目的 分析并探讨T细胞斑点技术诊断结核性胸膜炎的临床价值。方法 将2014年5月—2016年5月收治61例的结核性胸膜炎的患者和61例非结核性胸膜炎的患者同时行外周血和胸水的T细胞斑点实验,对各组数据进行统计学处理与分析。结果 结核组血T-SPOT的阳性率为88.52%,高于非结核组的16.39%,(P<0.001),其诊断结核性胸膜炎的灵敏度、特异度、阳性预测值、阴性预测值、阳性似然比、阴性似然比、youden指数分别为88.52％、83.61%、85.71%、86.44%、5.40、0.14、0.72。结核组胸水T-SPOT的阳性率为90.16%,高于非结核组的8.20%,(P<0.001),其诊断结核性胸膜炎的灵敏度、特异度、阳性预测值、阴性预测值、阳性似然比、阴性似然比、youden指数分别为90.16％、91.80%、91.67%、90.32%、11.00、0.11、0.82。联合外周血及胸水T-SPOT检测阳性率为91.80%,特异度为93.44%,阳性似然比为14.00,阴性似然比为0.09,youden指数为:0.85。结论 T细胞斑点实验检测对结核性胸膜炎有较高的临床诊断价值,外周血及胸水T-SPOT联合检测可提高诊断率,可作为结核性胸膜炎快速早期诊断的有效辅助手段。

Objective To analysis the value of T-SPOT.TB in the diagnosis of tuberculous pleurisy. Methods A total of 61 patients with clinically suspected tuberculous pleurisy (group A) and 61 cases of other diseases caused by pleural effusion (group B)admitted from May 2014 to May 2015 from the Guangzhou Chest Hospital had peripheral blood T-SPOT.TB and pleural effusion T-SPOT.TB,for each group of data for statistical processing and analysis. Results The positive rate of T-SPOT in the blood of the group A was 88.52%, which was higher than group B, (P<0.001) (16.39.0%). And the sensitivity, specificity, positive predictive value and negative predictive value, positive likelihood ratio(LR+), and negative likelihood ratio(LR－), Youden index were 88.52%, 83.61%,85.714%, 86.44%, 540, 0.14, 0.72 respectively by peripheral blood T-SPOT.TB in diagnosis of tuberculosis pleurisy. The positive rate of pleural effusion T-SPOT in the group A was 90.16%, which was also higher than group B (P<0.001). The sensitivity, specificity, positive predictive value and negative predictive value, positive likelihood ratio(LR+), and negative likelihood ratio(LR－), Youden index were 90.16％、91.80%、91.67%、90.32%、11.00、0.11、0.82 respectively by pleural effusion T-SPOT.TB in diagnosis of tuberculosis pleurisy. The positive rate of the joint tests of T-SPOT in peripheral blood and in peripheral blood was 91.80%, the specificity was 93.44%, the positive likelihood ratio was 14, the negative likelihood ratio was 0.09, and the youden index was 0.85. Conclusion T-SPOT.TB for the diagnosis of tuberculous pleurisy has a higher clinical value in the diagnosis. The joint tests of T-SPOT in peripheral blood and in pleural effusion may improve the positive rate in the diagnosis of tuberculous pleurisy. The joint tests of T-SPOT in peripheral blood and in pleural effusion have an important reference for diagnosing suspected tuberculous pleurisy quickly, early and accurately.

目的 研究鸟胞内分枝杆菌复合群(MAC)MAV3104基因编码蛋白与药物外排的关系。方法 以MAC标准株基因组为模板,扩增MAV3104基因,构建pMV261-MAV3104c重组质粒;测序正确后,转化重组质粒到大肠杆菌DH5并在MAC标准株中诱导表达, Western Blot鉴定MAV3104表达;按照CLSIM24-A2的操作要求检测MAC标准株对克拉霉素的敏感性及外排泵抑制试验。结果 经基因测序及Western Blot蛋白表达验证重组质粒构建成功;MAV3104过表达能提高鸟分枝杆菌对克拉霉素的MIC,且硫利达嗪能抑制该作用;MAV3104过表达也能提高胞内分枝杆菌对克拉霉素的MIC,但硫利达嗪对其没有抑制效应。结论 MAV3104转运蛋白介导的药物外排在鸟分枝杆菌耐克拉霉素中起重要作用。

Objective To investigate the association of the protein coded by Mycobacterium avium-Mycobacterium intracellulare complex in MAV3104 in drug efflux of Clarithromycin. Methods According to the Mycobacterium avium-Mycobacterium intracellulare complex standard strain, the MAV3104 gene was amplified by PCR and cloned into expression vector pMV261 to generate the recombinant plasmids. After verification by sequence analysis, the recombinant plasmids was transformed into E. coli DH5 andMycoba ctenum avium-Mycobacterium intracellulare complex standard strain. To identify the protein expression by western blotting and investigate the sensitivity of clarithromycin and efflux pump inhibition test in the light of CLSIM24-A2 protocol. Results It was verified by sequence analysis and western blotting that the recombinant plasmids were successfully constructed. Over expression of Mycobacterium avium MAV3104 gene could enhance clarithromycin MIC, which could be inhibited by thioridazine. Over expression of Mycobacterium intracellulare MAV3104 gene also could enhance clarithromycin MIC, but it could not inhibited by thioridazine. Conclusion This study demonstrates that efflux pumps mediated by MAV3104 protein play an important role in Mycobacterium avium resistance to clarithromycin.

目的 探讨拉米夫定治疗对乙肝活动的肺结核病人抗结核治疗中的临床价值。方法 通过回顾性分析159例初治肺结核乙肝表面抗原(HBsAg)阳性、HBV-DNA定量阳性病人,所有病例在抗结核前查肝功能均正常,分为两组,治疗组:在抗结核及护肝治疗过程中,加用拉米夫定抗乙肝病毒治疗;对照组:没用任何抗乙肝病毒药物;分别在抗结核治疗前、治疗2周、4周及8周复查肝功能,对比两组间肝损发生率,及肝损发生时间及严重程度。抗结核治疗4周后复查HBV-DNA定量,对比两组间HBV-DNA定量下降例数。结果 治疗组的肝损发生率仅20.5%,对照组病人抗结核治疗后肝损的发生率为53.1%,两者间差异有统计学意义。治疗组肝损出现时间多为大于4周,而且多数是轻度肝损。治疗组出现肝损中断抗结核治疗的病例数低于对照组。抗乙肝病毒治疗后复查HBV-DNA定量降低例数高于对照组。结论 拉米夫定抗乙肝病毒治疗,能抑制乙肝病毒复制,降低乙肝合并肺结核病人的肝损发生率并减轻肝损严重程度,提高病人对抗结核药物的耐受性。

目的 评价实时荧光定量PCR分析低氧性肺动脉高压大鼠肺动脉平滑肌基因表达时12个候选内参基因表达的稳定性,获得最适合的内参基因。方法 以低氧性肺动脉高压大鼠肺动脉平滑肌为研究对象,选择文献报道的常用12种内参基因为候选内参基因,利用geNorm和NormFinder程序分析实时荧光定量PCR数据,筛选出最适内参基因。结果 12个候选内参基因在低氧性肺动脉高压大鼠肺动脉平滑肌表达稳定性由强到弱顺序为:TBP>B2M>HPRT1>HMBS>RPL13a>18sRNA>PPIA>ACTB>GUSB>TFRC>GAPDH>PGK1,平均表达稳定度(M值)均<0.5,geNorm和NormFinder评估后推荐使用TBP和B2M一起作为该研究时的内参基因。结论 同时使用TBP和B2M是实时荧光定量PCR分析低氧性肺动脉高压大鼠肺动脉平滑肌基因表达的最适合内参基因,为低氧性肺动脉高压相关基因研究提供最优内参基因。

Objective To compare and select the suitable reference genes in real-time quantitative PCR analysis of rat pulmonary artery smooth muscle cells mRNA expression level of pulmonary hypertension. Methods To choose appropriate reference gene, the expression of twelve commonly use housekeeping genes were examined in rat pulmonary artery smooth muscle cells of hypoxia-induced pulmonary hypertension by using geNorm and NormFinder programs. Results The expression consistency of 12 genes was (from high to low): TBP>B2M>HPRT1>HMBS>RPL13a>18sRNA>PPIA>ACTB>GUSB>TFRC>GAPDH>PGK1. The average expression stability(M) values of them were low than 0.5. TBP and B2M reference genes were recommended to use in the same condition. Conclusion TBP and B2M reference genes were the most suitable combination of the reference genes for real-time quantitative PCR analysis in rat pulmonary artery smooth muscle cells of hypoxia-induced pulmonary hypertension.