慢性腹泻患者肠道微生物群失调的调查及应用微生态学治疗的意义

本室对57例慢性腹泻患者肠道微生态的菌群变化作了初步探讨,现将结果与52例健康人的材料作了比较。实验中我们挑选肠道微生物群中具有代表性的需氧和厌氧菌10种做了定性和定量分析。健康人组10种细菌的检出率为94％～100％;病人组的检出率则为75％～96％。二组间“t”检验的结果:说明包括肠杆菌科,葡萄球菌,类杆菌、双歧杆菌、消化球菌和真杆菌属在内都表现有非常显著性差异(P<0.01),梭菌也有显著性差异(P<0.05)实验结果表明慢性腹泻患者的肠道微生物群发生了显著失调。本文从4个方面讨论了微生态失调发生的原因:1.慢性腹泻与肠道微生物群间的相互关系;2.慢性腹泻影响了肠道微生物群的生长、繁殖、以及它们的定植受阻;3.肠粘膜牯液丢失可使肠道微生物群发生紊乱;4.抗生素与肠道内微生物群的微生态失调有关。我们认为发生肠道微生物群失调时应尽快重建其间关系的平衡。治疗时不宜使用以抗生素为主的方案,而应尽可能地使用生物制剂,此外,还应减少肠道蠕动,避免肠粘膜粘液的大量排出。     

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     

双歧三联活菌胶囊对结肠癌术后化疗相关性腹泻的疗效

目的 探讨双歧三联活菌胶囊对结肠癌术后化疗相关性腹泻患者的疗效,为该病的治疗提供参考。 方法 选择2017年5月至2019年5月在本院接受治疗的90例结肠癌术后化疗相关性腹泻患者为研究对象,回顾性分析其治疗方案并分为对照组(48例,接受常规治疗)和研究组(42例,接受双歧三联活菌胶囊联合常规治疗)。两组患者持续治疗1周后对比其治疗效果,肠黏膜屏障功能指标[二胺氧化酶(DAO)、D乳酸(DLA)、内毒素(Endotoxin)],营养相关指标[血红蛋白(Hb)、白蛋白(ALB)、转铁蛋白(TRF)、前白蛋白(PA)]水平和药物相关不良反应发生情况的差异。 结果 治疗1周后,研究组患者治疗总有效率(100.00%)大于对照组(89.58%)患者,同时其疗效分级差异也有统计学意义(均P结论 双歧三联活菌胶囊辅助治疗可有效增加结肠癌术后化疗相关性腹泻的治疗效果,安全性良好。     

Proteome analysis of porcine epidemic diarrhea virus (PEDV)‐infected Vero cells

Porcine epidemic diarrhea virus (PEDV) causes an acute, highly contagious, and devastating viral enteric disease with a high mortality rate in suckling pigs. A large‐scale outbreak of PED occurred in China in 2010, with PEDV emerging in the United States in 2013 and spreading rapidly, posing significant economic and public health concerns. In this study, LC–MS/MS coupled to iTRAQ labeling was used to quantitatively identify differentially expressed cellular proteins in PEDV‐infected Vero cells. We identified 49 differentially expressed cellular proteins, of which 8 were upregulated and 41 downregulated. These differentially expressed proteins were involved in apoptosis, signal transduction, and stress responses. Based on these differentially expressed proteins, we propose that PEDV might utilize apoptosis and extracellular signal regulated kinases pathways for maximum viral replication. Our study is the first attempt to analyze the protein profile of PEDV‐infected cells by quantitative proteomics, and we believe our findings provide valuable information with respect to better understanding the host response to PEDV infection.     

菌群失调腹泻抗生素造模对小鼠肠黏膜的影响

目的研究抗生素造模对菌群失调腹泻小鼠肠黏膜的影响。方法正常组给予无菌生理盐水0.35m L/(只·次)灌胃,其余各组采用头孢拉定胶囊和硫酸庆大霉素注射液用无菌生理盐水配成浓度为62.5g/L抗生素混合液0.35 m L/(只·次)灌胃。每天2次,连续5 d。造模成功后,采集小鼠空肠、回肠和结肠,分别测量绒毛高度、隐窝深度、淋巴细胞数和肠黏膜厚度,观察肠黏膜情况。结果菌群失调腹泻抗生素造模后,空肠、回肠和结肠的隐窝深度及肠黏膜厚度与正常组相比,差异无统计学意义(P0.05);造模后的结肠淋巴细胞数与正常组相比较,高于正常组,差异有统计学意义(P0.05),造模后的空肠绒毛高度与正常组相比,明显低于正常组,差异有统计学意义(P0.01)。结论菌群失调腹泻抗生素造模使结肠淋巴细胞数增高,空肠绒毛高度降低。     

Giraffe strain of pestivirus: its taxonomic status based on the 5'-untranslated region

The 5'-untranslated region (5'-UTR) of the 'Giraffe' strain of pestivirus was sequenced for comparison with those of other pestiviruses from cattle, sheep, goats, and swine. A phylogenetic tree constructed with these strains suggested that the 'Giraffe' strain was allocated to a new taxon. This observation was also confirmed by a newly proposed method based on palindromic nucleotide substitutions (PNS) at the three variable regions in the 5'-UTR. Other reported pestivirus strains isolated from deer were assigned as bovine viral disease virus (BVDV)-1 according to the PNS as well as phylogenetic analysis, suggesting that BVDV-1 strains can cross-infect deer as well as cattle, sheep, goats, and swine, and that wild deer may serve as a reservoir of BVDV-1. We also identified the genovar of a deer isolate, SH9/11, as BVDV-1c by the PNS method.     

Intestinal lactoflora in Estonian and Norwegian patients with antibiotic associated diarrhea

The disruption of intestinal microbiota is an important risk factor for the development of Clostridium difficile caused antibiotic associated diarrhea (AAD). The role of intestinal lactoflora in protection against C. difficile is unclear. Fecal samples (n = 74) from AAD patients were investigated for C. difficile and lactobacilli by culture and real-time PCR. Lactobacilli were identified by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and sequencing of 16S rRNA. In C. difficile negative cases we found somewhat higher counts of intestinal Lactobacilli (5.02 vs. 2.15 CFU log₁₀/g; p = 0.053) by culture and more frequently Lactobacillus plantarum (33.3% vs. 9.4%; p = 0.03) as compared with positive ones. Results of total counts of lactobacilli comparing Estonian and Norwegian samples were conflicting by culture and PCR. We found higher colonization of Norwegian AAD patients with L. plantarum (21% vs. 5%, p = 0.053) and Estonians with Lactobacillus gasseri (19% vs. 2%, p = 0.023). Particular lactobacilli (e.g. L. plantarum) may have a role in protection against C. difficile, whereas the meaning of total counts of lactobacilli remains questionable. In different persons and nations, different lactobacilli species may have a protective role against C. difficile.     

Circannual Incidence of Giardia Lamblia

A circannual rhythm of Giardia lamblia positive stools was found by examination of records from three clinical laboratories in central Arkansas for the period 1980-1986. Cosinor analysis of monthly Giardia incidence based on stool specimen records from approximately 12,000 patients over the 7-year period revealed a circannual rhythm (P< 0.001) on the basis of percent positive patients/month, with a computive acrophase occurring in late summer and minimum values in the winter. Patients involved in the study were primarily from the central Arkansas metropolitan areas, southern delta regions and northern mountainous regions of the state. Analysis of the data on the basis of total positive Giardia patients/month also revealed a circannual rhythm with the acrophase again occurring in late summer. The overall mean for percent positive stool specimens for the 7-year period was 5.3%, compared with the national average of 3.8% for G. lamblia positive stools. The data indicate that there may be a “Giardia season” in Arkansas since they could not be explained on the basis of day-care age distribution, or geographic origin. Awareness by epidemiologists, public health officials and other health care professionals of this circannual incidence of giardiasis is important for the prevention, diagnosis and treatment of this infectious disorder.     

粪便分离大肠埃希菌CRISPR系统在耐药及毒力中的作用

目的 探索粪便分离大肠埃希菌成簇规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)在耐药及毒力中的作用。 方法 收集某院2018年8-12月分离自慢性腹泻患者及健康体检者粪便样本的大肠埃希菌。采用纸片扩散法检测其药物敏感性,PCR法检测并比较分析腹泻患者及健康体检者粪便样本分离大肠埃希菌的系统发育群、耐药基因及CRISPR系统。 结果 共收集到142株大肠埃希菌,源于63例慢性腹泻患者(疾病组)和79例健康体检者(健康组)。药敏结果显示,氨苄西林耐药率为48.0%,其余抗菌药物敏感率为73.1%～100.0%。63株源于慢性腹泻患者粪便样本分离大肠埃希菌中CRISPR系统的检出率显著低于79株源于健康体检者(3.2% vs 46.8%,χ2=33.538,P2=8.656,P=0.003)。 结论 粪便分离大肠埃希菌对常用抗菌药物仍保持较高敏感性。CRISPR系统可能在粪便分离大肠埃希菌毒力及耐药基因的传播方面发挥重要作用。