孤独症谱系障碍儿童肠道菌群与行为症状的相关研究

目的 探索孤独症谱系障碍（ASD）儿童肠道菌群的构成及其与行为症状严重程度之间的关系。方法 选取30名ASD儿童为ASD组,20名年龄、性别与之匹配的健康儿童为健康对照组,统计两组相关基本信息。对两组儿童粪便样本中细菌16S rRNA基因的V3-V4高变区进行测序,并采用孤独症治疗评估量表评估ASD儿童行为症状的严重程度。应用Spearman秩相关分析ASD儿童肠道菌群与行为症状严重程度的相关性。结果 研究表明两组儿童肠道微生态菌群结构存在显著差异。与健康对照组相比,ASD组儿童α多样性指数（Shannon和Shannoneven）显著降低（P < 0.05）;粪便Firmicutes比例显著降低,而Acidobacteria比例显著增高（P < 0.05）。ASD组儿童粪便的优势菌群为Megamonas、Megasphaera、Barnesiella等;而健康对照组儿童粪便的优势菌群为Eubacterium_rectale_group、Ezakiella、Streptococcus等。且Megamonas丰度与ASD儿童健康/身体/行为和语言评分均呈正相关（P < 0.05）。结论 ASD的发展及其行为症状的严重程度与肠道菌群结构密切相关。     

支气管肺炎患儿治疗过程中肠道双歧杆菌和大肠杆菌的变化

目的 研究支气管肺炎患儿治疗过程中肠道双歧杆菌和大肠杆菌的变化,并与同年龄段同性别的健康儿童进行比较.为采用微生态制剂防治抗生索引起的肠道菌群变化提供数据参考.方法 收集30例支气管肺炎患儿及30例健康儿童的粪便标本,提取目标细菌DNA,测量和比较肺炎患儿和健康儿童标本细菌的DNA-D(260)值;应用细菌的16S rRNA/DNA序列设计双歧杆菌和大肠杆菌的引物,行常规PCR完成细菌定性;然后取标准定量的两种细菌DNA经一序列稀释后行荧光定量PCR,并制成标准曲线;将待测标本同样行荧光定量PCR后与标准曲线进行对比,得出标本中两种细菌含量.结果 支气管肺炎患儿治疗后的肠道菌群与健康儿童比较发生了明显的变化(F=50.78,P＜0.01);肠道中双歧杆菌数量(拷贝数/克湿便)的对数值,患儿治疗第1天为7.10±0.59,治疗第3天为6.03±0.51,治疗第7天为6.34±0.52,健康儿童为9.48±0.44,经统计学处理差异有统计学意义(P＜0.01).而大肠杆菌数量的对数值,患儿治疗第1天为6.74±0.38,治疗第3天为6.13±0.35,治疗第7天为7.08±0.40.健康儿童为7.54±0.45,经统计学处理差异有统计学意义(P＜0.01).结论 支气管肺炎患儿治疗过程中肠道菌群比健康组低.因此在治疗支气管肺炎患儿时采用微生态制剂,调整细菌比例.改善肠道内环境,使肠道内益生菌的数量尽快恢复正常.     

Features of intestinal flora in children with food protein-induced proctocolitis based on high-throughput sequencing北大核心CSCD

目的 通过高通量测序分析食物蛋白诱导性直肠结肠炎（food protein-induced proctocolitis，FPIP）患儿肠道菌群特征。 方法 选取2018年10月至2021年2月遵义医科大学第三附属医院门诊就诊的小于6月龄纯母乳喂养发生FPIP患儿31例纳入FPIP组，正常健康婴儿31例纳入健康对照组；采集两组粪便样本提取DNA并通过PCR扩增，采用高通量测序分析粪便样品中的16S rDNA V3~V4片段并进行相关生物信息学分析。 结果 肠道菌群多样性分析示FPIP组菌群多样性Shannon指数低于健康对照组，但差异无统计学意义（P>0.05）；菌群丰富度Chao指数高于健康对照组（P<0.01）。在门水平上，两组菌群组成均主要由厚壁菌门、放线菌门、变形菌门、拟杆菌门4个菌门构成；与健康对照组相比，FPIP组中放线菌门构成比显著降低（P<0.001），变形菌门构成比明显增加（P<0.05）。在属水平上，FPIP组主要由大肠埃希氏菌属、梭状芽孢杆菌属、肠球菌属、克雷伯氏菌属和双歧杆菌属组成；健康对照组主要由双歧杆菌属、链球菌属组成；与健康对照组相比，FPIP组中双歧杆菌属和瘤胃球菌属构成比明显降低（P<0.05），梭状芽孢杆菌属和志贺氏菌属构成比明显增加（P<0.05）。 结论 FPIP组与健康对照组相比，肠道菌群多样性下降，丰富度升高；在菌群组成结构上某些细菌菌属存在差异。提示在FPIP的发生发展过程中，肠道微生物群在属水平上的构成改变可能起到了重要作用。     

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目的以轮状病毒(RV)肠炎患儿和健康婴幼儿为研究对象,探讨荧光定量PCR技术在检测肠道菌群变化的实用性和可行性。方法应用荧光定量PCR技术测定细菌的16SrRNA,对RV肠炎患儿和健康婴幼儿粪便中双歧杆菌、乳酸杆菌和大肠杆菌进行定量检测和分析,并和其他专家用传统方法所获得的结果进行比较。结果RV肠炎患儿的肠道菌群与健康儿童比较发现肠道中双歧杆菌和乳酸杆菌的数量变化差异有显著性(P<0·05),而大肠杆菌的数量差异无显著性(P>0·05)。其结果与其他文献报道的用细菌培养的方法所得结果一致。结论RV肠炎患儿肠道中益生菌的数量较正常对照组明显减少。荧光定量PCR技术比传统方法特异性高、敏感性强,更省时和省力,临床上可用此方法对患儿肠道细菌变化进行定量分析。     

肠道微生态与益生菌

除口腔外，人体消化道自上而下的细菌数量逐渐增多。成人肠道菌群主要集中在结肠和末端小肠，有多达10^13个不同类型、不同含量的细菌，其细菌数是人体体细胞的10倍。在正常生理状态下，肠道含有近500个细菌品种，近200万个编码基因。宿主和细菌之间相互作用、相互影响，共同维系着肠道微生态的动态平衡。宿主的许多因素会影响肠道微生态的构成，如宿主的年龄、饮食结构、是否使用益生菌和／或抗生素、婴儿所处的外部环境、母亲的肠道菌群、分娩方式等。     

过敏性紫癜患儿肠道菌群的变化

目的研究过敏性紫癜(HSP)患儿的肠道菌群变化。方法提取64例HSP患儿和25例健康儿童粪便标本,采用16S rRNA/DNA荧光定量PCR技术分析并比较粪便标本中乳酸杆菌、双歧杆菌和大肠杆菌。结果 HSP患儿粪便中的乳酸杆菌、双歧杆菌和大肠杆菌的含量与健康儿童比较,差异均无统计学意义(P0.05)。其中伴腹痛、呕吐等消化道症状HSP患儿的乳酸杆菌、双歧杆菌含量均低于无消化道症状的HSP患儿以及健康儿童,差异均有统计学意义(P均0.05)。结论 HSP患儿急性期可能存在肠道菌群结构失调的情况,尤其是伴有消化道症状的HSP患儿。     

过敏性紫癜伴幽门螺杆菌感染患儿肠道菌群变化的研究

目的 研究过敏性紫癜(HSP)伴有幽门螺杆菌(H.pylori)感染患儿肠道菌群的变化.方法 随机收集40例HSP患儿及40例正常儿童的粪便标本,先用快速免疫检测卡进行粪便幽门螺杆菌抗原(HpSA)检测,判定有无H.pylori感染.然后提取两组粪便标本目标细菌DNA,采用16SrRNA荧光定量PCR技术对两组粪便标本中的双歧杆菌和大肠杆菌进行定量分析和比较.并计算双歧杆菌/大肠杆菌(B/E)比值.结果 HSP患儿H.priori检出率为50.0%,正常儿童为27.5%(x2=4.266.P<0.05).双歧杆菌在HSP伴H.Pylori感染组和HSP非H.pylori感染组分别与正常儿童H.pylori感染组和非H.pylori感染组相比,数鼍明显减少(P<0.008 3);大肠杆菌在正常儿童H.pylori感染组和非H.pylori感染组分别与HSP非H.pylori感染组相比,数量明显升高(P<0.008 3),在HSP伴H.pylorii感染组与HSP非H.pylori感染组相比,数量明显升高(P<0.008 3).B/E值在HSP伴H.pylori感染组分别与HSP非H.pylori感染组、正常儿童H.priori感染组和正常儿童非H.pylon感染组相比,数值明显降低(P<0.008 3).结论 HSP患儿H.pylori检出率较正常儿童明显增多,HSP发病可能与H.pylori感染有关.HSP伴H.pylori感染和HSP非H.pylori感染患儿肠道双歧杆菌均较正常儿童减少,HSP伴H.pylori感染患儿肠道大肠杆菌较HSP非H.pylori感染息儿升高,HSP伴H.pylori感染患儿B/E值明显降低,提示HSP患儿肠道菌群失调明显.     

基于QIIME 2平台对先天性巨结肠患儿与正常儿童肠道菌群的比较分析

目的比较先天性巨结肠患儿与正常儿童肠道菌群差异, 阐明先天性巨结肠对肠道菌群组成的影响。方法收集2019年4月至2020年12月来河北省儿童医院就诊的先天性巨结肠患儿与体检的正常儿童粪便样本。10例先天性巨结肠患儿纳入先天性巨结肠患儿组(HDB_Nor组), 其中男7例, 女3例;年龄为(1.22±0.65)岁;14例正常儿童纳入对照组, 即正常儿童组(Normal组), 其中男8例, 女6例;年龄为(1.67±0.84)岁。利用细菌16S rRNA基因序列测序, 通过QIIME 2平台对测序数据进行统计分析并绘图, 对比分析两组肠道菌群组成、多样性、菌群差异、菌群物种间相关性以及菌群与疾病之间联系。结果在肠道菌群组成、Beta多样性方面先天性巨结肠患儿与正常儿童肠道菌群差异有统计学意义;LEfSe差异分析结果显示假小链双歧杆菌、两歧双歧杆菌、瘤胃菌属等在先天性巨结肠患儿组中物种显著增加;放线菌属、粘滑罗斯菌属及弗氏柠檬酸杆菌等三个细菌种属在先天性巨结肠患儿组中显著减少。通过肠道菌群与先天性巨结肠相关性分析发现AKK菌属、大肠埃希-志贺氏菌属、肠球菌属和伯克霍尔德菌...     

婴儿肠道菌群的形成及其与食物过敏的关系

目的　观察健康婴儿肠道菌群的定植过程及其在食物过敏症患儿的变化 ,分析肠道菌群形成与婴幼儿食物过敏的相互关系。方法　采用荧光定量PCR技术测定细菌 16SrRNA ,经与标准曲线对照计算细菌数量 ,对 71例健康无过敏症母乳喂养婴儿和 10 0例食物过敏婴儿粪便乳酸杆菌、双歧杆菌和大肠杆菌行定量检测。结果　婴儿肠道菌群处于动态定植过程 ,随生长发育 ,双歧杆菌和乳酸杆菌在肠道定植增加 ,大肠杆菌数量减少。食物过敏婴幼儿肠道乳酸杆菌、双歧杆菌数量较健康婴幼儿低 ,而大肠杆菌数量较健康婴幼儿高。结论　婴儿期肠道菌群仍处于动态演替过程。食物过敏婴儿肠道菌群与健康婴儿不同     

局灶性癫痫患儿治疗前后肠道菌群的变化及意义北大核心CSCD

目的研究局灶性癫痫患儿与健康儿童肠道菌群是否不同,以及癫痫患儿治疗前后肠道菌群的变化。方法招募未经治疗的初诊局灶性癫痫患儿10例为病例组,且均用单药奥卡西平治疗。收集患儿临床资料,以及治疗前和治疗3个月后的粪便标本;另招募相同年龄段的健康儿童14例为对照组。提取粪便标本中细菌总DNA,进行16S rDNA测序,并行生物信息学分析。结果病例组经奥卡西平治疗3个月后癫痫发作频率减少>50%。门水平方面,治疗前组放线菌门丰度高于治疗后组和对照组(P<0.05);属水平方面,治疗前组埃希菌/志贺菌属、链球菌属、柯林斯菌属和巨单胞菌属丰度高于对照组(P<0.05),经治疗后其丰度降低(P<0.05)。结论与健康儿童相比,局灶性癫痫患儿的肠道菌群存在显著差异;奥卡西平可以显著改善癫痫症状,重塑癫痫患儿的肠道菌群。     

益生菌与口服免疫球蛋白辅助治疗儿童轮状病毒肠炎的疗效比较

目的：比较益生菌及口服免疫球蛋白对儿童轮状病毒肠炎的疗效。方法：150例轮状病毒肠炎患儿随机分为3组,每组50例。3组患儿在给予基础治疗的同时给予不同的干预治疗方案。对照组予安慰剂治疗,益生菌组予双歧杆菌乳杆菌三联活菌治疗,免疫球蛋白组予抗轮状病毒鸡卵黄IgY口服治疗。每日记录大便次数、大便性状等临床症状,并于治疗后1、3、5、7、9、11?d收集新鲜大便标本,镜检行大便菌群紊乱分度,放射免疫法检测大便SIgA水平,双抗体夹心酶联免疫法检测大便轮状病毒排泄量。结果：与对照组比较,益生菌治疗组干预用药3 d后菌群失调改善,大便频次减少,继发肠道细菌感染发生率降低（P0.05）。免疫球蛋白组用药1 d后大便SIgA水平显著高于对照组（P<0.05）,治疗3 d后腹泻频次及大便病毒排泄量均低于对照组（P<0.05）。免疫球蛋白组病程较对照组显著缩短（4.5±1.0?d vs 5.8±1.7?d,P<0.05）。结论：对儿童轮状病毒肠炎,益生菌能缓解菌群紊乱及预防继发感染,但缓解临床症状较慢,且不能缩短病程。口服免疫球蛋白显效快,能快速清除病毒及促进机体SIgA生成,且能缩短病程。     

Survival of rotavirus antibody activity derived from bovine colostrum after passage through the human gastrointestinal tract

BACKGROUND: Rotavirus is a major cause of infectious diarrhea in infants and young children. Several studies have shown that hyperimmune bovine colostrum, derived from cows immunized with rotavirus, can prevent rotavirus diarrhea when given passively. The objective of this study was to determine whether colostral antibody activity survived transit through the gut by measuring the level of rotavirus antibody activity in the feces. METHODS: Hyperimmune colostrum containing different levels of rotavirus antibody was administered to 105 children attending nine Adelaide childcare centres. Subjects were asked to drink 100 ml of whole milk supplemented with colostrum 3 times a day, for a period of 6 days. Stool samples were collected from the subjects before, during, and after consumption of the study product. Rotavirus activity was determined using a novel virus reduction enzyme-linked immunosorbent assay (ELISA) and a virus neutralization assay. RESULTS: Rotavirus antibody activity was detected in 521 (86%) of 602 fecal specimens obtained during the study using the virus reduction ELISA. The antibody activity was detected as early as 8 hours after ingestion of hyperimmune colostrum and up to 72 hours after consumption had ceased. There was a strong relation (r = 0.81) between the titer of rotavirus antibody administered to subjects and the level of antibody activity detected in the feces. CONCLUSIONS: The results show that antirotavirus activity survived passage through the gut. Therefore, passive immunotherapy may be used to prevent or treat infectious diseases that affect the entire length of the gastrointestinal tract.     

A prospective evaluation of community acquired gastroenteritis in paediatric practices: impact and disease burden of rotavirus infection.

AIMS: To examine the disease burden and epidemiology of community acquired rotavirus gastroenteritis in Austrian children treated in a paediatric practice. METHODS: A prospective, population based, multicentre study in four paediatric practices and two children's hospitals (Innsbruck and Leoben). Children 相似文献   

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??Necrotizing enterocolitis??NEC?? is considered to be the most common gastrointestinal emergency among neonates. Although the pathogenesis of NEC is incompletely understood??there are several established risk factors??including prematurity??altered intestinal blood flow/oxygen delivery??formula feeding and bacterial infection. Recently??a large number of studies showed that intestinal flora imbalance had been implicated as key risk factor in the pathogenesis of NEC. After birth??the neonatal gut must acquire a healthy complement of commensal bacteria??which leads to deficient or abnormal microbial colonization of the gut??may protect the immature gut from inflammation and injury. Providing a healthy complement of commensal bacteria can maintain the intestinal microflora balance??shift the balance of intestinal microbiota from a pathegenic to protective complement of bacteria??protect the gut from inflammation and subsequent injury??and prevent NEC. We review the relationship between intestinal microbiota and NEC in preterm infants??the mechanism of probiotics in preventing NEC??and the efficacy and safety of probiotics in preterm infants.     

Probiotika in der Prävention und Behandlung der akuten Gastroenteritis bei Kindern

Probiotics are live microbial food supplements (bacteria, yeast) with a beneficial effect on health. They have been studied mainly for the prevention and treatment of acute infectious diarrhea in children. In recent years there have been preliminary reports that probiotic bacteria may also play a role in the prevention and treatment of inflammatory bowel diseases as well as allergic conditions. Most randomized controlled clinical studies as well as a recently published meta-analysis and a systematic review confirm a modest but significant effect of probiotics, in particular of Lactobacillus GG, in the treatment of acute infectious diarrhea in children. However, it is premature to give practical recommendations for the use of probiotics for several reasons such as limited numbers of controlled studies as well as heterogeneity concerning study design und probiotic strains.     

儿童呼吸道轮状病毒感染的临床调查

了解小儿急性呼吸道感染中人轮状病毒的致病情况。方法采用单克隆抗体酶联免疫吸附一步法,对68例5a以下呼吸道感染患儿与46例正常儿童的口咽分泌物进行了轮状病毒抗原检测。结果患病组8例阳性,感染率11．8％,正常组全部阴性,经统计学处理两组有显著性差异。结论口咽分泌物的轮状病毒可作为呼吸道感染轮状病毒的一个参考指标。轻症呼吸道轮状病毒感染,通过自然免疫作用,有可能对严重轮状病毒再感染有保护作用。     

Prenatal administration of Lactobacillus rhamnosus has no effect on the diversity of the early infant gut microbiota

We have recently shown that maternal administration of Lactobacillus rhamnosus GG (LGG) during late pregnancy can have beneficial effects on the early development of infant gut microbiota, promoting a bifidobacteria profile similar to that of a healthy breastfed infant. It is uncertain, however, whether such probiotic supplementation could influence the diversity of infant gut microbiota. We investigated the effect of pre-natal LGG on gut microbial diversity in the early post-natal period. Day-7 faecal samples were collected from 98 infants at high risk of allergic disease, whose mothers participated in a pre-natal probiotic eczema prevention study. Faecal microbial diversity was assessed by terminal restriction fragment length polymorphism using restriction enzymes Sau96I and AluI. A greater number of peaks represent greater diversity of bacterial communities. Administration of LGG to mothers during late pregnancy had no effects on the mean number of peaks in faecal samples from 1-wk-old infants as compared to placebo (AluI 14.4 vs. 15.5, p?=?0.17, 95% CI -0.4, 2.5; Sau96I 17.3 vs. 15.8, p?=?0.15, 95% CI -3.5, 0.5). Prenatal LGG failed to modulate diversity of early infant gut microbiota despite promoting a beneficial bifidobacteria profile.     

A prospective evaluation of community acquired gastroenteritis in paediatric practices: impact and disease burden of rotavirus infection

AIMS—To examine the disease burden and epidemiology of community acquired rotavirus gastroenteritis in Austrian children treated in a paediatric practice.
METHODS—A prospective, population based, multicentre study in four paediatric practices and two children''s hospitals (Innsbruck and Leoben). Children ⩽ 48 months of age presenting with gastroenteritis during a six month period of rotavirus peak between December 1997and May 1998 were included. Prospective testing of stool samples for rotavirus was performed using ELISA.
RESULTS—A total of 6969 children were enrolled; 171 (2.4%) had community acquired gastroenteritis. Of 144 children who could be included in further analysis, 49 (34%; median age 16.7 months) were rotavirus positive, and 95 (66%; median age 17.0 months) were rotavirus negative. Three of the rotavirus positive children (median age 14.6 months) were hospitalised. The severity of rotavirus positive gastroenteritis was significantly higher than that of rotavirus negative gastroenteritis. The incidence of community acquired gastroenteritis was 4.67 per 100 children per year, and of rotavirus positive gastroenteritis 1.33 per 100 children per year.
CONCLUSION—Rotavirus is a relevant cause of community acquired gastroenteritis in children aged 4 years and younger treated by a paediatrician. The data can be used as a basis for developing strategies to prevent infection.

     

Prebiotics in infant milk formulas: new perspectives

In recent years it has become accepted that healthy human intestinal microflora may play an important part in priming the infants' systemic and mucosal immunity. Dietary modulation of the gut microbiota is a topical area of nutritional sciences and the main focus of many current functional foods such as non-digestible oligosaccharides (NDOs). Fructo-oligosaccharides (FOS) and trans-beta-galacto-oligosaccharides (TOS) have been claimed to benefit the health of the colon by selectively stimulating the growth of bifidobacteria and lactobacilli (prebiotic effect). It could be of clinical interest to manipulate colonic flora because it is supposed that specific bacteria in the gut microbial microflora could promote potentially antiallergenic processes and play a key part in atopic disease prevention. Supporting this view is the finding that analysis of the composition of the intestinal bacterial populations showed different microbial patterns between healthy and allergic individuals. Assuming that non-digestible TOS and FOS can affect the intestinal ecosystem beneficially, the opportunity for gut flora manipulation arises in bottle-fed infants. New preterm and term infant milk formulas, supplemented with a mixture of TOS and FOS as prebiotic ingredients induced a significantly higher colonization of bifidobacteria and lactobacilli. In the future, selective manipulation of the intestinal microbiota might be an approach to novel prophylactic and therapeutic intervention strategies of atopy, by redirecting allergic Th-2 responses in favour of Th-1 responses.