Background Following the approval of KRAS G12C inhibitors, the necessity of serial KRAS testing remains debated. This study evaluates the molecular concordance and dynamic evolution of KRAS mutations across different clinical scenarios and explores their diagnostic utility in differentiating multiple primary lung cancers (MPLCs) from intra-pulmonary metastases. Methods A total of 108 patients with paired NGS tests and ≥ 1 KRAS-positive result were categorized into three groups: Group A (same lesion sampled longitudinally, n = 16; stratified by neoadjuvant therapy status), Group B (synchronous MPLCs, n = 30), and Group C (longitudinal monitoring of distinct lesions, n = 62). Group C was further stratified into three subgroups based on lesion relationships: metastasis-to-metastasis (C1), primary-to-metastasis (C2), and primary-to-new pulmonary lesion (C3). Results In Group A, the intra-lesional KRAS retention rate was 100% in upfront surgery without prior treatment, but decreased to 83.3% after neoadjuvant therapy. In Group B, 80.0% (24/30) of multifocal cases exhibited inter-lesion molecular discordance, including 7 cases with distinct KRAS amino acid subtypes that confirmed synchronous MPLCs. In Group C, molecular concordance was significantly lower between primary and new pulmonary lesions (C3) than between primary and metastatic lesions (C2) (65.5% vs. 88.5%, P = 0.046). Baseline variant allele frequency (VAF) served as a strong predictor of subsequent KRAS loss (median VAF: 8.4% vs. 22.4%, P = 0.031). Furthermore, patients with co-mutations of EGFR and KRAS were more prone to experience KRAS loss than those with isolated KRAS mutations (70.0% vs. 12.0%, P < 0.001). Conclusions KRAS mutations display scenario-dependent concordance, remaining clonally stable in true metastases while exhibiting divergence in independent primary lesions or under therapeutic selection. Repeat NGS testing should be prioritized for patients presenting with new or multifocal pulmonary lesions, lower baseline KRAS VAF, or co-occurring EGFR mutations.

BACKGROUND:Next-generation sequencing (NGS)-based driver gene testing has become a routine component of molecular subtyping and precision therapy for non-small cell lung cancer (NSCLC). Dynamic genomic monitoring facilitates early detection of resistance-related molecular alterations and informs timely therapeutic adjustments. However, standardized criteria for evaluating the stability of serial NGS testing are currently lacking, and the applicability of NGS using formalin-fixed paraffin-embedded (FFPE) specimens for dynamic monitoring remains poorly defined. This study aims to establish a stability grading system for driver gene status alterations based on repeated NGS testing, and to provide evidence-based support for clinical repeat biopsy strategies. METHODS:Data from 1232 patients with NSCLC who underwent two or more NGS tests on FFPE tissue specimens at Beijing Chest Hospital between June 2019 and April 2026 were collected retrospectively. Patients with an interval of ≥4 months between the initial and last tests were included to ensure the representativeness of temporal analysis, resulting in a main analysis cohort of 942 patients. The Kappa consistency test was used to evaluate the state stability of nine core driver genes [epidermal growth factor receptor (EGFR), Kirsten rat sarcoma viral oncogene homolog (KRAS), anaplastic lymphoma kinase (ALK), ROS proto-oncogene 1, receptor tyrosine kinase (ROS1), mesenchymal‑epithelial transition factor (MET), rearranged during transfection (RET), v-raf murine sarcoma viral oncogene homolog B1 (BRAF), erb‑b2 receptor tyrosine kinase 2 (ERBB2), and phosphatidylinositol‑4,5‑bisphosphate 3‑kinase catalytic subunit alpha (PIK3CA)] and to construct a five‑level grading system. Paired variant allele frequency (VAF) differences were compared using the Wilcoxon signed‑rank test. Independent influencing factors for mutation accumulation were identified by binary Logistic regression. RESULTS:The state stability of the nine genes was classified into five levels: EGFR showed high stability (Kappa=0.838), ROS1/ALK/KRAS good stability, BRAF/PIK3CA/RET moderate stability, and ERBB2 low stability, and MET showed high instability. MET exhibited the highest rate of state change (9.3%) with a raw observed agreement of 90.7%. Its Kappa value (0.172) was influenced by the low prevalence (3.7%) compression effect and should therefore be interpreted alongside the observed agreement (90.7%) and the prevalence-adjusted and bias-adjusted Kappa (PABAK). The VAF of PIK3CA increased significantly (P=0.005). T790M positivity increased from 5.8% to 10.8%, and 30 new C797S mutations were detected at the last test (13 with T790M, 17 without). The overall rate of new driver gene variants in the main cohort was 18.0%. Binary Logistic regression showed that a lower number of initial mutated genes was the only independent predictor of new variants [odds ratio (OR)=0.399, P<0.001], while sex and detection interval showed no independent association. CONCLUSIONS:A five level stability grading system for state changes of driver genes in NSCLC based on repeated NGS testing has been established. MET showed the most frequent state changes, which should be interpreted in conjunction with the prevalence effect. The VAF increase of PIK3CA is an observational finding, and its clinical significance requires further prospective validation. A lower initial mutation burden may reflect tumor clonal complexity and was associated with a higher likelihood of subsequent acquisition of new variants. FFPE based NGS is applicable for repeated testing at clinical treatment decision nodes.

Henoch-Schönlein purpura (HSP) or immunoglobulin A vasculitis (IgAV) is the most common systemic vasculitis in children. Rutin is a flavonoid with anti-inflammatory and antioxidant effects. This study aimed to investigate the protective effects of rutin in a rat model of HSP induced by ovalbumin. Results showed that 12.05 mg/kg of rutin significantly alleviated skin purpura, renal injury and systemic inflammation. It ameliorated vascular endothelial injury by reducing IgA deposition, modulating vasoactive substances including nitric oxide and endothelin-1, as well as vascular endothelial growth factor, and inhibiting the transforming growth factor-β1/SMAD family member 3 signaling pathway. These therapeutic effects were mediated through multiple mechanisms, including increasing the expression of zonula occludens-1 and occludin to maintain intestinal barrier integrity, modulating gut microbiota composition, and regulating the metabolism of short-chain fatty acids and amino acids. This study provides novel mechanistic insights and evidence supporting the clinical application of rutin in HSP.

Intussusception and intestinal perforation are surgical severe complications in children with IgA vasculitis (IgAV). Early diagnosis and timely appropriate surgical intervention can reduce damage to the children. We analyzed retrospectively the clinical characteristics, auxiliary examinations, diagnosis, treatment, and prognosis of 28 children with IgAV accompanied by surgical complications (including intussusception in 21 patients and intestinal perforation in seven patients) who were admitted to the Beijing Children's Hospital of the Capital Medical University from May 2016 to December 2020. Within one year after the data was collected, the parents of the children were interviewed by telephone about their treatment. Increased peripheral leukocytes were observed in 60.7% of children. Serum C -reactive protein (CRP) and D-dimer were elevated in 53.3% and 75% of children, respectively. Gastrointestinal bleeding was identified in 39% of children. Of the children with intussusception, the symptoms subsided spontaneously in four children and after air enema in another four. Four children underwent laparotomy and manual reduction. Nine patients underwent enterectomy and anastomosis. Enterectomy and anastomosis were performed in the seven patients with intestinal perforation, two of whom underwent enterostomy concurrently. Increased inflammation indexes, elevated D-dimer, and persistent abdominal pain without relief may be risk factors for surgical complications in children with IgAV.

Background: Tumor genomic profiling has a significant impact on the selection of targeted therapy. Circulating tumor DNA (ctDNA) has emerged as a noninvasive, and reproducible assay compared with tissue biopsy. We aimed to evaluate its utility in identifying mutations and guiding targeted therapy for lung cancer. Methods: A total of 173 lung cancer patients underwent next-generation sequencing (NGS) using a targeted enrichment panel covering 20 lung cancer-related genes. The performance of the ctDNA NGS assay in identifying genetic mutations or alterations was compared with tissue biopsy and droplet digital PCR (ddPCR). The treatment response to epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) therapies based on the ctDNA assay results was also assessed. Results: The ctDNA was detected in 61.85% of patients. Tissue mutations were detected in paired ctDNA in 38.57% of cases, while ctDNA mutations were detected in paired tissues in 89.1% of cases. The ctDNA increased the number of advanced non-small cell lung cancer (NSCLC) patients who received NCCN-recommended genetic testing by 12%. The concordance between ddPCR and ctDNA was relatively high reaching 99.43%. EGFR T790M/C797S c.G2390C and EGFR T790M/C797S c.T2389A were detected in tissue and ctDNA, respectively, in patient 01015. Moreover, ctDNA assay identified the EGFR T790M mutation, which was missed by tissue biopsy in patient 01149, who developed drug resistance after 1 year of EGFR-TKI therapy. Of the 17 patients who received EGFR-TKI targeted therapies based on the ctDNA NGS results, 12 patients achieved a partial response and two patients had stable disease. Conclusions: The results demonstrated that the ctDNA assay could partially overcome tumor heterogeneity in detecting mutations and provide complementary information on tumor genomic profiles. Moreover, the presence of EGFR mutations in ctDNA could offer valuable guidance for selecting appropriate EGFR-TKI treatment for advanced lung cancer patients. However, it is important to note that the ctDNA NGS assay has certain limitations in fully identifying all genomic alterations present in the tumor.

Abstract Purpose The concordance between tumor tissue and liquid biopsies has been extensively investigated in previous studies. Among the influential factors, the time interval between tissue and blood draw has received significant attention, but its impact on the concordance between tissue and liquid biopsies has yielded inconsistent results. In this study, we aimed to evaluate the effect of the time interval between tissue and blood draw on the concordance between tissue and liquid biopsies while adjusting potential confounding factors such as clinical stage, smoking history, and treatment. Methods A total of 116 paired tissue and plasma lung cancer samples were collected from Beijing Chest Hospital and sequenced by hybridization capture-based next-generation sequencing (NGS) using a targeted enrichment panel covering 20 lung cancer-related genes. Mutation profiles obtained from tissue and circulating tumor DNA (ctDNA) were compared. The impact of time interval between tissue and blood draw on the concordance between tissue and liquid biopsies was assessed, stratifying the analysis based on clinical stage, smoking history, and treatment using the Cochran-Mantel-Haenszel test. Results ctDNA was detected in 49.14% (57/116) of clinical lung cancer samples. TP53 and EGFR carried the most mutations, both in tissue and ctDNA. Among the tissue mutations, 37.39% (83/222) were detected in paired ctDNA samples, while 89.25% (83/93) of ctDNA mutations were detected in paired tissue samples. The stratified analysis results demonstrated a significant correlation between concordance rates and the time interval between tissue and blood draw when stratified by clinical stage (P < 0.001, Cochran-Mantel-Haenszel test). Additionally, the genomic characteristics observed in tissue and ctDNA samples from two clinical lung cancer patients exhibited tumor heterogeneity. Conclusion The time interval between tissue and blood draw had a significant impact on the concordance between tissue and liquid biopsies when stratified by clinical stage. Furthermore, ctDNA demonstrated the ability to overcome tumor heterogeneity to some extent, indicating its potential as a viable alternative to tissue biopsies when tumor tissue is not accessible.

Immunoglobulin A vasculitis (IgAV) is a systemic small vessel vasculitis common in children. Pancreatic involvement in IgAV is rare. The purpose of this study was to analyze the clinical characteristics of IgAV-related acute pancreatitis in children. Records of patients with IgAV-related acute pancreatitis admitted to our institution between January 2016 and December 2019 were reviewed. We summarized the clinical characteristics, laboratory characteristics, imaging findings, treatment, and outcomes of 15 children with IgAV-related acute pancreatitis. The patients’ median age was 9.6 years. Pancreatitis was the initial manifestation of IgAV in 3 patients. All patients had abdominal manifestations, including abdominal pain (15/15), vomiting (10/15), and gastrointestinal bleeding (7/15). Serum amylase and lipase levels were elevated in all patients. Serum amylase in 4 cases reached more than three-fold elevation and serum lipase in 14 cases reached more than three-fold elevation. Morphological abnormalities and abnormal signals of the pancreas were observed in 13 cases by magnetic resonance imaging. Pancreaticobiliary maljunction was seen in 3 cases by magnetic resonance cholangiopancreatography. Glucocorticoid therapy and intravenous immunoglobulins were used to treat acute pancreatitis in IgAV. All patients showed clinical improvement after treatment. During the follow-up period of 6–12 months, all 15 cases with pancreatitis were cured without recurrence. Pancreatic involvement is rare in IgAV; however, this should be considered in IgAV patients with severe abdominal pain. The timely application of steroid therapy is important for IgAV-associated pancreatitis.

目的:回顾性分析脓液标本细菌学检测技术和组织标本病理学检测技术诊断骨关节结核的检测效能.方法:以2016年1月至2018年12月在北京胸科医院住院治疗的213例送检同部位脓液和组织标本进行检测的疑似骨关节结核患者作为研究对象,取新鲜脓液标本进行涂片染色镜检、GeneXpert MTB/RIF(GeneXpert)检测、实时荧光定量PCR(FQ-PCR)检测和BACTEC MGIT 960液体培养(MGIT 960液体培养);组织标本经石蜡包埋后进行抗酸染色镜检和FQ-PCR检测.结果:根据骨结核临床诊断综合参考标准(CRS),186例纳入患者诊断为骨关节结核,其中124例为确诊结核患者,23例为高度疑似结核患者,39例为疑似结核患者;27例患者排除骨关节结核.与CRS比较,脓液标本涂片镜检、MGIT 960液体培养、FQ-PCR、GeneXpert以及组织标本抗酸染色和FQ-PCR 检测的敏感度分别为28.7％(49/171)、49.0％(48/98)、58.1％(25/43)、87.5％(161/184)、48.6％(90/185)和85.4％(146/171);特异度分别为 100.0％(26/26)、11/11、8/9、96.2％(25/26)、96.3％(26/27)和100.0％(27/27).结论:脓液标本GeneXpert检测和组织标本FQ-PCR检测的敏感度高,检测用时短,可用于骨关节结核的早期诊断.

Introduction Anaplastic lymphoma kinase ( ALK ) gene rearrangements, have been identified in approximately 2-7% of patients with lung adenocarcinoma (LUAD). However, co-occurrence of double ALK fusions in one patient was rare. Herein, we reported two Chinese female LUAD patients with confirmed double ALK fusion variants by next generation sequencing. Case presentation Case 1, a 38-year-old female was diagnosed as peripheral LUAD in left upper lobe with synchronous multiple intrapulmonary metastases (pT2N0M1b, stage IVa). And case 2, a 58-year-old female had left lower lobe primary LUAD and synchronous multiple lung metastases (pT4N2M1b, stage IVa). In both patients, tumor cells displayed strong expression of ALK protein. Genetic profiling by next generation sequencing showed both patients concurrently harbored two types of ALK rearrangements. Case 1 had an unreported ALK - SSH2 / EML4 - ALK double fusions, and case 2 had an another novel ARID2 - ALK / EML4 ‐ ALK double fusions. Both of these patients responded to ALK inhibitor crizotinib. Conclusions Our study reported two novel ALK fusion partners never reported, which expands the knowledge of ALK fusion spectrum and provides insight into therapeutic options for patients with double ALK fusions.

To analyze and summarize the clinical features, diagnosis, and treatment of children with Henoch-Schönlein purpura (HSP) complicated by overt gastrointestinal bleeding (GI bleeding) for achieving early identification, prevention, and treatment in terms of severe GI bleeding. A retrospective analysis was conducted on children with HSP complicated by overt GI bleeding who were admitted to the Department of Traditional Chinese Medicine of Beijing Children’s Hospital from January 2017 to December 2019. According to the severity of GI bleeding, the patients were divided into mild bleeding group (61 cases) and moderate and severe bleeding group (38 cases). Inflammatory parameters, coagulation function, GI ultrasound findings, and clinical features were compared. Logistic regression analysis was used to determine the related variables affecting the severity of GI bleeding, and the ROC curve was used to determine the variable test efficacy. Onset in summer, wide distribution of skin rash with facial involvement, the elevation of D-dimer and high neutrophil-to-lymphocyte ratio (NLR) had significant effects on the severity of GI bleeding. ROC curve analysis showed that the optimal cut-off points of NLR and D-dimer for predicting severe GI bleeding in children with HSP were 10.56 and 0.89 mg/L, respectively. Facial rash may be a warning sign of GI bleeding. Enhanced monitoring of NLR and D-dimer is helpful for early recognition of GI bleeding as well as assessment of severity.

OBJECTIVES:The MeltPro MTB assays for detection of resistance to antituberculosis (TB) drugs perform well in genotypic drug susceptibility testing (DST) of clinical samples, but their effectiveness with formalin-fixed, paraffin-embedded (FFPE) tissues is unknown.METHODS:FFPE tissues were obtained from 334 patients with TB. Susceptibility to rifampicin (RIF), isoniazid (INH), and fluoroquinolones was examined using the MeltPro MTB assays, with Xpert MTB/RIF (Xpert) and/or phenotypic DST (pDST) results as references. Samples with discordant results were analyzed by multiplex polymerase chain reaction-targeted amplicon sequencing (MTA-seq).RESULTS:With pDST as the reference, the MeltPro MTB assays sensitivity for RIF, INH, levofloxacin (LVX), and moxifloxacin (MXF) was 95.00%, 96.00%, 100%, and 100%, respectively, and the specificity was 95.15%, 95.92%, 94.69%, and 89.92%, respectively. Concordance was 99.08% between the MeltPro MTB and Xpert (κ = 0.956) for RIF and 95.12% (κ = 0.834), 95.93% (κ = 0.880), 95.12% (κ = 0.744), and 90.24% (κ = 0.367) between the MeltPro MTB and pDST for RIF, INH, LVX, and MXF, respectively. MTA-seq confirmed the discordancy between the MeltPro MTB and pDST for 26 (89.66%) of 29 samples.CONCLUSIONS:The MeltPro MTB assays rapidly and efficiently predict Mycobacterium tuberculosis resistance to the main first- and second-line anti-TB drugs in FFPE tissues.

AZD9291 (osimertinib) is the third-generation EGFR-TKI treat for EGFR mutated NSCLC patients. Despite its encouraging efficacy in clinical, acquired resistance is still inevitable. The mechanism of drug resistance needs to be further explored. In a previous study, we established an AZD9291-resistant cell strain named HCC827/AZDR. We found that insulin-like growth factor binding protein 7 (IGFBP7) expression was markedly increased in HCC827/AZDR cells and AZD9291-resistant patients by RNA sequencing and immunohistochemical analysis, respectively. Reduced IGFBP7 in HCC827/AZDR cells by si-RNA interference recovered the sensitivity to AZD9291 partially and increased AZD9291-induced cell apoptosis. Enhancing IGFBP7 expression in EGFR-mutated non-small cell lung cancer (NSCLC) cells using lentiviruses infection reduced their sensitivity to AZD9291. This study is the first to discover that high IGFBP7 expression could occur following treatment with AZD9291. This might be one of the mechanisms underlying AZD9291 resistance and a potential therapeutic target following AZD9291 resistance.

我国是全球排名第二的结核病高负担国家,2018年估算新发结核病例86.6万[1],其中约10％为复治患者[2].近年来,随着我国结核病控制工作的进展,复治结核病患者患病率有所下降,但复治结核病患者的总数量仍处于较高的水平.复治结核病不仅治疗时间长、病情相对严重,而且耐药的发生率明显高于初治结核病患者(21％与7.1％)[1].2010年我国开展的第五次结核病流行病学调查显示,复治患者对抗结核药物的任一耐药率为38.5％[2].如此高的耐药率严重影响了复治肺结核患者的治疗效果.因而,快速而准确的诊断复治肺结核并从中识别和确诊耐药结核,对结核病的防控具有重要的意义.

目的 探讨肺结核患者肺部局灶性磨玻璃样(focal ground-glass opacity,fGGO)病灶的CT特征,以提高对肺结核少见征象的认识.方法 收集2009 2019年北京胸科医院确诊的11例具有fGGO病灶的肺结核患者(肺结核组,11个fGGO病灶),其中7例经临床综合诊断、3例经手术组织标本病理检查证实、1例经皮肺穿刺活检病理证实.同期按照1∶4比例选取经我院手术病理证实的44例具有fGGO病灶的肺癌患者(肺癌组,44个fGGO病灶).分析两组患者fGGO病灶的CT扫描特征.结果 肺结核组和肺癌组患者fGGO病灶分布在上叶[分别为63.6％(7/11)和65.9％ (29/44)]、中叶[分别为9.1％ (1/11)和11.4％(5/44)]、下叶[分别为27.3％(3/11)和22.7％(10/44)]的差异无统计学意义(x2=2.608,P=0.106).肺结核组fCGO病灶呈圆或椭圆形[36.4％(4/11)]、边缘与肺交界面清晰[9.1％(1/11)]、有分叶[0.0％(0/11)]的比例均明显低于肺癌组[77.3％(34/44)、93.2％(41/44)、43.2％(19/44)](x2=5.114,P=0.024;x2 =29.974,P=0.000;x2 =5.473,P=0.019).肺结核组fGGO病灶内部密度均匀[9.1％(1/11)]、有支气管影或空泡[0.0％(0/11)]、出现增粗或扭曲血管影[0.0％(0/11)]的比率均明显低于肺癌组[分别为54.5％(24/44)、38.6％(17/44)、77.3％ (34/44)](x2=7.333,P=0.007;x2 =4.475,P=0.034;x2=19.108,P=0.000),而多发结节样病灶[90.9％(10/11)]的比率高于肺癌组[0.0％(0/44)] (x2=42.969,P=0.000).结论 肺结核fGGO病灶分布与肺癌无特异性差别;依据fGGO边缘模糊、无分叶、无胸膜凹陷征、病灶内部密度不均匀、无支气管影或空泡、无增粗或扭曲血管影、多发簇状分布结节样病灶的CT表现,在与肺癌鉴别诊断时有一定的参考价值.

Delta-like homolog 1 (DLK1) regulates noncanonical Notch signaling pathway as ligand. DLK1 was abnormally expressed in a variety of tumors, affecting tumorigenesis and developments. The biological function of DLK1 toward cell proliferation and signaling activation was controversial across different cell types. Two currently known isoforms of DLK1, which are membrane-tethered isoform and soluble isoform, are believed to be the key of DLK1 dual behaviors. While these isoforms are not enough to explain the phenomena, our observations offer the possibility of a third isoform of DLK1. In the present study, we verified the nuclear localization of DLK1 in lung cancer cells. The nuclear localized DLK1 was observed in 107 of 351 non-small cell lung cancer (NSCLC) samples and was associated with tissue differentiation and tumor size. Through co-immunoprecipitation (co-IP) combined mass spectrometry (MS), we identified nuclear receptor corepressor 1 (NCOR1) as DLK1's novel interaction protein and confirmed their interaction in nuclear. We analyzed the expression of NCOR1 in two independent cohorts and demonstrated that NCOR1 is a tumor suppressor and has prognosis potential in lung squamous carcinomas. At last, we analyzed the colocalization of DLK1 and NCOR1 in 147 NSCLC samples by immunohistochemistry (IHC). The result indicated NCOR1 might participate with nuclear localized DLK1 in regulating cell differentiation.

目的 应用非标记label-free蛋白质组学方法筛选出活动性肺结核患者外周血CD4+T淋巴细胞差异表达蛋白质,为阐明结核病的发病机制和早期诊断提供理论依据.方法 收集分离9例活动性肺结核患者和9例健康人外周血CD4+T淋巴细胞,应用label-free非标记蛋白组学方法检测外周血CD4+T淋巴细胞蛋白质谱,筛选出差异表达蛋白质.并用基因本体论(GO)及京都基因与基因组百科全书(KEGG)生物信息学分析软件对差异表达蛋白质进行分析.结果 活动性肺结核与健康人外周血CD4+T淋巴细胞中的蛋白质组分布的基本框架很相似,但可发现二者之间有38个明显差异表达的蛋白点,其中有26种蛋白质在肺结核组中表达上调(差异倍数＞1.5,P＜0.05),12种蛋白质在肺结核组中表达下调(差异倍数＜0.67,P＜0.05).GO分析结果表明,大部分的差异蛋白质主要定位于胞内区域,结核感染相关差异蛋白质功能体现在结合、调控以及代谢过程中.KEGG分析表明,磷酸戊糖途径、cGMP/PKG信号通路、磷脂酰肌醇信号系统等与结核分枝杆菌感染密切相关.结论 对活动性肺结核患者外周血CD4+T淋巴的比较蛋白质组学研究一共鉴定出38种差异蛋白质,他们可能在肺结核发病过程中起着重要作用,也可能可做为诊断结核病的潜在生物学标志物.

目的 评价分子病理学方法诊断颈部淋巴结结核及其耐药性的临床应用价值.方法 搜集2010年3月至2013年10月首都医科大学附属北京胸科医院病理科收治的符合纳入标准(临床症状和体征均符合颈部淋巴结结核、抗结核药物治疗均有效)的全部97例颈部淋巴结结核患者(结核组);以及符合纳入标准(通过病理学或临床检测结果明确诊断为其他淋巴结疾病)的全部20例其他淋巴结病变患者(非结核组)的石蜡包埋标本.所有标本以萋-尼(Z-N)抗酸染色法查找抗酸杆菌,荧光定量聚合酶链式反应(FQ-PCR)检测结核分枝杆菌特异基因序列IS6110;以临床最后诊断为标准,比较两种方法的检测效能;并对FQ-PCR检查结果为阳性且结核分枝杆菌DNA含量满足耐药突变检测下限的标本,以探针熔解曲线法检测利福平、异烟肼耐药相关基因突变情况.结果 以临床最后诊断为标准,抗酸染色和FQ-PCR检测结核组的敏感度分别为22.7％(22/97)和67.0％(65/97);FQ-PCR技术检测敏感度明显高于抗酸染色法,差异有统计学意义(x2=38.53,P＜0.001).抗酸染色和FQ-PCR检测非结核组标本均为阴性,特异度均为100.0％(20/20).抗酸染色和FQ-PCR的阳性预测值分别为100.0％(22/22)和100.0％(65/65),阴性预测值分别为21.1％(20/95)和38.5％(20/52),符合率分别为35.9％(42/117)和72.6％(85/117).对41例FQ-PCR检查结果为阳性且结核分枝杆菌DNA含量满足耐药突变检测下限的患者标本进行结核分枝杆菌耐药基因突变检测,利福平和异烟肼可评估标本分别为10份(例)和27份(例),其中利福平耐药1份(例),异烟肼耐药13份(例).结论 分子病理学诊断技术在颈部淋巴结结核石蜡包埋标本中检测结核分枝杆菌DNA的敏感度和特异度较高,并且能筛查可能的耐药患者,可为颈部淋巴结结核的正确诊断与合理化治疗提供依据.

OBJECTIVE The influence of anti-tuberculosis (TB) treatment history on tuberculous lymphadenitis (TBLN) diagnosis is unclear. Therefore, this study aims to evaluate the diagnostic methods, including histology, microbiology, and molecular tests, used for TBLN. METHODS In this study, suspected patients with TBLN and having different anti-TB treatment background were enrolled. All the samples were tested simultaneously by histology, Ziehl-Neelsen (ZN) staining, mycobacterial culture (culture), Xpert MTB/RIF (xpert), real-time PCR, and high-resolution melting curve PCR (HRM). Thereafter, the performance of these methods on samples with different anti-TB treatment background was assessed. RESULTS In our study, 89 patients were prospectively included 82 patients with TBLN and 7 with other diseases. The overall sensitivities of Xpert, real-time PCR, histology, ZN staining, and culture were 86.6%, 69.5%, 58.5%, 43.9%, and 22.0%, respectively. The anti-TB treatment history revealed dramatic influences on the sensitivity of culture (P < 0.0001). In fact, the treatment that lasted over 3 months also influenced the sensitivity of Xpert (P < 0.05). However, the treatment history did not affect the performance of remaining tests (P > 0.05). For rifampicin drug susceptibility test (DST), the anti-TB treatment showed only significant influence on the success rate of culture DST (P = 0.001), but not on those of Xpert and HRM tests (P > 0.05). CONCLUSION Other tests as well as culture should be considered for patients with TBLN having retreatment history or over 1-month treatment to avoid false negative results.

结核病是一种严重危害人类健康的慢性呼吸道传染性疾病.传统病理学诊断结核病主要依靠形态学特征和抗酸染色法查找结核分枝杆菌(Mycobacterium tuberculosis,MTB).近年来,精准医学理念对结核病的病理学诊断提出新的挑战,基于核酸扩增技术的分子诊断技术在结核病的病理学诊断中逐渐推广应用,在结核病的诊断和鉴别诊断,以及耐药相关基因突变检测中发挥着越来越重要的作用.结合传统病理学诊断方法,分子诊断技术有效提高了结核病诊断的准确性,并为临床治疗提供了重要依据.作为桥梁学科,病理学在结核病发病机制及治疗相关研究中也发挥着重要作用.笔者对传统病理学和分子病理学诊断在结核病诊断和鉴别诊断,以及发病机制等方面的研究进展做一综述.

Objective To investigate the clinicopathological feature and outcome of pulmonary adenocarcinoma with a micropapillary pattern (MPPAC) and to improve the understanding of the aggressiveness and poor prognostic influence of MPPAC.Methods 299 patients with invasive lung adenocarcinomas were analyzed retrospectively and tumor histological patterns were reassessed by two pathologists according to the Study of Lung Cancer/American Thoracic Society/European Respiratory Society (IASLC/ATS/ERS) criteria.Patients were divided into 2 major groups:MPP-positive group and MPP-negative group.The MPP-positive group was further subdivided according to presence and proportion of MPP:MPP1 +(5％ ～10％ of theMPP),MPP2+(10％ ～30％ of the MPP),and MPP3+ (＞30％ of the MPP).Results The progression-free survival (PFS) for patients with micropapillary predominant subtype,which was significantly shorter than those with acinar predominant subtypes (P ＜ 0.01) or those with papillary predominant subtype (P ＜ 0.01),not with those with solid predominant subtype (P ＞ 0.05) and those with invasive mucin adenocarcinoma (P ＞ 0.05).Compared with the MPP-negative group,the MPP-positive group was significantly correlated with vascular invasion (P ＜ 0.01),pleural effusion (P ＜ 0.01),lymph node metastasis (P ＜ 0.01) and tumor size (P ＜ 0.01).The extent of MPP showed significant association with vascular invasion (P ＜ 0.01).MPP-positive group had poorer prognosis than MPP-negative group (12.0 months vs 24.0 months,P ＜ 0.01).However,stratification analysis showed that MPP positive group had worse outcome comparing with MPP-positive group at stage Ⅰ + Ⅱ + m A (for stage Ⅰ + Ⅱ,18.0 months vs.37.0 months,P ＜ 0.01;for stage Ⅲ a,8.3 months vs 13.6 months,P ＜ 0.01).But yet,there was no significant difference between the 2 group at stage mb + Ⅳ (7.7 months vs.11.3 months,P ＞ 0.05).The extent of MPP was not significantly correlated with PFS (P ＞ 0.05).By multivariate analysis,MPP (HR,1.467;95％ CI,1.089-1.975;P ＜ 0.05) was independent prognostic factors influencing progression-free survival.Conclusions MPP in lung adenocarcinoma is more aggressive and a poor prognostic factor.Patients with MPPAC,especially the early stage patients,have a high risk for disease recurrence and metastasis.For patients with MPPAC we should provide proper treatment to prolong the survival time.