目的 分析对比肺部受累的惠普尔病与初治菌阴肺结核患者的临床特征,提高对肺部受累的惠普尔病的认识、诊断及鉴别水平。方法 回顾性收集20例肺部受累的惠普尔病为病例组,并随机选取同期56例初治菌阴肺结核患者为对照组进行对比,探讨肺部受累的惠普尔病的临床特征。结果 病例组中老年（>37岁）、急性或亚急性的病程比例高于对照组（P<0.05）。CT影像学中,从病灶分布来看,病例组的病灶分布在下叶的病例比例高于对照组（P<0.05）,病灶分布在上叶的病例比例低于对照组（P<0.05）;从形态上看,病例组中表现为网状病灶、间质性肺炎的比例高于对照组（P<0.05）,病例组中表现为空洞、纤维条索、实性影、结节影、树芽征、肺门淋巴结肿大的比例低于对照组（P<0.05）,而性别、合并免疫力低下疾病、咳嗽、咳痰、发热、咳血、关节痛、腹泻等在组间比较差异均无统计学意义。病例组全部病例的肺泡灌洗液宏基因测序结果显示：惠普尔养障体是唯一菌19例（95.0%）或主要菌1例（5.00%,合并结核菌1序列）,而对照组肺泡灌洗液宏基因测序结果显示：21例（37.5%）检测出结核分枝杆菌（TB）复合群是唯一菌或主要致病菌,18例TB-RNA（+）,15例TB-LAMP（+）。结论 惠普尔病在临床可表现为急性或亚急性病程,好发中老年男性,以发热和（或）呼吸道症状为主,可伴或不伴有腹泻、关节痛;肺部影像学以下肺网状、间质性肺炎改变为主,可以单独累及肺部;与初治菌阴肺结核患者临床症状极其相似。应尽快完善支气管镜检查,肺泡灌洗液的宏基因检测对早期、快速诊断此病尤为重要。

Objective To analyze and compare the clinical features of patients with pulmonary involvement of Whipple disease and primary treated bacteriological negative pulmonary tuberculosis,so as to improve the recognition,diagnosis and differentiation ability of pulmonary involvement of Whipple disease．Methods Clinical features of Whipple disease with pulmonary involvement were investigated by retrospectively collecting 20 cases as a case group,comparing with 56 randomly selected patients with primary treated bacteriological negative pulmonary tuberculosis as a control group during the same period．Results The case group had a significantly higher proportion of older patients（>37 years）,acute or subacute disease courses than the control group （P<0.05）．On CT imaging,in terms of lesion distribution,the proportion of cases with lesions in the lower lobe was significantly more in the case group than in the control group（P<0.05）,and the ratio of cases with lesions in the upper lobe was considerably less than in the control group（P<0.05）．Regarding morphology,significantly more of the case group showed reticular lesions and interstitial pneumonia than the control group（P<0.05）．Significantly fewer of the case group showed cavities,fibrous bands,solid shadows,nodular shadows,tree-bud signs,and enlarged hilar lymph nodes than the control group（P<0.05）．No statistically significant differences existed between the two groups in gender,combined immunocompromising diseases,cough,sputum,fever,coughing up blood,arthralgia or diarrhea. At the same time,the differences of gender,combined immunocompromised disease,cough,sputum,fever,coughing up blood,arthralgia,and diarrhea were not statistically significant．The metagenomic sequencing of alveolar lavage fluid in the case group revealed Tropheryma whipplei as the sole pathogenic bacteria in 19 cases（95%）or the primary pathogenic bacteria in 1 case（5.00%,combined TB 1 sequence）．In contrast,metagenomic sequencing of alveolar lavage fluid in the control group detected Mycobacterium tuberculosis complex as the sole or primary pathogenic bacteria in 21 cases（37.5%）,TB-RNA（+）in 18 cases and TB-LAMP（+）in 15 cases．Conclusions Clinical presentation in Whipple disease can be of an acute or subacute course,preferably in middle-aged and older men,with fever or/and respiratory symptoms,either with or without diarrhea and arthralgia．Its pulmonary imaging dominates with reticular,interstitial pneumonia changes in the lower lungs,which can involve the lungs alone in the disease．At the same time,the clinical symptoms are highly similar to those patients with primary treated bacteriological negative pulmonary tuberculosis．Therefore,it is vital to complete bronchoscopy and metagenomic sequencing of alveolar lavage fluid as soon as possible for early and rapid diagnosis of this Whipple disease．Treatment with sensitive antibacterial drugs can result in significant improvement and save patients' lives．

目的 探讨肠道菌群多样性及丰度在抗结核治疗强化期的变化。方法 收集广州市胸科医院20例初治菌阳肺结核患者使用强化方案治疗1周及强化期结束的粪便标本,所有标本进行16S rDNA测序,进行生物信息学分析。结果 强化期结束治疗组的Alpha多样性指标Chao1和observed_otus指数分别为(97.8±28.3)和(97.6±28.2),高于治疗1周组的(81±34.7)和(81±34.7),差异有统计学意义(P=0.04;P=0.038)。基于加权和未加权Unifrac距离的PCoA分析显示两组整体菌群结构无明显差异。在相对丰度较高的菌科中,强化期结束治疗组的拟杆菌科、肠球菌科、肠杆菌科、丹毒菌科、卟啉单胞菌科、双歧杆菌科等相对丰度下降,毛螺菌科、梭杆菌科、普氏菌科、瘤胃球菌科、韦氏菌科等相对丰度增加。通过线性判别分析效应量LEfse分析两组数据,发现在属水平上,厚壁菌门的瘤胃球菌、乳杆菌及放线菌门的柯林氏菌为组间显著差异菌种。结论 强化期治疗结束与治疗1周相比,肠道菌群多样性增加,但肠道菌群结构无明显差异。肠道拟杆菌科丰度减少,毛螺菌科丰度增加。

Objective To investigate the changes in the diversity and abundance of intestinal flora during the intensive period of anti-tuberculosis treatment. Methods Stool specimens from 20 patients with newly treated bacteria-positive pulmonary tuberculosis in Guangzhou Chest Hospital, which were treated with the intensive treatment for 1 week and whole intensive treatment, were collected. All the specimens were subjected to 16S rDNA sequencing and bioinformatics analysis. Results The Alpha diversity index Chao1 and observed_otus index of the treatment group with whole process were (97.8±28.3) and (97.6±28.2) respectively, which were higher than (81±34.7) and (81±34.7) of the 1-week treatment group, which had statistical differences (P=0.04;P=0.038). PCoA analysis based on weighted and unweighted Unifrac distance showed that there was no significant difference in the overall flora community structure between the two groups. Among the bacteria families with higher relative abundance, the relative abundance of Bacteroidaceae,Enterococcaceae,Enterobacteriaceae,Erysipelotrichaceae,Porphyromonadaceae and Bifidobacteriaceae in the treatment group with whole process decreased, the relative abundance content of Lachnodoiraceae,Fusobacteriaceae,Prevotellaceae, Rumincoccaceae, and Veillonellaceae increased. By using linear discriminant analysis effect size (LEfse) to analyze the two sets of data, it was found that at the genus level,Rumincoccaceae of Firmicutes and Lactobacillus;Collinsella of Actinobacteria were significantly different species between the groups. Conclusion Compared with 1 week of treatment, the diversity of intestinal flora increased at the end of intensive treatment, but there was no significant difference in the community structure of intestinal flora. The abundance of intestinal Bacteroidaceae decreased, and the abundance of Lachnodoiraceae increased.

目的 探讨继发性肺结核合并肺部真菌感染的临床特点及相关高危因素。方法 收集广州市胸科医院2017年7月—2019年10月收治的继发性肺结核患者资料,病程均大于3个月,分为真菌感染组106例和非真菌感染组100例进行回顾性分析。结果 单因素分析结果显示,合并肺部其他疾病、非初治、咯血、空洞、应用广谱抗生素>l周、侵袭性操作存在统计学差异(P<0.05)。Logistic多因素分析结果显示,广谱抗生素使用>l周、侵袭性操作为真菌感染的独立危险因素(P<0.05)。结论 对于应用广谱抗生素、进行侵袭性操作的肺结核患者应警惕真菌感染风险,及早预防及诊治。

Objective To investigate the clinical characteristics and related high risk factors of secondary pulmonary tuberculosis complicated with pulmonary fungal infection. Methods Data of patients with secondary tuberculosis admitted to Guangzhou Chest Hospital from July 2017 to October 2019 were collected. All patients with a course of disease longer than 3 months were divided into the fungal infection group (n =106) and the non-fungal infection group (n =100) for retrospective analysis. Results Univariate analysis results showed that there were statistical differences in combined other pulmonary diseases, non-initial treatment, hemoptysis, cavity, application of broad-spectrum antibiotic > for 1 week, and invasive operation (P<0.05). Logistic multivariate analysis showed that >1 week of broad-spectrum antibiotics and invasive procedures were independent risk factors for fungal infection (P<0.05). Conclusion Patients with tuberculosis who are treated with broad-spectrum antibiotics and invasive procedures should be alert to the risk of fungal infection, early prevention and treatment should be undertaken.

目的 探讨近期大咯血对支气管镜诊疗操作相关出血的影响。方法 回顾性分析2015年12月—2019年12月我院支气管镜操作导致出血超过30 mL患者的临床资料,根据近1周是否发生大咯血,分为观察组(大咯血组)和对照组(无大咯血组),比较2组间的出血量以及大出血、血凝块、窒息的发生率。结果 观察组的出血量以及大出血的发生率高于对照组,2组间差异有统计学意义,血凝块、窒息的发生率两组间差异无统计学意义。结论 近期大咯血的患者支气管镜操作发生大出血的风险高。

Objective To explore the effect of short-term massive hemoptysis on the procedure-related bleeding of bronchoscopy. Methods The clinical data of patients with more than 30 mL bleeding caused by bronchoscopy operation in Guangzhou Chest Hospital from December 2015 to December 2019 were analyzed retrospectively. According to whether massive hemoptysis occurred in the past week, the patients were divided into observation group (massive hemoptysis group) and control group (no massive hemoptysis group). The bleeding volume, the incidence of massive hemorrhage, blood clot and asphyxia were compared between the two groups. Results The amount of bleeding and the incidence of massive hemorrhage in the observation group were higher than those in the control group, that there was a statistical significance between the two groups. There was no statistical significance between the two groups in the incidence of blood clots and asphyxia. Conclusion Patients with recent massive hemoptysis have a higher risk of massive haemorrhage due to bronchoscopy.

目的 探讨以肺间质改变为主的不典型肺结核的CT特征。方法 收集2018 年1月—2018 年12月期间44例经我院临床及实验室确诊的以肺间质改变为主的不典型肺结核的CT影像学资料,进行归纳总结,分析其影像特点。结果 44例以肺间质改变为主的不典型肺结核其中14例患者在肺叶的分布呈局限性分布,其中主要分布于右肺上叶占13例;30例呈弥漫性分布;以间质改变为主的局限性分布及弥漫性分布的不典型肺结核均以细网织影、微结节、细支气管壁增厚、小叶间隔增厚为主要表现,树芽征、磨玻璃影在弥漫性分布中相对常见,其合并空洞、支气管扩张、纵膈淋巴结肿大钙化、胸膜炎、肺大疱的比例高于局限性分布患者;以肺间质改变为主的不典型肺结核呈局限性分布(14例)的患者预后良好,13例呈显著吸收,1例吸收良好;弥漫性分布(30例)的患者中,18例显著吸收,其中6例未吸收或吸收欠佳。结论 以肺间质为改变为主的非典型肺结核具有其特征性,了解其CT特点有助于提高临床诊断及更好的评价治疗预后。

Objective To discuss value of the CT features of atypical pulmonary tuberculosis with pulmonary interstitial changes and improve the early diagnosis of pulmonary tubeculosis. Methods The CT imaging data of 44 patients with atypical pulmonary tuberculosis with interstitial changes was collected from January 2018 to December 2018,and the imaging characteristics were analyzed. Results In 44 cases of atypical pulmonary tuberculosis,14 cases of atypical pulmonary tubeculosis were the localized distribution,13 cases were mainly in the right upper lobe,and 30 cases were diffuse distribution. The atypical pulmonary tuberculosis of the localized and diffuse distribution mainly with interstitial changes was characterized by tine reticular shadow,micnodule,thickening of bronchiole wall and thickening of interlobular Septa. Tree-bud sign and ground-glass shadow were common in diffuse distribution,and the proportions of cavity,bronchiectasis,mediastinal lymphadenopathy calcification,pleurisy and bullae were higher than that in localized distribution. The atypical pulmonary tuberculosis patients with localized pulmonary interstitial changes (14 cases) have a favourable prognosis,13 cases with significant absorption,1 case with favourable absorption,and 18 cases with the atypical pulmonary tuberculosis patients with diffuse pulmonary interstitial changes (30 cases) with significant absorption,6 cases with no absorption or poor absorption. Conclusion The atypical pulmonary tuberculosis with interstitial changes has its characteristics,and the CT features is helpful to improve the clinical diagnosis and better evaluation of treatment prognosis.

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 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.