高分辨率熔解曲线分析技术检测结核分枝杆菌的耐药性

目的:探讨高分辨率熔解曲线分析(High resolution melting,HRM)技术检测结核分枝杆菌耐药突变位点的可行性。方法:对218株结核分枝杆菌进行利福平(RFP)和异烟肼(INH)的药物敏感性测定,并进行耐药基因位点的PCR扩增和测序,同时采用HRM方法检测RFP和INH耐药基因位点情况,分析HRM的敏感性和特异性。结果:218株结核分枝杆菌药敏试验结果显示,有106株(48.6%)对RFP耐药,100株(45.9%)对INH耐药,81株(37.4%)对RFP和INH均耐药。测序发现,101株(46.3%)存在RFP耐药基因的突变,107株(49.1%)存在INH耐药基因的突变。HRM检测结果显示,100株(45.9%)存在RFP耐药基因的突变,103株(47.2%)存在INH耐药基因的突变。分别以药敏试验和测序结果为标准,HRM检测RFP耐药的敏感性为94.3%(100/106)和99.0%(100/101);特异性为97.3%(109/112)和100%(117/117);INH耐药的敏感性为97.0%(97/100)和98.1%(103/105);特异性为97.3%(109/112)和100%(113/113)。结论:HRM快速检测结核分枝杆菌耐药具有较高的特异性和灵敏度,能够满足临床需求。     

Interactions of pathogenic mycobacteria with host macrophages

Tuberculosis, caused by the bacterium Mycobacterium tuberculosis, is one of the most deadly infectious diseases across the globe. The success of M. tuberculosis is related to its capacity to survive and replicate in macrophages, cells of the host innate immune system that are designed to detect and eliminate pathogens [1,2]. In this review, we will focus on the mechanisms used by the innate system of the host to detect and eliminate mycobacteria and the strategies used by M. tuberculosis to overcome host responses to establish a successful infection.     

Solid-phase synthesis and biological evaluation of a uridinyl branched peptide urea library

A 1000-member uridinyl branched peptide library was synthesized on PS-DES support using IRORI technology. High-throughput screening of this library for anti-tuberculosis activity identified several members with a MIC₉₀ value of 12.5 μg/mL.     

结核杆菌可溶性细胞壁抗原与重组38Kd蛋白用于结核病血清学诊断的价值

目的探讨结核杆菌CW抗原和rTPA38蛋白用于结核病血清学诊断的价值。方法以CW和rTPA38蛋白为抗原,LAM为对照,用DICFA检测血清中的抗结核抗体。结果191例肺结核病人血清,用CW、rTPA38和LAM检测的敏感性分别为78．0％、65．5％和72．3％,特异性分别为95．9％、98．4％和95．9％。统计分析显示CW和rTPA38检测肺结核病人血清抗结核抗体的敏感性差异有非常显著性（χ^2=16．230,P〈0．01）。两者检测健康人和非结核组病人血清的特异性差异有显著性（χ^2=3．972,P〈0．05）。检测痰涂片阳性血清86例,发现CW和rTPA38与痰阳的一致率分别为84．9％和69．8％,CW抗原与痰涂片的阳性反应明显高于rTPA38。结论CW抗原有较好的敏感性和特异性,且与痰涂片有较高的符合率,有助于结核病的血清学诊断。     

Molecular Basis of Phosphatidyl-myo-inositol Mannoside Biosynthesis and Regulation in Mycobacteria

Phosphatidyl-myo-inositol mannosides (PIMs) are unique glycolipids found in abundant quantities in the inner and outer membranes of the cell envelope of all Mycobacterium species. They are based on a phosphatidyl-myo-inositol lipid anchor carrying one to six mannose residues and up to four acyl chains. PIMs are considered not only essential structural components of the cell envelope but also the structural basis of the lipoglycans (lipomannan and lipoarabinomannan), all important molecules implicated in host-pathogen interactions in the course of tuberculosis and leprosy. Although the chemical structure of PIMs is now well established, knowledge of the enzymes and sequential events leading to their biosynthesis and regulation is still incomplete. Recent advances in the identification of key proteins involved in PIM biogenesis and the determination of the three-dimensional structures of the essential phosphatidyl-myo-inositol mannosyltransferase PimA and the lipoprotein LpqW have led to important insights into the molecular basis of this pathway.     

MTB新疆临床分离株MDR-TB和XDR-TB耐药情况初步调查

目的:分析新疆喀什地区结核分枝杆菌(MTB)临床分离株对4种一线和7种二线抗结核药物的耐药情况,初步探讨本地区耐多药结核病(MDR-TB)和广泛耐药结核病(XDR-TB)的流行情况。方法:收集2008.11.1~2009.10.31日期间新疆喀什地区结核病患者痰液标本,进行分离培养及菌种鉴定,并应用比例法对所分离得到的菌株进行耐药性检测。结果:102株分枝杆菌中,88株(86.3%)属于结核分枝杆菌复合群,7株(6.9%)属于牛型分枝杆菌,7株(6.9%)属于非结核分枝杆菌。对一种以上的抗结核药物具有耐药性的有20株,总耐药率为22.7%(20/88),耐多药率为6.8%(6/88),pre-XDR为33.3%(2/6),无XDR-TB病例。结论:新疆喀什地区结核病患者耐药率较高,在结核病治疗工作中应给予重视,尤其应加强对Pre-XDR的重视,以免发展成XDR-TB。     

On the Mathematical Reconstruction of Two Dimensional Plants

A set of 10, chosen medicinal plants (some of them with a reputation as remedies for tuberculosis) has been investigated through Partitioned Iterated Function Systems-Semi Fractals with Angle (PIFS-SFA) coding, Lempel, Ziv, Welch with quantization and noise (LZW-QN) compression, and surface density statistics (f(α)-SDS) discrimination techniques. The final outcomes of this quantitative analysis were, firstly: the linear ordering of the plants in question accompanied by the hope that it reflects their medical significance, secondly: the mathematical representation of each of the plants, and thirdly: the impressive compression achieved, leading to remarkable computer memory saving, and still permitting successful pattern recognition i.e., proper identification of the plant from the compressed image.     

An unusual pro-inflammatory role of interleukin-10 induced by arabinosylated lipoarabinomannan in murine peritoneal macrophages

Various species of Mycobacteria produce a major cell wall-associated lipoglycan, called Lipoarabinomannan (LAM), which is involved in the virulence of Mycobacterial species. In this study, we tried to establish the role of the increased IL-10 secretion under Arabinosylated-LAM (Ara-LAM) treatment, the LAM that induces apoptosis in host macrophages or PBMC. We have studied the survival and apoptotic factors by western blotting, and estimated nitrite generation by Griess reaction, quantified iNOS mRNA by semi-quantitative RT-PCR, and ultimately the fate of the cells were studied by Flow Cytometric Analysis of AnnexinV-FITC binding. As per our observations, neutralization of released IL-10 in C57BL/6 peritoneal macrophages prior to Ara-LAM treatment, as well as macrophages from IL-10 knockout (KO) mice treated with Ara-LAM, showed significant down regulation of pro-apoptotic factors and up regulation of survival factors. These effects were strikingly similar to those when peritoneal macrophages were subjected to TNF-α and IL-12 neutralization followed by Ara-LAM-treatment. However, under similar conditions virulent Mannosylated-LAM (from Mycobacterium tuberculosis) treatment of macrophages clearly depicts the importance of IL-10 in the maintenance of pathogenesis, proving its usual immunosuppressive role. Thus, from our detailed investigations we point out an unusual pro-inflammatory action of IL-10 in Ara-LAM treated macrophages, where it behaves in a similar manner as the known Th1 cytokines TNF- α and IL-12. This work is financed by the Council of Scientific and Industrial Research (CSIR), Govt. of India.     

Human treponematosis and tuberculosis: evidence from the New World.

The purpose of this study has been first, to critically review the evidence for the presence of human treponematosis and tuberculosis in the skeletal remains of prehistoric natives in the New World, and second, to report on nine new cases dated to before contact and suggesting the presence of these two disease conditions. A review of the medical history and findings by human paleopathologists leaves little doubt that both diseases originated in the Old World. The findings of this study lend further support to the fact that, although rare, human treponematosis and tuberculosis were indeed endemic in the pre-Columbia New World before contact. There is no evidence that these two diseases could have arisen independently and de novo, especially during the relatively short time since man's arrival in the New World. Where a disease has been endemic for quite some time as appears to be the case with human treponematosis and tuberculosis, milder forms of the disease and improved host response could have developed in which only the most severe cases would be observable. This explains the rarity of skeletal lesions suggestive of these two human disease conditions in prehistoric human populations.