新疆维、哈两民族粪样中韦荣球菌属和梭菌属水平与2型糖尿病的相关性研究

目的探讨目标差异肠道菌群与新疆维吾尔族、哈萨克族2型糖尿病(T2DM)的相关性。方法采用16S r DNA实时荧光定量PCR对新疆维吾尔族、哈萨克族正常糖耐量及2型糖尿病患者粪样中韦荣球菌属和梭菌属水平进行检测。两组间均数比较采用t检验,运用Pearson分析方法对差异肠道菌群与新疆维、哈两民族正常糖耐量及2型糖尿病人群的体检及生化指标进行相关性分析。结果 (1)16S r DNA实时定量PCR结果显示,韦荣球菌属及梭菌属水平在哈萨克族2型糖尿病组粪样中显著高于该民族正常糖耐量组(P=0.035,P=0.028)。(2)新疆维吾尔族、哈萨克族正常糖耐量人群和2型糖尿病人群粪样中韦荣球菌属水平与体重及体重指数(BMI)显著正相关(r=0.469,P=0.009;r=0.623,P=0.002),与高密度脂蛋白胆固醇(HDL-C)水平显著负相关(r=-0.466,P=0.025);梭菌属水平与体重、空腹血糖(FBG)、甘油三酯(TG)水平显著正相关(r=0.375,P=0.049;r=0.398,P=0.033;r=0.375,P=0.041)。结论肠道中Veillonella spp和C.coccoides subgroup水平的变化可能与新疆哈萨克族、维吾尔族2型糖尿病的发病有关,需进一步深入探讨。     

新疆维吾尔族2型糖尿病人群肠道中Faecalibacterium prausnitzii的定量研究

目的定量检测新疆维吾尔族2型糖尿病(T_2DM)和糖耐量正常(NGT)人群肠道中的普拉梭菌(Faecalibacterium prausnitzii)水平。方法提取上述人群粪便细菌总DNA后,采用16SrDNA基因实时荧光定量PCR对普拉梭菌水平进行定量检测;运用Pearson分析普拉梭菌水平与研究对象的空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)、体重和体重指数(BMI)的相关性。结果 (1)16S rDNA基因实时定量PCR结果显示:与新疆维吾尔族NGT组相比,普拉梭菌水平在T_2DM中较低(t=2.590,P=0.014),差异有统计学意义。(2)新疆维吾尔族上述人群肠道中普拉梭菌水平与FBG呈负相关(r=-0.434,P=0.012),体重呈负相关(r=-0.359,P=0.044),TG呈负相关(r=-0.410,P=0.034)。结论 Faecalibacterium prausnitzii水平在肠道中降低可能与2型糖尿病的发病有关,其机制需进一步研究探讨。     

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。     

新疆维吾尔族、哈萨克族血浆LBP，sCD14的含量测定及其与2型糖尿病的相关性研究

目的通过分析新疆维吾尔族、哈萨克族正常糖耐量人群和2型糖尿病患者血浆中脂多糖结合蛋白（LBP）、可溶性白细胞分化抗原14（sCDl4）及LBP／sCDl4水平的变化,探讨LBP、sCDl4、LBP／sCDl4与新疆维吾尔族、哈萨克族2型糖尿病（T2DM）的相关性。方法采用酶联免疫吸附法（Elisa）检测LBP、sCDl4在血浆中的浓度。两组问均数比较采用t检验,运用Pearson分析方法对血浆LBP、sCDl4、LBP／sCDl4与新疆维吾尔族、哈萨克族2型糖尿病和糖耐量正常人群的空腹血糖（FBG）、年龄、身高、体重、体重指数（BMI）、腰围、胆固醇（Tc）、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL—C）、低密度脂蛋白胆固醇（LDL—C）做相关性分析。结果与哈萨克族糖耐量正常组相比较,该民族T2DM组中LBP和LBP／sCDl4的表达量升高且差异均有统计学意义（P=0．012,P=0．003）;与哈萨克族糖耐量正常组相比,维吾尔族糖耐量正常组血浆LBP和LBP／sCDl4的表达量升高且差异均有统计学意义（P=0．005,P=0．006）。血浆LBP的含量与相对应的FBG、体重、BMI、TC、TG、HDL—C存在一定的相关性及LBP／sCDl4与FBG、TG显著正相关性。结论与哈萨克族糖耐量正常组相比较,LBP和LBP／sCDl4的表达量在哈萨克族2型糖尿病组和维吾尔族糖耐量正常组均显著升高。     

新疆维、哈两民族人群血浆中谷氨酰胺的含量及其与2型糖尿病关系的研究

目的 探讨谷氨酰胺与2型糖尿病的关系.方法 采用高效液相色谱仪(HPLC)来测定新疆维、哈两民族血浆中谷氨酰胺的含量;使用放免法测定空腹血清胰岛素(FINS)含量;采用全自动生化分析仪来测定空腹血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDLC)、低密度脂蛋白(LDLC)的含量.结果 维吾尔族正常人与糖尿病患者之间血浆中谷氨酰胺含量差异有统计学意义(P＜0.01);哈萨克族正常人与糖尿病患者之间血浆中谷氨酰胺含量差异无统计学意义;维哈两民族的正常人血浆中谷氨酰胺含量差异有统计学意义(P＜0.01);维哈两民族的糖尿病患者血浆中谷氨酰胺的含量差异有统计学意义(P＜0.05).维哈两民族人群血浆中的谷氨酰胺含量与空腹血糖(FBG)水平呈负相关(r=-0.4858,P＜0.001),与FINS水平呈正相关(r =0.236,P＜0.05),与胰岛素分泌指数(HOMA-IS)呈正相关(r=0.52,P=0.001),与TG呈负相关(r=-0.4330,P＜0.01),与LDLC呈负相关(r=-0.4077,P＜0.01),与HDLC呈正相关(r=0.2553,P＜0.01).结论 谷氨酰胺可能是2型糖尿病的一个保护性的影响因素.     

2型糖尿病患者代谢清除率与血清游离脂肪酸的相关性

目的:探讨2型糖尿病(type 2 diabetes mellitus,T2DM)患者代谢清除率(metabolic clearance rate,MCR)与血清游离脂肪酸(nonesterified fatty acid,NEFA)、甘油三酯(triglyceride,TG)和胆固醇的相关性。方法:选择2014年10月至2016年12月我院收治的127例T2DM患者,测量患者身高、体重,并计算身体质量指数(BMI),进行口服糖耐量试验、胰岛素释放试验,检测血清脂质的水平。将T2DM患者按MCR值分为低MCR组(63例)、高MCR组(64例),比较两组间临床指标的差异,评价MCR、HOMA-IR与变量间的相关性。结果:T2DM患者BMI、TG、空腹血糖和糖化血红蛋白(Hb A1C)均高于参考值;T2DM患者低MCR组Hb A1C、空腹血糖、总胆固醇(total cholesterol,TC)、TG、低密度脂蛋白胆固醇(LDL-C)和NEFA明显升高(P0.05),高密度脂蛋白胆固醇(HDL-C)明显降低(P0.05);相关分析显示:MCR与HDL-C呈显著正相关(P0.05,r=0.215),与TC、TG、LDL-C、Hb A1C、NEFA均呈显著负相关(P0.05;r=-0.191,-0.380,-0.216,-0.587,-0.356)。结论:2型糖尿病患者MCR降低,与Hb A1C与血清NEFA水平呈负相关。MCR不仅能评价胰岛素抵抗,而且能反映机体糖脂代谢水平。     

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。     

母乳和新生儿粪便细菌与母乳性黄疸的相关性分析

目的探讨母乳和新生儿粪便细菌与母乳性黄疸(BMJ)的相关性。方法以BMJ患儿为研究对象,并与健康新生儿对照。RT-PCR法检测母乳和新生儿粪便中细菌含量,比较BMJ组和对照组细菌含量的差别,分析血总胆红素与细菌含量的相关性。结果 (1)BMJ组母乳和新生儿粪便中青春双歧杆菌、两歧双歧杆菌和长双歧杆菌浓度显著低于对照组,差异均有统计学意义(均P0.01);(2)母乳两歧双歧杆菌与血清TSB显著负相关(r=-534,P=0.000);新生儿粪便青春双歧杆菌(r=-0.675,P=0.000)、两歧双歧杆菌(r=-0.598,P=0.000)和长双歧杆菌(r=-0.472,P=0.000)与血清TSB显著负相关。结论母乳中双歧杆菌含量下降可能与新生儿BMJ相关。     

过敏性鼻炎患者鼻腔菌群特征及其 与血清IgE和黏膜嗜酸性粒细胞水平的关系

目的探究过敏性鼻炎(AR)患者鼻腔菌群特征及其与血清IgE和黏膜嗜酸性粒细胞(Eos)水平的关系。方法选取2019年3月至6月在首都医科大学北京友谊医院平谷医院诊断及治疗的AR患者28例作为过敏性鼻炎组,选取同时期体检的健康个体28例作为对照组,检测两组患者鼻腔微生物特征,同时对比两组血清IgE及粘膜Eos%,使用Pearson相关性分析探究各指标间的相关性。结果两组样本微生物构成存在显著差异,样本PCA1及PCA2分别为27.67%及14.23%,过敏性鼻炎组链球菌属、金黄色葡萄球菌、嗜血杆菌属、梭杆菌属、差异球菌属、变形菌门的相对丰度显著高于对照组,对照组痤疮丙酸杆菌、放线菌门、丙酸杆菌属、气球菌属、棒杆菌属、嗜胨菌属的相对丰度显著高于过敏性鼻炎组(Ps0.05),过敏性鼻炎组IgE水平(Z=6.005,P=0.000)及Eos%(t=10.776,P=0.000)显著高于对照组,变形菌门与IgE水平呈显著正相关(r=0.391,P=0.017),痤疮丙酸杆菌与IgE(r=-0.421,P=0.009)及Eos%(r=-0.328,P=0.048)呈显著负相关。结论 AR患者鼻腔变形菌门、痤疮丙酸杆菌分别与IgE水平、鼻粘膜Eos水平具有相关性。     

培菌白蚁菌圃和粪便微生物多样性分析

【背景】培菌白蚁是属于白蚁科的一类与鸡枞菌属真菌共生的高等白蚁,其与体内肠道微生物和体外菌圃微生物形成三维共生体系。【目的】分析培菌白蚁菌圃和粪便的微生物多样性,并与肠道微生物进行比较。【方法】通过Illumina MiSeq高通量测序方法对培菌白蚁菌圃和粪便样品进行细菌16S rRNA基因和真菌ITS测序分析。【结果】高通量测序获得培菌白蚁菌圃和粪便样品细菌和真菌的有效序列和OTU数目。5个样品细菌OTU数目在90-199之间,而真菌OTU在10-58之间,细菌的种类多样性明显大于真菌。不论是细菌还是真菌,粪便样品的OTU数目多于菌圃样品。经物种分类分析,菌圃样品主要优势细菌是变形菌门(Proteobacteria),其相对含量超过82.4%;其次是拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes);粪便样品中优势细菌为拟杆菌门,其次是变形菌门,粪便优势菌属为别样杆菌属和营发酵单胞菌属,这与培菌白蚁肠道菌多样性组成一致。培菌白蚁菌圃和粪便样品共生真菌主要为担子菌门(Basidiomycota)和子囊菌门(Ascomycota)。菌圃优势真菌为鸡枞菌属(Termitomyces),相对含量在51.83%以上,菌圃中还鉴定到炭角菌属(1%,Xylaria)。【结论】为今后培菌白蚁-体内外微生物共生关系研究以及微生物的分离培养提供了依据和参考。     

Degradation and conversion of somatostatin in normal and diabetic rats in vivo and in vitro

Plasma somatostatin-like immunoreactivity in the portal and jugular veins of streptozotocin diabetic rats was compared with that in normal control rats. In the diabetic group, somatostatin levels in the portal (p less than 0.05) and jugular (p less than 0.01) veins were both elevated compared with those in the control group. Moreover, the degree of elevation was greater in the jugular vein than in the portal vein. To further investigate the role of the liver in the clearance of somatostatin-28 in vivo, 2 micrograms of somatostatin-28 was administered as a bolus into the external jugular vein of intact and functionally hepatectomized rats. The mean half-time of somatostatin-28 was significantly longer in intact diabetic rats than in controls (p less than 0.05). The functional hepatectomy did not cause a significant difference in the half-time in diabetic rats but made it longer in control rats. These results suggest that the longer half-time of somatostatin-28 in diabetic rats in vivo is due to its slower hepatic clearance. The hepatic clearance of somatostatin-28 and somatostatin-14 was further studied in vitro using a recirculating liver perfusion method. The hepatic clearance of 1.2 nM of either somatostatin-28 or somatostatin-14 was significantly lower in diabetic rats than in controls (p less than 0.01). This indicates that elevated plasma somatostatin levels in diabetic rats are caused at least in part by decreased hepatic clearance of somatostatin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Production of arsine and methylarsines in soil and in culture

Arsenate, arsenite, monomethylarsonate, and dimethylarsinate were added to different soils, and evolution of gaseous arsenical products was determined over 3 weeks. Arsine was produced in all three soils from all substrates, whereas methylarsine and dimethylarsine were produced only from methylarsonate and dimethylarsinate, respectively. At least three times more arsine than dimethylarsine was produced in soil incubated with dimethylarsinate. Resting cell suspensions of Pseudomonas and Alcaligenes produced arsine as the sole product when incubated anaerobically in the presence of arsenate or arsenite. In all instances, no trimethylarsine was observed, nor could any evidence be shown for the methylation of any arsenical substrate in soil or in culture. It was concluded that reduction to arsine, not methylation to trimethylarsine, was the primary mechanism for gaseous loss of arsenicals from soil.     

Development and comparison of in vivo and in vitro models for endometritis in cows and mares

In order to investigate pathogenic mechanisms of acute endometritis in cows and mares, we established an in vivo model in both species. Based on the results of an in vitro transmigration system, human recombinant interleukin-8 (rhIL-8; 1.25 microg per mare and 5 microg per cow in 50 ml phosphate-buffered saline) was used to attract polymorphonuclear neutrophil granulocytes (PMNs) into the uteri. Peak numbers of uterine neutrophils were attracted after 6h, in both cows and mares. On average, mares responded more sensitively than cows, with 15 times higher numbers of rhIL-8-attracted uterine neutrophils (72+/-8 x 10(7)cells). In contrast to in vitro studies, in vivo migrated neutrophils (uterine neutrophils) of both species displayed a significantly reduced MHC class I expression. Expression of the CD11a molecule was significantly enhanced on equine uterine neutrophils but downregulated on bovine cells. Compared with untreated autologous peripheral neutrophils, both uterine and in vitro migrated neutrophils showed no alteration of phagocytic capacity. The ability to generate reactive oxygen species (ROS) was significantly upregulated in bovine and equine uterine neutrophils. This was also observed after in vitro migration of equine neutrophils, whereas ROS generation by bovine neutrophils was significantly depressed. In summary, the concept of inducing endometritis directly by local application of human interleukin-8 has been reliably successful in cows and mares. The model permits the analysis of PMN migration into the uterus under defined and controlled conditions. The observed differences between cows and mares with respect to phenotypical and functional characteristics of in vivo attracted uterine cells point to species-related features of neutrophil migration. In vitro transmigrated bovine and equine cells partially differ in phenotype and function from uterine neutrophils. Therefore, the in vitro transmigration assay cannot completely represent the in vivo endometritis model described here.     

Importance of guanine nitration and hydroxylation in DNA in vitro and in vivo

Guanine (Gua) modification by nitrating and hydroxylating systems was investigated in DNA. In isolated calf thymus DNA, 8-NO(2)-Gua and 8-oxo-Gua were dose-dependently formed with peroxynitrite, and 8-NO(2)-Gua was released in substantial amounts. Myeloperoxidase (MPO) with H(2)O(2) and NO(2)(-) reacted with calf thymus DNA to form 8-NO(2)-Gua dose dependently without release of 8-NO(2)-Gua. The frequency of strand breaks was higher than the sum of 8-NO(2)-Gua and 8-oxo-Gua, particularly in the MPO-treated DNA, indicating the importance of other types of damage. The activation of human neutrophils and lymphocytes with phorbol ester did not induce 8-NO(2)-Gua and 8-oxo-Gua in their nuclear DNA. However, 8-NO(2)-Gua was found in calf thymus DNA co-incubated with activated neutrophils in the presence of NO(2)(-). No significant formation of 8-NO(2)-Gua was found in liver DNA from mice treated with Escherichia coli lipopolysaccharide. The incubation of peroxynitrite or MPO-H(2)O(2)-NO(2)(-)-treated DNA with formamidopyrimidine glycosylase (Fpg) released 8-oxo-Gua, but not 8-NO(2)-Gua, indicating that 8-NO(2)-Gua is not a substrate for Fpg. Although 8-NO(2)-Gua was generated in isolated DNA by different nitrating systems, other types of damage were formed in abundance, and the lesion could not be found reliably in nuclear DNA, suggesting that the biological importance is limited.     

Activity of native and dextran-modified hyaluronidase in in vitro and in vivo systems

Modified hyaluronidase derivatives have been obtained. Covalent coupling of the enzyme with aldehyde dextran results in 65-85% protein binding to the carrier, residual catalytic activity accounting for 90-100% of the baseline. Modified hyaluronidase is more thermostable than the native enzyme. The data on intravenous drug distribution in the mouse organs are promising and ensure effective use of modified hyaluronidase for the treatment of pulmonary diseases.     

Interactions of cadmium and selenium in rat plasma in vivo and in vitro

⁷⁵Se and ¹⁰⁹Cd tracers were used to study the binding of Se and Cd to plasma proteins at various SeO₃^2? doses and times up to 24 h after the simultaneous subcutaneous administration of SeO₃^2? and CdCl₂ to adult male rats. The simultaneous injection of CdCl₂ and SeO₃^2? markedly increased both Se and Cd plasma levels over that in control animals. Gel permeation chromatography of plasma indicated that at all times up to 24 h Cd and Se were bound in an atomic ratio of approx. 1 : 1 in 330 000 and 130 000 dalton fractions. From 4 to 24 h, Cd and Se appeared in the 420 000 dalton fraction, also with an atomic ratio of approx. 1 : 1. The 330 000 dalton molecules appeared to have a maximal binding capacity for the Cd-Se complex at a concentration of approx. 30 μmol/ml of plasma, while the 130 000 and 420 000 dalton molecules show a higher binding capacity. Studies in vitro revealed that SeO₃^2? does not interact directly with Cd and plasma proteins. It is metabolized by erythrocytes to a form that interacts in an atomic ratio of 1 : 1 with Cd to form a protein-bound complex of 130 000 daltons.     

Biosynthesis of beta-glucuronides of retinol and of retinoic acid in vivo and in vitro

After the intraportal injection of retinol-6,7-(14)C to rats, the O-ether derivative of retinol, retinyl -glucosiduronate, appears in the bile. Both retinoyl -glucuronide and retinyl -glucosiduronate are also synthesized in vitro when washed rat liver microsomes are incubated with uridine diphosphoglucuronic acid (UDPGA) and either retinoic acid or retinol, respectively. The synthesis of retinoyl -glucuronide was also demonstrated in microsomes of the kidney and in particulate fractions of the intestinal mucosa. The glucuronides were characterized by their UV absorption spectra, by their quenching of UV light or fluorescence under it, by their thin-layer chromatographic behavior in two solvent systems, and by the identification of products released during their hydrolysis by -glucuronidase. With retinoic acid as the substrate, the UDP glucuronyl transferase of rat liver microsomes had a pH optimum of 7.0, a temperature optimum of 38 degrees C, and a marked dependence on the concentrations of both retinoic acid and UDPGA, but was unaffected by a number of possible inhibitors, protective agents, and competitive substrates. The conversion of retinal to retinoic acid and the synthesis of retinoyl -glucuronide from retinoic acid could not be detected in whole homogenates, cell fractions, or outer segments of the bovine retina.