中药复方对抗生素相关性腹泻微生态药理的研究进展

抗生素相关性腹泻指应用抗生素后继发腹泻,为一种严重的药源性腹泻病,具传染性和普遍性,对人们的健康危害严重,中药复方在该病的治疗中疗效显著且具更合理的治疗理念。基于目前中药复方对AAD的微生态药理学尚无综合报道,本文综述了研究领域的最新进展,并结合本人正开展的工作对未来的研究进行了展望。     

320例腹泻患者病原菌分离及流行病学分析

目的了解广州市2009年至2010年肠道门诊就诊的细菌性腹泻患者病原谱的分布情况,为制定针对重点人群肠道传染病防治策略提供依据。方法收集2009年5月至2010年5月暨南大学附属第一医院腹泻患者的粪便标本,使用卡-布运送培养基,增菌培养后,采用生化反应和氧化酶试验进行菌株鉴定,并用梅里埃API生化板条进行验证,用病原菌诊断血清进行细菌分型。结果从320份粪便标本中分离到38株菌株,其中沙门菌15株,产毒大肠埃希菌12株,致病大肠埃希菌6株,志贺菌2株,出血性大肠埃希菌、霍乱弧菌、气单胞菌各1株。0—20岁年龄段高发,以1岁以内婴幼儿为主;7—10月为发病高峰期。结论来该院肠道门诊就诊的细菌性腹泻患者,其病原体以沙门菌为主,其次为产毒大肠埃希菌。因此．广州地区细菌件腹泻的预防．廊右针对件的面向雷占人群和重占病厢莴．     

凝结芽孢杆菌活菌片预防小儿肺炎继发性腹泻疗效观察

目的观察和评价凝结芽孢杆菌活菌片（商品名：爽舒宝）与抗菌药联用预防小儿肺炎继发性腹泻的临床疗效。方法将164例肺炎患儿随机分为预防组和对照组,预防组84例,对照组80例,两组均给予抗菌药及对症支持治疗。其中预防组在治疗的同时序贯（间隔2～3h）应用凝结芽孢杆菌活菌片,出现腹泻继续服用;对照组单用抗菌药,出现腹泻用凝结芽孢杆菌活菌片治疗。对两组继发性腹泻的发生率、腹泻持续天数、肺炎治疗的总疗程、腹痛和肠鸣音异常等症状和体征进行统计分析。结果预防组继发腹泻10例,发生率11．9％,对照组继发腹泻41例,发生率51．3％,两组相比差异具有统计学意义（P〈0．01）;预防组腹泻持续时间和治疗疗程显著短于对照组（P〈0．05）;预防组患儿腹痛、肠鸣音异常和脱水等症状和体征发生率均少于对照组（P〈0．05）。结论凝结芽孢杆菌活菌片与抗菌药序贯应用治疗肺炎,能显著降低继发性腹泻的发生率,缩短疗程,值得临床推广。     

An overview of atypical enteropathogenic Escherichia coli

The enteropathogenic Escherichia coli (EPEC) pathotype is currently divided into two groups, typical EPEC (tEPEC) and atypical EPEC (aEPEC). The property that distinguishes these two groups is the presence of the EPEC adherence factor plasmid, which is only found in tEPEC. aEPEC strains are emerging enteropathogens that have been detected worldwide. Herein, we review the serotypes, virulence properties, genetic relationships, epidemiology, reservoir and diagnosis of aEPEC, including those strains not belonging to the classical EPEC serogroups (nonclassical EPEC serogroups). The large variety of serotypes and genetic virulence properties of aEPEC strains from nonclassical EPEC serogroups makes it difficult to determine which strains are truly pathogenic.     

表面表达猪流行性腹泻病毒核蛋白蛋白的重组干酪乳杆菌诱导产生的免疫应答

为探讨表达猪流行性腹泻病毒(PEDV)核蛋白(N)基因的重组干酪乳杆菌口服免疫小鼠后诱导特异性免疫应答,本研究制备表达流行性腹泻病毒核蛋白的重组干酪乳杆菌,应用Western blotting、间接免疫荧光和全细胞ELISA鉴定目的蛋白的表达。然后用该重组干酪乳杆菌口服免疫BALB/c小鼠,分别测定了免疫后不同时间血清中特异性IgG、粪便中特异性的sIgA水平以及血清的中和活性;并测定免疫小鼠脾淋巴细胞增殖情况和细胞因子水平。结果显示,目的蛋白表达在细胞表面,可被阳性血清所识别。免疫小鼠后,可分别在血清中和粪便中检测到较高水平特异性IgG、sIgA(P<0.01),但血清并没有中和活性;淋巴细胞增殖试验和细胞因子测定结果显示,免疫组可产生明显的细胞免疫应答。结果表明,该重组干酪乳杆菌表达系统可诱导小鼠产生黏膜免疫应答和系统免疫应答,具有作为口服疫苗潜在的应用价值。     

Bovine viral diarrhea virus (BVDV) associated with single in vivo-derived and in vitro-produced preimplantation bovine embryos following artificial exposure

The objective was to determine the average amount of bovine viral diarrhea virus (BVDV) associated with single in vivo-derived and in vitro-produced bovine embryos following recommended processing procedures for embryos. In vivo-derived and in vitro-produced bovine embryos at 7 d post-fertilization were exposed (for 2 h) to 2 × 10^5-7 cell culture infective dose (CCID₅₀)/mL of SD-1 (a noncytopathic, Type 1a strain of BVDV), and then washed according to International Embryo Transfer Society (IETS) guidelines prior to testing. Of the 87 in vivo-derived embryos tested, 27% were positive for virus by quantitative polymerase chain reaction (qPCR). The range in amount of virus associated with 99% of the contaminated embryos was ≤6.62 ± 1.57 copies/5 μL; 90% of the contaminated embryos had ≤4.64 ± 1.57 viral copies/5 μL of embryo-associated virus, using tolerance intervals (P < 0.05). The SEM was 0.33 and the mean of averages was 1.12/5 μL. Of the 87 in vitro-produced embryos, 42% were positive for virus. The range in amount of virus associated with 99% of the contaminated embryos was ≤3.44 ± 0.89 copies/5 μL; 90% of the contaminated embryos had ≤2.40 ± 0.89 viral copies/5 μL of embryo-associated virus using tolerance intervals (P < 0.05; S.E.M. was 0.14 and the mean of averages was 0.55/5 μL). Therefore, although many embryos were positive for virus, there were limited numbers of copies, thereby posing doubt regarding their potential for contamination following embryo transfer.     

猪流行性腹泻抗原基因在乳酸菌中的表达及优化

目的：优化猪流行性腹泻抗原基因COE在乳酸乳球菌中的表达。方法：将表达猪流行性腹泻抗原基因COE的重组乳酸菌活化,酶切鉴定其稳定性,然后设计实验分别从pH值、温度、Nisin浓度、诱导时间、菌体密度等条件对COE表达进行优化,SDS-PAGE检测表达效果。结果：COE在乳酸乳球菌中的最佳表达条件为pH 7.0、T（温度）=30℃、Nisin=2ng/mLt、（诱导时间）=4h和OD600=0.5,在以上条件下相对表达量分别达到了15.48%、15.05%、15.82%、14.72%和20.47%。在最佳表达条件下得到SOE的相对表达含量达到21%。结论：COE重组质粒稳定,其在乳酸菌中经优化表达后可为今后研制猪流行性腹泻乳酸菌疫苗提供数据。     

牛病毒性腹泻病毒Erns蛋白的中华仓鼠卵巢细胞表达及免疫原性分析

本研究利用中华仓鼠卵巢(Chinese hamster ovary,CHO)细胞表达系统制备牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV) E^rns蛋白,并分析其免疫原性。以BVDV-1 NADL标准毒株基因序列为基础,构建BVDV E^rns蛋白重组真核表达质粒pcDNA3.1-BVDV-E^rns,转染悬浮培养的CHO细胞,进行上清分泌表达。SDS-PAGE分析E^rns蛋白的表达和纯化,并用抗His单克隆抗体和BVDV阳性血清进行Western blotting鉴定纯化蛋白;进一步使用纯化的E^rns蛋白免疫新西兰大白兔,通过间接酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)和细胞间接免疫荧光(indirect immunofluorescence,IFA)实验检测血清抗体水平及其免疫反应活性,用病毒中和实验测定免疫兔血清的中和抗体滴度。BCA蛋白定量试剂盒检测纯化的E^rns     

An Occurrence of Diarrheal Cases Associated with Group C Rotavirus in Adults

Six of the 23 college students who joined a group trip in February of 1991 developed acute nonbacterial gastroenteritis with severe diarrhea. The causal agent was identified as group C rotaviruses by electron microscopy (EM), immune-EM (IEM) and the molecular examinations including polyacrylamide gel electrophoresis (PAGE) and polymerase chain reaction (PCR) on virus particles detected in the extract of watery fecal specimens of the patients. The patients positive for virus isolation showed significant increase in IEM antibody to the isolated virus in their paired sera. These findings suggest that the group C rotavirus is an important etiological agent of diarrhea and may also cause serious food-borne diarrheal disease in adults.     

Identification and Characterization of Heat-Stable Enterotoxin II-Producing Escherichia coli from Patients with Diarrhea

Stock strains of Eschericia coli isolated from patients with traveller's diarrhea were examined for production of heat-stable enterotoxin II (STII). Of 400 strains examined, 3 were found to produce STII. The nucleotide sequence of the STII gene of these human strains was shown to be identical to that of porcine strains. Cultured cells of these strains induced fluid accumulation in ligated mouse intestinal loops and the activity was neutralized by anti-STII antiserum. These results suggest that STII-produciing enterotoxigenic E. coli can cause human diarrhea.