果胶甲酯酶的结构与功能研究进展

果胶甲酯酶(PME)是一种重要的果胶酶,其水解果胶中的甲酯基从而释放甲醇并降低果胶的甲酯化程度。目前在食品加工、茶饮料、造纸等生产工艺中有着广泛的应用前景。随着对PME的深入研究,已报道了几种不同来源的酶晶体结构,对这些已获得的晶体结构进行分析发现,PME属于右手平行β-螺旋结构,其催化残基为2个保守的天冬氨酸和1个谷氨酰胺残基,并且在催化过程中分别起到了一般酸碱、亲核试剂以及稳定中间体的作用。同时对其底物特异性进行分析,初步了解其底物与活性位点的识别机制。文中针对这几个相关方面进行了系统的综述。     

A baseline epidemiological study of the co-infection of enteric protozoans with human immunodeficiency virus among men who have sex with men from Northeast China

BackgroundHuman immunodeficiency virus (HIV) and enteric parasite co-infection not only aggravates the clinical symptoms of parasites but also accelerates acquired immunodeficiency syndrome (AIDS) progression. However, co-infection research on men who have sex with men (MSM), the predominant high-risk population of HIV/AIDS in China, is still limited. In this study, we investigated the epidemiology of enteric parasites, risk factors, and associations with clinical significance in an MSM HIV/AIDS population in Heilongjiang Province, northeast China.MethodsWe recruited 308 MSMs HIV/AIDS patients and 199 HIV-negative individuals in two designated AIDS hospitals in Heilongjiang between April 2016 and July 2017. Fresh stool samples were collected. DNA extraction, molecular identification, and genotyping of Cryptosporidium species, Entamoeba histolytica, Cyclospora cayetanensis, Enterocytozoon bieneusi, and Blastocystis hominis were performed. Fourteen diarrhea-related pathogens were examined to exclude the influence of other bacterial pathogens on diarrhea incidence.Results31.5% of MSM HIV/AIDS participants were infected with at least one parasite species, a significantly higher proportion than that found in the HIV-negative individuals (2.5%). E. bieneusi presented the highest prevalence, followed by B. hominis, E. histolytica, Cryptosporidium spp., and C. cayetanensis. Warm seasons were the risk factor for parasitic infections in this population [odds ratio (OR) = 2.6, 95% CI: 1.47–4.57]. In addition, these individuals showed a higher proportion (35.8%) of present diarrhea (PD) compared with men who have sex with women (MSW) with HIV/AIDS (16.7%). The infection proportions of both Cryptosporidium spp. and E. histolytica were significantly higher in the PD. E. bieneusi infection was more prevalent in the historic diarrhea (HD) group. CD4⁺ T cell counts in the MSM patients with the above three parasites were significantly lower. New species and genotypes were found, and MSM patients had a wider range of species or genotypes.ConclusionsEnteric parasitic infection was prevalent in the MSM HIV/AIDS population, especially in patients with present diarrhea during warm seasons. E. histolytica and B. hominis should also be considered high-risk parasites for opportunistic infections in AIDS patients in addition to Cryptosporidium spp.     

青藏高原1979-2007年间的积雪变化

利用雪深被动微波遥感数据产品,对青藏高原1979—2007年积雪深度、积雪日数的分布变化及其趋势进行了分析。结果表明:青藏高原积雪日数、积雪深度和海拔三者之间在空间上具有显著正相关;青藏高原积雪在1988年发生突变,该年前后积雪分布有显著不同,这与20世纪80年代中后期青藏高原由暖干时期进入暖湿时期有关;将青藏高原按夏季水汽来源不同将其分为南北两部分,发现29年来北部积雪日数变化与全国积雪变化相反呈极显著增加趋势(R2=0.39,P0.01),以1.40 d/a的趋势增加,主要原因是西北部地区冬季积雪日数增加;南部积雪深度与全国积雪变化一致呈极显著减少趋势(R2=0.24,P0.01),以-0.04 cm/a的趋势减少,主要原因是东南部春、夏、秋三季积雪深度减少。     

Direct competitive chemiluminescence immunoassays based on gold‐coated magnetic particles for detection of chloramphenicol

Direct competitive chemiluminescence immunoassays (CLIA) based on gold‐coated magnetic nanospheres (Au‐MNPs) were developed for rapid analysis of chloramphenicol (CAP). The Au‐MNPs were modified with carboxyl groups and amino groups by 11‐mercaptoundecanoic acid (MUA) and cysteamine respectively, and then were respectively conjugated with CAP base and CAP succinate via an activating reaction using 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide hydrochloride (EDC) and N‐hydroxysuccinimide (NHS). NSP‐DMAE‐NHS, a new and effective luminescence reagent, was employed to label anti‐CAP antibody (mAb) as a tracer in direct CLIA for CAP detection using a ‘homemade’ luminescent measurement system that was set up with a photomultiplier tube (PMT) and a photon counting unit linked to a computer. The sensitivities and limits of detection (LODs) of the two methods were obtained and compared according to the inhibition curves. The 50% inhibition concentration (IC₅₀) values of the two methods were about 0.044 ng/mL and 0.072 ng/mL respectively and LODs were approximately 0.001 ng/mL and 0.006 ng/mL respectively. To our knowledge, they were much more sensitive than any traditional enzyme‐linked immunosorbent assay (ELISA) ever reported. Moreover, the new luminescence reagent NSP‐DMAE‐NHS is much more sensitive and stable than luminol and its derivatives, contributing to the sensitivity enhancement. Copyright © 2015 John Wiley & Sons, Ltd.     

利用SRAP标记分析中国主栽孔雀草品种的遗传多样性

为了解中国主栽孔雀草品种的遗传背景,采用相关序列扩增多态性(SRAP)分子标记分析了28份孔雀草材料的遗传多样性。14对SRAP引物组合共获得271个位点,其中多态位点151个,占55.72%。每对引物可扩增出14~24条DNA片段,平均19.4条。引物的多态信息含量PIC值在0.693~0.967之间,平均为0.909;每个材料得到的多态性条带比例介于38.78%与51.42%之间,平均46.38%,说明SRAP分子标记可有效鉴别孔雀草种质在分子水平上的遗传变异。品种间的遗传距离值在0.047~0.198之间,平均为0.126;Shannon多样性指数变化于0.178~0.217之间,平均0.201,表明参试的孔雀草材料总体的遗传多样性水平较低。UPGMA聚类后,在遗传距离阈值为0.146处,可将28份材料分为4大类群,与花色表现基本相符,花色可考虑作为孔雀草基于表型分类的主要因子。本研究结果对孔雀草品种鉴定、杂交育种中亲本选配和分子标记辅助选择具有重要意义。     

Rapid isolation and characterization of polymorphic microsatellite loci in the mud crab, Scylla paramamosain (Portunidae)

Scylla paramamosain is a widespread and commercially important species of coastal marine crab. We identified 13 polymorphic microsatellite loci from a genome library constructed with 5'-anchored PCR method. Thirty-two S. paramamosain from the East China Sea were used to analyze the characteristics of these loci. The number of alleles per locus ranged from 3 to 8, with a mean of 5.923. Observed and expected heterozygosities ranged from 0.500 to 0.875 and from 0.500 to 0.859, respectively. Eleven of the 13 loci were highly polymorphic (polymorphic information content >0.5). All of the 13 novel loci were in Hardy-Weinberg equilibrium after Bonferroni's correction (P < 0.0038). There was no null allele, stuttering errors or evidence of allelic dropout in any of the loci analyzed by MICRO-CHECKER. According to pairwise tests, no significant linkage disequilibrium was found among the 13 loci (P < 0.0038, adjusted value). These novel developed microsatellites will be useful for studies of genetic variation, population structure, conservation genetics, and molecular-assisted selective breeding of S. paramamosain.     

2010年山东省环境污水中脊髓灰质炎病毒的监测及其基因特征分析

本研究在山东省开展了脊髓灰质炎病毒(Poliovirus,PV)的外环境监测,从济南、临沂两地采集污水标本,浓缩处理后进行病毒分离,对分离到的PV采用中和试验进行血清定型,并对其VP1及3D区进行序列测定,分析其基因突变和重组情况。2010年,共采集污水标本32份,PV阳性10份,阳性率31.3%;分离到18株PV(PV1型3株,PV2型9株,PV3型6株),均为疫苗相关株,VP1完整编码区核苷酸变异数在0~4个之间,在3株PV2型病毒和4株PV3型病毒的基因组中发现重组;对VP1区影响神经毒力的减毒位点分析发现,PV1型病毒中有1株在nt 2 749发生突变(A→G),PV2型病毒中有1株在nt2 908发生A→G突变,3株在nt2 909发生U→C突变,6株PV3型病毒全部在nt2 493发生C→U突变。环境污水中可以分离到PV,其基因重组率和主要减毒位点的回复突变率较高,未发现脊灰野毒株和疫苗衍生株脊灰病毒(Vaccine-derived poliovirus,VDPV)。     

微核技术研究进展

微核试验作为检测染色体损伤最常用而有效的细胞遗传学检测方法之一,经济、简便、快速,在敏感性、特异性和准确性方面,与经典的染色体畸变分析方法基本相当。主要综述国内外微核检测技术的最新研究进展,尤其是微核的自动化检测技术。其中流式细胞仪自动化检测和激光扫描细胞仪自动化检测,以及微核试验高内涵筛选方法由于其特有的优势,应用和发展前景广阔。     

一株广谱中和抗原性出血热病毒株的发现

一株分离自杭州市褐家鼠的出血热病毒Gou_3株的免疫血清对10株I型病毒的中和滴度除二株为160外均为320,而对4株Ⅱ型病毒的滴度为320—640,说明Gou_3株免疫血清对两型毒株中和效价大多数无差异或只差2倍,是一株中和抗原广谱的毒株。用I型和Ⅱ型毒株免疫血清对Gou_3株进行型别检定结果表明Gou_3株是Ⅱ型病毒。