儿童细菌感染性腹泻临床特点及病原体分析

目的:分析儿童细菌感染性腹泻的临床特点、病原菌分布及耐药性,指导临床诊治。方法:回顾性分析本院2017年1-12月诊治的296例细菌感染性腹泻患儿的临床资料。结果:296例患儿中,男171例,女125例,男女比为1.37∶1;5岁以下儿童239例,占80.74%;发病时间集中于5~9月(79.39%);临床表现为腹泻、发热、腹痛、黏液糊便,部分表现为血便,大便次数以5~10次居多;沙门菌166例(56.08%),大肠埃希菌73例(24.66%),类志贺邻单胞菌41例(13.85%),副溶血弧菌15例(5.07%),志贺菌1例(0.34%)。所有细菌对亚胺培南、哌拉西林/他唑巴坦均敏感(敏感率100%);沙门菌和致泻性大肠埃希菌对头孢他啶的敏感率分别为75.00%、93.55%,对头孢曲松的敏感率分别为67.31%、80.65%,对三代头孢菌素药物有不同程度耐药;副溶血弧菌药物敏感性较高。结论:儿童细菌感染性腹泻发病率高,传播快,耐药率高。临床应尽早完善病原学检查,合理选用抗生素。     

Antimicrobial resistance pattern and in vivo activity of azithromycin in Salmonella isolates

We evaluated antimicrobial susceptibility pattern of 42 Salmonella isolates from February 2012 through January 2013. We also determined the minimum inhibitory concentrations (MICs) of azithromycin against Salmonella isolates and compared them with corresponding disc diffusion sizes. Entire 42 isolates were sensitive to chloramphenicol, 41 (97.6%) were sensitive to cotrimoxazole and amoxicillin each. MICs for azithromycin ranged from 2 μg/ml to 24 μg/ml, corresponding zone diameters ranged from 15 mm to 33 mm and the two were significantly correlated (P = 0.001). Our results indicate that whereas, azithromycin is a potential therapeutic option, the sensitivity to the first line drugs and absence of multidrug resistance reinforce the concept of antimicrobial recycling.     

Poly(Ethyleneimine)‐Immobilized‐Cloth Enzyme Immunoassay for the Detection of Salmonella Lipopolysaccharide

Poly(ethyleneimine) (PEI) was immobilized on non‐woven polyester cloth and examined for application on a simple, rapid and economical “cloth enzyme immunoassay (CEIA)” which was developed originally as polymyxin‐CEIA for the detection of Salmonella lipopolysaccharide (LPS). PEI‐cloth regardless of the PEI molecular weight, but with the amine group contents of 0.1 ～ 0.35 meq/g immobilized either in a physisorption‐like or chemisorption‐like manner, adsorbed LPS rapidly, preferentially and effectively. The captured LPS was then able to be detected qualitatively and quantitatively as an antigen by enzyme immunoassay. PEI‐CEIA had a detection limit for Salmonella LPS of 10 ng/ml, which was equivalent to 1.6 × 10⁵ cell/ml and was ten times more sensitive than polymyxin‐CEIA. It was possible to detect Salmonella LPS in the presence of a 100‐fold excess of E. coli LPS. PEI‐CEIA was found to be more sensitive and much easier to carry out than polymyxin‐CEIA but had the same advantages as polymyxin‐CEIA.     

Salmonella reduces tumor metastasis by downregulation C-X-C chemokine receptor type 4

Tumor metastasis is the main reason for the death of most cancer patients. C-X-C chemokine receptor type 4 (CXCR4) has been demonstrated to be overexpressed in numerous types of cancer. CXCR4 selectively binds with stromal cell-derived factor 1 (SDF1), also known as C-X-C family chemokine ligand 12 (CXCL12) (CXCL12/SDF-1), which induced tumor proliferation and metastasis. Recently, the use of conventional cancer treatments had some limitation; bacteria treatment for cancer becomes a trend that overcomes these limitations. Plenty of studies show that Salmonella has anti-tumor and anti-metastatic activity. The current study aimed to investigate Salmonella suppresses CXCR4 protein expression and tumor cell migration ability in B16F10 melanoma and LL2 lung carcinoma cells. Salmonella reduced CXCR4 protein expression through downregulating Protein Kinase-B (Akt)/Mammalian Target of Rapamycin (mTOR) signaling pathway. In cells transfected with constitutively active Akt plasmids, a reverse effect of Salmonella-induced inhibition of CXCR4 was observed. Tumor cells have chemotactic response to CXCL12 in migration assay, and we found that Salmonella reduced tumor chemotactic response after CXCL12 treatment. The C57BL/6 mice were intravenously injected with B16F10 and LL2 cells pre-incubated with or without Salmonella, the tumor size and lung weight of Salmonella group had obviously decreased, indicating anti-metastatic effect that confirmed the findings from the in vitro experiments.     

Salmonella alters heparanase expression and reduces tumor metastasis

Salmonella causes salmonellosis, is a facultative anaerobe and is one of the common Gram-negative bacteria. Salmonella has anti-tumor potential and tumor-targeting activity. The heparin sulfate on cell surfaces can be cleaved by heparanase that is an endo-β-D-glucuronidase. Heparanase can destroy the extracellular matrix and is involved in tumor metastasis and angiogenic activity. Previously, Salmonella was demonstrated to inhibit tumor metastasis. It remains unclear whether Salmonella inhibits metastasis by regulating heparanase. The expression of heparanase in Salmonella-treated tumor cells was found to be decreased. Transwell and wound-healing assays demonstrated the inhibition of cell migration after Salmonella treatment. Salmonella was found to influence the levels of phosphate-protein kinase B (P-AKT) and phosphate-extracellular regulated protein kinases (P-ERK), which are involved in heparanase expression. Salmonella reduced the heparanase expression induced upregulating PERK and PAKT signaling pathways. The mice bearing an experimental metastasis tumor model was used to evaluate the anti-tumor metastatic effects of Salmonella. Compared with the control group, Salmonella significantly reduced the number of metastatic nodules and enhanced survival. The results of our study indicate that Salmonella plays a vital role in the inhibition of tumor metastasis through the downregulation of heparanase.     

The Contribution of Cytolethal Distending Toxin to Bacterial Pathogenesis

Cytolethal distending toxin (CDT) is a bacterial toxin that initiates a eukaryotic cell cycle block at the G₂ stage prior to mitosis. CDT is produced by a number of bacterial pathogens including: Campylobacter species, Escherichia coli, Salmonella enterica serovar Typhi, Shigella dystenteriae, enterohepatic Helicobacter species, Actinobacillus actinomycetemcomitans (the cause of aggressive periodontitis), and Haemophilus ducreyi (the cause of chancroid). The functional toxin is composed of three proteins; CdtB potentiates a cascade leading to cell cycle block, and CdtA and CdtC function as dimeric subunits, which bind CdtB and delivers it to the mammalian cell interior. Once inside the cell, CdtB enters the nucleus and exhibits a DNase I-like activity that results in DNA double-strand breaks. The eukaryotic cell responds to the DNA double-strand breaks by initiating a regulatory cascade that results in cell cycle arrest, cellular distension, and cell death. Mutations in CdtABC that cause any of the three subunits to lose function prevent the bacterial cell from inducing cytotoxicity. The result of CDT activity can differ somewhat depending on the eukaryotic cell types affected. Epithelial cells, endothelial cells, and keratinocytes undergo G₂ cell cycle arrest, cellular distension, and death; fibroblasts undergo G₁ and G₂ arrest, cellular distension, and death; and immune cells undergo G₂ arrest followed by apoptosis. CDT contributes to pathogenesis by inhibiting both cellular and humoral immunity via apoptosis of immune response cells, and by generating necrosis of epithelial-type cells and fibroblasts involved in the repair of lesions produced by pathogens resulting in slow healing and production of disease symptoms. Thus, CDT may function as a virulence factor in pathogens that produce the toxin.     

The “Atypical” Mycobacteria: Recognition and Disease Association

Abstract

Current advances in the understanding of the pathogenicity of the agents of diarrheal infections, Vibrio cholerae, diarrheagenic E. coli, Shigella, Salmonella, and enteropathogenic Yersinia, have, to a great extent, become possible due to morphological studies of host-pathogen interactions in natural and experimental infections. Despite a multigenic nature and a diversity of pathogenic features in the bacterial species and even in serogroups of the same species, it is now possible to delineate four major patterns of interaction of enteric pathogens with their cellular targets, the enterocytes, and with the immune apparatus of the gut. These patterns, epicellular cytotonic, epicellular restructuring cytotonic, invasive intraepithelial cytotonic and cytotoxic, and invasive transcellular cytotonic and cytotoxic bacteremic, underlie early pathogenesis and clinical manifestations in the respective diarrheal diseases. In this review, the results of the morphological analyses of these patterns over the last 3 decades as well as some methodological problems encountered in the interpretation of morphological observations are discussed.     

Molecular methods for serovar determination of Salmonella

Abstract

Salmonella is a diverse foodborne pathogen, which has more than 2600 recognized serovars. Classification of Salmonella isolates into serovars is essential for surveillance and epidemiological investigations; however, determination of Salmonella serovars, by traditional serotyping, has some important limitations (e.g. labor intensive, time consuming). To overcome these limitations, multiple methods have been investigated to develop molecular serotyping schemes. Currently, molecular methods to predict Salmonella serovars include (i) molecular subtyping methods (e.g. PFGE, MLST), (ii) classification using serovar-specific genomic markers and (iii) direct methods, which identify genes encoding antigens or biosynthesis of antigens used for serotyping. Here, we reviewed reported methodologies for Salmonella molecular serotyping and determined the “serovar-prediction accuracy”, as the percentage of isolates for which the serovar was correctly classified by a given method. Serovar-prediction accuracy ranged from 0 to 100%, 51 to 100% and 33 to 100% for molecular subtyping, serovar-specific genomic markers and direct methods, respectively. Major limitations of available schemes are errors in predicting closely related serovars (e.g. Typhimurium and 4,5,12:i:-), and polyphyletic serovars (e.g. Newport, Saintpaul). The high diversity of Salmonella serovars represents a considerable challenge for molecular serotyping approaches. With the recent improvement in sequencing technologies, full genome sequencing could be developed into a promising molecular approach to serotype Salmonella.     

基于上转发光免疫层析技术的常见食源性致病菌快速检测方法研究与评价

目的：研制针对4种常见食源性致病菌：甲型副伤寒沙门菌（ Salmonella paratyphi A）、乙型副伤寒沙门菌（Salmonella paratyphi B）、大肠埃希菌O157∶H7（Escherichia coli O157∶H7）、副溶血弧菌（Vibrio parahaemolyticus）的基于上转发光免疫层析技术（ UPT－LF）的快速定量检测试纸,并对其检测性能进行评价。方法以上转发光纳米颗粒（ UCP－NPs）作为示踪物,基于双抗体夹心检测模型,研制针对上述4种靶标菌的UPT－LF试纸;以4种靶标菌的系列浓度标准菌悬液作为标准样品,评价试纸的敏感性、特异性、线性和精密性,并进行模拟染菌食品的检测,评价模拟样品阳性检出率。结果针对4种靶标菌的UPT－LF试纸均具有良好的线性,相关系数r为0．985～0．996;检测敏感性达105～106 CFU／ml,与其他近缘菌株无交叉反应;对乳制品、饮料、小食品、水产、肉类等细菌污染的食品检出率较高。结论该研究建立的对4种常见食源性致病菌进行快速检测的UPT－LF,简便快速,具有良好的敏感性、特异性和线性定量能力,操作性能可满足食品安全检测的要求。