Background The aim of the study was to investigate whether the expression of CD27 − CD38 + in interferon (IFN)-γ + CD4 + T cells stimulated by the specific antigen early secreted antigenic target-6 (ESAT-6)/culture filter protein-10 (CFP-10) could be a potential new therapeutic evaluation indicator for anti-tuberculosis (TB) treatment. Methods Newly diagnosed active pulmonary TB patients, latent TB infection (LTBI) and healthy controls were enrolled from January 2021 to December 2021. PTB patients were treated by standard anti-TB regimen 2HREZ/4HR (2 months of isoniazid (H), rifampin (R), ethambutol (E), and pyrazinamide (Z) followed by 4 months of isoniazid (H) and rifampin (R)). The difference of CD27 − CD38 + expression in IFN-γ + CD4 + T cells before treatment, 2 months after treatment, and 6 months after treatment were compared. Results Total 45 PTB patients, 38 LTBI cases and 43 healthy controls were enrolled. The expression of CD27 − CD38 + decreased significantly after anti-TB treatment and was comparable with that in LTBI and healthy controls when the 6-month anti-TB treatment course was completed. The decline rate of CD27 − CD38 + between 6 months after treatment and baseline was positively correlated with erythrocyte sedimentation rate ( r = 0.766, P < 0.0001), C-reactive protein ( r = 0.560, P = 0.003) and chest computerized tomography severity score ( r = 0.632, P = 0.0005). The area under receiver operator characteristic curve of CD27 − CD38 + in distinguish pulmonary TB patients before and after treatment was 0.779. Conclusion The expression of CD27 − CD38 + in ESAT-6/CFP-10 stimulated IFN-γ + CD4 + T cells can well reflect the changes of the disease before and after anti-TB treatment, which is expected to be a potential new therapeutic evaluation index. Clinical Registry number chiCTR1800019966.

Background Leptomeningeal metastasis (LM) of small cell lung cancer (SCLC) is a highly detrimental occurrence associated with severe neurological disorders, lacking effective treatment currently. Proteolysis-targeting chimeric molecules (PROTACs) may provide new therapeutic avenues for treatment of podophyllotoxin derivatives-resistant SCLC with LM, warranting further exploration. Methods The SCLC cell line H128 expressing luciferase were mutated by MNNG to generate H128-Mut cell line. After subcutaneous inoculation of H128-Mut into nude mice, H128-LM and H128-BPM (brain parenchymal metastasis) cell lines were primarily cultured from LM and BPM tissues individually, and employed to in vitro drug testing. The SCLC-LM mouse model was established by inoculating H128-LM into nude mice via carotid artery and subjected to in vivo drug testing. RNA-seq and immunoblotting were conducted to uncover the molecular targets for LM. Results The SCLC-LM mouse model was successfully established, confirmed by in vivo live imaging and histological examination. The upregulated genes included EZH2, SLC44A4, VEGFA, etc. in both BPM and LM cells, while SLC44A4 was particularly upregulated in LM cells. When combined with PROTAC EZH2 degrader-1, the drug sensitivity of cisplatin, etoposide (VP16), and teniposide (VM26) for H128-LM was significantly increased in vitro. The in vivo drug trials with SCLC-LM mouse model demonstrated that PROTAC EZH2 degrader-1 plus VM26 or cisplatin/ VP16 inhibited H128-LM tumour significantly compared to VM26 or cisplatin/ VP16 alone (P < 0.01). Conclusion The SCLC-LM model effectively simulates the pathophysiological process of SCLC metastasis to the leptomeninges. PROTAC EZH2 degrader-1 overcomes chemoresistance in SCLC, suggesting its potential therapeutic value for SCLC LM.

Objectives Histological transformation to small cell lung cancer (SCLC) has been identified as a mechanism of TKIs resistance in EGFR-mutant non-small cell lung cancer (NSCLC). We aim to explore the prevalence of transformation in EGFR-wildtype NSCLC and the mechanism of SCLC transformation, which are rarely understood. Methods We reviewed 1474 NSCLC patients to investigate the NSCLC-to-SCLC transformed cases and the basic clinical characteristics, driver gene status and disease course of them. To explore the potential functional genes in SCLC transformation, we obtained pre- and post-transformation specimens and subjected them to a multigene NGS panel involving 416 cancer-related genes. To validate the putative gene function, we established knocked-out models by CRISPR-Cas 9 in HCC827 and A549-TP53(-/-) cells and investigated the effects on tumor growth, drug sensitivity and neuroendocrine phenotype in vitro and in vivo. We also detected the expression level of protein and mRNA to explore the molecular mechanism involved. Results We firstly reported an incidence rate of 9.73% (11/113) of SCLC transformation in EGFR-wildtype NSCLC and demonstrated that SCLC transformation is irrespective of EGFR mutation status (P = 0.16). We sequenced 8 paired tumors and identified a series of mutant genes specially in transformed SCLC such as SMAD4, RICTOR and RET. We firstly demonstrated that SMAD4 deficiency can accelerate SCLC transition by inducing neuroendocrine phenotype regardless of RB1 status in TP53-deficient NSCLC cells. Further mechanical experiments identified the SMAD4 can regulate ASCL1 transcription competitively with Myc in NSCLC cells and Myc inhibitor acts as a potential subsequent treatment agent. Conclusions Transformation to SCLC is irrespective of EFGR status and can be accelerated by SMAD4 in non-small cell lung cancer. Myc inhibitor acts as a potential therapeutic drug for SMAD4-mediated resistant lung cancer.

Cancer cells lacking functional p53 exhibit poor prognosis, necessitating effective treatment strategies. Inhibiting WEE1, the G2/M cell cycle checkpoint gatekeeper, represents a promising approach for treating p53-deficient NSCLC. Here, we investigate the connection between p53 and WEE1, as well as explore a synergistic therapeutic approach for managing p53-deficient NSCLC. Our study reveals that p53 deficiency upregulates both protein levels and kinase activity of WEE1 by inhibiting its SUMOylation process, thereby enhancing the susceptibility of p53-deficient NSCLC to WEE1 inhibitors. Furthermore, we demonstrate that the WEE1 inhibitor Adavosertib induces intracellular lipid peroxidation, specifically in p53-deficient NSCLC cells, suggesting potential synergy with pro-oxidant reagents. Repurposing Disulfiram (DSF), an alcoholism medication used in combination with copper (Cu), exhibits pro-oxidant properties against NSCLC. The levels of WEE1 protein in p53-deficient NSCLC cells treated with DSF-Cu exhibit a time-dependent increase. Subsequent evaluation of the combination therapy involving Adavosertib and DSF-Cu reveals reduced cell viability along with smaller tumor volumes and lighter tumor weights observed in both p53-deficient cells and xenograft models while correlating with solute carrier family 7-member 11 (SLC7A11)/glutathione-regulated ferroptosis pathway activation. In conclusion, our findings elucidate the molecular interplay between p53 and WEE1 and unveil a novel synergistic therapeutic strategy for treating p53-deficient NSCLC.

Background:Lung squamous cell cancer in situ (LSCIS) is preinvasive squamous tumor and generally overlooked as a potential subtype of pathological and clinical significance, which has seldom been investigated systematically. This study sought to explore the clinical features, prognostic factors, and optimal treatments for LSCIS patients.Methods:Patients diagnosed with LSCIS (n=449), lung adenocarcinoma in situ (LAIS; n=1,132), stage IA lung squamous cell cancer (LSQCC; n=22,289) and stage IA lung adenocarcinoma (LUAD; n=68,523) were identified in the Surveillance Epidemiology and End Results (SEER) database. Additionally, 512 patients from the Shanghai Pulmonary Hospital diagnosed with LSCIS (n=34), LAIS (n=248), stage IA LSQCC (n=118) and stage IA LUAD (n=112) were included in the study. Kaplan-Meier survival curves were constructed, and Cox proportional hazards regression analyses were performed to examine the overall survival (OS), lung cancer-specific survival (LCSS), and progression-free survival (PFS) of the patients.Results:The univariate and multivariate analyses showed the patients with LSCIS had significantly worse survival than those with LAIS. Although, the univariate analysis revealed that the LSCIS patients had significantly worse OS and LCSS than the stage IA LSQCC patients, the multivariate analyses showed that the prognosis of the LSCIS was similar to that of the stage IA LSQCC in the SEER cohort. The prognosis of the LSCIS was similar to that of the stage IA LSQCC in the Shanghai Pulmonary Hospital cohort. The univariate and multivariate analyses showed that age (>70 years) and chemotherapy were negative prognostic factors, and surgery was a favorable prognostic factor for the LSCIS patients. The survival of the LSCIS patients who underwent local tumor destruction or excision was similar to that of those who did not receive surgery. Lobectomy was the surgical procedure associated with the highest OS and LCSS in LSCIS patients.Conclusions:The survivals of the LSCIS were similar to those of the stage IA LSQCC, but significantly worse than those of the LAIS. Surgery was an independent favorable prognostic factor for the LSCIS patients. Lobectomy was a superior choice of surgical procedure, and significantly improved the current outcomes of the LSCIS patients.

Abstract Background: Traditional bacteriological tests are time-consuming and of low sensitivity for the diagnosis of active tuberculosis (ATB). The purpose of this study is to investigate the accuracy of CD27-CD38+IFN-γ+CD4+/CD4+ for active tuberculosis (ATB) diagnosis. Methods: This prospective diagnostic accuracy study was conducted at Shanghai Pulmonary Hospital between January 2019 and December 2021. Patients with ATB, non-tuberculosis mycobacterium infection (NTM), latent tuberculosis infection (LTBI), other respiratory diseases (OD), and healthy individuals (HC) were enrolled. The accuracy of CD27-CD38+IFN-γ+CD4+/CD4+ and other phenotypic markers for ATB diagnosis was assessed.Results: A total of 376 patients (237 ATB, 38 LTBI, 8 NTM, 50 OD, and 43 HC) were enrolled. The ratios of CD4+IFN-γ+CD27- and CD4+IFN-γ+CD27-CD38+ profiles in CD4+IFN-γ+ cells and the ratios of CD4+IFN-γ+CD38+, CD4+IFN-γ+CD27-, and CD4+IFN-γ+CD38+CD27- profiles in CD4+ cells in the ATB group were significantly higher than in the other groups. The area under the curve (AUC) of CD27-CD38+IFN-γ+CD4+/CD4+ for the diagnosis of ATB was the highest, with a value of 0.890. With the optimal cutoff value of 1.34´10-4, the sensitivity and specificity of CD27-CD38+IFN-γ+CD4+/CD4+ for ATB diagnosis was 0.869 and 0.849, respectively. Conclusion: CD27-CD38+IFN-γ+CD4+/CD4+ might be an effective biomarker for active tuberculosis diagnosis.

Sepsis is a global health care issue that affects millions of people. DNA methyltransferase I (DNMT1)-mediated DNA methylation is involved in a number of human diseases by affecting many types of cellular progression events. However, the role and underlying molecular mechanism of DNMT1 in development of sepsis remain largely unknown. Lipopolysaccharide (LPS) induced lung fibrosis in the sepsis mouse model, and DNMT1 was upregulated in lung tissues of a sepsis mouse model compared with lung tissues from control mice. Then, this study demonstrated that LPS induced the production of interleukin (IL)-7 and tumor necrosis factor (TNF)-α and promoted DNMT1 expression in primary type II alveolar epithelial cells (AECII cells). Knockdown of DNMT1 inhibited IL-7 and TNF-α secretion in AECII cells exposed to LPS. Further study demonstrated that DNMT1 repressed the expression of miR-130a in AECII cells with or without LPS exposure. Next, this study demonstrated that miR-130a inhibited ZEB1 expression in AECII cells exposed to LPS. Ultimately, this study revealed the role of the DNMT1/miR-130a/ZEB1 regulatory pathway in AECII cells exposed to LPS. Overall, our data revealed that LPS induced the secretion of inflammatory factors by modulating the DNMT1/miR-130a/ZEB1 regulatory pathway in AECII cells, thus providing a novel theoretical basis that might be beneficial for establishment of diagnostic and therapeutic strategies for sepsis.

Background Chronic obstructive pulmonary disease (COPD) is a progressive disorder that causes airway obstruction and lung inflammation. The first-line treatment of COPD is the bronchodilators of β2-agonists and antimuscarinic drugs, which can help control the airway obstruction, but the long-term use might render the drug tolerance. Bisphosphonates are widely used in osteoclast-mediated bone diseases treatment for decades. For drug repurposing, can delivery of a third generation of nitrogen-containing bisphosphonate, risedronate (RIS) ameliorate the progression of COPD? Methods COPD rats or mice models have been established through cigarette-smoking and elastase injection, and then the animals are received RIS treatment via nebulization. Lung deposition of RIS was primarily assessed by high-performance liquid chromatography (HPLC). The respiratory parameters of airway obstruction in COPD rats and mice were documented using plethysmography method and resistance-compliance system. Results High lung deposition and bioavailability of RIS was monitored with 88.8% of RIS input dose. We found that RIS could rescue the lung function decline of airspace enlargement and mean linear intercept in the COPD lung. RIS could curb the airway obstruction by suppressing 60% of the respiratory resistance and elevating the airway’s dynamic compliance, tidal volume and mid-expiratory flow. As an inhibitor of farnesyl diphosphate synthase (FDPS), RIS suppresses FDPS-mediated RAS and RhoA prenylation to obstruct its membrane localization in airway smooth muscle cells (ASMCs), leading to the inhibition of downstream ERK-MLCK and ROCK1-MLCP pathway to cause ASMCs relaxation. Additionally, RIS nebulization impeded pro-inflammatory cell accumulation, particularly macrophages infiltration in alveolar parenchyma. The NF-κB, tumor necrosis factor-alpha, IL-1β, IL-8, and IL-6 declined in microphages following RIS nebulization. Surprisingly, nebulization of RIS could overcome the tolerance of β2-agonists in COPD-rats by increasing the expression of β2 receptors. Conclusions Nebulization of RIS could alleviate airway obstruction and lung inflammation in COPD, providing a novel strategy for treating COPD patients, even those with β2-agonists tolerance.

Objective: Lynch syndrome (LS) pre-screening methods remain under-investigated in colorectal cancers (CRCs) in Asia. Here, we aimed to systematically investigate LS pre-screening and comprehensively characterize LS CRCs. Methods: Microsatellite instability (MSI) and germline variants of DNA mismatch repair (MMR) genes were examined in 406 deficient MMR (dMMR) and 250 proficient MMR CRCs. The genetic differences between LS and sporadic CRCs were studied with whole exome sequencing analysis. Results: The incidence of dMMR in Chinese patients with CRCs was 13.8%. Consistency analysis between MMR immunohistochemistry (IHC) and MSI testing showed the kappa value was 0.758. With next-generation sequencing (NGS), germline variants were detected in 154 CRCs. Finally, 88 patients with CRC were identified as having LS by Sanger sequencing. Among them, we discovered 21 previously unreported pathogenic germline variants of MMR genes. Chinese patients with LS, compared with sporadic CRCs, tended to be early-onset, right-sided, early-stage and mucinous. Overall, the performance of MMR IHC and MSI testing for LS pre-screening was comparable: the area under the ROC curve for dMMR, MSI-H, and MSI-H/L was 0.725, 0.750, and 0.745, respectively. dMMR_MSI-H LS and sporadic CRCs showed substantial differences in somatic genetic characteristics, including different variant frequencies of APC, CREBBP, and KRAS, as well as different enriched pathways of VEGF, Notch, TGFβR, mTOR, ErbB, and Rac protein signal transduction. Conclusions: MMR IHC and MSI testing were effective methods for LS pre-screening. The revealed clinical and somatic genetic characteristics in LS CRCs may have the potential to improve the performance of LS pre-screening in combination with dMMR/MSI.

Objectives Air temperature has been considered a modifiable and contributable variable in COVID-19 transmission. Implementation of non-pharmaceutical interventions (NPIs) has also made an impact on COVID-19 transmission, changing the transmission pattern to intrahousehold transmission under stringent containment measures. Therefore, it is necessary to re-estimate the influence of air temperature on COVID-19 transmission while excluding the influence of NPIs. Design, setting and participants This study is a data-based comprehensive modelling analysis. A stochastic epidemiological model, the ScEIQR model (contactable susceptible-exposed-infected-quarantined-removed), was established to evaluate the influence of air temperature and containment measures on the intrahousehold spread of COVID-19. Epidemic data on COVID-19, including daily confirmed cases, number of close contacts, etc, were collected from the National Health Commission of China. Outcome measures The model was fitted using the Metropolis-Hastings algorithm with a cost function based on the least squares method. The LOESS (locally weighted scatterplot smoothing) regression function was used to assess the relationship between air temperature and rate of COVID-19 transmission within the ScEIQR model. Results The ScEIQR model indicated that the optimal temperature for spread of COVID-19 peaked at 10℃ (50℉), ranging from 5℃ to 14℃ (41℉−57.2℉). In the fitted model, the fitted intrahousehold transmission rate (β’) of COVID-19 was 10.22 (IQR 8.47–12.35) across mainland China. The association between air temperature and β’ of COVID-19 suggests that COVID-19 might be seasonal. Our model also validated the effectiveness of NPIs, demonstrating that diminishing contactable susceptibility (Sc) and avoiding delay in diagnosis and hospitalisation (η) were more effective than contact tracing (κ and ρ). Conclusions We constructed a novel epidemic model to estimate the effect of air temperature on COVID-19 transmission beyond implementation of NPIs, which can inform public health strategy and predict the transmission of COVID-19.

Abstract Background: The aim of the study was to investigate whether the CD27-CD38+ in IFN-γ+CD4+T cells stimulated by the specific antigen ESAT-6/CFP-10 could be a potential new therapeutic evaluation indicator for anti-tuberculosis (TB) treatment. Methods: Newly diagnosed active pulmonary TB patients, latent TB infection (LTBI) and healthy controls were enrolled from January 2021 to December 2021. PTB patients were treated by standard anti-TB regimen 2HREZ/4HR. The difference of CD27-CD38+ in IFN-γ+CD4+ T cells before treatment, 2 months after treatment, and 6 months after treatment were compared. Results: The CD27-CD38+ in IFN-γ+CD4+ T cells decreased significantly after anti-TB treatment and was comparable with that in LTBI and healthy controls when the 6-month anti-TB treatment course was completed. The decline rate of CD27-CD38+ between 6 months after treatment and base line was positively correlated with ESR (r=0.766, P<0.0001) and chest CT severity score (r=0.632, P=0.0005). The area under ROC curve of CD27-CD38+ in distinguish PTB patients before and after treatment was 0.779. Conclusion: The CD27-CD38+ in ESAT-6/CFP-10 stimulated IFN-γ+CD4+T cells can well reflect the changes of the disease before and after anti-TB treatment, which is expected to be a potential new therapeutic monitoring biomarker. Clinical Registry number：chiCTR1800019966

Abstract Background Active tuberculosis (ATB) originates from primary Mycobacterium tuberculosis (MTB) infection or reactivation of latent tuberculosis. Besides bacteriological examination, MTB-reactive immunocytes detection can be an alternative testing for discrimination of active tuberculosis. The purpose of this study is to investigate the accuracy of peripheral blood CD27−CD38+IFN-γ+CD4+T cells in ATB diagnosis. Methods This prospective diagnostic accuracy study was conducted at Shanghai Pulmonary Hospital between January 2019 and December 2021. Patients with ATB, non-tuberculosis mycobacterium infection (NTM), latent tuberculosis infection (LTBI), other respiratory diseases (OD), and healthy individuals (HC) were enrolled. The accuracy of CD27−CD38+IFN-γ+CD4+/CD4+ and other phenotypic markers for ATB diagnosis was assessed. Results A total of 376 patients (237 ATB, 38 LTBI, 8 NTM, 50 OD, and 43 HC) were enrolled. The ratios of CD4+IFN-γ+CD27− and CD4+IFN-γ+CD27−CD38+ profiles in CD4+IFN−γ+ cells and the ratios of CD4+IFN-γ+CD38+, CD4+IFN-γ+CD27−, and CD4+IFN-γ+CD38+CD27− profiles in CD4+ cells in the ATB group were significantly higher than in the other groups. The area under the curve (AUC) of CD27−CD38+IFN-γ+CD4+/CD4+ for the diagnosis of ATB was the highest, with a value of 0.890. With the optimal cutoff value of 1.34 × 10–4, the sensitivity and specificity of CD27−CD38+IFN-γ+CD4+/CD4+ for ATB diagnosis was 0.869 and 0.849, respectively. Conclusion CD27−CD38+IFN-γ+CD4+/CD4+ might be a potential biomarker for active tuberculosis diagnosis.

Lung adenocarcinoma, the most prevalent lung cancer subtype, is characterized by its high propensity to metastasize. Despite the importance of metastasis in lung cancer mortality, its underlying cellular and molecular mechanisms remain largely elusive. Here, we identified miR-200 miRNAs as potent suppressors for lung adenocarcinoma metastasis. miR-200 expression is specifically repressed in mouse metastatic lung adenocarcinomas, and miR-200 decrease strongly correlates with poor patient survival. Consistently, deletion of mir-200c/141 in the KrasLSL-G12D/+ ; Trp53flox/flox lung adenocarcinoma mouse model significantly promoted metastasis, generating a desmoplastic tumor stroma highly reminiscent of metastatic human lung cancer. miR-200 deficiency in lung cancer cells promotes the proliferation and activation of adjacent cancer-associated fibroblasts (CAFs), which in turn elevates the metastatic potential of cancer cells. miR-200 regulates the functional interaction between cancer cells and CAFs, at least in part, by targeting Notch ligand Jagged1 and Jagged2 in cancer cells and inducing Notch activation in adjacent CAFs. Hence, the interaction between cancer cells and CAFs constitutes an essential mechanism to promote metastatic potential.

Background Patients with stage I lung adenocarcinoma (LUAD) have varying postoperative prognosis. This study aimed to investigate the prognostic significance of postoperative longitudinal change of serum carcinoembryonic antigen (CEA) level in patients with stage I LUAD. Methods The study cohort comprised 241 patients with stage I LUAD completely resected with single-port video-assisted thoracic surgery (VATS). The patients were categorized into 4 groups according to the postoperative longitudinal change of serum CEA levels measured in the third and sixth months after surgery: the NN group (continuously normal), HN group (increase first and then decrease), NH group (decrease first and then increase), and HH group (continuously high). Recurrence-free survival (RFS) was analyzed by the Kaplan-Meier method and compared by log-rank test. A nomogram was developed to predict recurrence in the stage I LUAD patients. Results In univariate analysis, differentiation (P<0.001), visceral pleural invasion (VPI) (P=0.025), tumor diameter (P<0.001), tumor-node-metastasis (TNM) stage (P=0.008), preoperative CEA levels (≥10.0 vs. <10.0 ng/mL, P<0.001), and postoperative CEA grouping (NH/HH vs. NN/HN, P<0.001) were significant prognostic factors for stage I LUAD patients. Multivariate analysis showed that tumor diameter (P=0.009) and postoperative CEA grouping (P<0.001) were considered to be independent prognostic factors of postoperative recurrence of stage I LUAD. Tumor diameter (≥20 mm) and postoperative CEA (NH/HH vs. NN/HN) were associated with worse RFS. Receiver operating characteristic (ROC) curve analysis showed that postoperative CEA (NH/HH vs. NN/HN) have high sensitivity (64.7%) and specificity (83.2%) for early prediction of postoperative recurrence of stage I LUAD. The area under curve (AUC) value was 0.745. The nomogram based on multivariate Cox regression had a concordance index (value of 0.789). The calibration plot showed that the predicted probabilities closely matched the observed probabilities. Conclusions Longitudinal change in serum CEA level after surgery was found to be an independent unfavorable prognostic factor in completely resected stage I LUAD patients. The NH group and HH group were significantly associated with worse RFS. A nomogram was established to predict the postoperative recurrence of patients with stage I LUAD.

Background Acute lung injury (ALI) is a common clinical complication with a high mortality rate. Resveratrol (Res) has been shown to protect against ALI, but the role of long noncoding RNAs (lncRNAs) in this process is still unclear. Methods Male rats (n=20) aged 7–8 weeks were randomly divided into four groups: control, lipopolysaccharide (LPS), LPS + Res, and LPS + dexamethasone (Dexa). Intragastric administration of Res (0.5 mg/kg) or Dexa (1.5 mg/kg) was performed 1 h before intraperitoneal injection of LPS (5 mg/kg). Lung tissue, serum, and bronchoalveolar lavage fluid were sampled 6 h after LPS treatment for inflammatory factor detection, pathological detection, lncRNA sequencing and bioinformatical analysis, and TdT-mediated dUTP Nick-End Labeling. Quantitative real time polymerase chain reaction and western blotting were used to verify the sequencing results. LPS, Res, and RNA interference were used in rat alveolar epithelial cells experiments to confirm the protective of Res/lncRNA against ALI. Results Res pretreatment inhibited lung injury and the increase of inflammatory cytokines induced by LPS. The differentially expressed lncRNAs and mRNAs (P<0.05 and |fold change| >2) were mainly involved in the signaling pathway of immunity, infection, signaling molecules and interactions. Among the lncRNAs and mRNAs, 26 mRNAs and 23 lncRNAs had high levels in lungs treated with LPS but decreased with Res, and 17 mRNAs and 27 lncRNAs were at lower levels in lungs treated with LPS but increased with Res. lncRNA and adjacent mRNA analysis showed that lncRNAs XLOC_014869 and the adjacent gene Fos, and the possible downstream genes Jun and Faslg were increased by LPS, but these changes were attenuated by Res. Pretreatment with Res reduced LPS-induced lung tissue apoptosis. Similarly, Res treatment and knockdown of lncRNA XLOC_014869 reduced LPS-induced apoptosis and the levels of Fos, c-Jun, and Fas-L. Conclusions Res can inhibit the increase of lncRNAs XLOC_014869 caused by LPS stimulation and inhibit lung cell apoptosis. These effects may be due to lncRNA XLOC_014869 mediation of the pro-apoptotic factors (Fos, c-Jun, and Fas-L).

MAIT cells are unconventional innate-like T lymphocytes contributing to host immune protection against Mycobacteria tuberculosis (Mtb) infection. CD4- MAIT cells play a major role in immune protection against tuberculosis (TB), however, the role of CD4+ MAIT cells was elusive due to their low abundance. We firstly investigated the frequency and functions of CD4+ MAIT cells in pulmonary tuberculosis (PTB) patients before and after anti-TB treatment. We found that the frequency of Mtb-reactive CD4+ MAIT cells and IFN-γ, granzyme B (GrzB), CD69 expression on them were increased while LAG-3+ cells of them were decreased in PTB patients. After the treatment, the frequency of Mtb-reactive CD4+ MAIT cells and CD69, IFN-γ, GrzB expression on them were decreased while LAG-3 increased. The results indicated the expression profile is distinct between CD4+ MAIT cells and CD4- MAIT cells in PTB patients, the increased IFN-γ and GrzB expression of CD4+ MAIT cells play a role in anti-TB immunity.

BACKGROUNDPulmonary large cell carcinoma (LCC) is a poorly differentiated and rare tumor with dismal outcome, and there are no recommended treatments for LCC. Little is known about the efficacy of postoperative chemotherapy in patients with early stage LCC.METHODSThe patients with early stage I/II LCC in the Surveillance, Epidemiology and End Results (SEER) database between 2004 and 2015 were retrospectively reviewed. The overall survival (OS) of patients with LCC at different stages and treatments were evaluated by Kaplan-Meier analysis with log-rank test. Univariate and multivariate Cox proportional risk regression analysis were employed to determine the independent risk factors of OS. Finally, a nomogram was constructed to predict the 1 -, 3- and 5-year OS of early stage LCC patients.RESULTSA total of 1,099 pulmonary LCC cases were included in this study. 71.8% of patients were over 60 years old, and 66.7% of the tumor lesions located in the upper lobe, followed by the lower lobe (25.7%). Meanwhile, the majority of tumors showed poor differentiation (96.1%). The median OS of surgical patients with or without post-operative adjuvant chemotherapy was 61 and 47 months, respectively. Post-operative chemotherapy was associated with better OS (HR: 0.805; 95% CI: 0.676-0.959, P=0.020). For patients with tumor size >3 cm or IB stage tumor, the prognosis of postoperative chemotherapy was better than that of patients without chemotherapy. Multivariate Cox analysis revealed the age, stage and treatments were independent risk factors of OS for early stage LCC. The nomogram had a calibration index of 0.581.CONCLUSIONSThe incidence of LCC was high in the elderly, and it generally had poor differentiation. Post-operative chemotherapy is strongly recommended for patients with LCC at stage IB or higher.

Near-infrared (NIR) fluorescence-guided surgery is a developing field in precision medicine for cancer treatment. However, traditional NIR fluorescent reagents have limitations including lower sensitivity and specificity for tumor delineation, as well as insufficient photostability and tissue penetration depth. Herein, an improved dual-targeting lanthanide-doped NIR-to-visible upconversion nanoprobe is constructed for fluorescent imaging of lung cancer. The nanoprobes (UCNP@P-RGD-NGR) are synthesized by preparing polydopamine-coated NaYF4:Yb/Tm@NaYF(4)upconversion nanoparticles (UCNP) and conjugating them with dual-targeting peptides RGD10-NGR9, which are designed to target both integrin alpha v beta 3/alpha v beta 5 and aminopeptidase N receptors on tumor cells. The monodispersed core-shell nanoprobe is rod-shaped (approximate to 32 nm in minor axis) with ultrathin polydopamine shell (approximate to 2 nm) under transmission electron microscopy. The absorption spectrum and upconversion luminescence properties of the nanoprobes are identified by spectrometry. Toxicity analysis on cellular and animal experiments suggests the nanoprobes are well biocompatible. UCNP@P-RGD-NGR can specifically target A549 cancer cells as revealed by competitive cell binding assay and ICP-MS. Imaging studies in vivo with BALB/c nude mice bearing tumor xenografts demonstrate that UCNP@P-RGD-NGR can delineate tumor from surrounding normal tissues with strong upconversion blue-violet luminescence. This dual-targeting nanoprobe shows great potential for clinical application for NIR fluorescence surgical navigation of lung cancer.

BACKGROUND AND OBJECTIVE: Chemotherapy remains the basis of the treatment of lung cancer, and screening biomarkers with predictive value for chemotherapy is of great interest. The present study focused on status of genes methylation in NSCLC patients receiving pemetrexed- or gemcitabine-based chemotherapy. PATIENTS AND METHODS: Promoter methylation of Ras association domain family (RASSF1A) and short stature homeobox 2 (SHOX2) was examined in bronchoalveolar lavage (BAL) from 117 NSCLC patients treated with chemotherapy. Multivariate analysis was used to identify the predictive value of gene methylation. Progression-free survival (PFS) rather than overall survival (OS) was used as the clinical outcome to minimize the impact of chemotherapy on gene methylation. RESULTS: The methylation of RASSF1A and SHOX2 was significantly associated with shorter PFS (RASSF1A: HR = 2.355, 95% CI: 1.533-3.617, P < 0.0001; SHOX2: HR = 2.123, 95% CI: 1.392-3.236, P = 0.0004). After adjusting for confounding factors, RASSF1A methylation was still a predictive factor for PFS (HR = 1.765, 95% CI: 1.064-2.928, P = 0.0278). In the pemetrexed group, unmethylated RASSF1A could be used to predict longer PFS (P = 0.0001), and no predictive value was found in the gemcitabine group. CONCLUSION: Unmethylated RASSF1A is a favorable prognostic indicator for patients receiving pemetrexed doublets. Because of the promoting effect of most chemotherapeutic drugs on gene methylation, unmethylated RASSF1A is not suitable as a predictor for gemcitabine doublets.

Early non-pharmaceutical interventions (NPIs) are crucial to prevent and control of COVID-19 pandemic. We established a stochastic non-classical SEIR NPIs model (ScEIQRsh) which can quantify the three kinds of NPIs measures simultaneously to mimic the clustered intra-family or intra-acquaintance spreading pattern of COVID-19 under the effective integrated NPIs in Mainland China. Model simulation demonstrated that measures to diminish contactable susceptible (Sc), such as home confinement, travel constraint, social distancing etc. and measures to avoid delay of diagnosis and hospitalized isolation (η) were more effective but consumptive than contact tracing (κ, ρ). From fitted model by MCMC method, the proportion of asymptomatic infectors was 14.88% (IQR 8.17%, 25.37%). The association between air temperature and the fitted transmission rate (β) of COVID-19 suggests that COVID-19 pandemic would be seasonal with the optimal temperature range of 5°C-14°C and peak of 10°C for spreading, and vaccine is indispensable to ultimate prevention COVID-19. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement No funding ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Shanghai Pulmonary Hospital, Tongji University School of Medicine All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes No additional data avaiable.