低出生体重儿肠道微生态构建影响因素的 动态监测

目的动态监测低出生体重儿肠道菌群,分析不同体重、不同喂养方式及疾病状态等因素对患儿肠道微生态的影响,为规范临床低出生体重儿宫外营养支持措施及治疗手段提供依据。方法应用16SrRNA荧光定量PCR技术检测正常新生儿和低出生体重儿生后第1、3、7天粪便中大肠埃希菌、肠球菌、乳杆菌及双歧杆菌的含量。结果 (1)在生后7d内,无论正常新生儿还是低出生体重儿,其粪便中大肠埃希菌、肠球菌、乳杆菌和双歧杆菌的含量均随日龄的增加而增加,且生后7d内正常新生儿的粪便中大肠埃希菌、肠球菌、乳杆菌和双歧杆菌的含量均显著高于低出生体重儿(P0.05),正常新生儿生后7d内粪便中各细菌的增长率均高于低出生体重儿。(2)体重2 000~2 500g的低出生体重儿粪便中大肠埃希菌和肠球菌在各日龄中的含量明显高于体重2 000g的新生儿(P0.05);同时其粪便中双歧杆菌和乳杆菌含量在3日龄和7日龄阶段明显高于体重2 000g的新生儿(P0.05)。(3)3日龄和7日龄母乳喂养组的低出生体重儿粪便中双歧杆菌和乳杆菌含量明显高于乳制品喂养组(P0.05);且母乳喂养组新生儿生后7日内粪便中大肠埃希菌、乳杆菌和双歧杆菌含量的增长率均高于乳制品喂养组,尤其是双歧杆菌的增长率(126.49%vs 54.81%)。(4)合并并发症的3日龄和7日龄的低出生体重儿,粪便中乳杆菌和双歧杆菌含量均明显低于无合并症的低出生体重儿(P0.05);且无并发症组的低出生体重儿其粪便中肠球菌、乳杆菌和双歧杆菌的增长率均高于有并发症组的低出生体重儿,大肠埃希菌增长率则低于有并发症组。结论低出生体重儿肠道菌群的定植时间晚且数量少,体重、喂养方式及有无并发症是影响新生儿肠道菌群丰度的重要因素。母乳喂养可促进低出生体重儿肠道中益生菌的定植。疾病因素会导致肠道菌群丰度的降低,使肠道菌群紊乱,其程度可能与病情的严重程度相关。     

新生鼠坏死性小肠结肠炎肠道菌群变化及意义

目的探讨新生鼠坏死性小肠结肠炎(NEC)时肠道菌群变化,旨在阐明肠道菌群在NEC发病中的作用,为探讨新生儿NEC的发病机制及寻求有效的防治措施提供理论依据。方法SD新生大鼠出生48 h开始给予鼠乳代用品人工喂养,100%氮气缺氧90 s,4℃冷刺激10 min,每天2次,连续3 d,建立新生SD大鼠NEC模型。40只新生SD大鼠随机分成NEC模型组(A组)和正常对照组(B组)。每组动物各20只。在最后一次缺氧、冷刺激后24 h空腹断头处死大鼠,留取回盲部近端肠管组织进行肠组织损伤评分,组织学评分≥2确定为NEC;实验前后留取两组新生鼠粪便,按照张秀荣方法进行肠道菌群检测。实验前后比较采用配对t检验,组间比较采用方差分析,α=0.05为显著性检验标准。结果模型组新生鼠相继出现腹泻、腹胀、萎靡、活动减少,生长减慢,对照组新生大鼠进食及排便均正常,无腹胀及胃潴留,活动度良好,皮下脂肪丰满。模型组新生大鼠NEC的发生率为100%(20/20),对照组无1例发生NEC。实验组和对照组肠损伤病理评分(x±s)分别为:3.25±0.85、0.45±0.51,t=12.622,P<0.01。模型组和对照组新生鼠实验前肠道细菌总数,杆菌、球菌总数,G 杆菌、G 球菌,G-杆菌、G-球菌数差异均无显著性,肠道菌群中杆菌和球菌的比例都在正常范围中。实验结束时,正常对照组新生大鼠肠道菌群总数明显增多,其中以G 杆菌增加为主;G-杆菌属及G 球菌属菌数占肠道菌群的比率在实验前后差异无显著性;模型组新生大鼠肠道群总数亦明显增多,而且明显高于正常对照组。实验结束时肠道菌群数量,其中主要是G 球菌显著增加,而G 杆菌却较对照组明显减少,与实验前相比,明显下降,差异有显著性;模型组实验结束时肠道菌群中杆菌和球菌比值减少、倒置。结论NEC发病前正常肠道菌群已发生质和量的变化,肠道菌群紊乱在NEC发病机制中起关键作用;及时纠正肠道菌群紊乱可能会降低新生大鼠发生NEC危险性。     

血液恶性肿瘤患者肠道菌群分析

目的将血液恶性肿瘤患者肠道菌群与健康个体进行比较,观察血液肿瘤患者肠道菌群结构的变化,探讨肠道菌群与血液恶性肿瘤发生发展的联系。方法收集血液恶性肿瘤患者与健康志愿者粪便样品,提取样品中菌群总DNA,然后通过变性凝胶梯度电泳(DGGE)技术分析肠道菌群多样性和差异性。结果血液恶性肿瘤组与健康组肠道菌群的DGGE指纹图谱有明显差异。与正常组相比,患者组肠道大肠埃希菌呈现过度增长趋势,有益菌柔嫩梭菌减少或缺失,某些患者肠道内一些细菌呈现特异性增长,如粪肠球菌、硫磺肠球菌、约氏不动杆菌等。结论与健康对照组相比,血液恶性肿瘤患者肠道菌群结构与多样性发生改变,这可能为血液恶性肿瘤早期抗感染提供实验依据。     

不同季节腹泻病患儿粪便菌群变化和微生态治疗结果的分析

为观察不同季节腹泻病患儿粪便菌群变化和分析微生态制剂治疗的效果。方法：收集70例不同季节腹泻病患儿粪便,用革兰染色法检查菌群,根据细菌的形态分为杆菌和球菌两大类,根据杆菌或球菌菌群数量的多少分为 、 、 三种等级。按腹泻者的病程对急性腹泻病和迁慢性腹泻病患儿粪便菌群检查统计分析。应用微生态制剂治疗急性腹泻病和迁慢性腹泻病患儿。结果：容易发生腹泻病的小儿年龄在4—24月。腹泻患儿粪便菌群总的变化趋势为杆菌不是以 为主,而是以 — 为主,球菌 — 占有一定的比例。急性腹泻病住院治疗时间短,而迁慢性腹泻病住院治疗时间长。结论：腹泻病患儿粪便菌群总的变化趋势为杆菌明显减少,球菌增多或相对增多,提示腹泻患儿的确存在肠道菌群的紊乱。急性腹泻病患儿的肠道菌群紊乱,应用微生态制剂治疗易调节,病程短;而迁慢性腹泻病患儿的肠道菌群紊乱,应用微生态制剂治疗较难调节,病程长。     

老年房颤患者肠道菌群变化及其影响因素

目的探讨老年房颤患者肠道菌群变化及影响菌群变化的相关因素。方法回顾性收集20例老年房颤患者作为房颤组,并选取性别、年龄与患者匹配的30例老年健康体检者作为对照组,分析两组对象肠道优势菌的特点。同时收集房颤组患者C反应蛋白(CRP)、总胆汁酸(TBA)、同型半胱氨酸(Hcy)、B型纳尿肽(BNP)、左室射血分数数值(LVEF)等检测结果,分析上述因素与肠道优势菌群的关系。结果房颤组患者直肠真杆菌数量为(4.66±1.62)lg copies/g粪便,低于对照组的(6.10±1.44)lg copies/g粪便,差异有统计学意义(t=-3.286,P=0.002)。房颤组患者肠道普拉梭菌、肠球菌、类杆菌、乳杆菌、双歧杆菌、柔嫩梭菌、酪酸梭菌、Atopobium cluster(Ato)、肠杆菌数量及双歧杆菌/肠杆菌(B/E)值与对照组比较差异无统计学意义(均P0.05)。在房颤组中,CRP升高患者肠道双歧杆菌数量和B/E值显著高于CRP正常患者,CRP升高与双歧杆菌数量呈正相关(r=0.651,P=0.002);Hcy升高患者肠道肠杆菌数量显著高于Hcy正常患者,Hcy升高与肠杆菌数量呈正相关(r=0.567,P=0.009)。BNP、TBA、LVEF与优势菌群无显著相关性。结论老年房颤患者存在肠道菌群失调。CRP、Hcy和老年房颤患者肠道菌群具有一定的相关性。     

低出生体重儿肠道菌群及肠道屏障功能的研究

目的研究低出生体重儿的肠道菌群分布情况和肠道屏障功能的变化。方法以低出生体重儿(1 500g≤体重2 500g)为研究对象,采用16SrRNA荧光定量PCR技术和JY-DLT肠道屏障功能分析系统检测低出生体重儿出生后第7天粪便中双歧杆菌、乳杆菌、大肠埃希菌、肠球菌4种细菌的含量以及血清中的二胺氧化酶、D-乳酸和细菌内毒素的浓度,比较正常新生儿与低出生体重儿肠道菌群和肠道屏障功能的差异,分析不同喂养方式、并发症对低出生体重儿肠道菌群及肠道屏障功能的影响。结果 (1)低出生体重儿组粪便中大肠埃希菌、肠球菌、乳杆菌、双歧杆菌含量均明显低于健康新生儿组(P0.05),血清中二胺氧化酶、D-乳酸高于健康新生儿组(P0.05),细菌内毒素水平差异无统计学意义(P0.05)。(2)母乳喂养组低出生体重儿粪便中双歧杆菌和乳杆菌含量明显高于乳制品喂养组(P0.05),且血清中二胺氧化酶和和D-乳酸含量低于乳制品喂养组(P0.05),细菌内毒素水平差异无统计学意义(P0.05)。(3)无并发症组低出生体重儿粪便中乳杆菌和双歧杆菌含量明显高于有并发症组(P0.05),其血清中二胺氧化酶、D-乳酸和细菌内毒素水平均低于有并发症的低出生体重儿(P0.05)。结论低出生体重儿的肠道菌群和肠道屏障功能都与正常新生儿存在差异,母乳喂养有助于肠道有益菌的定植和肠道屏障功能的恢复。     

结肠清洗后肠道微生态平衡的重建

目的观察结肠清洗对人肠道菌群的影响和结肠清洗后肠道微生态平衡再建的过程。方法10名健康男性志愿者随机平均分为2组,一组为结肠清洗后食用普通膳食,另一组为结肠清洗后,先食用航天食品,后食用普通膳食,在选定的时相点收集粪便,选择性培养基分别对肠球菌、大肠埃希菌、类杆菌、乳杆菌及双歧杆菌定量测定。结果结肠清洗后,肠道所有的微生物数量均显著减少,7d后肠道菌群恢复正常。结论结肠清洗引起肠道微生态平衡紊乱,航天食品不利于肠道微生态重建。     

肠道菌群分析及粪便炎性标志物检测在 炎症性肠病活动度评估中的作用

目的探讨肠道菌群和粪便炎性标志物在炎症性肠病(IBD)活动度评估中的临床价值。方法共纳入120例IBD患者为研究组,其中溃疡性结肠炎(UC)患者68例,克罗恩病(CD)患者52例。选择30例经结肠镜检查正常的健康体检者为对照组。采集全部研究对象的新鲜粪便标本进行粪便细菌培养及炎性标志物检测,比较不同疾病活动度IBD患者的肠道菌群及粪便钙卫蛋白(FC)、乳铁蛋白(LF)、基质金属蛋白酶-9(MMP-9)、髓过氧化酶(MPO)水平的变化。结果与对照组比,UC和CD患者肠道中肠杆菌、肠球菌、拟杆菌、消化球菌及酵母菌数量均明显增加(P0.05),双歧杆菌、乳杆菌及真杆菌数量明显减少(P0.05)。UC患者梭菌数量较对照组增加(P0.05),CD患者梭菌数量较对照组减少(P0.05)。UC、CD活动期患者肠杆菌、肠球菌、拟杆菌、消化球菌及酵母菌数量明显多于缓解期患者(P0.05),双歧杆菌、乳杆菌及真杆菌数量明显少于缓解期患者(P0.05),且重度活动期患者肠道菌群改变较轻、中度活动期改变更明显(P0.05)。UC活动期患者梭菌数量明显多于缓解期(P0.05),CD活动期患者梭菌数量明显少于缓解期(P0.05)。UC和CD患者粪便中FC、LF、MMP-9及MPO水平均显著高于对照组(P0.05)。UC、CD活动期患者FC、LF、MMP-9及MPO水平显著高于缓解期患者(P0.05),且重度活动期患者高于轻、中度活动期患者(P0.05)。结论肠道菌群变化和粪便中FC、LF、MMP-9及MPO水平可作为IBD患者疾病活动性评估的辅助指标。     

用16S rRNA荧光定量PCR技术分析低出生体重儿肠道菌群变化及影响因素

目的监测低出生体重儿肠道细菌分布情况,并分析影响低出生体重儿肠道微生态平衡的因素。方法以低出生体重儿(1 500g≤体重2 500g)为研究对象,采用16SrRNA荧光定量PCR技术检测新生儿出生后第1天、3天、7天粪便中双歧杆菌、乳杆菌、大肠埃希菌和肠球菌4种细菌的含量,比较正常新生儿与低出生体重儿肠道菌群构建的差异;分析不同胎龄、体重、喂养方式、疾病状态等因素对低出生体重儿肠道微生态平衡的影响。结果 (1)低出生体重儿组和健康新生儿组粪便中大肠埃希菌、肠球菌、乳杆菌和双歧杆菌含量与婴儿日龄呈明显正相关关系,且低出生体重儿组婴儿粪便4种细菌含量均明显低于健康对照组(P0.05)。(2)2 000g≤体重2 500g组低出生体重儿大肠埃希菌和肠球菌含量在各日龄明显高于体重2 000g组新生儿(P0.05),双歧杆菌和乳杆菌含量在3日龄和7日龄阶段明显高于体重2 000g组(P0.05),而1日龄阶段差异无统计学意义(P0.05)。(3)母乳喂养组在3日龄和7日龄阶段双歧杆菌和乳杆菌含量明显高于乳制品喂养组(P0.05)。(4)无并发症患儿组在3日龄和7日龄阶段乳杆菌和双歧杆菌含量明显高于有并发症组(P0.05)。结论低出生体重儿肠道菌群构建规律异于正常新生儿,尤其是乳杆菌和双歧杆菌的定植差异更为突出;低出生体重儿的出生体重与肠道乳杆菌和双歧杆菌的含量呈正相关;母乳喂养对低出生体重儿肠道中益生菌的定植有明显的优势;新生儿相关疾病直接影响低出生体重儿肠道微生态的构建,可导致其胃肠道生态系统的异常;16S rRNA荧光定量PCR技术适用于评价婴幼儿肠道微生态状况。     

加替沙星对小鼠肠道菌群的影响

目的 了解加替沙星正常剂量用药和非正常剂量用药方式对小鼠肠道菌群的影响.方法 24只昆明小鼠随机分为3组,每组8只,对照组只灌胃生理盐水,另外2组分别按正常剂量和正常剂量的一半给小鼠灌胃加替沙星溶液,7 d后无菌采取小鼠粪便,观察各组小鼠粪便中双歧杆菌、类杆菌、乳酸杆菌、大肠埃希菌及肠球菌数量.结果 加替沙星灌胃7 d后,与给药前相比,肠杆菌和肠球菌药敏结果无明显变化.灌胃前后肠杆菌对CIP、LEV、GAT、AM、CRO、AMK、NOR均敏感,肠球菌对AM、LEV、VA、GAT均敏感,对NOR、CIP、CRO敏感性不同.2种用药方式可导致大肠埃希菌数量显著减少(P＜0.05),肠球菌数量稍有增加(P＞0.05).而双歧杆菌、乳酸杆菌和类杆菌数量无明显变化(P＞0.05).结论 加替沙星2种用药方式短时期对肠道菌群影响不大(肠杆菌除外),不易产生耐药性.其杀灭肠杆菌的作用远大于对厌氧菌的作用.     

Rhesus monkeys (Macaca mulatta) in the Taihang mountains,Jiyuan county,Henan, China

A population of rhesus macaques in the Taihang mountains on the Henan-Shanxi border is probably the northernmost population of the species. A more northerly population north of Beijing was locally extirpated in 1988. In the Taihang mountains, 26 natural groups, with >2000 monkeys, have been found. Six of them have been studied for 8 years. They live in a deciduous forest with temperate climate and cold snowy winters. The average group size is 82 individuals, and the population density is 3.1–7.2 individuals per km². Home rangs are very large, varying from 11 to 22 km². A natural reserve for the rhesus was established at Jiyuan in 1982, and one group, monitored and provisioned since 1984, has increased from 47 to 83 individuals, an increase of 76.5%.     

Four-year study of controlled timed breeding of rhesus monkeys (Macaca mulatta)

As part of the timed breeding colony at Tulane National Primate Research Center, exogenous progesterone administration (5 mg/day for 10 days) has been used to select conception dates by inducing artificial luteal phases in female rhesus monkeys. A retrospective analysis of data obtained during four breeding seasons (1998-2001) revealed that conceptions occurred an average of 18 days after the last administration of progesterone. The age of the female to be bred, previous pregnancy history, and timing of breeding during the breeding season were determined to be critical factors in the success of the procedure. The benefit of this method of timed breeding is that it does not require tracking of menstrual cycles, which can be labor-intensive and requires that animals be monitored several months in advance of breeding to determine each female's individual cycle length. It also provided an efficient use of breeding-age males.     

Self-agency in rhesus monkeys

Rhesus monkeys (Macaca mulatta) have shown the ability to monitor their own mental states, but fail the mirror self-recognition test. In humans, the sense of self-agency is closely related to self-awareness, and results from monitoring the relationship between intentional, sensorimotor and perceptual information. Humans and rhesus monkeys were trained to move a computer icon with a joystick while a distractor icon partially matched their movements. Both humans and monkeys were able to monitor and identify the icon they were controlling, suggesting they have some understanding of self-agency.     

Experimental infection of rhesus macaques with Streptococcus pneumoniae: a possible model for vaccine assessment

BACKGROUND: We explored the possibility of using normal adult rhesus macaques for the preclinical assessment of safety, immunogenicity, and efficacy of newly developed vaccines against Streptococcus pneumoniae infection of the lung. METHODS: Our primary objective was to determine whether an intra-bronchial inoculum of at least 10(6)S. pneumoniae colony-forming units, or one as high as 10(8)-10(9) organisms, could detectably survive in rhesus macaques for a period longer than 1-2 weeks. If so, we hypothesized, it would be possible to observe signs of pneumonia commonly observed in humans, and discriminate between vaccinated/protected animals and controls. Infection was detectable in bronchoalveolar lavage fluids 3-5 weeks post-inoculation. RESULTS: The clinical course of disease mimicked aspects of that of human pneumococcal pneumonia. Signs of inflammation typical of the disease in humans, such as elevated concentrations of neutrophils and of pro-inflammatory cytokines in bronchoalveolar lavage fluids were also observed. CONCLUSIONS: These findings underscore the utility of this model to assess the safety, immunogenicity, and efficacy of newly developed S. pneumoniae vaccines.     

Extinction of rhesus monkeys (Macaca mulatto) in Xinglung,North China

An isolated population of rhesus monkeys living in Xinglung County, north of Beijing (latitude 40⪤gree N), was a natural population at the northernmost range of the species in Asia. Rhesus had been reported there at the end of the last century, based on a few specimens, but were not well known by zoologists until the beginning of the 1960s. The population remained approximately 50–60 individuals in the 1960s but then declined to only 4 or 5 monkeys about 5 years ago. It has been confirmed by local hunters and two recent field trips that the population is now probably extinct. As a result, the northern extent of the remaining natural range of the rhesus macaque in Asia has dropped 650 km southward to the Jiyuan Nature Reserve on the border of Hunan and Shanxi provinces, just north of the Yellow River.     

Electromagnetic dosimetry in a sitting rhesus model at 225 MHz

Dosimetric measurements in a 9.5-kg tissue-equivalent rhesus model were conducted at 225 MHz using a nonperturbing temperature probe and a gradient-layer calorimeter. Temperature probe measurements showed deep penetration of electromagnetic energy, and calorimeter experiments showed an average SAR (0.285 W/kg per mW/cm²) that was nearly three times greater than that observed for the same model at 1.29 GHz.     

Postthaw survival of rhesus macaque sperm: variation in the response of individual males to different freezing protocols

This study was designed to assess the effects of various cryopreservation protocols on the postthaw survival of rhesus macaque sperm. Multiple ejaculates of five mature males were collected and frozen by each of four different methods. Cryopreservation significantly reduced motility in all samples, regardless of the method, and the response of different ejaculates of the same male to each method proved to be consistent. The freezing method was a significant factor in determining the immediate postthaw motility, although this effect disappeared after 4 hr. Moreover, there was a significant interaction between individual males and freezing method suggesting that developing a single freezing protocol that is universally suitable may be difficult.     

A non-invasive technique for the control of intestinal parasites in rhesus macaques (Macaca mulatta).

Control of intestinal parasites is a major concern in maintaining the health and well-being of captive primates. This study established that percutaneous application of ivermectin was as effective as subcutaneous administration for the control of nematodes in captive rhesus macaques. The method is less intrusive than the standard subcutaneous or oral delivery routes and did not adversely affect the health of any of the study subjects.