慢性胃炎患者胃黏膜菌群定位及定量分析

目的对慢性胃炎患者胃黏膜菌群进行定位及定量分析,探究其与胃炎及幽门螺杆菌(Helicobacter pylori,H.pylori)感染的相关性。方法收集58例慢性胃炎患者胃黏膜标本,提取胃黏膜菌群DNA,行荧光定量PCR定量胃黏膜总细菌及H.pylori,并进行相关性分析;另收集12例慢性胃炎患者胃黏膜标本石蜡包埋切片行荧光原位杂交定位胃黏膜菌群;慢性胃炎、肠化生程度的分类依据新悉尼分类系统。结果慢性胃炎患者胃黏膜细菌主要分布于胃黏膜表面、胃小凹及腺体中,细菌单个散在分布或聚集成团。多元线性回归分析显示胃黏膜总细菌数与性别、年龄、肠化生程度无关,与H.pylori感染、慢性胃炎程度有关(P0.05)。H.pylori阳性组胃黏膜总细菌数与H.pylori细菌数目呈明显正相关(r=0.536,P0.01)。不同胃炎程度之间胃黏膜总细菌数差异有统计学意义(P0.05),其中重度胃炎组胃黏膜总细菌数明显高于轻、中度胃炎组(P0.05、0.01)。不同肠化生程度之间胃黏膜总细菌数差异无统计学意义(P0.05)。H.pylori阳性组胃黏膜总细菌数明显高于阴性组(P0.01)。结论慢性胃炎患者胃黏膜菌群主要分布于胃黏膜表面、胃小凹及腺体中,细菌单个散在分布或聚集成团。胃黏膜菌群与慢性胃炎程度、H.pylori感染有关,与性别、年龄、肠化生程度无关,提示胃黏膜菌群的改变参与慢性胃炎的发展,H.pylori感染可改变胃黏膜菌群。     

胃黏膜菌群与幽门螺杆菌的相关性

除幽门螺杆菌之外,胃黏膜内还定居着大量细菌,占主导地位的是厚壁菌门、变形菌门、拟杆菌门、放线菌门和梭杆菌门。幽门螺杆菌和胃黏膜菌群之间可通过竞争营养和空间、扰乱抑菌肽的分泌以及改变宿主胃生理环境等直接或间接相互影响。本研究总结了胃内正常菌群的组成特征,分析了胃黏膜菌群与幽门螺杆菌之间的相互关系及其潜在机制,并进一步探讨了胃黏膜菌群对幽门螺杆菌相关胃部疾病的影响,有利于深入理解慢性胃病的发病机制,为疾病预防及治疗提供理论依据。     

一次性菌群分析培养瓶的应用

用一次性菌群分析培养瓶对５０ 例小鼠及３５ 人粪便标本菌群分析结果比较可知,我们开发的一次性菌群分析培养基与光岗介绍的经典的菌群分析培养基效果相当,两种方法重复性都好,结果也比较稳定准确,只是经典方法必须把平板培养置厌氧环境３５ ℃培养３６ｈ４８ｈ 观察结果,而一次性菌群分析培养瓶只需放通温箱培养,不仅效果较好而且比传统的经典方法更简易型,易于推广使用。     

蜜蜂肠道菌群的培养方法及特性研究进展

肠道菌群在蜜蜂的消化、营养和抗病性等方面发挥了很多潜在的益生作用。为了加深对以蜜蜂为主的传粉昆虫肠道菌群的了解,本文综述了蜜蜂肠道菌群的人工厌氧培养方法及特性,重点综述了Snodgrassella属、Gilliamella属、Frischella属、Lactobacillus属、Bifidobacterium属和Alpha-1,Alpha-2厌氧细菌类群。旨在通过特定的培养方法获取纯培养的细菌,以供进一步研究特定肠道菌群与特定功能的直接联系。希望这些培养技术能帮助大家提升对蜜蜂肠道共生菌在蜜蜂营养和健康方面所起的作用有所了解。     

微生态制剂在老年人肠道菌群失衡中的应用

人类从青年到老年这个过程,随着年龄的增长,退行性和感染性疾病的易感性也增加,其可能与人体肠道菌群失衡有着密切的关系。人体肠道内生理菌群对机体健康具有重要的作用,其变化与宿主的免疫功能、食物、疾病和年龄等有关。了解老年人肠道菌群特点,且在老年人肠道菌群失调相关疾病中合理介入微生态制剂,将大大有利于老年人身体健康。本文就老年人肠道菌群失衡相关疾病以及微生态制剂在这些疾病中的应用进行综述。     

少量贴壁培养细胞电镜原位包埋方法的探讨

该文探讨了对少量贴壁培养细胞较易操作且能保存较好超微结构的透射电镜样品包埋的方法。将Hela细胞分为三组:(1)不使用环氧丙烷,将树脂胶囊直接倒扣包埋于塑料培养皿;(2)不使用环氧丙烷,将细胞爬片倒扣包埋于胶囊;(3)使用环氧丙烷并将细胞爬片倒扣包埋于胶囊。将三组带有细胞的树脂胶囊进行超薄切片,电镜观察后发现,第一种方法包埋简便,超薄切片上无细胞缺失孔洞,且超微结构保存较好。     

一次性肠道菌群分析培养瓶的研制和应用

目前国内外进行肠道菌群分析多一般使标本称重连续１０倍稀释后用选择性培养基和非选择性培养基,选择一定稀释度滴种（接种）于厌氧菌和需氧菌的有关培养基上分别进行培养,最后以活菌计数方法分析肠道菌群基本情况。这种常规的肠道菌群分析方法,不仅操作繁锁,且每次菌...     

老年性便秘肠道菌群的微生态学研究

目的 :了解老年性便秘患者肠道菌群构成特点。方法 :通过活菌定量培养计数法 ,动态观察 30例老年性便秘患者粪菌群的变化规律 ,并与 32例健康老年人比较。结果 :老年性便秘组每克粪便的活菌数(x± s)分别为 :大肠埃希菌 9.90± 1.4 9,肠球菌 7.81± 2 .6 8,乳酸杆菌 7.2 7± 1.0 1,双歧杆菌 6 .6 8± 1.2 9,类杆菌 6 .92± 0 .91,梭杆菌 10 .0 5± 1.2 1。老年健康组每克粪便各类活菌数 (x± s)分别是 :大肠埃希菌8.6 1± 2 .2 9,肠球菌 7.5 4± 2 .5 2 ,乳酸杆菌 8.2 2± 0 .6 3,双歧杆菌 9.86± 0 .4 4 ,类杆菌 9.4 1± 0 .35 ,梭杆菌7.73± 1.32。两组比较 ,老年性便秘组肠杆菌显著增高 (P<0 .0 5 ) ,梭杆菌非常显著性增高 (P<0 .0 1) ;类杆菌和双歧杆菌非常显著性减低 (均 P<0 .0 1) ;乳酸杆菌显著性减低 (P<0 .0 5 )。结论 :老年性便秘病人肠道菌群正常结构的生态平衡受到破坏 ,主要表现在以双歧杆菌为主的有益菌数量非常显著性减低 ,腐败梭菌等条件致病菌数量呈非常显著性或显著性增高。     

涂片法与培养法在咽部菌群检测研究方面的应用比较

目的 探讨涂片法在咽部菌群研究方面的实际应用。方法 分别应用涂片法和培养法对26例健康成人的咽部菌群进行细菌密集度和多样性的检测。以秩序和检验等统计分析两种方法的差异。结果 两种方法用于密集度检测时无差异,（P＞0．05）;用于多样性检测时,结果显示出非优势菌多样性明显不同。而以优势菌的种类结果相比较时,两种方汉检测结果基本一致（P＞0．05）。结论 要了解人体某部位微生态的改变,依据统一的换算标准,在对细菌密集度和多样性进行检测的两项指标中,涂片法是能够替代培养法的,且作片法的确是一种实用、便捷且又较为可靠的检查方法。     

微生态制剂和肠道菌群与亚健康

本研究简述了亚健康的普遍性及其危害,提出用微生态制剂和肠道正常菌群来调理亚健康的理念,即调理消化系统、呼吸系统;增强免疫功能;调理代谢和其他病前的亚健康;调理心理亚健康。     

Gastric mucosal integrity: gastric mucosal blood flow and microcirculation. An overview

The stomach is in a state of continuous exposure to potentially hazardous agents. Hydrochloric acid together with pepsin constitutes a major and serious threat to the gastric mucosa. Reflux of alkaline duodenal contents containing bile and pancreatic enzymes are additional important injurious factors of endogenous origin. Alcohol, cigarette smoking, drugs and particularly aspirin and aspirin-like drugs, and steroids are among exogenous mucosal irritants that can inflict mucosal injury. The ability of the stomach to defend itself against these noxious agents has been ascribed to a number of factors constituting the gastric mucosal defense. These include mucus and bicarbonate secreted by surface epithelial cells, prostaglandins, sulfhydryl compounds and gastric mucosal blood flow. The latter is considered by several researchers to be of paramount importance in maintaining gastric mucosal integrity. The aim of this paper is to review the experimental and clinical data dealing with the role of mucosal blood flow and in particular the microcirculation in both damage and protection of the gastric mucosa.     

Nocistatin and nociceptin given centrally induce opioid-mediated gastric mucosal protection

Nociceptin (N/OFQ) and nocistatin (NST) are two endogenous neuropeptides derived from the same precursor protein, preproN/OFQ. The aim of the present work was to study the effect of NST on the ethanol-induced mucosal damage compared with that of N/OFQ following intracerebroventricular (i.c.v.) administration in the rat and to analyze the mechanism of the gastroprotective action. It was found that both NST and N/OFQ reduced the mucosal lesions in the same dose range (0.2–1 nmol i.c.v.), but in higher doses (2–5 nmol i.c.v.) the gastroprotective effect of both peptides was highly diminished. The gastroprotective effect of N/OFQ (1 nmol), but not that of NST (1 nmol), was reduced by the selective nociceptin receptor antagonist J-113397 (69 nmol i.c.v.). Similarly, decrease of the gastroprotective effect was observed after the combination of NST (1 nmol) with N/OFQ (0.6 or 1 nmol). However, addition of the gastroprotective effects was observed, when lower dose (0.2 nmol) of NST was given prior to N/OFQ (0.6 nmol). The gastroprotective effect of both N/OFQ and NST was antagonized by naloxone (27 nmol), β-funaltrexamine (20 nmol), naltrindole (5 nmol) and norbinaltorphimine (14 nmol), the μ-, δ- and κ-opioid receptor antagonists, respectively, given i.c.v. The mucosal protection was significantly decreased after bilateral cervical vagotomy. The present findings suggest that NST similar to N/OFQ, may also induce gastric mucosal protective action initiated centrally in a vagal-dependent mechanism. Opioid component is likely to be involved in the gastroprotective effect of both NST and N/OFQ.     

Gastric mucosal blood flow changes in Helicobacter pylori infection and NSAID-induced gastric injury

Background. The impact of H. pylori infection on gastric mucosal blood flow and NSAID‐induced gastric damage is unclear. Aim. To study the effects of H. pylori infection on gastric mucosal blood flow, both at basal conditions and after NSAID exposure, and its relation with mucosal damage and nitric oxide production. Methods. Gastric mucosal blood flow, nitric oxide production and gastric damage were assessed in time after H. pylori SS1 or E. coli inoculation in mice. Experiments were conducted in basal conditions or after oral exposure to indomethacin (20 mg/kg). Results. H. pylori infected mice exhibited a significant increase in gastric blood flow and gastric nitric oxide production 1 week after infection, but those parameters returned to basal levels by 4 weeks. NSAID challenge elicited a similar reduction in gastric blood flow [25–35%] in H. pylori‐infected and control animals. However, only 1 week H. pylori‐infected mice, which exhibited a significant baseline hyperemia, were able to maintain gastric blood flow values within the normal range after NSAID exposure. NSAID‐induced gastric damage was increased in H. pylori‐infected mice by 4 weeks, but not 1 week after infection. Conclusions. Underlying H. pylori infection aggravates acute NSAID‐induced gastric damage. However, at early phases, gastric hyperemia associated with increased nitric oxide production may exert some protective role.     

Effect of peripherally-injected glucagon-like peptide-1 on gastric mucosal blood flow

The aim of this study was to investigate the mechanisms involved in the effect of glucagon-like peptide-1 (GLP-1) on the decrease in gastric mucosal blood flow (GMBF) induced by intragastric ethanol.After preparation of the stomach for GMBF recording, a probe was placed to the gastric mucosa and basal GMBF recordings were obtained by a laser Doppler flowmeter after a 30-minute stabilization period. Following GLP-1 (1000 ng/kg; i.p.) injection, 1 ml of absolute ethanol was applied to the gastric chamber and GMBF was recorded continuously during a 30-minute period. GLP-1 (1000 ng/kg; i.p.) prevented the decrease in GMBF induced by ethanol. Nitric oxide (NO) synthase inhibitor L-NAME, (30 mg/kg; s.c.), calcitonine gene-related peptide (CGRP) receptor antagonist CGRP-(8–37) (10μg/kg; i.p.), and cyclooxygenase inhibitor indomethacin (5 mg/kg; i.p.) all inhibited the GMBF-improving effect of GLP-1.We concluded that, NO, CGRP and prostaglandins may be involved in the effect of peripherally-injected GLP-1 on GMBF reduction induced by intraluminal ethanol.     

Enhancement of gastric mucosal integrity by Lactobacillus rhamnosus GG

The gastric mucosa is frequently exposed to different exogenous and endogenous ulcerative agents. Alcoholism is one of the risk factors for the development of mucosal damage in the stomach. This study aimed to assess if a probiotic strain Lactobacillus rhamnosus GG (LGG) is capable of protecting the gastric mucosa from acute damage induced by intragastric administration of ethanol. Pre-treatment of rats with LGG at 10(9) cfu/ml twice daily for three consecutive days markedly reduced ethanol-induced mucosal lesion area by 45%. LGG pre-treatment also significantly increased the basal mucosal prostaglandin E(2) (PGE(2)) level. In addition, LGG attenuated the suppressive actions of ethanol on mucus-secreting layer and transmucosal resistance and reduced cellular apoptosis in the gastric mucosa. It is suggested that the protective action of LGG on ethanol-induced gastric mucosal lesions is likely attributed to the up-regulation of PGE(2), which could stimulate the mucus secretion and increase the transmucosal resistance in the gastric mucosa. All these would protect mucosal cells from apoptosis in the stomach.     

Changes of gastric mucosal biochemistry in ethanol-treated rats with and without acute surgical vagotomy

The biochemical background of ethanol-(ETOH) induced gastric mucosal damage was studied in rats with intact vagus and after acute surgical vagotomy. Observations were carried out on Sprague-Dawley (CFY) strain rats of both sexes. Gastric mucosal lesions were produced by intragastric administration of 1 ml 96% ethanol. Bilateral truncal surgical vagotomy was carried out 30 min before ETOH administration. The number and severity of gastric mucosal lesions was noted 1 h after ETOH administration. Biochemical measurements (gastric mucosal level of ATP, ADP, AMP, cAMP and lactate) were carried out from the total homogenized gastric mucosa. The adenylate pool (ATP + ADP + AMP), energy charge ((ATP + 0.5 ADP)/(ATP + ADP + AMP)) and ratio of ATP/ADP were calculated. It was found that: 1) ATP transformation into ADP increased, while ATP transformation in cAMP decreased in ethanol-treated animals with intact vagus nerve, while these transformations were quite the opposite in vagotomized animals; 2) no significant changes were found in the tissue level of lactate: and 3) the extent of biochemical changes was significantly less after surgical vagotomy. It is concluded that an intact vagus is basically necessary for the metabolic adaptation of gastric mucosa.     

Protective effects of oral administration of yeast thioredoxin against gastric mucosal injury

Thioredoxin (TRX) is a redox regulating protein which has protective effects against oxidative stress-induced damage to cells and tissues. In this study, we investigated the effects of orally administered TRX derived from edible yeast, Saccharomyces cerevisiae, on gastric mucosa. First, we examined the digestibility of orally administered yeast TRX in mice, and detected yeast TRX in the stomach for 4?h after administration. Next, we investigated the mitigation of gastric mucosal injury after the oral administration of yeast TRX in water-immersion restraint stress and HCl/ethanol-induced gastric ulcer models. Furthermore, we conducted DNA microarray analysis, using the HCl/ethanol-induced model, which revealed that several groups of genes related to tissue repair were upregulated in ulcer regions in the stomachs of rats administered with yeast TRX. These results demonstrated the viability of the use of oral administrations of yeast TRX to protect the gastric mucosa.