抑郁症患者肠道菌群研究

目的研究抑郁症患者肠道微生物群落分布情况。方法选择2017年1月-2018年1月在本院临床心理科就诊的确诊为抑郁症的患者79例,以及同时期在本院体检的健康者80例。收集患者新鲜粪便,利用16SrRNA基因测序技术分析患者肠道菌群。结果对159例粪便标本进行16SrRNA基因测序,共获得1 276 841条有效16SrRNA基因序列,抑郁组Shannon指数明显高于对照组。在门水平,共发现20个细菌门,抑郁患者肠道菌群丰度前3位的分别是厚壁菌门、拟杆菌门、梭杆菌门;在科水平,前3位主要为拟杆菌科、普雷沃氏菌科和瘤胃菌科;在属水平,前3位分别是多形杆菌属、普氏菌属、另枝菌属。对照组肠道菌群丰度前3位的分别是厚壁菌门、拟杆菌门、梭杆菌门;在科水平,前3位主要是拟杆菌科,普雷沃氏菌科和瘤胃菌科;在属水平,前3位分别是多形拟杆菌属、普氏菌属、另枝菌属。抑郁组多形杆菌属、普氏菌属、另枝菌属、栖粪杆菌属、考拉杆菌属丰度明显低于对照组,抑郁组毛螺菌属、副杆菌属、布劳特氏菌属、巨单胞菌属丰度明显高于对照组。抑郁组拟杆菌属和栖粪杆菌属丰度与SDS评分成负相关,毛螺菌属丰度与SDS评分成正相关。结论抑郁症患者病情严重程度与抗炎性细菌拟杆菌属和栖粪杆菌属丰度成反比,与毛螺菌属丰度成正比。     

乌鲁木齐市3~5岁儿童口腔微生物与肠道微生物的构成差异

目的探索乌鲁木齐市3～5岁儿童口腔微生物与肠道微生物的菌群构成及多样性等差异。方法从本团队前期流行病学调查的乌鲁木齐市3～5岁儿童中,按调查时间顺序随机抽取12名儿童。分别收集唾液及粪便样本共计24份,分为口腔微生物组和肠道微生物组。利用16S V3-V4区设计引物来进行PCR扩增,使用MiSeq测序仪进行二代测序,比较两组的微生物构成及多样性差异。结果门水平上:放线菌门(t=5.98,P0.001)、变形菌门(z=20.0,P=0.005)、TM7(z=78.0,P0.001)在口腔中丰度较高;厚壁菌门(z=134.0,P0.001)在肠道中丰度较高。厚壁菌门/拟杆菌门的值在肠道中较大(z=113.0,P=0.017)。属水平上:罗斯菌属、普雷沃菌属、链球菌属为口腔中丰度较高的菌属;粪杆菌属、Ruminococcaceae、拟杆菌属为肠道中丰度较高的菌属(P0.001)。奈瑟菌属、卟啉单胞菌属等仅存在于口腔中。Catenibacterium、肠杆菌属等仅存在于肠道中。菌群功能显示:在细胞运动(z=136.0,P0.001)、碳水化合物代谢(t=-4.71,P0.001)等方面表现出肠道组占优势;在遗传信息的翻译(t=8.17,P0.001)、神经退行性疾病(z=78.0,P0.001)等方面表现出口腔组占优势。结论放线菌门、变形菌门、TM7在口腔中丰度较高;厚壁菌门在肠道中丰度较高。口腔和肠道微生物在菌群功能上差异较大。奈瑟菌属、卟啉单胞菌属、Catenibacterium等可能为某些全身系统性疾病的标志性菌属。     

高通量测序检测肺炎婴幼儿抗生素治疗前后肠道菌群变化

目的了解抗生素对肺炎婴幼儿肠道菌群多样性和结构的影响。方法选取5例1岁以内因轻度社区获得性肺炎住院的患儿,收集抗生素治疗前及治疗后第3~5天的粪便标本。利用Illumina MiSeq测序平台,对细菌的16SrRNA基因V3~V4区进行高通量测序,并对测序结果进行生物信息学分析。结果抗生素治疗后第3~5天婴幼儿肠道菌群的多样性明显下降。变形菌门的相对丰度在3例婴幼儿的肠道中下降;厚壁菌门的相对丰度在4例婴幼儿的肠道中下降;而拟杆菌门的相对丰度在4例婴幼儿的肠道中上升;厚壁菌门与拟杆菌门的比例下降。肠道内不动杆菌属、喜胆汁菌属、埃格特菌属、肠球菌属、支原体、颤螺菌属、紫单胞菌属、普雷沃菌属和真杆菌属增长1倍以上;奇异菌属、双歧杆菌属、黄杆菌属、乳酸菌属、链球菌属、萨特菌属和韦荣球菌下降超过50%。结论婴幼儿短期使用抗生素治疗肺炎后,肠道菌群的多样性下降,肠道微生态失衡。     

胃癌患者化疗前后口腔菌群与肠道菌群变化特征

目的研究胃癌患者化疗前后口腔菌群与肠道菌群的变化特征,为该类患者的治疗提供参考。方法选取2017年2月至2020年2月于我院接受治疗的96例胃癌患者为观察组,另外选取同期我院健康体检者90例为对照组,收集两组对象唾液样本和粪便样本,其中观察组患者需收集化疗前及化疗后两个时间点的样本。采用16S rRNA基因高通量测序法检测菌群构成,分析两组对象组间及观察组患者化疗前后口腔菌群与肠道菌群变化特征。结果化疗前,观察组患者唾液样本、粪便样本菌群Simpson指数、Shannon指数、厚壁菌门、拟杆菌门丰富度与对照组比较差异均无统计学意义(均P>0.05)。化疗前,观察组患者唾液样本及粪便样本中放线菌门丰富度显著高于对照组,变形菌门丰富度显著低于对照组(均P<0.05)。化疗后,观察组患者唾液样本中厚壁菌门、放线菌门、变形菌门丰富度较化疗前显著下降,拟杆菌门丰富度显著上升(均P<0.05);同时粪便样本中厚壁菌门、变形菌门丰富度较化疗前显著上升,拟杆菌门、放线菌门丰富度显著下降(均P<0.05)。化疗后,观察组患者唾液样本中韦荣球菌属、罗氏菌属、普氏菌属丰富度较化疗前显著增加,放线菌属、链球菌属、奈瑟菌属、粪梭杆菌属丰度显著下降(均P<0.05);同时粪便样本中链球菌属、肠志贺菌属、乳杆菌属丰富度较化疗前显著增加,粪梭杆菌属、双歧杆菌属、拟杆菌属丰富度显著下降(均P<0.05)。结论化疗对胃癌患者口腔菌群及肠道菌群构成有一定程度影响,患者口腔菌群中韦荣球菌属丰度上升,而肠道中正常菌群的生长受抑制。     

WHBE兔与日本大耳白兔腹泻型肠易激综合征模型的肠道菌群研究

目的比较分析WHBE兔和日本大耳白(Japanese white rabbit,JW)兔腹泻型肠易激综合征(irritable bowel syndrome,IBS)模型肠道菌群的差异性。方法 WHBE兔和JW兔各16只,按品系随机分为两组,即正常对照(NC)组和IBS模型组,每组8只。采用湿热应激复合番泻叶方法建立腹泻型IBS兔模型,观察实验兔的腹围指数、粪便含水量、结肠转运功能,处死后分别取结肠组织和结肠内容物进行病理组织学观察和肠道菌群多样性分析。结果与NC组比,IBS模型兔均出现腹围指数和粪便含水量增加,结肠转运时间缩短,但结肠组织未见病理性改变;同时IBS模型兔Shannon指数和Chao1指数均显著下降(P0.05)。根据OTU分类分析的结果,厚壁菌门和拟杆菌门为兔肠道菌群的绝对优势菌群。与NC组比较,WHBE兔IBS模型组厚壁菌门、疣微菌门、绿弯菌门、Akk菌属、链球菌属均明显下降(P0.05,P0.01),拟杆菌门和rc4-4菌属丰度显著上升(P0.05,P0.01);而JW兔IBS模型组优杆菌属和罕见小球菌属均显著上升(P0.05),乳酸杆菌属、粪杆菌属、韦荣球菌属和链球菌属均显著下降(P0.05)。与JW兔NC组比,WHBE兔NC组厚壁菌门、Odoribacter菌属、韦荣球菌属、链球菌属、颤螺旋菌属、Pseudoflavonifractor菌属较低(P0.05,P0.01),拟杆菌门、疣微菌门、优杆菌属、粪杆菌属、Akk菌属较高(P0.05,P0.01)。与JW兔IBS模型组比,WHBE兔IBS模型rc4-4菌属、粪杆菌属、梭菌属的丰度较高(P0.05,P0.01),而厚壁菌门、多尔氏菌属、粪球菌属和罕见小球菌属的丰度则较低(P0.05,P0.01)。结论 IBS模型兔存在肠道菌群失调,导致菌群多样性降低;WHBE兔和JW兔IBS模型肠道菌群变化均具有其自身的特点,且具有明显的差异性。     

老年阿尔茨海默病患者肠道微生态改变及其与认知功能的关系探讨

目的探讨老年阿尔茨海默症患者肠道微生态结构与认知功能的关系。方法将2016年7月-2019年5月医院收治的86例老年阿尔茨海默病患者和73例健康志愿者,分别记为研究组和对照组。采用简易精神状态评价量表(MMSE)进行评分评价两组认知功能,采用高通量测序检测两组肠道微生物多样性。比较两组肠道微生物多样性与MMSE评分,并比较研究组中认知功能正常者与障碍者肠道微生物结构;采用Pearson相关性分析法探讨肠道微生物结构与认知功能障碍者的相关性。结果研究组肠道菌群Chao1指数、Shannon指数、放线菌门、拟杆菌门、放线菌科、乳杆菌科、拟杆菌科、双歧杆菌属、乳杆菌属、拟杆菌属、MMSE评分均低于对照组(均P0.05);研究组厚壁菌门、变形菌门、子囊菌门、梭菌科、假单胞菌科、葡萄球菌科、肠杆菌科、隐球酵母科、梭状芽孢杆菌属、假单胞菌属、葡萄球菌属、变形杆菌属、假丝酵母菌属相对丰度均高于对照组(均P0.05);研究组中认知功能障碍发生率为70.93%,且认知功能障碍者肠道菌群Chao1指数、Shannon指数,门、科、属水平相对丰度与认知功能正常者对比和上述结果一致;研究组与对照组肠道拟杆菌门、子囊菌门、拟杆菌科、梭状芽胞杆菌属、假丝酵母菌属、放线菌门、乳杆菌属、拟杆菌属相对丰度差异具有统计学意义(均P0.05);研究组认知功能障碍与认知功能正常者肠道未分类拟杆菌门、子囊菌门、假单胞菌属、拟杆菌属、变形杆菌属、梭状芽孢杆菌属、拟杆菌门、拟杆菌科、假丝酵母菌属相对丰度差异具有统计学意义(均P0.05);研究组中肠道菌群Chao1指数、Shannon指数,放线菌门、拟杆菌门、放线菌科、乳杆菌科、拟杆菌科、双歧杆菌属、乳杆菌属、拟杆菌属相对丰度与MMSE评分均呈正相关,厚壁菌门、变形菌门、子囊菌门、梭菌科、假单胞菌科、葡萄球菌科、肠杆菌科、隐球酵母科、梭状芽孢杆菌属、假单胞菌属、葡萄球菌属、变形杆菌属、假丝酵母菌属相对丰度与MMSE评分均呈负相关。结论老年阿尔茨海默病患者肠道微生物结构存在异常,且认知功能障碍者更加严重,认知功能与肠道微生物相关。     

基于16S rDNA测序绝经综合征抑郁患者肠道菌群结构与功能性分析

目的 通过16S rDNA测序技术对绝经综合征抑郁患者肠道菌群分布、多样性和基因功能进行分析,探讨肠道菌群与绝经综合征抑郁发生的关联。方法 选取2021年6月至2022年3月就诊于我院妇科门诊的绝经综合征抑郁患者19例为观察组（A组）,同期绝经综合征患者10例为对照组（C组）。利用16S r DNA基因测序法对患者肠道菌群进行物种注释分析和功能比较,统计两组肠道菌群分布、多样性和基因功能的变化。结果 两组肠道菌群差异明显。GraPhlAn物种组成图显示,患者肠道菌群总体以厚壁菌门、拟杆菌门、放线菌门和变形菌门组成。在门水平,与C组相比,A组放线菌门、变形菌门相对丰度较低,拟杆菌门相对丰度较高。在属水平,与C组相比,A组拟杆菌属、小杆菌属、巨单胞菌属、Lachnospiracea＿incertae＿se相对丰度较高,而Gemmiger、布劳特菌属、普雷沃菌属、粪球菌属、粪杆菌属、双歧杆菌属相对丰度偏低。进一步在属水平上选取了10种相对丰度差异具有统计学意义的菌群（P<0.05）：韦氏球菌属、消化球菌属、巨单胞菌属、史雷克菌属在A组的相对丰度较高（P<0.05）,柯林斯菌属、C...     

强直性脊柱炎患者咽部菌群变化

目的探讨强直性脊柱炎患者的咽部菌群变化。方法筛选入组7例强直性脊柱炎患者和7例健康者咽拭子样本,提取咽部DNA,扩增16SrRNA基因,在Illumina平台测序,对测序结果进行生物信息学分析。结果从ACE指数、Chao1指数、Shannon指数和Simpson指数综合来看强直性脊柱炎患者的咽部菌群Alpha多样性差异不大。Beta多样性分析显示两组研究对象咽部菌群样本可被区分。强直性脊柱炎患者咽部菌群组成和含量发生显著改变,主要变化包括:拟杆菌门(Bacteroidetes)和放线菌门(Actinobacteria)显著降低。拟杆菌门中普雷沃杆菌属(Prevotella)相关的纲目科属水平都显著降低。放线菌门变化落实到属水平,放线菌属(Actinomyces)显著降低,丙酸杆菌属(Propionibacterium)和棒状杆菌属(Corynebacterium)显著增高。厚壁菌门(Firmicutes)中,芽胞杆菌纲(Bacilli)所属的与链球菌属(Streptococcus)相关的纲目科属水平显著增加,而梭状芽胞杆菌纲(Clostridia)包含的韦荣球菌属(Veillonella)、消化球菌属(Peptococcus)显著下降。此外,变形菌门中出现弧菌属(Vibrio)的增加和弯曲菌属(Campylobacter)的降低等变化。结论强直性脊柱炎患者(本次研究样本)的咽部菌群出现紊乱,以普雷沃杆菌属、放线菌属、韦荣球菌属、消化球菌属和弯曲菌属等显著降低,丙酸杆菌属、棒状杆菌属、链球菌属和弧菌属等显著增加为主要特征。     

2型糖尿病与糖尿病肾病患者微炎症及肠道微生物多样性分析

目的分析2型糖尿病(T2DM)及糖尿病肾病(DKD)患者微炎症情况和肠道微生物多样性。方法将2016年4月至2019年7月在我院进行治疗的68例T2DM患者(T2DM组)和57例DKD患者(DKD组)纳入研究,选择同期于我院进行健康体检的36例志愿者作为对照组。收集3组对象一般资料、血液标本和粪便标本,测定血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDLC)、高密度脂蛋白胆固醇(HDLC)、糖化血红蛋白(HbAlc)、空腹血糖(FBG)、超敏C反应蛋白(hsCRP)、白细胞介素6(IL6)水平,并对肠道细菌进行16S rDNA序列测序。比较3组对象一般资料,血液指标,肠道菌群门水平构成情况,肠道菌群多样性,肠道菌属差异性,并对患者炎性指标与菌群种类进行相关性分析。结果DKD组患者糖尿病病程长于T2DM组(P<0.05)。与对照组比较,T2DM组和DKD组患者BMI、舒张压和收缩压增加,TC、TG、LDLC、HbAlc、FBG、hsCRP、IL6水平升高,HDLC水平降低(均P<0.05)。3组对象肠道菌群以厚壁菌门、拟杆菌门和变形菌门为主。T2DM组和DKD组患者拟杆菌门/厚壁菌门、蓝藻菌门及放线菌门比例均低于对照组(均P<0.05)。与对照组比较,T2DM和DKD患者肠道毛螺菌属、罗氏菌属、粪球菌属丰度升高,双歧杆菌属丰度降低(t=34.269 2、23.157 8、3.589 7、12.364 8,P<0.000 1、<0.000 1、=0.012 2、=0.036 1)。DKD组患者毛螺菌属和粪球菌属丰度高于T2DM组(t=65.349 2、13.573 1,P=0.021 3、0.001 3)。T2DM组和DKD组患者血清hsCRP水平与肠道毛螺菌属、瘤胃菌科、粪球菌属、梭杆菌属呈正相关,与红蝽菌科呈负相关;血清IL6水平与瘤胃菌科呈正相关,与红蝽菌科、双歧杆菌属、乳杆菌属呈负相关(均P<0.05)。结论2型糖尿病患者普遍存在微炎症和肠道菌群失衡,微炎症程度与肾脏病变和肠道菌群数量密切相关。T2DM与DKD患者在肠道菌群结构上具有一致性和差异性,肠道菌群检测有可能成为预测T2DM患者发生肾脏病变的风险指标。     

川崎病患儿肠道菌群构成及分布与其冠状动脉病变的相关性分析

摘要 目的：分析川崎病患儿肠道菌群构成及分布与其冠状动脉病变的相关性。方法：选择我院自2020年1月至2023年2月接诊的86例川崎病患儿作为研究对象,根据是否出现冠状动脉病变,分为冠状动脉病变组（35例）和非冠状动脉病变组（51例）。检测所有患儿的肠道菌群多样性[肠道菌群丰度（Ace指数）、肠道菌群多样性（Shannon指数）]、肠道菌群构成比例[门水平（变形菌门、厚壁菌门、拟杆菌门）、属水平（乳杆菌属、拟杆菌属、韦荣球菌属）],使用多因素Logistic回归分析肠道菌群构成及分布与冠状动脉病变的关系。结果：冠状动脉病变组Ace指数大于非冠状动脉病变组（P＜0.05）;冠状动脉病变组与非冠状动脉病变组的Shannon指数比较无差异（P＞0.05）;冠状动脉病变组肠道厚壁菌门占比低于非冠状动脉病变组,拟杆菌门占比高于非冠状动脉病变组（P＜0.05）;冠状动脉病变组与非冠状动脉病变组的肠道变形菌门占比比较无差异（P＞0.05）;冠状动脉病变组肠道乳杆菌属占比、韦荣球菌属占比均低于非冠状动脉病变组（P＜0.05）;冠状动脉病变组与非冠状动脉病变组的肠道拟杆菌属占比比较无差异（P＞0.05）;经多因素Logistic回归分析,肠道Ace指数、厚壁菌门、拟杆菌门、乳杆菌属、韦荣球菌属均是川崎病患儿发生冠状动脉病变的独立影响因素（P＜0.05）。结论：川崎病患儿肠道菌群构成及分布与其冠状动脉病变密切相关,为改善肠道菌群失调、增加患儿的临床获益提供依据,应引起临床重视。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.