2010年上海市食源性沙门菌菌型分布和药敏分析以及快速检测方法的建立

目的：通过对2010年上海市市售食品中食源性沙门菌污染状况、菌型分布及药敏试验进行分析,初步确定该地区食源性沙门菌的菌型分布及耐药性,为防治因沙门菌感染引起的食源性疾病提供科学依据,并摸索建立了针对沙门菌的快速检测方法。方法：对农贸市场和超市的5类840份市售食品进行沙门菌分离鉴定及耐药性分析,并根据沙门菌的侵袭蛋白A（invA）基因序列设计保守引物,特异性快速检测沙门菌。结果：本次市售食品沙门菌阳性率检出率为4．29％,共36株沙门菌,生禽畜肉所占比例高达91％。主要血型为鼠伤寒沙门菌,德尔卑沙门菌和都柏林沙门菌。36株沙门菌药敏分析显示：所有菌株对头孢类等抗生紊敏感较高,但对氨苄西林、麦迪霉素、环丙沙辛、呋喃妥因等存在普遍较多耐药菌株,并在此基础上通过PCR法成功地特异性检测出沙门菌。结论：上海市市售食品沙门菌污染以生禽畜肉为主,对抗菌药物的耐药性较高。目前治疗市售食品中食源性沙门菌引起的食源性疾病应首选头孢内等抗生素。另外PCR快速检测方法也操作简单,特异性强,灵敏度高,对食品中沙门菌污染能起到快速检测和监控     

PAX3基因及其蛋白的生物信息学分析

目的：对PAX3基因和PAX3蛋白进行生物信息学分析,更多的了解该基因的相关信息,为进一步研究PAX3与神经管畸形的相关性研究提供基础。方法：运用生物信息学方法对PAX3基因的基因结构、单核苷酸多态性位点（SNP）、PAX3基因与其他基因的相互作用网络、PAX3蛋白结构域、蛋白二级结构、蛋白间相互作用网络、以及PAX3蛋白所调控和影响的靶基因进行分析。结果：PAX3基因有9中可变剪切形式,编码区存在14个SNP位点,其中错意突变13个,移码突变1个。PAX3蛋白由479个氨基酸组成,分子量52968Da,PAX3蛋白可能调控和影响151个靶基因的转录和表达,与PAX3基因存在相互作用的基因和与PAX3蛋白存在相互作用的蛋白多数与发育相关。结论：通过对PAX3基因和PAX3蛋白的生物信息学分析获得了其相应的分子生物学特征,为进一步研究提供基础。     

Epidemiology and Mutational Analysis of Global Strains of Crimean-Congo Haemorrhagic Fever Virus

Crimean-Congo hemorrhagic fever (CCHF) is a severe illness with high fatality.Cases are reported in several countries in Africa,Europe,the Middle East,and Asia.Phylogenetic analyses based on the virus S (nucleocapsid),M (glycoprotein),and L (polymerase) genome segments sequences indicate distinct geographic lineages exist but their specific genetic characteristics require elucidation.In this work we collected all full length S segment sequences and generated a phylogenetic tree based on the alignment of these 62 samples.We then analyzed the alignment using entries from AAIndex,the Amino Acid Index database,to identify amino acid mutations that performed significant changes in charge,pka,hydropathy and side chain volume.Finally,we mapped these changes back to the tree and alignment to identify correlated mutations or sites that characterized a specific lineage.Based on this analysis we are able to propose a number of sites that appear to be important for virus function and which would be good candidates for experimental mutational analysis studies.     

Shift in chicken intestinal gene association networks after infection with Salmonella

A primary infection of Salmonella enteritidis causes a spatial-temporal dependent change in the gene expression patterns in the intestine of chickens (Gallus gallus). This is the result of a dynamic intestinal response to adapt to the altered environment and to optimize its ‘health’ and functionality under the new circumstances. By inferring gene association networks (GANs), the complexities of and changes in biological networks can be uncovered. Within such GANs highly interacting (hub) genes can be identified, which are supposed to be high-level regulators connected to multiple processes. By exploring the intestinal expression of genes differing between control and Salmonella infected chicken in a time-dependent manner differences in GANs were found. In control chickens more developmental processes were observed, whereas in infected chickens relatively more processes were associated to ‘defense/pathogen response’. Moreover the conserved protein domains of the identified hub genes in controls were nuclear-associated, whereas hub genes in infected chickens were involved in ‘cellular communication’. The shift in topology and functionality of the intestinal GANs in control and Salmonella infected animals and the identification of GAN-specific hubs is a first step to understand the complexity of biological networks and processes regulating intestinal health and functionality under normal and disturbed conditions.     

Comparative proteomics using 2-D gel electrophoresis and mass spectrometry as tools to dissect stimulons and regulons in bacteria with sequenced or partially sequenced genomes

We propose two-dimensional gel electrophoresis (2-DE) and mass spectrometry to define the protein components of regulons and stimulons in bacteria, including those organisms where genome sequencing is still in progress. The basic 2-DE protocol allows high resolution and reproducibility and enables the direct comparison of hundreds or even thousands of proteins simultaneously. To identify proteins that comprise stimulons and regulons, peptide mass fingerprint (PMF) with matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS) analysis is the first option and, if results from this tool are insufficient, complementary data obtained with electrospray ionization tandem-MS (ESI-MS/MS) may permit successful protein identification. ESI-MS/MS and MALDI-TOF-MS provide complementary data sets, and so a more comprehensive coverage of a proteome can be obtained using both techniques with the same sample, especially when few sequenced proteins of a particular organism exist or genome sequencing is still in progress.     

Chromatographic analysis and sequencing approach of heparin oligosaccharides using cetyltrimethylammonium dynamically coated stationary phases

C(18) and C(8) bonded stationary phases dynamically coated with cetyltrimethylammonium (CTA) and strong anion exchange (SAX) were developed to obtain separations of oligosaccharide mixtures resulting from chemical or enzymatic depolymerization of heparin. With this method, the retention of sulfated oligosaccharides is directly adjustable depending on the amount of CTA adsorbed into the column. Oligosaccharides containing up to 20 sulfates were separated with a resolving power superior to that of conventional SAX analysis. The stability of the column coating enables hundreds of injections. Using ammonium methane sulfonate aqueous solutions as ultraviolet transparent mobile phases, it was possible to set up double detection, including selective detection of acetylated oligosaccharides. Analytical gel permeation chromatography was directly coupled to CTA-SAX, obtaining a two-dimensional profile of analyzed oligosaccharidic mixtures. A sequencing method of heparin oligosaccharides using partial depolymerization by heparinases according to their size and sulfation pattern and digest analysis by CTA-SAX was developed. A direct application of this method to the analysis of oligosaccharide mixtures obtained by complete digestion of heparins by heparinases I, II, and III was done. It allowed a reliable quantification of heparin building blocks. We also focused our attention on di- and tetrasaccharidic species containing the 3-O-sulfated glucosamines taken as markers of the active sites for antithrombin III. The method was also applied to more complex mixtures resulting from porcine heparin partially depolymerized with heparinase I. The specificity of the reaction was studied up to decasaccharidic fractions.     

Relationships between inhibitory activity against a cancer cell line panel, profiles of plants collected, and compound classes isolated in an anticancer drug discovery project

In an attempt to determine the relationships between the plant profiles (country of collection, taxonomy, plant part) and the compound classes isolated with cytotoxic activity against a panel of human tumor cell lines, the data compiled from a 15-year anticancer drug-discovery project were subjected to an analysis of variance (ANOVA). The results indicate significant trends in cytotoxic activity relative to collection location, taxonomy, plant part, and compound classes isolated. Plant collections were made in tropical forests in six countries, with collections from Ecuador resulting in higher activity than those from Indonesia and Peru. Interestingly, collections from Florida were not statistically different than those from the countries with higher biodiversity. One hundred and forty-five families were represented in the collections, with the Clusiaceae, Elaeocarpaceae, Meliaceae, and Rubiaceae having low ED50 (half maximal effective dose) values. Especially active genera included Aglaia, Casearia, Exostema, Mallotus, and Trichosanthes. Roots and below-ground plant materials were significantly more active than above-ground materials. Cucurbitacins, flavaglines, anthraquinones, fatty acids, tropane alkaloids, lignans, and sesquiterpenoids were significantly more active than xanthones and oligorhamnosides. The results from this study should serve as a guide for future plant collection endeavors for anticancer drug discovery.     

黄酮化合物色谱保留时间与其三维结构的关系研究

利用比较分子相似性指数分析（CoMSIA）方法,结合黄酮类化合物含有较多羟基、易形成较强分子内氢键的特点,建立了黄酮类化合物色谱保留时间与其三维结构的关系模型,以探讨黄酮类化合物色谱保留时间预测的新方法。模型交叉验证相关系数q2值为0．705,非交叉验证相关系数r2为0．981,表明模型具有较好的预测能力。该研究结果对进一步开展黄酮类化合物液相色谱保留参数与三维结构关系的研究提供了思路和方法。     

The fate of sheep-dispersed seeds: Plant species emergence and spatial patterns

Sheep epizoochory has often been proposed as an important vector which can help to overcome the dispersal limitation of plants in fragmented landscapes. In order to evaluate the contribution of herbivores to recruitment especially of target species, the dispersal and post-dispersal fate of such seeds must be known. In a field experiment sheep with seeds of mainly target species (experimentally attached to their coats) were present at three sand plots for 24 h. Natural epizoochorous dispersal was already shown for most of the species in our study area. Seed detachment, trampling intensity and seed shadow were measured; seedling emergence and survival were recorded over an 8-month period. In addition, the effect of sheep trampling on seedling emergence and survival of two threatened species, Jurinea cyanoides and Koeleria glauca, were studied.     

基于飞行质谱分析技术对小鼠肠道菌群蛋白组分分析的初步探讨

目的应用飞行质谱分析技术对小鼠肠道菌群蛋白组分进行分析,探讨肠道菌群定性、定量和蛋白组学分析方法。方法实验分为抗生素干预组与正常对照组。抗生素干预组小鼠用400 mg/ml头孢曲松钠灌服,每次0.2 ml,每日2次,间隔6 h,连续8 d。收集2组小鼠盲肠内容物菌群细胞,采用飞行质谱分析技术进行肠道菌群蛋白组分分析,采用传统培养法对2组优势菌群进行定量分析。结果抗生素干预组的肠道菌群菌细胞的有效蛋白峰出峰的数量与正常对照组比对差异有统计学意义,2组小鼠的肠道菌群蛋白成分有明显的不同。抗生素干预组的肠道优势菌群数量显著低于正常对照组。结论应用飞行质谱分析技术可以快速明显区分抗生素干预小鼠与正常小鼠的肠道菌群,对肠道菌群研究和临床合理应用抗生素具有指导意义。