GC‐MS metabolomics identifies novel biomarkers to distinguish tuberculosis pleural effusion from malignant pleural effusion

BackgroundTuberculous pleural effusions (TBPEs) and malignant pleural effusions (MPEs) are two of the most common and severe forms of exudative effusions. Clinical differentiation is challenging; however, metabolomics is a collection of powerful tools currently being used to screen for disease‐specific biomarkers.Methods17 TBPE and 17 MPE patients were enrolled according to the inclusion criteria. The normalization gas chromatography‐mass spectrometry (GC‐MS) data were imported into the SIMCA‐P + 14.1 software for multivariate analysis. The principal component analysis (PCA) and orthogonal partial least‐squares discriminant analysis (OPLS‐DA) were used to analyze the data, and the top 50 metabolites of variable importance projection (VIP) were obtained. Metabolites were qualitatively analyzed using the National Institute of Standards and Technology (NIST) databases. Pathway analysis was performed by MetaboAnalyst 4.0. The detection of biochemical indexes such as urea and free fatty acids in these pleural effusions was also verified, and significant differences were found between these two groups.Results1319 metabolites were screened by non‐targeted metabonomics of GC‐MS. 9 small molecules (urea, L‐5‐oxoproline, L‐valine, DL‐ornithine, glycine, L‐cystine, citric acid, stearic acid, and oleamide) were found to be significantly different (p < 0.05 for all). In OPLS‐DA, 9 variables were considered significant for biological interpretation (VIP≥1). However, after the ROC curve was performed, it was found that the metabolites with better diagnostic value were stearic acid, L‐cystine, citric acid, free fatty acid, and creatinine (AUC > 0.8), with good sensitivity and specificity.ConclusionStearic acid, L‐cystine, and citric acid may be potential biomarkers, which can be used to distinguish between the TBPE and the MPE.     

利奈唑胺片治疗结核性脑膜炎的临床研究

目的观察利奈唑胺片联合常规抗结核治疗方案治疗结核性脑膜炎(TBM)的临床疗效及安全性。方法将65例初治TBM患者随机分为对照组35例和试验组30例。对照组予以异烟肼片每次0.4~0.6 g,qd,口服+吡嗪酰胺片每次0.5 g,tid,口服+乙胺丁醇片每次0.75 g,qd,口服+利福平胶囊Ⅱ每次0.45~0.60 g,qd,口服,总抗结核治疗疗程为12~18个月;试验组在对照组治疗的基础上,予以利奈唑胺片每次600 mg,qd,口服(重度昏迷患者予以鼻饲给药),疗程6个月。比较2组患者的临床疗效和药物不良反应发生情况,并分析利奈唑胺药物谷浓度的变化情况。结果治疗后,试验组和对照组的总有效率分别为90.00%(27例/30例)和65.71%(23例/35例),差异有统计学意义(P<0.05)。2组患者在周围神经炎、肝功能损害、视神经炎、胃肠道不适、骨髓抑制等方面的药物不良反应发生率相似。利奈唑胺在通过血脑屏障到达脑脊液谷浓度值与血浆中谷浓度值之比方面,在TBM治疗前12周明显高于后期,此比值与脑脊液各项生化指标无明显相关性。结论利奈唑胺片联合常规抗结核治疗方案能显著改善初治TBM患者的临床症状,提高临床疗效,缩短治疗疗程,且安全性较高;利奈唑胺的应用疗程以12周为宜。     

Potential cross‐reactivity of monoclonal antibodies against clinically relevant mycobacteria

Tuberculosis is a disease caused by the Mycobacterium tuberculosis complex (MTb). In 2011, global mortality due to tuberculosis was 1·4 million individuals. The only available vaccine is the attenuated M. bovis [bacillus Calmette–Guérin (BCG)] strain, which confers variable protection against pulmonary tuberculosis. Some widely distributed non‐tuberculous mycobacteria (NTM), such as M. avium and M. arupense, are also potential pathogens for humans. This work aimed to produce and characterize monoclonal antibodies against the M. bovis BCG Mexico strain of the MTb, M. avium subs. hominissuis and the M. arupense strain from NTM. Hybridomas were produced from splenocytes of BALB/c female mice immunized with radiation‐inactivated mycobacteria, and the immunoglobulin (Ig)G2a antibody‐producing clones with the highest antigenic recognition were selected. The selected clones, Mbv 2A10 for M. bovis BCG Mexico, Mav 3H1 for M. avium and Mar 2D10 for M. arupense, were used in further studies. Enzyme‐linked immunosorbent assay (ELISA) and immune proteomics analyses characterized the clones as having the highest cross‐reactivity with mycobacteria. Using mass spectrometry, a number of proteins recognized by the monoclonal antibody (mAb) clones were identified. These proteins had roles in metabolic processes, hypoxia, cell cycle and dormancy. In addition, a Clustal W and Immune Epitope Database (IEDB) in‐silico analysis was performed in protein sequences that result in the conserved regions within probability epitopes that could be recognized for Mbv2A10 and Mav3H1 clones.     

结核感染T细胞斑点试验联合胸腔积液Xpert MTB/RIF检测诊断结核性胸腔积液的价值

目的评估结核感染T细胞斑点试验（T-SPOT.TB）联合胸腔积液结核分枝杆菌/利福平耐药实时荧光定量核酸扩增检测技术(Xpert MTB/RIF) 在结核性胸腔积液的诊断价值。方法收集胸腔积液患者112例,其中确诊或临床诊断为结核性胸腔积液76例,非结核性胸腔积液36例。所纳入患者治疗前均行胸腔积液T-SPOT.TB、血T-SPOT.TB、胸腔积液Xpert MTB/RIF、胸腔积液腺苷脱氨酶（ADA）检测,并对其检测结果进行分析。结果胸腔积液T-SPOT.TB诊断结核性胸腔积液的敏感度（97.37%,74/76),高于外周血T-SPOT.TB （76.32%,58/76）、胸腔积液Xpert MTB/RIF（65.79%,50/76）和胸腔积液ADA（28.95%,22/76）,差异均有统计学意义（χ2=14.74、25.22、76.45,P<0.01）。胸腔积液Xpert MTB/RIF检测的特异度（100%,36/36）均高于胸腔积液T-SPOT.TB（77.78%,28/36）、外周血T-SPOT.TB（55.56%,20/36）和胸腔积液ADA（50.00%, 18/36）,差异均有统计学意义（χ2=6.89、20.57、24.00,P均＜0.01）。胸腔积液T-SPOT.TB和胸腔积液Xpert MTB/RIF联合检测的敏感度（64.47%, 49/76）低于胸腔积液T-SPOT.TB单独检测（97.37%,74/76）,差异有统计学意义（χ2=26.63,P<0.01）,但特异度由77.78%(28/36)提高至100%(36/36),差异有统计学意义（χ2=45.82,P<0.01）。结论胸腔积液T-SPOT.TB检测结核性胸腔积液具有较高的敏感度和特异度,胸腔积液T-SPOT.TB和胸腔积液Xpert Mtb/ＲIF联合检测可明显提高诊断的特异度。     

临床决策树法与积分模型法对结核性胸膜炎的诊断效能比较

目的 探讨临床决策树法与积分模型法对结核性胸膜炎的诊断效能。方法 采用前瞻性随机对照研究方法,以2019年1月1日—2019年6月30日武汉市肺科医院收治的胸腔积液患者为研究对象。符合入组标准的胸腔积液患者随机入组临床决策树法组和积分模型法组,分别采用两种综合诊断模型进行诊断,对比两组方法的诊断效能。结果 临床决策树组共纳入127例患者,其中男性90例,女性37例,年龄18~88岁,平均（49.33±19.63）岁,最后诊断结核性胸膜炎108例,非结核性胸膜炎19例;积分模型组共纳入120例患者,其中男性89例,女性31例,年龄13~84岁,平均（46.41±20.01）岁,最后诊断结核性胸膜炎94例,非结核性胸膜炎26例。临床决策树法诊断结核性胸膜炎的敏感度57.4%,特异度78.9%,阳性预测值93.9%,阴性预测值24.6%,诊断准确率60.6%;积分模型法诊断结核性胸膜炎的敏感度75.5%,特异度96.2%,阳性预测值98.6%,阴性预测值52.1%,诊断准确率80.0%。结论 积分模型法诊断结核性胸膜炎敏感度、特异度及诊断准确率均高于临床决策树法,方法简单,可推广应用于结核性胸膜炎的临床诊断。     

电视辅助胸腔镜手术在慢性结核性脓胸治疗中的应用价值

目的 探讨电视辅助胸腔镜手术(video-assisted thoracoscopic surgery,VATS)在慢性结核性脓胸治疗中的应用效果和安全性。方法 回顾性搜集2015年1月至2017年12月西安市胸科医院胸外科治疗的82例慢性结核性脓胸患者的年龄、性别、病程、病变部位等临床资料,并根据患者施行的手术方式分为VATS组(42例)和开胸手术组(40例)。比较两组患者手术时间、术中出血量、术后引流量、住院时间、拔管时间及并发症发生情况,肺功能恢复状况[主要指标包括用力肺活量(forced vital capacity,FVC)、第1秒用力肺活量(forced expiratory volume in one second,FEV₁)、肺总量(total lung capacity,TLC)],随访2年后的复发情况。结果 VATS组的术中出血量[(284.70±40.47)ml]、术后引流量[(687.40±78.58)ml],均明显低于开胸手术组[分别为(309.90±58.82)ml和(748.00±65.22)ml],差异均有统计学意义(t值分别为2.27和3.79,P值分别为0.03和0.00);VATS组和开胸手术组的手术时间[(130.54±14.95)min vs (135.30±13.09)min]、住院时间[(12.30±2.54)d vs (11.70±2.06)d] 及拔管时间[(6.80±1.32)d vs (6.56±1.03)d]差异均无统计学意义(t值分别为1.53、1.17、0.92,P值分别为0.13、0.25、0.36)。VATS组术后肺功能指标中FVC[(3.41±0.64)L]、FEV₁[(3.45±0.56)L]、TLC[(5.82±0.87)L]与开胸手术组比较[分别为(3.26±0.89)L、(3.34±0.73)L、(5.78±0.35)L],差异均无统计学意义(t值分别为0.79、0.50、0.26,P值分别为0.43、0.60、0.81)。VATS组术后并发症发生率为11.9%(5/42),明显低于开胸手术组的30.0%(12/40),差异有统计学意义(χ²=4.08,P<0.05)。随访2年后,开胸手术组复发率为5.0%(2/40),VATS组无复发,两组比较差异无统计学意义(χ²=2.15,P=0.14)。结论 对慢性结核性脓胸患者实施VATS,术中出血量、术后引流量及术后并发症发生率低,值得临床推应用。     

血清-腹水白蛋白梯度和腹水ADA、CA125在结核性腹膜炎诊治中的临床价值

[目的]探讨血清-腹水白蛋白梯度(SAAG)和腹水腺苷脱氨酶(ADA)、CA125对结核性腹膜炎的临床诊断价值。[方法]选取132例腹水患者进行回顾性分析,并将其分为4组:单纯性结核性腹膜炎组(A组)75例,肝硬化合并结核性腹膜炎组(B组)14例,肝硬化组(C组)17例,其他原因腹水组(D组)26例。对所有患者同一天的血清白蛋白及腹水白蛋白浓度进行检测,计算出SAAG;并测定腹水中的ADA、CA125浓度。[结果]以SAAG11g/L为临界值,诊断结核性腹膜炎的敏感度为96.6%(86/89)、特异性为74.4%(32/43)、准确率为89.4%(118/132),A、B组SAAG浓度显著低于C、D组(P0.05),A、B组间比较差异无统计学意义(P0.05);以ADA40U/L为临界值,诊断结核性腹膜炎的敏感性为92.1%(82/89)、特异性为97.7%(42/43)、准确率为93.9%(124/132),A、B组腹水ADA浓度与C、D组比较,差异有统计学意义(P0.05),A、B组间比较差异无统计学意义(P0.05),A、B组抗结核治疗前后腹水ADA浓度比较差异有统计学意义(P0.05);以CA12535U/ml为临界值,诊断结核性腹膜炎的敏感性为100%(89/89)、特异性为34.9%(15/43)、准确性为78.8%(104/132),各组腹水CA125浓度比较均差异无统计学意义(P0.05),A、B组抗结核治疗前后腹水CA125浓度比较差异有统计学意义(P0.05)。[结论]腹水ADA诊断结核性腹膜炎的敏感性、特异性、准确性均较高,并可作为判断结核性腹膜炎抗结核治疗疗效的观察指标;SAAG诊断结核性腹膜炎的敏感性较高,但特异性较低;腹水CA125对结核性腹膜炎无诊断价值,但可作为治疗疗效追踪的观察指标。     

糖尿病合并结核性胸膜炎24例临床分析

目的 回顾性分析糖尿病合并结核性胸膜炎患者的临床特征及其药物治疗的效果.方法 选择2009年1月至2011年6月在我院内分泌科住院的糖尿病合并结核性胸膜炎24例患者与同期住院的非糖尿病的结核性胸膜炎26例作对比分析.结果 相对于非糖尿病的结核性胸膜炎患者,糖尿病合并结核性胸膜炎患者胸水吸收慢,结核中毒症状缓解时间长.糖尿病合并结核性胸膜炎患者血糖控制良好患者的胸水吸收快.结论 糖尿病合并结核性胸膜炎应坚持两病同时治疗,在正规抗结核治疗同时,积极控制血糖.