Data on the 10-year survival of incident IPAH in China is lacking. The changing trend of characteristics and survival of IPAH over the past decade has yet been demonstrated. The 4-stratum risk model needs further validation in different cohorts. Incident cases of IPAH identified in the national multicenter prospective registry of PAH from 2009 to 2019 were analyzed. Patients with at least two of the three acquired variables (WHO-FC, 6MWD, NT-pro BNP/BNP) were included for risk stratification. 605 incident patients with IPAH were included (age: 35 ± 12 years; 75.2

Objective To compare the diagnostic value between mNGS and conventional tests in suspected pulmonary tuberculosis (PTB) patients with scarce sputum or with negative sputum etiological test results. Methods We enrolled eligible patients admitted to our department from 2018 to 2021. Their bronchoalveolar lavage fluid (BALF) and lung biopsy tissue samples were sent for mNGS and conventional tests. The diagnostic value of mNGS was compared respectively with that of conventional tests. Results 94 of 226 enrolled patients were diagnosed PTB. The diagnostic concordance rate of mNGS was significantly higher than that of acid-fast staining in all samples (p<0.001), as well as that of culture in BALF samples (p=0.011). mNGS in parallel with conventional tests had significantly higher AUC than mNGS alone (p=0.013). Conclusions mNGS has higher diagnostic value than some conventional tests. mNGS in parallel with conventional tests is more reliable to accurately diagnose these suspected PTB patients.

Industrialization and urbanization produce hazardous particulate matter (PM), including ammonium compounds like (NH4)2SO4 4 ) 2 SO 4 and NH4NO3, 4 NO 3 , which comprise around 50 % of PM. Despite this, the influence of ammonium on intestinal inflammation remains unclear. We studied an ammonium mixture's effects on the intestine, finding elevated pro-inflammatory cytokines and oxidative stress in exposed mice, causing small intestine morphological changes. Investigating mitigation strategies, we assessed silibinin, an antioxidant from milk thistle seeds. Silibinin treatment in (NH4)2SO4- 4 ) 2 SO 4- and NH4NO3-exposed 4 NO 3-exposed mice significantly reduced inflammatory markers, alleviated oxidative stress, and preserved intestinal tissue integrity via the PI3K/AKT pathway. Our findings elucidate ammonium's potential impact on intestinal inflammation and highlight silibinin's therapeutic impact against PM-induced small intestine damage.

Fine particulate matter (PM) 2.5 contains metals, ions, and carbon compounds, varying dynamically due to environmental factors. This study identified five major metals (Ca, Fe, Al, Zn and Pb) in urban PM2.5 via meta-analysis and evaluated their hazardous effects on health. Mice were exposed to these metals through intratracheal instillation, revealing changes in white blood cell count and serum substances (bilirubin, BUN and LDH), along with lung inflammation. Increased levels of proinflammatory cytokines (IL-1b, IL-6 and TNF-alpha) were observed, indicating adverse effects on the respiratory system. These results suggest that further studies on the health effects of the major metals of PM2.5, development of indicators for determining the PM2.5 exposure levels, and methods for reducing the metals together with other components in PM2.5 are needed.

Epidermal growth factor receptor (EGFR) is a validated target for non-small-cell lung cancer (NSCLC). However, the treatment for EGFR-C797S mutation induced by third-generation EGFR inhibitors remains a concern. Therefore, the development of the fourth-generation EGFR inhibitors to overcome the EGFR-C797S mutation has great potential for clinical treatment. In this article, we designed and synthesized a series of diphenyl ether substituted quinazolin-4-amine derivatives that simultaneously occupy the ATP binding pocket and the allosteric site of EGFR. Among the newly synthesized compounds, 9d displayed excellent kinase activity against EGFR(L858R/T790M/C797S) with an IC50 value of 0.005 mu M, and exhibited anti-proliferation activity in BaF3-EGFR(L858R/T790M/C797S) cells with the IC50 value of 0.865 mu M. Furthermore, 9d could suppress phosphorylation of EGFR and induce cell apoptosis and cycle arrest at G2 phase in a dose-dependent manner in BaF3-EGFR(L858R/T790M/C797S) cells. More importantly, 9d displayed significant antitumor effects in BaF3-EGFR(L858R/T790M/C797S) xenograft mouse model (30 mg/kg, TGI = 71.14 %). All the results indicated compound 9d might be a novel fourth-generation EGFR inhibitor for further development in overcoming the EGFR-C797S resistance mutation.

This review explores the concept of life-on-a-chip, which involves the creation of miniaturized biological systems, such as organs, tissues, and model organisms, on microscale platforms called microfluidic chips. These chips consist of intricately etched channels, wells, and chambers that enable precise control and observation of fluids, cells, and biochemical reactions, facilitating the simulation of various aspects of human or animal physiology and the study of responses to different stimuli, drugs, or disease conditions. The review highlights the application of a novel technology, "Beyond Limit Manufacturing" (BLM), in the development of sophisticated three-dimensional cell models and model organism microchips. Model-organism-on-a-chip and organ-on-a-chip (OoC) are among the thriving developments in the field of microfluidics, allowing for the reconstruction of living microenvironments and implementation of multiple stimuli. The review discusses the latest advancements in life-on-a-chip technology using BLM and outlines potential future research directions, emphasizing the significant role of these chips in studying complex biological processes in a controlled and scalable manner.

Background and ObjectiveOff-label pulmonary arterial hypertension (PAH)-targeted drugs are commonly prescribed for non-operated chronic thromboembolic pulmonary hypertension (CTEPH), but their effect on the long-term prognosis of CTEPH remains unknown. This study investigated the effect of off-label PAH-targeted drugs on the long-term survival of CTEPH patients.MethodsCTEPH patients were enrolled from a prospective multicentre national registry. Except for licensed riociguat and treprostinil, other PAH-targeted drugs were off-label. In the original and propensity score-matched (PSM) samples, five-year survival was compared in two groups: (a) patients not receiving off-label PAH-targeted drugs (control) versus (b) patients receiving off-label PAH-targeted drugs (treatment). The latter group was investigated for the effect of started off-label PAH-targeted drugs at baselines (initial) or during follow-up (subsequent).ResultsOf 347 enrolled patients, 212 were treated with off-label PAH-targeted drugs initially (n = 173) or subsequently (n = 39), and 135 were untreated. The 1-, 2-, 3- and 5-year survival of the treatment group was significantly higher than that of the control group (97.1% vs. 89.4%, 92.3% vs. 82.1%, 83.2% vs. 75.1% and 71.1% vs. 55.3%, respectively, log-rank test, p = 0.005). Initial treatment was correlated with better 5-year survival after excluding patients with subsequent treatment to reduce the immortal-time bias (hazard ratio: 0.611; 95% CI: 0.397-0.940; p = 0.025). In PSM samples, patients given initial treatment showed significantly better 5-year survival than untreated patients (68.9% vs. 49.3%, log-rank test, p = 0.008).ConclusionOff-label targeted drugs contributed to improved long-term survival in CTEPH patients receiving pharmacotherapies. imageConclusionOff-label targeted drugs contributed to improved long-term survival in CTEPH patients receiving pharmacotherapies. image The study demonstrates that off-label use of pulmonary arterial hypertension medication improves long-term survival in patients with chronic thromboembolic pulmonary hypertension. This is a national, multicentre, prospective registry with a large sample size and low rate of loss to follow-up. The efficacy is more transparent after rigorously reducing bias. image

Background: There is limited information on the effect of different distant metastases on the prognosis of malignant pleural mesothelioma (MPM). This study aimed to investigate the clinical value of subdividing the M1 stage in predicting the prognosis of MPM.Methods: The clinical data from 338 patients diagnosed with distant metastatic MPM between 2010 and 2015 were collected from the Surveillance, Epidemiology and End Result database. An M1 subdivision system was established based on the independent prognostic factors obtained from a multivariate Cox regression model and further validated using clinicopathologic parameters by multivariate analysis.Results: Brain, bone and multi-site metastases were identified as independent prognostic factors for overall survival (OS) and cancer-specific survival (CSS) in patients with distant metastatic MPM. M1 stage patients were subdivided into three groups: M1a, single-site metastasis except for the brain and bone; M1b, only bone metastasis or multi-site metastasis except for the brain and bone; and M1c, brain metastasis regardless of metastasis at other sites or metastases of bone and other sites except for the brain. Both OS [M1b vs M1a: HR = 1.460, 95% CI = 1.114 – 1.912 and M1c vs M1b: HR = 1.494, 95% CI = 1.074 – 2.078] and CSS (M1b vs M1a: HR = 1.626, 95% CI = 1.174 – 2.252 and M1c vs M1b: HR = 1.434, 95% CI = 0.9751 – 2.109) were decreased in all M1 subcategories.Conclusions: The involvement of brain, bone and multi-site metastases was associated with poor prognosis in patients with distant metastatic MPM. Subdivision of the M1 stage facilitated better prediction of prognosis in patients with distant metastatic MPM.

BACKGROUND: Registry-based studies of pediatric pulmonary arterial hypertension (PPAH) are scarce in developing countries, including China. The PPAH risk assessment tool needs further evaluation and improvement.RESEARCH QUESTION: What are the characteristics and long-term survival of PPAH in China and what is the performance of the PPAH risk model in Chinese patients?STUDY DESIGN AND METHODS: Patients with PAH were enrolled in the national prospective multicenter registry from August 2009 through December 2019. Children 3 months to 18 years of age at the time of PAH diagnosis were analyzed.RESULTS: A total of 247 children with PAH were enrolled. The median patient age was 14.8 years, and 58.3% of patients were female. Most patients had a diagnosis of PAH associated with congenital heart disease (CHD; 61.5%) and idiopathic or heritable PAH (37.7%). The median time from symptom onset to PAH diagnosis was 24 months. The mean pulmonary artery pressure and pulmonary vascular resistance index were 70.78 + 19.80 mm Hg and 21.82 + 11.18 Wood Units$m2, respectively. Patients with CHD-associated PAH experienced a longer diagnostic delay and demonstrated higher pulmonary artery pressure, but better cardiac performance, than those with idiopathic or heritable PAH. An increased number of patients received targeted therapy at the last follow-up compared with baseline. The 5-and 10-year survival rates of the entire cohort were 74.9% and 55.7%, respectively, with better survival in patients with CHD-associated PAH than in those with idiopathic or heritable PAH. Patients with low risk had better survival than those with high risk according to the simplified noninvasive risk score model with weight, function class, and echocardiographic right ventricular size, both at baseline and follow-up.INTERPRETATION: Patients with PPAH in China showed severely compromised hemody-namics with marked diagnostic delay. The long-term survival of PPAH is poor despite the increased usefulness of targeted drugs. The simplified noninvasive risk model demonstrated good performance for predicting survival in Chinese children with PAH.TRIAL REGISTRY: ClinicalTrials.gov; No.: NCT01417338; URL: www.clinicaltrials.gov CHEST 2023; 163(6):1531-1542

Background As a member of the Ephrin protein family that elicits short distance cell-cell signaling, EphrinA3 has been shown to promote or inhibit tumorigenesis depending on tumor types, but its roles and the underlying mechanisms in lung adenocarcinoma (LUAD) have not been reported. Materials and Methods The TCGA database and Kaplan-Meier Plotter database were used to analyze the differential expression of EphrinA3 between LUAD and para-carcinoma tissues, and its effect on overall survival of LUAD patients. CCK-8 assay, Edu assay, and flow cytometry were used to probe the effect of EphrinA3 on the proliferation of LUAD cells, and transwell assay was employed to examine its effect on migration and invasion. In addition, the effect of EphrinA3 on the growth of LUAD was further evaluated using a xenograft tumor model. Results EphrinA3 was expressed highly in LUAD, and its expression level was negatively correlated with the prognosis of LUAD patients. In addition, EphrinA3 promoted proliferation, migration, and invasion of LUAD cells, and accelerated tumor growth in a xenograft LUAD model. The reported EphrinA3 receptors, EphA1 and EphA10, were expressed in clinical LUAD tissues and co-localized with EphrinA3 in LUAD cells. Mechanistically, EphrinA3/Eph signaling activated AKT, ERK, and p38MAPK, induced epithelial-mesenchymal transition (EMT), and upregulated matrix metalloproteases-2 and -9 (MMP-2/-9). Conclusion EphrinA3 expression was negatively correlated with prognosis of patients with LUAD. EphrinA3 promoted proliferation, migration, and invasion of LUAD cells. EphrinA3 enhanced the phosphorylation of ERK and AKT, and potentiates EMT and MMP expression in LUAD cells.

Conventional etiological detection and pathogenic antibody methods make it challenging to identify the atypical pathogens among the community‐acquired pneumonia (CAP). Metagenomic next‐generation sequencing (mNGS) could rapidly detect all potentially infectious diseases and identifies novel or potential pathogens.

PDF file - 145K, The MAPK protein spectrum activated by UVA, UVB or SUV. (A) N/TERT-1 cells were irradiated with UVA (60 kJ/m2), UVB (3.6 kJ/m2) or SUV (60 kJ UVA/m2/3.6 kJ UVB/m2). Protein extracts (500 μg) were used for MAPK array analysis. Array spots were visualized using an ECL kit. The density of the each duplicated array spot in Fig. 2A was measured as described in Materials and Methods. The graph shows the fold change in SUV-induced phosphorylation of all 21 proteins compared to untreated control (i.e., value of 1). Data are shown as an average of duplicate samples. (B) Identification of proteins comprising the human phospho-MAPK array.

PDF file - 309K, The protein phosphorylation spectrum activated by UVA, UVB or SUV irradiation. N/TERT-1 cells were irradiated with UVA (60 kJ/m2), UVB (3.6 kJ/m2) or SUV (60 kJ UVA/m2/3.6 kJ UVB/m2). Protein extracts (500 μg) were used for phospho-kinase protein array analysis. Array spots were visualized by ECL kit. The density of the each duplicated array spot in Figure 3A was measured as described in Materials and Methods. The table shows the fold change in SUV-induced phosphorylation of all 46 kinases and transcription factors compared to untreated control (i.e., value of 1). Data are shown as an average of duplicate samples.

The diagnosis of invasive pulmonary aspergillosis (IPA) diseases in non-neutropenic patients remains challenging. It is essential to develop optimal non-invasive or minimally invasive detection methods for the rapid and reliable diagnosis of IPA. Metagenomic next-generation sequencing (mNGS) in bronchoalveolar lavage fluid (BALF) can be a valuable tool for identifying the microorganism. Our study aims to evaluate the performance of mNGS in BALF in suspected IPA patients and compare it with other detection tests, including serum/BALF galactomannan antigen (GM) and traditional microbiological tests (BALF fungal culture and smear and lung biopsy histopathology). Ninety-four patients with suspicion of IPA were finally enrolled in our study. Thirty-nine patients were diagnosed with IPA, and 55 patients were non-IPA. There was significance between the IPA and non-IPA groups, such as BALF GM (P < 0.001), history of glucocorticoid use (P = 0.004), and pulmonary comorbidities (P = 0.002), as well as no significance of the other demographic data including age, sex, BMI, history of cigarette, blood GM assay, T-SPOT.TB, and NEUT#/LYMPH#. The sensitivity of the BALF mNGS was 92.31%, which was higher than that of the traditional tests or the GM assays. The specificity of BALF mNGS was 92.73%, which was relatively similar to that of the traditional tests. The AUC of BALF mNGS was 0.925, which presented an excellent performance compared with other traditional tests or GM assays. Our study demonstrated the important role of BALF detection by the mNGS platform for pathogen identification in IPA patients with non-neutropenic states, which may provide an optimal way to diagnose suspected IPA disease.

BACKGROUND:Hypoxia-induced pulmonary hypertension (HPH) represents a severe pulmonary disorder with high morbidity and mortality, which necessitates identifying the critical molecular mechanisms underlying HPH pathogenesis.METHODS:The mRNA expression microarray GSE15197 (containing 8 pulmonary tissues from HPH and 13 normal controls) was downloaded from Gene Expression Omnibus (GEO). Gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) were executed by RStudio software. The Protein-Protein Interaction (PPI) network was visualized and established using Cytoscape, and the cytoHubba app from Cytoscape was used to pick out the hub modules. The infiltration of immune cells in HPH was analyzed using the CIBERSORTx. To confirm the potential hub genes, real-time quantitative reverse transcription PCR (qRT-PCR) was conducted using lung tissues of rat HPH models and controls.RESULTS:A total of 852 upregulated and 547 downregulated genes were identified. The top terms in biological processes were apoptosis, proliferation, and regulation of the MAPK cascade, including ERK1/2. Cytoplasm, cytosol, and membrane were enriched in cellular component groups. Molecular functions mainly focus on protein binding, protein serine/threonine kinase activity and identical protein binding. KEGG analysis identified pathways in cancer, regulation of actin cytoskeleton and rap1 signaling pathway. There was significantly different immune cell infiltration between HPH and normal control samples. High proportions of the memory subsets of B cells and CD4 cells, Macrophages M2 subtype, and resting Dendritic cells were found in HPH samples, while high proportions of naive CD4 cells and resting mast cells were found in normal control samples. The qRT-PCR results showed that among the ten identified hub modules, FBXL3, FBXL13 and XCL1 mRNA levels were upregulated, while NEDD4L, NPFFR2 and EDN3 were downregulated in HPH rats compared with control rats.CONCLUSION:Our study revealed the key genes and the involvement of immune cell infiltration in HPH, thus providing new insight into the pathogenesis of HPH and potential treatment targets for patients with HPH.

PDF file - 45K, Identification of transgenic mice. The transgenic mice were identified by the tail test method. The tail (1 cm) was cut when the mouse was 12 d old. The genomic DNA was extracted using lysis buffer (100 mM Tris-HCl pH 8.5, 5 mM EDTA, 0.2% SDS, and 200 mM NaCl) with Proteinase K. K14 forward and reverse primers were used for PCR and ras was amplified as a control.

Nationally representative reports on the characteristics and long‐term survival of pulmonary arterial hypertension (PAH) from developing countries are scarce. The applicability of the current main risk stratifications and the longitudinal changes in goal‐oriented treatments have yet to be elucidated in real‐world settings. Therefore, we aimed to provide insights into the characteristics, goal‐oriented treatments and survival of PAH in China and to explore the applicability of the main risk stratifications in our independent cohort.

OBJECTIVE:The efficacy of rapid on-site evaluation (ROSE) combined with computed tomography-guided transthoracic core needle biopsy (CT-guided TCNB) is rarely investigated. This study aimed to evaluate the diagnostic efficiency and safety of ROSE combined with CT-guided TCNB for suspected lung cancer patients.MATERIALS AND METHODS:Clinical data from 285 patients who received CT-guided TCNB for suspected lung cancer in Huashan Hospital from 2015 to 2018 were retrospectively analysed. Of these 163 patients underwent CT-guided TCNB combined with ROSE (ROSE group), while the remaining 122 patients underwent without ROSE (non-ROSE group). The smears from TCNB were quickly processed with Diff-Quick staining and analysed by a skilled cytologist on-site. The consistency of ROSE with the final clinicopathological diagnosis and the diagnostic efficiency and safety of ROSE combined with CT-guided TCNB in suspected lung cancer patients were evaluated.RESULTS:ROSE was highly concordant with pathological diagnosis (κ = 0.791; P < 0.001), with an accuracy of 95.7%. Diagnostic accuracy was significantly higher in the ROSE compared with the non-ROSE group (96.3% vs 86.1%; P = 0.002), with overall incidences of complications of 36.8% and 23.8%, respectively. Minor pneumothorax without drainage was slightly greater in the ROSE compared with the non-ROSE group (14.1% vs 6.6%; P = 0.046). However, there was no significant difference in serious complications between the two groups.CONCLUSION:ROSE was highly consistent with the final clinicopathological diagnosis for suspected lung cancer. ROSE further improved the diagnostic efficiency of CT-guided TCNB with no increased incidence of serious complications.

OBJECTIVES:To validate that dexlansoprazole, an anti-acid drug, can prevent pulmonary artery hypertension (PAH) in preclinical animal models and find the possible mechanism of action of dexlansoprazole for this new indication.METHODS:The efficacy of dexlansoprazole to attenuate PAH in vivo was evaluated in PAH animal models. Plasma guanosine 3', 5'-cyclic phosphate (cGMP) in PAH rats was measured by enzyme linked immunosorbent assay (ELISA). To investigate the anti-PAH effect of dexlansoprazole in vitro, proliferation and migration assays of primary cultured pulmonary artery smooth muscle cells (PASMCs) were performed. Furthermore, dexlansoprazole's function on fibroblast transition of vascular smooth muscle cells (VSMC) was explored by single cell ribonucleic acid (RNA) sequencing and RNAscope.RESULTS:Dexlansoprazole could attenuate the pathologic process in monocrotaline (MCT)-, hypoxia-induced PAH rats and SU5416/hypoxia (SuHy)-induced PAH mice. The intervention with dexlansoprazole significantly inhibited elevated right ventricular systolic pressure (RVSP), right ventricular hypertrophy, and pulmonary vascular wall thickness. Furthermore, plasma cGMP in MCT-induced PAH rats was restored after receiving dexlansoprazole. In vitro, dexlansoprazole could inhibit PASMCs' proliferation and migration stimulated by platelet derived growth factor-BB (PDGF-BB). Moreover, dexlansoprazole significantly ameliorated pulmonary vascular remodeling by inhibiting VSMC phenotypic transition to fibroblast-like cells in a VSMC-specific multispectral lineage-tracing mouse.CONCLUSIONS:Dexlansoprazole can prevent PAH through promoting cGMP generation and inhibiting pulmonary vascular remodeling through restraining PASMCs' proliferation, migration, and phenotypic transition to fibroblast-like cells. Consequently, PAH might be a new indication for dexlansoprazole.