MiR‐140 modulates the inflammatory responses of Mycobacterium tuberculosis‐infected macrophages by targeting TRAF6

This study aimed to examine miR‐140 expression in clinical samples from tuberculosis (TB) patients and to explore the molecular mechanisms of miR‐140 in host‐bacterial interactions during Mycobacterium tuberculosis (M tb) infections. The miR‐140 expression and relevant mRNA expression were detected by quantitative real‐time PCR (qRT‐PCR); the protein expression levels were analysed by ELISA and western blot; M tb survival was measured by colony formation unit assay; potential interactions between miR‐140 and the 3′ untranslated region (UTR) of tumour necrosis factor receptor‐associated factor 6 (TRAF6) was confirmed by luciferase reporter assay. MiR‐140 was up‐regulated in the human peripheral blood mononuclear cells (PBMCs) from TB patients and in THP‐1 and U937 cells with M tb infection. Overexpression of miR‐140 promoted M tb survival; on the other hand, miR‐140 knockdown attenuated M tb survival. The pro‐inflammatory cytokines including interleukin 6, tumour necrosis‐α, interleukin‐1β and interferon‐γ were enhanced by M tb infection in THP‐1 and U937 cells. MiR‐140 overexpression reduced these pro‐inflammatory cytokines levels in THP‐1 and U937 cells with M tb infection; while knockdown of miR‐140 exerted the opposite actions. TRAF6 was identified to be a downstream target of miR‐140 and was negatively modulated by miR‐140. TRAF6 overexpression increased the pro‐inflammatory cytokines levels and partially restored the suppressive effects of miR‐140 overexpression on pro‐inflammatory cytokines levels in THP‐1 and U937 cells with M tb infection. In conclusion, our results implied that miR‐140 promoted M tb survival and reduced the pro‐inflammatory cytokines levels in macrophages with M tb infection partially via modulating TRAF6 expression.     

5-Chloro-2-thiophenyl-1,2,3-triazolylmethyldihydroquinolines as dual inhibitors of Mycobacterium tuberculosis and influenza virus: Synthesis and evaluation

This study describes synthesis and evaluation of novel 5-Chloro-2-thiophenyl-1,2,3-triazolylmethyldihydroquinolines 7a-o as dual inhibitors of Mycobacterium tuberculosis and influenza virus. Huisgen’s [3+2] dipolar cycloaddition of 6-(azidomethyl)-5-chloro-2-(thiophen-2-yl)-7,8-dihydroquinoline 5 with various alkynes 6a-o using sodium ascorbate and copper sulphate gave new dihydroquinoline-1,2,3-triazoles 7a-o in good to excellent yields. The new compounds were evaluated for in vitro antimycobacterial against M. tuberculosis H37Rv (Mtb) and antiviral activity against influenza virus A/Puerto Rico/8/34 (H1N1). Among the fifteen new analogs, compounds 7a (MIC: 3.12 µg/mL), 7j and 7k (MIC: 6.25 µg/mL) were identified as potent antitubercular agents. The virus-inhibiting activity of all the fifteen compounds was found to be moderate, and among them the compound 7l, bearing thiophene moiety appeared the most active with good selectivity index (IC₅₀ = 19.5 µg/mL; SI = 15). The results presented here will help developing newer dual inhibitors of tuberculosis and influenza virus.     

结核分枝杆菌多表位诊断抗原的制备

将结核分枝杆菌(Mycobacterium tuberculosis,Mtb)多个B细胞预测表位串联表达(命名为B102),并初步评价其作为诊断抗原的血清学诊断价值。将MtbPstS1、ESAT6、CFP10、Ag85B、Ag85A及PPE54等6个蛋白的11个B细胞预测表位串联,加入合适的连接臂后全基因合成;将多表位片段插入带有TRX标签的表达质粒中,在大肠杆菌BL21(DE3)中诱导表达,并利用Ni~(2+)-Chelating亲和层析和DEAE阴离子交换层析纯化目的蛋白;利用Western blotting (WB)技术对目的蛋白抗原性进行鉴定,并建立Mtb抗体检测竞争法ELISA技术,初步评价此方法对阴阳血清样本的鉴别能力。目的蛋白以包涵体形式存在,其表达量约占菌体总蛋白的31.25%,经纯化及复性后蛋白B102可溶性存在,浓度为3.124mg/mL,纯度为96.71%;WB实验表明目的蛋白能与Mtb阳性血清相应抗体发生反应。对60份Mtb阳性血清及60份Mtb阴性血清进行检测得出其灵敏度为90.00%,特异性为93.33%,阳性预测值为93.10%,阴性预测值为90.32%,符合率为91.67%,McNemer检验的结果提示与"金标准"诊断结果无差异,Kappa=0.833,提示两种方法诊断结果一致性优异。原核表达与层析纯化可以获取抗原性优异的Mtb多表位诊断抗原,作为诊断抗原可以应用于Mtb的血清学检测中。     

Dispersal patterns in a medium‐density Irish badger population: Implications for understanding the dynamics of tuberculosis transmission

European badgers (Meles meles) are group‐living mustelids implicated in the spread of bovine tuberculosis (TB) to cattle and act as a wildlife reservoir for the disease. In badgers, only a minority of individuals disperse from their natal social group. However, dispersal may be extremely important for the spread of TB, as dispersers could act as hubs for disease transmission. We monitored a population of 139 wild badgers over 7 years in a medium‐density population (1.8 individuals/km²). GPS tracking collars were applied to 80 different individuals. Of these, we identified 25 dispersers, 14 of which were wearing collars as they dispersed. This allowed us to record the process of dispersal in much greater detail than ever before. We show that dispersal is an extremely complex process, and measurements of straight‐line distance between old and new social groups can severely underestimate how far dispersers travel. Assumptions of straight‐line travel can also underestimate direct and indirect interactions and the potential for disease transmission. For example, one female disperser which eventually settled 1.5 km from her natal territory traveled 308 km and passed through 22 different territories during dispersal. Knowledge of badgers' ranging behavior during dispersal is crucial to understanding the dynamics of TB transmission, and for designing appropriate interventions, such as vaccination.     

Tuberculosis Exacerbates HIV-1 Infection through IL-10/STAT3-Dependent Tunneling Nanotube Formation in Macrophages

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上海口岸输入性传染性肺结核病患者中结核分枝杆菌北京型特征及耐药性分析

本研究旨在了解上海口岸输入性传染性肺结核病患者中结核分枝杆菌北京型特征及其对一线抗结核药物的耐药情况,从而正确评估输入性结核病给上海地区带来的公共卫生危害,同时为地区传染性结核病防治策略的制定提供依据。针对2009年1月～2013年5月上海口岸入境体检人员中胸部影像学诊断疑似活动性肺结核病的人群,采集其连续3 d的晨痰分离培养结核分枝杆菌,用MGIT960培养法分析其对抗结核药物（链霉素、异烟肼、利福平、乙胺丁醇、吡嗪酰胺）的耐药情况,用目标缺失多重聚合酶链反应（DTM-PCR ）进行结核分枝杆菌北京型分子分型,同时采集其人口学资料。期间共监测到入境外籍疑似活动性肺结核病者193例,其中50例痰液中分离培养到结核分枝杆菌,菌培阳率为25．9％,与我国结核病普查工作中活动性肺结核病菌培阳率（25．9％）相同。分离培养并成功传代、分型的40株结核分枝杆菌中,北京型占57．5％（23／40）,显著低于同期上海口岸出境人群中的90．2％（37／41）。输入性结核分枝杆菌北京型主要来自东南亚地区（71．4％,5／7）和西太平洋地区（57．1％,16／28）。一线5种抗结核药物的耐药性检测结果显示,输入性结核分枝杆菌北京型的总耐药率为30．4％（7／23）,接近上海口岸出境人群的34．1％。输入性耐多药菌株占2．4％（1／42）,分子分型结果显示为非北京型。输入性传染性肺结核病对吡嗪酰胺的耐药率为16．7％,显著高于上海口岸出境人群的2．4％。结果提示,上海口岸输入性肺结核病患者中结核分枝杆菌北京型所占比例显著低于同期出境人群,未发现与性别和年龄相关,未发现输入性结核分枝杆菌北京型对抗结核药物耐药性的显著变化。输入性耐药性结核病带来的公共卫生危害不容忽视,在直接督导下的短程化疗（DOTS）方案中需考虑其高吡嗪酰胺耐药特征。在口岸公共卫生安全风险评估中,需重视输入性结核分枝杆菌北京型与本地流行株的差异。     

椎弓根螺钉用于植骨融合术治疗胸腰椎结核的临床研究

目的:探讨椎弓根内固定联合植骨融合术治疗胸腰椎结核的临床效果。方法:回顾性分析2013年5月至2014年5月在我院接受治疗的50例胸腰椎结核患者的临床资料,结合影像资料评价患者手术前后的红细胞沉降率、后凸畸形矫正情况、Frankel分级及术后并发症的发生情况等。结果:所有患者术后病理均证实为脊柱结核,术中27例植入大块自体髂骨,23例植入自体肋骨捆绑植骨。24例采用椎体侧前方钉棒内固定,26例采用后路椎弓根螺钉系统内固定。术中未出现脊髓、神经、血管损伤及血气胸等并发症。患者术后红细胞沉降率获得改善,差异具有统计学意义(P0.05)。术后Cobb角明显获得矫正,差异具有统计学意义(P0.05)。患者术后脊柱损伤程度明显改善,差异具有统计学意义(P0.05)。结论:椎弓根内固定联合椎体间植骨融合术治疗胸腰椎结核具有良好的临床效果,不仅可以改善红细胞沉降率,而且可以矫正患者脊柱后凸畸形,值得临床推广应用。