Pulmonary fibrosis represents a progressive interstitial lung disease marked by excessive extracellular matrix deposition and architectural distortion. Vascular endothelial cells critically contribute to fibrogenesis through paracrine secretion of pro-fibrotic mediators, yet their mechanobiological regulation remains elusive. Using integrated single-cell multi-omics profiling of human pulmonary fibrosis specimens and experimental fibrosis models induced by bleomycin or silica, we identify mechanosensitive Piezo1 upregulation in Endothelial cells as a hallmark of fibrotic progression. Endothelial-specific Piezo1 knockout significantly attenuates Bleomycin-induced fibrotic remodeling in male mice, establishing its pathogenic necessity. Mechanistically, PIEZO1 activation promotes pulmonary fibrosis development via CAPN2-mediated STAT3 phosphorylation, which may regulate the secretion of the pro-fibrotic molecule interleukin-33. These findings suggest that the endothelial PIEZO1-CAPN2-STAT3-IL33 axis is a potential therapeutic target for PF intervention.

Eosinophilic asthma is a chronic inflammatory disease driven by Type 2 immune cells and cytokines. While cytokine-based immunotherapies have shown promising efficacy in asthma treatment, current approaches often fail to simultaneously target multiple cytokines. In this study, we present nanodecoys derived from the cellular membranes of M2 macrophages, which exhibit a high density of cytokine receptors, enabling efficient sequestration of multiple Type 2 cytokines. This mechanism significantly alleviates OVA-induced eosinophilic asthma inflammation in mice. To enhance therapeutic efficacy, D-mannose glycans, a potential immunoregulatory molecule for asthma, were modified onto the surface of the nanovesicles. This modification facilitates the phagocytic clearance of absorbed cytokines by targeting the mannose receptor (CD206). Consequently, the mannose-modified nanovesicles exhibit synergistic immunosuppressive effects on asthmatic inflammation by inhibiting M2 macrophage polarization, CD4+ T cell Th2 polarization, and eosinophil activation, as revealed by single-cell RNA sequencing and in vitro analyses. In summary, this study presents a safe and effective nanotechnology-based immunotherapy for asthma and potentially other Type 2 inflammatory diseases.

Purpose:This study aimed to explore factors affecting adherence to remote home-based pulmonary rehabilitation (PR) in patients with stable chronic obstructive pulmonary disease (COPD) and to develop a predictive model. Patients and Methods:This multicenter, cross-sectional survey study included 86 patients who underwent 12 weeks of health education-integrated, home-based PR with remote monitoring. Patients were stratified into high-completion (HC, ≥ 70%) and low-completion (LC, < 70%) groups. Demographic data, clinical features, and psychological parameters were analyzed. Receiver operating characteristic curve and area under the curve (AUC) analyses evaluated the predictive performance of key indicators. Binary logistic regression identified four predictors: Pulmonary Rehabilitation Adapted Index of Self-Efficacy (PRAISE), Outcome Expectations for Exercise Scale (OEE), Montreal Cognitive Assessment (MoCA), and Visual Analog Scale (VAS). These components formed an optimized predictive model with corresponding formula and cutoff values. Results:A cross-sectional survey of 71 patients, 44 in the HC group and 27 in the LC group, revealed significantly higher scores in the HC group in the following domains of the 36-Item Short Form Health Survey (SF-36), including physical functioning, role limitations due to physical health, role limitations due to emotional problems, energy/fatigue, mental health, and social functioning, as well as in the MoCA scores (all p-values < 0.05). Significant intergroup differences were also observed in PRAISE, OEE and VAS scores (all p < 0.001). PRAISE (AUC = 0.810), OEE (AUC = 0.784), MoCA (AUC = 0.719), and VAS (AUC = 0.801) demonstrated discriminatory power in assessing PR adherence. The combined predictive model achieved an AUC of 0.895 (95% confidence interval: 0.812-0.977, p < 0.05), with 77.8% sensitivity and 93.2% specificity. Conclusion:Social cognitive theory (SCT) originated from social learning theory. It explains human behavior through a triadic, dynamic, and reciprocal model. This model posits continuous interaction among an individual's behavior, cognitive factors, and environmental context. The four-variable predictive model, based on SCT, effectively evaluates adherence to home-based PR under remote monitoring in patients with COPD. Among the indicators in the four-variable model, PRAISE shows potential as a target for intervention to enhance PR completion rates.

BACKGROUND:Resveratrol has been found to have anti-inflammatory and anti-allergic properties. The effects of resveratrol on thymic stromal lymphopoietin (TSLP)-mediated atopic march remain unclear.PURPOSE:To explore the potential role of resveratrol in TSLP-mediated atopic march.METHODS:The atopic march mouse model was established by topical application of MC903 (a vitamin D3 analog). Following the treatment with resveratrol, airway resistance in mice was discovered by pulmonary function apparatus, and the number of total cells, neutrophils, and eosinophils in bronchoalveolar lavage fluid was counted. The histopathological features of pulmonary and ear skin tissues, inflammation, and cell infiltration were determined by hematoxylin and eosin staining. The messenger RNA (mRNA) levels of TSLP, immunoglobulin E, interleukin (IL)-4, IL-5, and IL-13 were measured by real-time quantitative polymerase chain reaction. The protein expression of nuclear factor kappa B (NF-κB)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling-associated molecules (p-p65, p65, p-I kappa B kinase alpha (IκBα), IκBα, Nrf2, and TSLP) in lung and ear skin tissues were assessed by Western blot analysis.RESULTS:Resveratrol attenuated airway resistance and infiltration of total cells, eosinophils, and neutrophils in both lung and ear skin tissues. Resveratrol ameliorates serum inflammatory markers in allergic mice. Moreover, the phosphorylation levels of NF-κB pathway-related proteins were significantly reduced by administration of resveratrol in allergic lung and ear skin tissues. Similarly, the protein expression of TSLP in both lung and ear skin tissues was reduced by resveratrol, and Nrf2, a protector molecule, was increased with resveratrol treatment.CONCLUSION:Resveratrol attenuates TSLP-reduced atopic march through ameliorating inflammation and cell infiltration in pulmonary and ear skin tissues by inhibiting the abnormal activation of NF-κB signaling pathway.

BACKGROUND:The pathogenesis and treatment strategies for chronic obstructive pulmonary disease (COPD) require further exploration. Abnormal neutrophil inflammation and the overexpression of neutrophil extracellular traps (NETs) are closely associated with acute exacerbations of COPD (AECOPD). Siglec-9, a specific receptor expressed on neutrophils that inhibits their function, prompted us to investigate its relationship with NETs found in induced sputum and the severity of the disease.METHODS:We collected clinical data from patients with AECOPD and assessed the expression of Siglec-9 in peripheral blood neutrophils and the presence of NETs in induced sputum. We then observed the correlation between Siglec-9, the inflammatory response, and the severity of AECOPD.RESULTS:We observed an increase in the expression of Siglec-9 in the peripheral blood neutrophils of patients with AECOPD. Concurrently, these patients exhibited more severe clinical symptoms, higher systemic inflammation levels, and a reduced quality of life compared to those with induced sputum NET expression. Further subgroup analysis of AECOPD patients with high Siglec-9 expression revealed worsened quality of life and more severe inflammation, particularly in indicators such as the BODE index, CRP, peripheral blood neutrophil count, IL-6, IL-8, TNF-α expression, and others. Furthermore, we noted a significant increase in NET-specific expression in the sputum of patients with high Siglec-9 expression levels. In comparison to patients with low Siglec-9 expression, those with high expression experienced more systemic inflammatory reactions and a lower quality of life. Correlation analysis of the aforementioned indicators revealed that the expression ratio of Siglec-9 in the peripheral blood of patients correlated with lung function, quality of life, and NETs in the induced sputum of patients with AECOPD.CONCLUSION:The increased expression of Siglec-9 in peripheral blood neutrophils of AECOPD patients leads to elevated NET expression in induced sputum, exacerbating the systemic inflammatory response and worsening lung function and quality of life in these patients.

Src-homology region 2 domain-containing phosphatase 1 (SHP-1) is considered an anti-inflammatory factor, but its role in chronic obstructive pulmonary disease (COPD) remains unknown. Herein, overexpression of SHP-1 was utilized to explore the functions of SHP-1 in COPD models established by stimulating 16HBE cells with cigarette smoke extracts (CSE) in vitro. SHP-1 was downregulated in both COPD patients and CES-treated 16HBE cells. SHP-1 overexpression reinforced cell viability and significantly prevented CSE-induced cell apoptosis in 16HBE cells. Furthermore, SHP-1 overexpression greatly reversed the CSE-induced migration, epithelial–mesenchymal transition (EMT), and pro-inflammatory factor production in 16HBE cells. In addition, CSE activated the P65 and PI3K/AKT pathways in 16HBE cells, which was also reversed by SHP-1 overexpression. Our findings indicated that SHP-1 alleviated CSE-induced EMT and inflammation in 16HBE cells, suggesting that SHP-1 regulated the development of COPD, and these functions may be linked to the inhibition of the PI3K/AKT pathway.

Pulmonary fibrosis (PF) is a lethal lung disease characterized by aberrant lung scarring. Vascular endothelial cells (VECs), which activate fibroblasts through secreting pro-fibrotic molecules, are vital for the PF development. However, the regulatory mechanisms are not fully understood. Here, by employing single-cell multiomics to analyze samples from PF patients and bleomycin (BLM) and SiO2-induced PF mouse models, we discovered that increased mechanical force and mechanosensitive PIEZO1 on VECs are closely associated with PF development, Subsequently, EC-specific deletion of PIEZO1 significantly reduced BLM-induced mouse PF, thereby further validated the critical role of endothelial PIEZO1 in PF. Mechanistically, PIEZO1 activation couples with a pro-fibrotic molecule interleukin-33 (IL-33) to drive the PF development via calpain-2 (CAPN2)-signal transducer and activator of transcription 3 (STAT3) axis. Additionally, PIEZO1 antagonist GsMTx4 effectively alleviated fibrotic outcome, while agonist YODA1 exacerbated the results in mouse model. Our findings reveals the therapeutic potential of mechanosensitive pathways for PF.

Cough is a common symptom of several respiratory diseases. However, frequent coughing from acute to chronic often causes great pain to patients. It may turn into cough variant asthma, which seriously affects people's quality of life. For cough treatment, it is dominated by over-the-counter antitussive drugs, such as asmeton, but most currently available antitussive drugs have serious side effects. Thus, there is a great need for the development of new drugs with potent cough suppressant. BALB/c mice were used to construct mice model with cough to investigate the pharmacological effects of pectolinarigenin (PEC). Hematoxylin-eosin and Masson staining were used to assess lung injury and airway remodeling, and ELISA was used to assess the level of inflammatory factor release. In addition, inflammatory cell counts were measured to assess airway inflammation. Airway hyperresponsiveness assay was used to assess respiratory resistance in mice. Finally, we used Western blotting to explore the potential mechanisms of PEC. We found that PEC could alleviate lung tissue injury and reduce the release of inflammatory factors, inhibit of cough frequency and airway wall collagen deposition in mice model with cough. Meanwhile, PEC inhibited the Ras/ERK/c-Fos pathway to exhibit antitussive effect. Therefore, PEC may be a potential drug for cough suppression.

Lung adenocarcinoma (LUAD) is one of the most common causes of cancer-related death. The role of pyroptosis in LUAD remains unclear. Our study aimed to identify a prognostic signature of pyroptosis-related genes (PRGs) and explore the connection of PRGs with the tumour microenvironment in LUAD. Gene expression and clinical information were obtained from The Cancer Genome Atlas database. Consensus clustering was applied to classify LUAD patients. The least absolute shrinkage and selection operator Cox and multivariate Cox regression models were used to generate a PRG-related prognostic signature. The correlations between PRGs and tumour-infiltrating immune cells or the tumour mutational burden were analysed by Spearman's correlation analysis. In this study, 44 PRGs significantly differed in expression between LUAD and normal tissues. Based on these genes, patients were clustered into three clusters with significantly different distributions of tumour-infiltrating immune cells and immune checkpoint regulators. A total of four PRGs (NLRP1, HMGB1, CYCS, and BAK1) were used to construct a prognostic model. Significant correlations were observed between these prognostic PRGs and immune cell infiltration or the tumour mutational burden. Predictive nomogram results showed that BAK1 could be an independent prognostic biomarker in LUAD. Additionally, the expression level of BAK1 was validated in two independent Gene Expression Omnibus cohorts. Our identified prognostic PRG signature may provide insight for future studies targeting pyroptosis and the tumour microenvironment in LUAD. Future studies are needed to verify our current findings.

Objective:To analyze the correlation between chronic obstructive pulmonary disease (COPD) and cognitive dysfunction.Methods:This is a case-control study. From February 2022 to October 2022, 32 COPD patients (inpatient and outpatient) from the Department of Respiratory and Critical Care Medicine and Rehabilitation Medical Center of the First Affiliated Hospital of Nanjing Medical University and 32 healthy subjects were recruited. All participants underwent a thorough evaluation, which included Montreal Assessment of Cognitive Function (MoCA), visuospatial n-back task included accuracy (ACC) and mean response time (RT), the pulmonary functions including forced vital capacity (FVC), forced expiratory volume in the first second (FEV 1), one-second rate (FEV 1/FVC) and maximum volume per minute (MVV), Health Survey Short Form (SF-36), and St. George′s Respiratory Questionnaire (SGRQ). The correlation between cognitive dysfunction and lung function, SF-36 and SGRQ in COPD patients were analyzed. Results:The prevalence of smoking, hypertension and cardiovascular disease in the two groups were significantly different (all P<0.05). MoCA score, 1-back ACC and 2-back ACC in COPD group were significantly lower than those in healthy control group [(23.86±4.50) vs (27.55±1.29) points, (76.82%±16.60%) vs (90.61%±7.40%), (67.93%±10.10%) vs (78.74%±10.38%), all P<0.001]; 2-back RT was significantly higher than that of healthy group [(316.43±108.17) vs (254.09±101.62) ms, P<0.05]; and the Physiological function (PF), physiological function (RP), emotional function (RE), energy (VT), social function (SF), physical pain (BP) in SF-36 were significantly worse than the healthy control group (all P<0.05). The MoCA score of COPD group was positively correlated with FEV 1/FVC ( r=0.501, P=0.018). The 1-back ACC was positively correlated with FEV 1 and FEV 1/FVC ( r=0.568, 0.634; both P<0.05). The 1-back RT was negatively correlated with FEV 1/FVC and MVV ( r=-0.452, -0.534; both P<0.05). The 2-back ACC was positively correlated with FEV 1/FVC ( r=0.426, P=0.048). The 2-back RT was negatively correlated with MVV ( r=-0.571, P=0.006). In COPD group, MoCA score was negatively correlated with activity, influence and total score in SGRQ ( r=-0.533, -0.466, -0.521; all P<0.05). The 1-back ACC was negatively correlated with activity, influence and total score ( r=-0.552, -0.517, -0.584; all P<0.05). The 1-back RT was positively correlated with activity, influence and total score ( r=0.430, 0.379, 0.417; all P<0.05). The 2-back ACC was negatively correlated with impact and total score ( r=-0.398, -0.412; both P<0.05). Conclusion:COPD patients have impaired cognitive function, which is mainly manifested by the decline of working memory and executive function, and is correlated with the lung function, general health condition and quality of life.

To the Editor: Asthma is an abnormal chronic inflammatory disease characterized by the involvement of a complex network of cells. Barrier epithelial cells (ECs) represent the first line of defense and express pattern recognition receptors to recognize type-2 cell-mediated immune insults. An exaggerated abnormal barrier function or inadequate immune response may contribute to the pathophysiology of asthma.[1,2] The alarmin high mobility group box 1 (HMGB1) is secreted into the extracellular environment by lung ECs during inflammation. HMGB1, an inflammatory mediator, promotes an immediate immune response to tissue damage in inflammatory pulmonary disease. Therefore, we aimed to determine whether HMGB1 levels are associated with asthma. This study was based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and was registered at Prospective Register of Systematic Review (PROSPERO, CRD42022329371). The Embase and PubMed databases were searched until April 28, 2022. The inclusion criteria were as follows: (1) case-control studies with more than three cases; (2) patients diagnosed with asthma; and (3) sufficient HMGB1 data. Literature screening and data extraction were performed independently by two investigators. Any disagreement was resolved through discussion with a third reviewer. The methodological quality of each risk-of-bias item for each eligible study was assessed using the Newcastle–Ottawa Scale (NOS). Review Manager software 5.2 (The Cochrane Collaboration, 2014) was used for data synthesis and analysis. After screening, 13 studies were included for qualitative and quantitative analyses in our meta-analysis [Supplementary Figure 1, https://links.lww.com/CM9/B340]. The characteristics of the eligible studies, which included 977 participants (359 healthy subjects in the control group and 618 patients with asthma in the case group), are presented in Supplementary Table 1, https://links.lww.com/CM9/B340. The percentage of females in the control group was 50.14% (180/359) and 49.84% (308/618) in the case group. The mean age range in the healthy subjects and patients with asthma was 11.07 to 54.23 years and 10.56 to 57.44 years, respectively. HMGB1 expression was detected by the enzyme-linked immunosorbent assay. The specimen sources were bronchial brushings, biopsies, serum, plasma, sputum, and bronchoalveolar lavage fluid. Four articles compared serum/plasma HMGB1 levels between patients with asthma and healthy subjects; the levels were significantly higher in patients with asthma (mean difference [MD] = 27.25, 95% confidence intervals [CI] = 17.73–36.77). However, heterogeneity (I2 = 95%, P < 0.001) was detected across studies; thus, a random-effects model was used [Figure 1A]. Furthermore, eight articles focused on sputum HMGB1 levels. HMGB1 levels were significantly elevated in patients with asthma (MD = 190.99, 95% CI = 127.08–254.91) compared to those in healthy subjects, and the results were highly heterogeneous (I2 = 100%, P < 0.001) [Figure 1B].Figure 1: Serum/plasma HMGB1 levels were significantly higher in asthma patients than those in healthy subjects (A). Sputum HMGB1 levels were significantly elevated in asthma patients compared to those in healthy subjects, and the results were highly heterogeneous (B). Serum/plasma HMGB1 levels were significantly elevated in patients with mild-to-moderate asthma and significantly higher in patients with severe asthma than in the control group. Furthermore, patients with severe asthma had significantly higher serum/plasma HMGB1 levels than those with mild-to-moderate asthma (C). Similar results were observed for sputum HMGB1 levels (D). HMGB1: High mobility group box 1.We also revealed a relationship between HMGB1 levels and disease severity. Eligible articles were pooled, and patients with asthma were classified as having mild-to-moderate asthma or severe asthma in accordance with the Global Initiative for Asthma. Serum/plasma HMGB1 levels were significantly elevated in patients with mild-to-moderate asthma (MD = 10.90, 95% CI = 2.16–19.65, I2=54%, P = 0.14) and significantly higher in patients with severe asthma (MD = 27.63, 95% CI = 7.92–47.33, I2 = 66%, P = 0.08) than in the control group. Furthermore, patients with severe asthma had significantly higher serum/plasma HMGB1 levels than those with mild-to-moderate asthma (MD = 13.31, 95% CI = 6.92–19.71, I2 = 0%, P = 0.38) [Figure 1C]. Similar results were observed for sputum HMGB1 levels. HMGB1 expression was significantly higher in patients with mild-to-moderate and severe asthma than in healthy subjects (mild-to-moderate asthma, MD = 176.80, 95% CI = 122.19–231.41, I2 = 97%, P < 0.001; severe asthma, MD = 265.80, 95% CI = 220.73–310.87, I2 = 95%, P < 0.001). Additionally, patients with severe asthma had significantly higher sputum HMGB1 levels than those with mild-to-moderate asthma (MD = 102.31, 95% CI = 78.04–126.57, I2 = 73%, P = 0.01) [Figure 1D]. Subgroup analyses based on country, sex, age, body mass index (BMI), and percent of forced expiratory volume in one second (FEV1%) predicted were performed to explore the source of heterogeneity. No significant heterogeneity was observed in the group with BMI < 18.5 kg/m2 (P = 1.000, I2 = 0%) and 18.5 ≤ BMI < 24.0 kg/m2 (P = 0.200, I2=39%); however, there was considerable heterogeneity in the BMI ≥ 24.0 kg/m2 groups (P = 0.002, I2 = 100%). No other variables were sources of heterogeneity [Supplementary Table 2, https://links.lww.com/CM9/B340]. The NOS scores are summarized in Supplementary Table 1, https://links.lww.com/CM9/B340, and each numbered item of all qualified studies is shown in the percentage chart [Supplementary Figure 2, https://links.lww.com/CM9/B340]. All included articles were high-quality studies (scores >7.0). New non-invasive biomarkers for airway inflammation are essential. Sputum HMGB1 levels were significantly higher in patients with severe asthma than in those with mild-to-moderate asthma, and HMGB1 levels increased with the degree of airflow limitation. Therefore, HMGB1 could be an independent risk factor affecting the predicted value of FEV1%.[3] Our results reveal a significant difference in HMGB1 levels in serum/plasma/sputum specimens between patients with asthma and healthy controls. Additionally, HMGB1 overexpression was more pronounced in patients with moderate-to-severe and severe asthma than in healthy subjects, suggesting that high HMGB1 levels correlated positively with disease severity. Therefore, elevated HMGB1 levels may be a potential biomarker of asthma severity. HMGB1, a damage-associated molecular pattern molecule, participates in the inflammatory response and releases inflammatory mediators by binding to specific receptors, including advanced glycation end product receptors (RAGEs) and Toll-like receptors (TLRs).[4] Airway ECs are direct participants and specific target cells in the development of asthmatic inflammation. The surface of these cells is enriched with TLRs and RAGEs, and this is one of the cell types that is the most responsive to HMGB1. Although the mechanism of HMGB1 in human asthma remains unclear, previous studies have demonstrated that HMGB1 is involved in the pathogenesis of airway inflammation, hyper-responsiveness, and remodeling in a mouse model of asthma, which was prevented or reversed by blocking HMGB1 activity. A recent study identified the G protein-coupled receptor P2Y13 as a novel gatekeeper of HMGB1, and targeting this receptor via genetic deletion or a small-molecule antagonist may inhibit the occurrence and development of experimental asthma.[5] However, long-term clinical trials are needed to ascertain whether the specific blockage of the receptor for HMGB1 can be achieved in an effective and safe manner in human asthma patients. In conclusion, our results suggest that HMGB1 could be a potential biomarker of asthma severity. Early detection of HMGB1 and blocking its specific receptors may be valuable therapeutic strategies for asthma. Given the limitations of this study, further investigations are warranted to confirm these findings and allow for efficient clinical translation. Funding This work was supported by grants from the National Key Research and Development Program of China (Nos. 2022YFF0710800 and 2018YFC1313600), Major International (Regional) Joint Research Project of China (No. 81820108001), National Natural Science Foundation of China (Nos. 81670029 and 82000038), Jiangsu Key Principal Investigator of Medicine (No. ZDRCA2016018), and Project 333 for Cultivation of High-Level Talents (Leading Talents of the Young and Middle-Aged) (No. BRA2019078), and the Nanjing Key Project of Science and Technology (No. 2019060002). Conflicts of interest None.

Jingxian Jiang1#, Shuanglan Xu1#, Zi Chen1#, Weihua Liu1, Liuchao Zhang1, Jianmin Li2, Zhou Zhu3, Linfu Zhou1,2,4 1Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China; 2Animal Core Facility, Key Laboratory of Model Animal, Nanjing Medical University, Nanjing 211166, Jiangsu Province, China; 3Department of Pediatrics and Department of Molecular Microbiology and Immunology, Brown University Warren Alpert Medical School, Providence, RI 02912, USA; 4Institute of Integrative Medicine, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China

Background: Lung cancer is one of the most prevalent cancers and the leading cause of cancer-related deaths worldwide; non-small cell lung cancer (NSCLC) comprises approximately 80% of all lung cancer cases. This study aimed to construct a competing endogenous RNA (ceRNA) network and identify prognostic signatures in elderly patients with NSCLC. Methods: We extracted data from elderly patients with NSCLC from The Cancer Genome Atlas and identified differentially expressed (DE) messenger RNAs (mRNAs), microRNAs (miRNAs), and long non-coding RNAs (lncRNAs). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to investigate the functions of DEmRNAs. The interactions between RNAs were predicted using starBase, TargetScan, miRTarBase, and miRanda. Cytoscape version 3.0 was used to construct and visualize the lncRNA-miRNA-mRNA ceRNA network. The association between the expression levels of DERNAs in the constructed ceRNA network and overall survival was determined using the survival package in R software. Furthermore, another Gene Expression Omnibus cohort was studied to externally validate the ceRNA network. Results: In total, 2865 DEmRNAs, 62 DEmiRNAs, and 131 DElncRNAs were identified. Dysregulated mRNAs are enriched in cancer-related processes and pathways. A ceRNA network was constructed using 38 miRNAs, 61 lncRNAs, and 164 mRNAs. Of these, 3 lncRNAs, 3 miRNAs, and 16 mRNAs were closely related to overall survival. The MIR99AHG-hsa-miR-31-5p-PRKCE axis has been identified as a potential ceRNA network involved in the development of NSCLC in elderly individuals. External validation of the MIR99AHG-hsa-miR-31-5p-PRKCE axis in the GSE19804 cohort showed that PRKCE was downregulated and that MIR99AHG was upregulated in the tumor tissues of elderly patients with NSCLC compared with normal lung tissues. Conclusions: This study provides novel insights into the lncRNA-miRNA-mRNA ceRNA network and reveals potential biomarkers for the diagnosis and prognosis of elderly patients with NSCLC.

Human interleukin-5 (IL-5) functions as an important pro-inflammatory factor by binding to its specific receptor, IL-5Rα, which has been implicated in the pathogenesis of asthma. Previously, a disulfide-bonded cyclic peptide AF17121 obtained from random library screening and sequence variation was found to competitively disrupt the cognate IL-5Rα/IL-5 interaction with moderate potency. In this study, the crystal complex of IL-5Rα with AF17121 was investigated at structural and energetic levels. It is revealed that the side-chain indole moiety of the AF17121 Trp5 residue is a potential site for a stem putative halogen bond (X-bond) with IL-5Rα, which is just located within the key 3 EXXR6 motif region recognized specifically by IL-5Rα. We systematically examined four halogen substitution types at five positions of the indole moiety; QM/MM calculations theoretically unraveled that only halogenations at 5 and 6 positions can form effective X-bonds with the side-chain hydroxyl oxygen of the IL-5Rα Thr21 residue and the backbone carbonyl oxygen of Ala66 residue, respectively. Binding assays observed that I-substitution at the 5 position and Br-substitution at the 6 position can result in two potent halogenated peptides, [5I]AF17121 and [6Br]AF17121, which are improved by 1.6-fold and 3.5-fold relative to the native AF17121, respectively. 5I/6Br-double substitution, resulting in [5I/6Br]AF17121, can further enhance the peptide affinity by 7.5-fold. Structural analysis revealed that the X-bond stemming from 6Br-substitution is also involved in an orthogonal interaction system with a H-bond; they share a common backbone carbonyl oxygen acceptor of IL-5Rα Ala66 residue and exhibit a significant synergistic effect between them.

Human epidermal growth factor receptor (EGFR) is involved in strong association with malignant proliferation, which has been shown to play a central role in the development and progression of non-small cell lung cancer and other solid tumors. The tumor-suppressor protein MIG6 is a negative regulator of EGFR kinase activity by binding at the activation interface of asymmetric dimer of EGFR kinase domain to disrupt EGFR dimerization and then inactivate the kinase. The protein adopts two discrete fragments 1 and 2 to directly interact with EGFR. It is revealed that the MIG6 fragment 2 is intrinsically disordered in free unbound state, but would fold into a well-structured β-hairpin when binding to EGFR, thus characterized by a so-called coupled folding-upon-binding process, which can be regarded as a compromise between favorable direct readout and unfavorable indirect readout. Here, a 23-mer F2P peptide was derived from MIG6 fragment 2, trimmed into a 17-mer tF2P peptide that contains the binding hotspot region of the fragment 2, and then constrained with an ordered hairpin conformation in free unbound state by disulfide stapling, finally resulting in a rationally stapled/trimmed stF2P peptide that largely minimizes the unfavorable indirect readout effect upon its binding to EGFR kinase domain, with affinity improved considerably upon the trimming and stapling/trimming. These rationally designed β-hairpin peptides may be further exploited as potent anti-lung cancer agents to target the activation event of EGFR dimerization.

BackgroundSevere neutrophilic asthma is often characterized by persistent airway inflammation and irreversible airway remodeling, which are overstimulated by the high-mobility group box protein 1 (HMGB1). Although wogonin, an O-methylated flavone, has been widely used to treat inflammatory and allergic diseases, its therapeutic effects and potential mechanisms on severe neutrophilic asthma remain elusive.ObjectiveTo evaluate whether wogonin alleviates airway neutrophilia through inducing neutrophil apoptosis and attenuates airway smooth muscle cells (ASMCs) proliferation and migration.MethodsThe effect of wogonin on reducing neutrophilic airway inflammation, including neutrophil infiltration and inflammatory mediators, was examined in a mouse model of severe neutrophilic asthma sensitized with ovalbumin and lipopolysaccharide. Also, the effect of wogonin on inducing human neutrophil apoptosis was manifested using cellular morphology, flow cytometry, and caspase inhibition assays. Furthermore, the effect of wogonin on inhibiting HMGB1-mediated ASMCs proliferation and migration was determined.ResultsWogonin reduced the frequency of neutrophils and inhibited the production of multiple inflammatory mediators, including ovalbumin-specific IgE, tumor necrosis factor-α, interleukin-6, and HMGB1, in bronchoalveolar lavage fluid and lung tissues of the neutrophilic asthmatic mouse model. These data strongly support a significantly suppressed neutrophilic airway inflammation, functionally consistent to the relieved airway hyperresponsiveness by wogonin in vivo. Wogonin induced human neutrophil apoptosis in a dose-dependent manner by activating caspase-8 and caspase-3 in vitro. Wogonin pretreatment abolished HMGB1-induced ASMCs proliferation and migration, which can be explained by the inhibition of phosphorylation in the mitogen-activated protein kinase (MAPK) /Akt singling pathways.ConclusionOur findings demonstrate that wogonin augments caspase-dependent apoptosis in neutrophils to alleviate neutrophilic inflammatory responses and regulates intracellular signaling to inhibit HMGB1-mediated ASMCs activation, providing a promising therapeutic agent for severe neutrophilic asthma.

Background: Airway neutrophilia has been associated with asthma severity and asthma exacerbations. This study attempted to identify biomarkers, pathogenesis, and therapeutic molecular targets for severe asthma in neutrophils using bioinformatics analysis. Methods: Fifteen healthy controls and 3 patients with neutrophilic severe asthma were screened from the Gene Expression Omnibus (GEO) database. Based on the analysis of differentially expressed genes (DEGs), functional and pathway enrichment analyses, gene set enrichment analysis, protein–protein interaction network construction, and analysis were performed. Moreover, small-molecule drug candidates have also been identified. Results: Three hundred and three upregulated and 59 downregulated genes were identified. Gene ontology function enrichment analyses were primarily related to inflammatory response, immune response, leukocyte migration, neutrophil chemotaxis, mitogen-activated protein kinase cascade, Jun N-terminal kinase cascade, I-kappaB kinase/nuclear factor-κB, and MyD88-dependent toll-like receptor signaling pathway. Pathway enrichment analyses and gene set enrichment analysis were mainly involved in cytokine-cytokine receptor interaction, the TNF signaling pathway, leukocyte transendothelial migration, and the NOD-like receptor signaling pathway. Furthermore, 1 important module and 10 hub genes (CXCL8, TLR2, CXCL1, ICAM1, CXCR4, FPR2, SELL, PTEN, TREM1, and LEP) were identified in the protein–protein interaction network. Moreover, indoprofen, mimosine, STOCK1N-35874, trapidil, iloprost, aminoglutethimide, ajmaline, levobunolol, ethionamide, cefaclor, dimenhydrinate, and bethanechol are potential drugs for the treatment of neutrophil-predominant severe asthma. Conclusion: This study identified potential biomarkers, pathogenesis, and therapeutic molecular targets for neutrophil-predominant severe asthma.

BACKGROUND:Baduanjin is a traditional Chinese exercise regimen used to treat various chronic illnesses and is associated with both psychological and physical benefits. However, its benefits for patients suffering from chronic obstructive pulmonary disease (COPD) are unclear. This study aims to assess the efficacy, safety, and underlying mechanisms of Baduanjin exercise in patients with moderate-to-severe COPD (BROCADE) by remote monitoring.METHODS:This study protocol describes a multicenter, open-label, prospective randomized computed tomography. A total of 150 individuals who meet the inclusion criteria after the screening and consent processes will take part in the study. All participants will be provided routine medication and lifestyle interventions. They will be randomly assigned to a control group, a classical pulmonary rehabilitation group, or a Baduanjin group, which will undergo remotely monitored Baduanjin exercises for a cumulative duration of 1 hour per day, three times per week for 12 weeks. The participants will be followed for 24 weeks. The primary outcomes will be a 6-minutes walking distance and St. George's Respiratory Questionnaire index. The secondary outcomes will be lung function, cross-sectional area of the pectoralis major and subcutaneous fat, modified Medical Research Council score, COPD assessment test questionnaire results, extremity muscle strength, and quality of life. Any adverse events that may occur will be monitored and recorded.RESULTS:This study is ongoing and will be submitted to a peer-reviewed journal for publication once completed.CONCLUSION:A novel neutrophil-related inflammatory mechanism will potentially be identified. In addition, the study results will provide a safe, effective, simple and operational Baduanjin exercise protocol for moderate-to-severe COPD patients aimed at improving prognosis and quality of life.