成都地区母乳双歧杆菌对婴儿肠道双歧杆菌构建的影响

目的 观察不同喂养方式婴儿肠道双歧杆菌的构建规律，初步探索母乳双歧杆菌与婴儿肠道双歧杆菌之间的相关性。方法 在四川大学华西第二医院纳入57对母婴志愿者，根据喂养方式分为母乳（BF）组（n=31）和混合喂养（MF）组（1月龄内添加奶粉时间>8 d,n=26），收集产妇产后0天、第30天的母乳，以及婴儿出生后0天、第7天、第30天的粪便。采用实时荧光定量PCR测定母乳和婴儿粪便中双歧杆菌属、角双歧杆菌（B. angulatum）、链双歧杆菌（B. catnulatum）、齿双歧杆菌（B. dentium）、短双歧杆菌（B.breve）、两歧双歧杆菌（B. bifidum）、长双歧杆菌长亚种（B. longum subsp. longum）、长双歧杆菌婴儿亚种（B.longum subsp. infantis）及青春双歧杆菌（B. adolescentis）的丰度。观察2组婴儿肠道双歧杆菌0―7―30天的构建规律；分析母乳双歧杆菌与婴儿肠道双歧杆菌之间的相关性。结果 0―7―30天2组婴儿肠道双歧杆菌构建过程存在差异。母乳双歧杆菌与婴儿肠道双歧杆菌存在相关：0 d时，BF组母乳B. a...     

母乳性黄疸患儿肠道菌群宏基因组学研究

目的探讨母乳性黄疸(BMJ)患儿的肠道菌群情况。方法采用鸟枪法宏基因组测序方法,对16例BMJ患儿及16例正常婴儿的肠道菌群进行研究。结果 BMJ组与对照组中样品菌种丰度及菌群种类总的水平上差异无统计学意义(P0.05);在部分菌种上有差异,双歧杆菌、脆弱拟杆菌及多形拟杆菌在BMJ组中较对照组少(P0.05);在菌群基因的功能通路上差异也有统计学意义(P≤0.05),尤其是在糖合成和代谢基因功能通路中,BMJ组比对照组表达较少些,差异有统计学意义(P0.05)。结论 BMJ患儿肠道菌群的种类、丰度与正常同龄婴儿总体上差异不大,在菌种上有部分差异,双歧杆菌、脆弱拟杆菌及多形拟杆菌在BMJ患儿肠道中含量较少,可补充双歧杆菌、拟杆菌制剂使BMJ患儿的肠道菌群平衡。同时,在基因功能通路表达上,BMJ患儿与正常同龄婴儿有显著差别,特别是糖合成和代谢通路,可深入研究。     

老年2型糖尿病患者 肠道双歧杆菌生长与血糖相关性

目的探讨老年2型糖尿病患者肠道双歧杆菌生长与血糖的相关性,从肠道菌群角度为2型糖尿病的防治与研究提供新的途径。方法根据纳入和排除标准,抽取2015年4月-10月福建某三甲医院糖尿病健康教育俱乐部的2型糖尿病患者90例,收集其粪便和血液标本,检测患者肠道双歧杆菌以及血糖状况,采用Spearman等级相关法分析两者之间的相关性。结果老年2型糖尿病患者肠道长双歧杆菌、短双歧杆菌、青春双歧杆菌、婴儿双歧杆菌和双歧杆菌总菌的定量结果分别是(0.0001±0.0006)、(4.405±8.475)、(4.752±2.779)、(0.003±0.005)和(8.069±8.206)logN/g。空腹血糖和早餐餐后血糖分别为(8.638±1.913)、(9.763±1.910)mmol/L。相关分析显示:青春双歧杆菌和婴儿双歧杆菌与早餐后2h血糖呈现负相关(r=-0.213、-0.217,P0.05),但相关性不显著;双歧杆菌总菌与空腹血糖和早餐后2h血糖有负相关关系(r=-0.302、-0.559,P0.01)。结论老年2型糖尿病患者肠道内双歧杆菌数量减少,血糖与肠道双歧杆菌的数量呈负相关,尤其是早餐后2h血糖,其相关机制有待进一步研究。     

母乳溶菌酶含量与婴儿生理性腹泻

观察５４例生理性腹泻的健康婴儿，分别在生后第４，１５天及１￣６月每月随访，共收集粪便标本２８７份及其母乳３２５份，同时观察其粪便性状及次数，测定母乳溶菌酶含量，用直接涂片加勒兰氏染色法，检测粪便中细菌的种类，探索溶菌酶对力群的影响及其与生理性腹泻的关系，发现母乳溶菌酶在生后１５￣３０天时最低，随着其水平的下降，生后１５－９０天，婴儿粪便中的类杆菌逐渐被双歧杆菌取代，腹泻发生率由几乎１００％降至１２     

益生菌对稳定性冠心病患者血清TMAO水平的影响及 与血清炎性因子水平的相关性

目的探究益生菌对稳定性冠心病(SCAD)患者血清氧化三甲胺(TMAO)水平的影响及其与血清炎性因子水平的相关性。方法选取我院2017年6月至2018年1月间诊断及治疗的SCAD患者74例,采用随机数表法分为益生菌组(37例)与对照组(37例),对照组予以稳定型冠心病标准治疗,益生菌组在对照组的基础上口服益生菌制剂,金双歧,2 g/次,3次/d,疗程均为2个月,比较治疗前后患者血液中TMAO、TNF-α、IL-6、IL-8、CRP水平及大便中双歧杆菌载量,使用Pearson相关性分析模型探究细菌载量与TMAO、TNF-α、IL-6、IL-8、CRP水平的相关性。结果治疗前两组TMAO、TNF-α、IL-6、IL-8及CRP水平无明显差异(Ps0.05),治疗后两组上述指标均显著下降(Ps0.05),其中益生菌组TMAO、TNF-α及CRP水平显著低于对照组(t=2.702,4.301,2.632;P=0.009,0.000,0.010)。治疗前两组患者大便中双歧杆菌载量无明显差异(P0.05),治疗后益生菌组双歧杆菌载量显著提高(t=4.382,P=0.000),且显著高于对照组(P0.05)。Pearson相关性分析显示,双歧杆菌载量与TMAO、TNF-α水平存在显著负相关(r=-0.706,-0.427;P=0.000,0.017)。结论益生菌可有效改善SCAD患者TMAO水平,其与TMAO、TNF-α水平呈显著负相关。     

孕晚期GDM孕妇阴道菌群与阴道上皮细胞的自噬和应激水平的相关性

目的探究孕晚期妊娠期糖尿病(GDM)孕妇阴道菌群与阴道上皮细胞的自噬和应激水平的相关性。方法选取陆军军医大学第一附属医院2018年9月至2019年1月诊断及治疗的孕晚期GDM孕妇23例作为GDM组,选取同时段、同年龄段产检正常的孕妇23例作为对照组。对两组孕妇阴道菌群及自噬和应激水平进行检测,使用Pearson相关性分析探究菌群指标与自噬和应激水平间的相关性。结果两组阴道细菌构成存在差异,GDM组在毛螺菌科、双歧杆菌目、普雷沃菌属、詹氏乳杆菌、双歧杆菌科、加德纳菌属的相对丰度大于对照组,对照组在乳杆菌属、卷曲乳杆菌、惰性乳杆菌、假单胞菌目、假单胞菌属、气球菌科的相对丰度大于GDM组。GDM组p62相对表达量显著低于对照组(t=3.903,P0.001),hsp70相对表达量显著高于对照组(t=5.704,P0.001)。Pearson相关性分析结果显示,双歧杆菌目、加德纳菌属与p62水平呈显著负相关(r=-0.407、-0.482,P=0.009、0.002),双歧杆菌科与hsp70呈显著正相关(r=0.448,P=0.004);乳杆菌属与p62呈显著正相关(r=0.437,P=0.011),与hsp70水平呈显著负相关(r=-0.475,P=0.005)。结论孕晚期GDM孕妇阴道菌群与阴道上皮细胞自噬和应激水平存在一定相关性。     

慢性肾病患者肠道菌群特征及与 微炎症因子水平的相关性

目的 探究慢性肾病(CKD)患者肠道菌群特征及与微炎症因子水平的相关性。方法 选取2018年6月至2018年12月在莆田学院附属医院诊断及治疗的CKD患者25例(CKD组)和健康体检者25例(对照组),对两组研究对象进行肠道微生物检测,同时比较两组研究对象炎症因子水平,应用Pearson模型探究患者肠道微生态特征与炎症因子水平的相关性。结果 CKD组与对照组研究对象的肠道菌群构成具有显著差异,其中对照组双歧杆菌、乳杆菌、产甲酸草酸杆菌、草酸杆菌属、草酸杆菌科和普拉梭菌相对丰度高于CKD组,CKD组克雷伯菌属、大肠埃希菌属、肠杆菌科、瘤胃菌科、毛螺菌科、梭杆菌属和拟杆菌目相对丰度高于对照组,CKD组患者的IL-6、TNF-α、CRP和LPa水平显著高于对照组(t=3.876、4.177、2.236、3.354,P=0.000、0.000、0.030、0.002),Pearson相关性分析显示,双歧杆菌与IL-6、TNF-α水平呈明显负相关(r=-0.272、-0.482,P=0.009、0.002),乳杆菌与IL-6、LPa水平呈明显负相关(r=-0.438、-0.384,P=0.005、0.014),普拉梭菌与TNF-α水平呈明显负相关(r=-0.407,P=0.009),瘤胃菌科与IL-6、CRP水平呈明显正相关(r=0.477、0.508,P=0.002、0.001)。结论 CKD患者肠道菌群与健康人群存在明显差异,其中益生菌水平与炎症因子呈负相关,瘤胃菌科与炎症因子呈明显的正相关。     

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。     

维吾尔族新发2型糖尿病患者肠道 乳杆菌属和多形拟杆菌的定量研究

目的定量研究维吾尔族新发2型糖尿病(T2DM)患者和糖耐量正常(NGT)人群肠道中乳杆菌属(Lactobacillus genus)和多形拟杆菌(Bacteroides thetaiotaomicron)的相对水平。方法严格按照纳入标准、排除标准收集维吾尔族新发T2DM患者96例,NGT 98例。提取所有研究对象的粪便细菌总DNA后,采用16SrDNA基因实时荧光定量PCR(Real-time PCR)对乳杆菌属和多形拟杆菌的水平进行定量检测;运用Pearson相关性分析乳杆菌属与研究对象的BMI、腰围、臀围、收缩压(SBP)、舒张压(DBP)、空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL-C)和低密度脂蛋白(LDL-C)的相关性。结果 16SrDNA基因Real-time PCR结果显示:(1)与新疆维吾尔族NGT组相比,乳杆菌属水平在新发T2DM中较低,差异有统计学意义(t=8.557,P=0.000)。但是多形拟杆菌在两组差异无统计学意义(t=0.524,P=0.601);(2)新疆维吾尔族上述人群肠道中乳杆菌属水平与FBG呈负相关(r=-0.334,P=0.000),腰围呈负相关(r=-0.170,P=0.018),TC呈负相关(r=-0.178,P=0.013),TG呈负相关(r=-0.157,P=0.030),收缩压呈负相关(r=-0.255,P=0.000)。结论 Lactobacillus genus水平在肠道中降低可能与2型糖尿病的发病,血糖和血脂代谢有关,其机制需进一步研究探讨。     

基于气相色谱法分析T2DM患者粪便中SCFAs水平与HbA1c的相关性

目的研究2型糖尿病患者粪便中6种短链脂肪酸水平与糖化血红蛋白的相关性。方法采用气相色谱法检测粪便中短链脂肪酸的含量,并对方法学进行考察。选取2018年在本院查体的2型糖尿病患者57例,根据糖化血红蛋白水平将其分为高糖化组(糖化血红蛋白7.0,28例)与低糖化组(糖化血红蛋白≤7.0,29例)。应用气相色谱法检测两组患者粪便中6种短链脂肪酸水平,并分析6种短链脂肪酸水平与糖化血红蛋白的相关性。结果低糖化组患者粪便中乙酸、丙酸、丁酸、戊酸水平显著高于高糖化组(均P0.05),异丁酸与异戊酸水平差异无统计学意义(均P0.05)。Pearson相关性分析结果表明:在2型糖尿病患者中,糖化血红蛋白与粪便中乙酸呈负相关(r=-0.540 1,P0.000 1),与丙酸呈负相关(r=-0.358 1,P=0.006 2),与丁酸呈负相关(r=-0.421 7,P=0.001 1),与戊酸呈负相关(r=-0.326 8,P=0.042 3),与异丁酸、异戊酸无相关性。结论 2型糖尿病患者粪便中短链脂肪酸水平与糖化血红蛋白存在一定的相关性,短链脂肪酸水平的下降可能是影响血糖控制不佳的主要原因。     

Haloperidol secreted in breast milk.

A nursing mother was given haloperidol 5 mg twice daily for puerperal psychosis and continued to breast feed under hospital supervision. Despite considerable amounts of haloperidol being secreted in the breast milk (up to 23.5 micrograms/l), the infant was apparently not sedated, fed well, and continued to thrive. The findings suggest that maternal ingestion of haloperidol for short periods has no deleterious effect on the infant''s development.     

Immunoregulation by breast milk cells.

Although B cells capable of synthesizing IgG and IgM have been identified in human milk, only IgA synthesis is measured in vitro. These data suggest that milk lymphocyte differentiation is a regulated process and that there may be a specific milk cell factor capable of stimulating differentiation of IgA-bearing B cells. To investigate this possibility lymphocyte/ macrophages from early (≤5 days) and late (≥8 days) milk were incubated and subsequently small aliquots of their cell-free culture media were added to peripheral blood lymphocyte cultures. The release of IgA, IgG, and IgM by the blood lymphocytes in culture was quantitated using double-antibody (Ab) competitive radioimmunoassays. The cell-free media from early (colostral) milk cell cultures significantly stimulated (P < 0.0001) IgA synthesis and had no effect on the production of IgG or IgM. There was no effect on immunoglobulin production when the milk cell supernate came from cells isolated from more mature milk. Therefore, it is postulated (i) that a soluble mediator(s) of immunologic regulation is released by human milk cells, (ii) that this factor(s) at least in part, explains the peculiar immunologic behavior of human milk cells in vitro, (iii) that this factor(s) is released in greater amounts by colostral cells than by cells in mature milk, and (iv) that human colostrum may play a role in affecting active local immunity in the gastrointestinal tract of the recipient newborn.     

n-acetyl-beta-hexosaminidase activity in human breast milk

1. The lysosomal enzyme, N-acetyl-beta-hexosaminidase (HEX) is present in human breast milk. It is composed predominantly of "A" (heat-labile) and "B" (heat-stable) isozymes which coelute with the corresponding major serum isozymes on DE-52 ion-exchange chromatography. 2. Total HEX activity in "early" milk obtained at 2.8 +/- 1.4 weeks post partum, is approx. 2.5-fold higher (87 +/- 29 nmol/60'/mg protein. n = 10) than that of pregnancy serum (35.7 nmol/60'/mg protein) prior to delivery. 3. These levels increase to greater than 3-fold (110 +/- 20 nmol/60'/mg protein, n = 13) as the milk matures (10.3 +/- 4.2 weeks). 4. The specific activity of HEX A in the milk changes little with time post partum, because absolute 5. In contrast, HEX B specific activity is increased, as absolute levels (per volume) remain constant in the face of decreasing milk protein content, 5. These changes result in a high degree of correlation (r = 0.81) between time of lactation and % HEX A observed.     

Concentrations and origin of oxytocin in breast milk

Samples of human milk obtained from lactating women in the early postpartum period were assayed for oxytocin concentrations by specific RIA, following extraction procedures with Florisil. Mean oxytocin concentrations in human milk at postpartum day 1 to 5 were 4.5 +/- 1.1, 4.7 +/- 1.1, 4.0 +/- 1.3, 3.2 +/- 0.4, 3.3 +/- 0.6 microunits/ml (+/- SE), respectively. Oxytocin levels in milk were significantly increased by nursing (3.1 +/- 0.6, 5.3 +/- 1.0 microunits/ml, respectively). 3H-oxytocin in human milk was stable even after incubation at 37 degrees C for 2 hours. The dilution curve for milk was parallel to the curve for the standard oxytocin. The chromatographic fraction of immunoreactive oxytocin was identical to that of 3H-oxytocin. 3H-oxytocin was administered to lactating rats. Radioactivity in the neonatal gastric contents and plasma were 12.8% and 4.4% of the counts in the maternal plasma. It was made clear that oxytocin is stable in milk and that oxytocin in maternal blood can be transferred to mik and then to neonates.     

Isolation and characterization of human breast milk lipoamidase

The mean lipoamidase activity in human breast milk was found to be 0.073 nmol/min per mg (S.D. = 0.028, range = 0.020-0.123, n = 44). The mean lipoamidase activity is approximately 3-fold higher in milk than that in serum (0.023 nmol/min per mg, S.D. = 0.016, range = 0.001-0.059, n = 32). Lipoamidase was purified to 4400-fold by a four-step procedure from 330 ml of human breast milk. The purified enzyme was identified as a single band (Mr = 135,000) by sodium dodecyl sulfate/polyacrylamide electrophoresis. Analysis by Edman degradation indicated that the N-terminal amino acid was glycine. These results strongly suggest that milk lipoamidase is composed of a single polypeptide chain. The enzyme is considered to be a glycoprotein since it reacted positively to periodate-Schiff (PAS) staining. The isoelectric point of the enzyme was 4.2. After treatment of lipoamidase with sialidase, its position on isoelectric focusing gel moved from pH 4.2 to 4.6. This is strongly indicative that lipoamidase contains sialic acid residues. The optimum pH for the enzyme activity is 7.0. The Michaelis constant (KM) for lipoyl p-aminobenzoate is calculated as 25 microM. The enzyme activity was completely lost by heating 60 degrees C for 5 min. The effects of thiol-reactive agents, such as 2-mercaptoethanol (ME) and p-chloromercuribenzoate, were not significant. However, the enzyme activity was completely inhibited by 50 microM diisopropylfluorophosphate. Thus, this enzyme seemed to contain an essential serine residue in the active site.