2023年8月31日，北京青年呼吸学者沙龙2023年第3次活动在北京顺利举行，本次沙龙主题是“分子生物学技术快速诊断非结核分枝杆菌病的是与非”。活动形式是病例分享和讨论。通过这次活动，与会学者对分子生物学技术快速诊断非结核分枝杆菌病的优势和局限性进行了深入讨论，加深了非结核分枝杆菌病快速诊断的认识。呼吸沙龙新颖的组织和活动形式，获得了与会中青年呼吸学者的好评。

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb) infection, is currently the deadliest infectious disease in human that can evolve to severe forms. A comprehensive immune landscape for Mtb infection is critical for achieving TB cure, especially for severe TB patients. We performed single-cell RNA transcriptome and T-cell/B-cell receptor (TCR/BCR) sequencing of 213,358 cells from 27 samples, including 6 healthy donors and 21 active TB patients with varying severity (6 mild, 6 moderate and 9 severe cases). Two published profiles of latent TB infection were integrated for the analysis. We observed an obviously elevated proportion of inflammatory immune cells (e.g., monocytes), as well as a markedly decreased abundance of various lymphocytes (e.g., NK and γδT cells) in severe patients, revealing that lymphopenia might be a prominent feature of severe disease. Further analyses indicated that significant activation of cell apoptosis pathways, including perforin/granzyme-, TNF-, FAS- and XAF1-induced apoptosis, as well as cell migration pathways might confer this reduction. The immune landscape in severe patients was characterized by widespread immune exhaustion in Th1, CD8+T and NK cells as well as high cytotoxic state in CD8+T and NK cells. We also discovered that myeloid cells in severe TB patients may involve in the immune paralysis. Systemic upregulation of S100A12 and TNFSF13B, mainly by monocytes in the peripheral blood, may contribute to the inflammatory cytokine storms in severe patients. Our data offered a rich resource for understanding of TB immunopathogenesis and designing effective therapeutic strategies for TB, especially for severe patients.

Objective: To assess the efficacy of different antibiotics combined on Mycobacterium abscessus subspecies abscessus pulmonary disease（M.abscessus-PD） and analyze the influencing factors of treatment outcomes for 6 months. Methods: Sixty-six patients in Beijing Chest Hospital, Capital Medical University diagnosed with M.abscessus-PD were enro1led prospectively from January 1st, 2016 to October 1st, 2022. Clinical information of the patients were collected, including gender, age, body mass index, clinical manifestations, history of past illness and anti-tuberculosis treatment, imaging examinations, laboratory examinations and medications. The factors influencing treatment outcomes of M.abscessus-PD for 6 months among different therapicregimens was compared. Results: After 6 months of treatment, 32（48.5%） achieved sputum culture conversion and 34（51.5%） didn’t among the 66 patients. The result shows that the 6-month sputum culture conversion rate of azithromycin users（57.4%, 27/47） was higher than clarithromycin users（26.3%, 5/19）, with statistically significant（χ~2=5.250,P=0.022）. In 24 cases（36.4%, 24/66） treated with imipenem, 17（70.8%, 17/24） achieved 6 months sputum culture conversion, which was higher than that of non-imipenem（35.7%, 15/42）, with statistically significant difference（χ~2=7.542, P=0.006）. The efficacy of multidrug therapy regimen showed that macrolide combined with amikacin, imipenem, linezolid and clofazimine achieved 68.4%（13/19） culture conversion rate after 6 months. Multivariate analysis shows that patients treated with macrolide combined with imipenem were more likely to achieve sputum culture conversion within 6 months（OR（95%CI）=0.229（0.077-0.676））. Conclusion: In the initial stage of treatment, at least 4 weeks of injection use（amikacin and imipenem）, and the combination of macrolides with amikacin, clofazimine, and linezolid in the continued treatment, may have good curative effect on M.abscessus-PD.

Objective: To understand the characteristics of nontuberculous mycobacteria（NTM） pulmonary disease, and provide evidence for clinical diagnosis and treatment of NTM pulmonary disease. Methods: The clinical data of 93 patients with NTM pulmonary disease admitted to Beijing Chest Hospital affiliated to Capital Medical University between January 2018 and December 2019 were analyzed retrospectively. Treatment effectivenesses were compared for different treatment. Results: Among 93 patients with NTM pulmonary disease, there were 49 cases infected with Mycobacterium intracellular, 28 cases with Mycobacterium abscesses, 4 cases with Mycobacterium avium, 4 cases with Mycobacterium kansasii, 3 cases with Mycobacterium xenopi, 2 case co-infected with Mycobacterium abscesses and Mycobacterium intracellular, one case with Mycobacterium gordonii, Mycobacterium fortuitum or Mycobacterium stutzeri each. 61 patients（65.59%） had received antituberculosis treatment before the diagnosis of NTM pulmonary disease. 90（96.77%） patients had complications, of which 52（57.78%） patients had bronchiectasis, followed by 12（13.33%） patients with chronic obstructive pulmonary disease（COPD）. 83 cases（89.25%） received anti-NTM treatment, and 47 cases（56.63%） completed the course of treatment among whom the success rate of the treatment of pulmonary disease with Mycobacterium avium complex was 51.85%（14/27） while the treatment success rate of Mycobacterium abscesses pulmonary disease was 76.92%（10/13）. 48 cases（57.83%） had adverse drug reactions of different degrees during the treatment, and 15 cases（18.07%） discontinued the treatment. 16 patients（19.28%） were lost to follow-up. Conclusion: Mycobacterium intracellular lung disease is the most common NTM pulmonary disease. Most patients were complicated with bronchiectasis, COPD and other basic lung diseases. The clinical characteristics of NTM pulmonary disease and pulmonary tuberculosis are similar, which are easily misdiagnosed as pulmonary tuberculosis. The cure rate of NTM pulmonary disease is low, and the rate of treatment interruption and loss to follow-up is high.

Background: Tuberculosis (TB), caused by Mycobacterium tuberculosis(Mtb), continues to be an important global health problem. Hence, gaining a comprehensive understanding of the immune characteristics in TB could aid the improvement of vaccines and therapeutics for TB, especially for severe TB.Methods: Single-cell RNA transcriptome and T-cell/B-cell receptor (TCR/BCR) sequencing were applied to dissect the peripheral immune responses of active TB and reveal the molecular mechanisms of TB pathogenesis. Single-cell transcriptional profiles were obtained from 6 healthy donors and 21 active TB patients with varying severity (6 mild, 6 moderate and 9 severe cases) and integrated with 2 published profiles of latent TB infection.Findings: We observed a decrease in multiple peripheral immune cells (e.g., NKs, DCs and MAIT), and an increase in monocytes and megakaryocytes in TB patients, particularly with severe disease. We also discovered a significant increase in monocytes resembling myeloid-derived suppressor cells in severe patients that may be involve in immune paralysis. The immune landscape in severe patients were characterized by widespread immune exhaustion in Th1, CD8+ T and NK cells, high cytotoxic state in CD8+ T and NK cells, and significant activation of cell apoptosis and migration pathways in T and NK cells. Systemic upregulation of pro-inflammatory cytokines and inflammatory response genes, mainly by monocytes, and similarly increased pro-inflammatory cytokine concentrations in the plasma may contribute to the inflammatory cytokine storms previously observed in severe patients.Interpretation: This comprehensive single-cell analysis of peripheral immune landscape will improve the understanding of TB immunopathogenesis, offering potential targets for therapies and vaccines for TB control.Funding Information: This work was supported by grants from National Key Research and Development Program of China (Grant Nos. 2021YFC2301101, 2021YFC2301102), Public service development and reform pilot project of Beijing Medical Research Institute (BMR2019-11), National natural science foundation of China (81970900, 82100011), Beijing Public Health Experts Project (2022-3-040), Beijing Social Science Foundation Project (19GLB033), Key Project of the Department of Science and Technology, Beijing, China (Grant Nos.D181100000418003, Z191100006619078).Declaration of Interests: The lead author and guarantor affirm that the manuscript is an honest, accurate, and transparent account of the study being reported; that no important aspects of the study have been omitted; and that any discrepancies from the study as planned and registered have been explained.Ethics Approval Statement: The ethical approval for this study was obtained from the Beijing Chest Hospital ethics committee (ethical approval No. YNLX-2022-006). Written informed consent was acquired from each participant.

Background and purpose: We investigated whether periodontal diseases, specifically, periodontitis and gingivitis, could be risk factors of the incidence of intracranial aneurysms (IAs).Methods: We performed a case–control study to compare the differences in the periodontal disease parameters of 281 cases that were divided into the IA and non-IA groups. All cases underwent complete radiographic examination for IAs and examination for periodontal health. Results: Compared with those in the non-IA group, the cases in the IA group were older (53.95 ± 8.56 vs 47.79 ± 12.33, p < 0.001) and had a higher incidence of hypertension (76 vs 34, p = 0.006). Univariate analysis revealed that age (>50 years) and hypertension were risk factors of aneurysm formation (odds ratio [OR] 1.047, 95% confidence interval [95%CI] 1.022–1.073, p < 0.001 and OR 2.047, 95%CI 1.232–3.401, p = 0.006). In addition, univariate and multivariate logistic regression analyses showed that the parameters of periodontal diseases, including gingival index, plaque index, clinical attachment loss, and alveolar bone loss, were significantly associated with the occurrence of IA (all p < 0.05). For further statistical investigation, the parameters of periodontal diseases were divided into four layers on the basis of their averages. Poor periodontal health condition was correlated with a high risk of IAs. Conclusion: Periodontal diseases, especially severe gingivitis or periodontitis combined with hypertension, were significantly associated with the incidence of IAs.

目的 探讨结核性胸膜炎患者化疗时是否加用糖皮质激素对肺功能的动态变化.方法 回顾性分析首都医科大学附属北京胸科医院2015年5月至2018年5月收治的结核性胸膜炎患者172例,对患者的肺通气、容积、弥散功能和呼吸肌力学指标在化疗前,以及化疗第1、6、12个月末进行4次检测;根据在抗结核药品化疗方案的基础上是否加用糖皮质激素分为两组,即加用组60例(醋酸泼尼松+2H-R-Z-E/10H-R-E)和未加用组112例(2H-R-Z-E/10H-R-E).肺通气功能的检测指标为患者用力肺活量检测值占正常预计值的百分比(FVC)、第1秒用力呼气容积检测值占正常预计值的百分比(FEV1％pred)、第1秒钟用力呼气容积/用力肺活量比值占正常预计值的百分比(FEV1/FVC％pred)、用力呼出75％肺总量时瞬间呼气流量检测值占正常预计值的百分比(FEF75)、最大分钟通气量检测值占正常预计值的百分比(MVV％pred);肺容积功能的检测指标为残气量检测值占正常预计值的百分比(RV％pred)、肺总量检测值占正常预计值的百分比(TLC％pred)、残气量/肺总量比值占正常预计值的百分比(RV/TLC％pred);弥散功能的检测指标为肺弥散量检测值占正常预计值的百分比(DLCO)和肺泡容量校正的肺弥散率检测值占正常预计值的百分比(DLCO/VA％pred);呼吸肌力学的检测指标为气道阻力检测值占正常预计值的百分比(Rtot％pred、呼气峰流量检测值占正常预计值的百分比(PEF％pred)、吸气峰值流量实测值(PIF),通过对检测指标的观察,了解肺功能的变化.肺功能测定结果按"测定值/正常预计值×100％"表示(除PIF为实测值),符合正态分布的计量资料采用"(x)±s"进行统计描述,统计学处理采用t检验;不符合正态分布的计量资料采用中位数(四分位数)[M(Q1,Q3)]表示,统计学处理采用Z检验,P＜0.05为差异有统计学意义.结果 (1)结核性胸膜炎患者化疗前,以限制性通气功能障碍为主,FVC％pred为(62.1±13.4)％,FEV1％pred为(64.4±15.5)％,FEV1/FVC％pred为(87.0±11.1)％,MVV％pred(65.7± 21.1)％,FEF75％pred为61.6(41.6,83.0)％,RV％pred为 111.3(89.8,131.4)％,TLC％pred为(77.0±16.9)％,RV/TLC％pred为(146.9±35.9)％,DLCO％pred为(62.6±18.3)％,DLCO/VA％pred为(92.3±16.6)％,PEF％pred为(64.1±18.13)％,PIF为3.2(2.3,4.1)L/s,Rtot％pred为96.0(69.3,118.9)％].(2)化疗过程中,第1、6、12个月末FVC＜80％的患者分别为80.2％(138/172)、75.0％(129/172)、0.0％(0/0);化疗第6个月末DLCO＜80％的患者为44.2％(76/172));(3)加用组和未加用组患者在化疗前、化疗第1、6、12个月末FEF75％pred[分别为(68.7(49.8,84.1)％和60.7(39.4,80.7)％);87.1(70.5,94.4)％和73.1(51.9,87.0)％;80.1(66.5,111.9)％和66.8(59.9,87.2)％;90.4(55.3,102.9)％和78.4(54.6,87.3)％],两组比较差异均无统计学意义值分别为-1.091、-0.111、—1.609、-1.171,P值均＞0.05);MVV％pred[分别为(65.8±19.4)％和(65.5±18.6)％);(86.9±18.6)％和(79.5±18.7)％;(90.3±16.0)％和(86.3±16.0)％;(96.8±11.1)％和(87.3±19.8)],两组比较差异均无统计学意义(t值分别为1.043、0.444、0.708、1.113,P值均＞0.05).结论 结核性胸膜炎患者治疗前、治疗第1个月末肺功能减退主要表现为限制性通气功能障碍和弥散功能减退,治疗第6个月末仅通气功能恢复正常,而弥散功能在治疗第12个月末恢复正常;早期糖皮质激素辅助治疗对结核性胸膜炎患者的肺功能无明显影响.

目的 评价经内科胸腔镜胸膜病变活检对结核性胸膜炎的诊断价值及安全性.方法 2015年6月至2018年10月,首都医科大学附属北京胸科医院、首都医科大学附属北京朝阳医院、北京积水潭医院、卫生部北京医院等4家临床中心采用前瞻性多中心诊断试验方法,对参照入组标准顺序纳入的229例不明原因胸腔积液的入院患者行内科胸腔镜检查,并对活检留取胸膜病变组织标本行结核分枝杆菌GeneXpert MTB/RIF(简称"GeneXpert")和BACTEC MGIT 960培养(简称"MGIT 960")及常规病原学检测和病理学检查.结果 229例患者经内科胸腔镜取胸膜活检组织行病理学和病原学检查结果显示,临床不能明确诊断者23例(10.0％),确诊者为206例(90.0％),其中129例(56.3％)确诊为结核性胸膜炎,77例(33.6％)诊断为其他原因所致的胸腔积液.胸膜活检组织经GeneXpert或MGIT 960检测结核感染的阳性率[分别为27.9％(64/229)和17.0％(39/229)]与病理学检测阳性率[(23.1％,53/229)]比较,差异无统计学意义(x2=1.32,P=0.251;x2=2.67,P=0.103);但GeneXpert+MGIT 960联合检测的阳性率[32.8％(75/229)]明显高于传统病理学检测(x2=5.25,P=0.022).结核性胸膜炎患者的胸腔镜镜下特征性表现在纤维粘连带[70.5％(91/129)]、弥漫性粟粒结节状病灶[41.1％(53/129)]、纤维素沉积[40.3％(52/129)]等方面均明显高于其他原因胸腔积液者[分别为32.5％(25/77)、6.5％(5/77)、15.6％(12/77)],但散在多发结节状病灶[26.4％(34/129)]明显低于其他原因胸腔积液者[53.2％ (41/77)](Z=28.41、28.52、24.42、15.06,P值均=0.000).229例患者胸腔镜术后均未出现严重不良事件,224例(97.8％)有轻中度胸痛,口服止痛药2～3 d后可缓解;134例(58.5％)活检部位有少量出血,107(46.7％)例局部皮下气肿,均未给予特殊处理,拔管后2～3 d内吸收;仅2例患者术后出现脓胸,引流管留置14d后行胸腔镜胸膜剥离术,随访6个月后均安全拔管.结论 内科胸腔镜镜下表现、胸膜病变活检组织标本病理和病原学检测均有助于结核性胸膜炎的诊断,而且安全性高,建议临床大力推广应用.

目的 利用临床及内科胸腔镜下特征对胸腔积液患者构建诺谟图(nomogram),探讨其诊断结核性胸膜炎的价值.方法 采用前瞻性研究的方法,收集2015-2018年首都医科大学附属北京胸科医院(99例)、首都医科大学附属北京朝阳医院(67例)、北京积水潭医院(16例)及北京医院(24例)收治的206例胸腔积液患者.其中确诊为结核性胸膜炎的患者129例,作为结核性胸膜炎组;确诊为其他原因胸腔积液的患者77例,作为其他原因胸腔积液组.收集两组患者的一般资料、胸腔积液实验室检查结果、胸腔镜下形态学特征和病理形态学特征信息,按照4∶1的比例通过SAS随机分组过程,将206例患者完全随机分为训练集(162例)和验证集(44例).利用套索(least absolute shrinkage and selection operator,LASSO)回归筛选结核性胸膜炎的诊断因素,构建辅助诊断诺谟图,分别验证模型的区分度、校准度及收益,综合评价诺谟图的辅助诊断能力.结果 训练集162例胸腔积液患者中,共104例(64.2％)确诊为结核性胸膜炎.经LASSO回归,共7个高诊断价值因素被筛选出,分别为病理形态学上的干酪样肉芽肿、坏死表现、不典型肉芽肿,胸腔镜观察下的粟粒样结节状病灶、胸膜粘连带、孤立结节状病灶,以及胸腔积液检查指标中的腺苷脱氨酶水平.训练集中ROC曲线下面积(AUC)为0.97,敏感度为92.31％,特异度为93.42％,在验证集中模型AUC为0.89,敏感度为80.44％,特异度为100.00％.结论 诺谟图能够在胸腔积液患者中有效诊断结核性胸膜炎,有一定的辅助诊断价值.

DESIGN:A prospective multicentre study was conducted to compare the diagnostic accuracy of the GeneXpert MTB/RIF Ultra (Xpert Ultra) and cell-free DNA (cfDNA) assay for tuberculous meningitis (TBM) in China.METHODS:A prospective cohort study was conducted among individuals with symptoms suggestive of TBM registered in three TB specialised hospitals in China between June 2018 and January 2019.RESULTS:Overall, 84 patients suggestive of TBM were included in this analysis between June 2018 and January 2019. Using microbiological evidence as reference, the sensitivity/specificity for the diagnostic tests were Xpert Ultra 93.3%/100%, cfDNA 93.3%/92.6% and mycobacteria growth indicator tube (MGIT) culture 13.3%/100%. In addition, the sensitivity of culture was 6.7% when using clinical diagnosis criteria as the gold standard. Xpert Ultra correctly identified 28 cases as TBM, indicating a sensitivity of 46.7%. Notably, four additional TBM cases were detected by cfDNA compared with Xpert Ultra, yielding an overall sensitivity of 53.3%. Statistical analysis revealed that the sensitivity of Xpert Ultra and cfDNA was significantly higher than that of culture.CONCLUSIONS:The data demonstrate that Xpert Ultra and cfDNA assay showed optimal sensitivity compared with MGIT culture. In addition, there was no significant correlation between bacterial load and TBM severity in the participants.

结核性脑膜炎(tuberculous meningitis,TBM)是致残、致死率最高的结核病,临床发病率有逐渐升高趋势.近年来,对于TBM的治疗有较多进展,笔者对TBM的化学治疗、辅助治疗和并发症的治疗等领域相关研究进行综述,以期为TBM的临床治疗提供帮助.