Determination of Metoprolol in Human Plasma

Abstract

A sensitive method for the determination of metoprolol in plasma has been developed. The procedure is based on gas chromatographic measurements of derivatized metoprolol, using 9-bromophenanthrene as internal standard. Metoprolol is derivatized with pentafluoropropionic anhydride. The resulting derivative gives a four-fold increase in sensitivity compared to the published methods where trifluoroacetic anhydride was used for derivatization.     

Fluorometric Method for Determination of Transketolase Activity

Abstract

A new method for rapid and accurate determination of transketolase is studied. The reaction is monitored by coupling the system with the glyceraldehyde-3-phosphate dehydrogenase system and flurometrically measureing the production of NADH. The assay is rapid and convenient. It responds linearly to transketolase activity as low as 0.1 units in three ml of reaction mixture.     

Improvement of the SOS/UMU Test for Preliminary Screning of Mutagens

Abstract

A short-term system to detect environmental mutagens is presented by using a plasmid (pSK1002) carrying a fused gene umuC′-′lacZ introduced into Salmonella typhimurium TA1535. Higher sensitivity, repeatability, and simplicity were achieved when (1) the culture volume taken from 2hrs after treatment with test compound was increased to 1.5ml diluted with distilled water to 2.5ml, (2) the 15μ 1 of 3% SDS solution saturated with chloroform was used as the disruptor of the cell membrane. and (3) an incubation -temperature of 37°C was used. At the above experimental conditions, the time required for the mutagen test was shortened to 3hrs. Based on the results of the present study, it seems that the improved SOS/umu test is useful for screening of mutagenic complex environmental mixtures.     

Fast immobilization of probe beads by dielectrophoresis-controlled adhesion in a versatile microfluidic platform for affinity assay

The use of probe beads for lab-on-chip affinity assays is very interesting from a practical point of view. It is easier to handle and trap beads than molecules in microfluidic systems. We present a method for the immobilization of probe beads at defined areas on a chip using dielectrophoresis (DEP)-controlled adhesion. The method is fast, i.e., it takes between 10 and 120 s--depending on the protocol--to functionalize a chip surface at defined areas. The method is versatile, i.e., it works for beads with different types of probe molecule coatings. The immobilization is irreversible, i.e., the retained beads are able to withstand high flow velocities in a flow-through device even after the DEP voltage is turned off, thus allowing the use of conventional high-conductivity analyte buffers in the following assay procedure. We demonstrate the on-chip immobilization of fluorescent beads coated with biotin, protein A, and goat-antimouse immunoglobulin G (IgG). The number of immobilized beads at an electrode array can be determined from their fluorescence signal. Further, we use this method to demonstrate the detection of streptavidin and mouse IgG. Finally, we demonstrate the feasibility of the parallel detection of different analyte molecules on the same chip.