Yu Fu

Objective: People undergoing lung transplantation may develop lung ischemia-reperfusion injury (LIRI), in which neutrophils contribute to the inflammatory response. However, the molecular features associated with distinct neutrophil states remain incompletely understood. This study aimed to identify candidate molecular features associated with the immunosuppressive-II (IS-II) neutrophil state, with particular attention to NFKBIE, an NF-κB inhibitor. Methods: We integrated bulk transcriptomic and single-cell RNA-sequencing data. Candidate genes were identified using co-expression analysis and machine-learning methods. We further examined cell-cell communication and the association of NFKBIE with neutrophil states using pseudotime analysis and computational gene perturbation. We validated NFKBIE expression in a mouse model of LIRI using RT-qPCR and Western blotting and assessed lung injury by histological examination. Results: A total of 553 differentially expressed genes and a key disease-associated module containing 4,456 genes were identified, with predominant enrichment in inflammation- and immunity-related pathways (eg, TNF and NF-κB). Five overlapping candidate transcriptomic markers, CXCL3, CYLD, NFKBIE, SLC30A1, and KBTBD8, were identified. The five-gene model showed strong discrimination in the discovery cohort (AUC = 0.986) and also showed strong discrimination in the separate GSE127003 validation dataset (AUC = 0.999, 95% CI 0.996– 1.000). Single-cell analysis showed that neutrophils were the largest annotated cell population in both CIT and post-reperfusion samples, whereas the IS-II neutrophil subtype was markedly enriched after reperfusion; NFKBIE was most highly expressed in IS-II neutrophils. Pseudotime trajectory analysis further indicated that elevated NFKBIE expression was associated with a later differentiation state toward the IS-II phenotype, while computational NFKBIE perturbation predicted alterations in several perturbation-associated genes. In vivo validation confirmed that both NFKBIE mRNA and protein levels were upregulated in lung tissue after reperfusion. Conclusion: The five-gene transcriptomic model showed reproducible discriminatory performance in a separate validation dataset. NFKBIE was consistently associated with the IS-II neutrophil state across multiple computational analyses, while in vivo experiments confirmed its increased expression after reperfusion. These findings nominate NFKBIE as a candidate gene associated with IS-II neutrophil differentiation and provide a hypothesis-generating basis for further functional investigation.

Background: The impact of mildly elevated pulmonary artery systolic pressure (PASP) on perioperative hemodynamic stability and myocardial injury during thoracoscopic lobectomy remains inadequately characterized despite its prevalence in the surgical population. Methods: This retrospective cohort study analyzed 3579 patients who underwent thoracoscopic lobectomy. Patients were categorized as having normal or mildly elevated PASP. Propensity score matching (PSM) was used to balance baseline characteristics between groups. The primary outcome was intraoperative hypotension. Secondary outcomes included myocardial injury after noncardiac surgery (MINS), hypoxemia, new-onset atrial fibrillation, postoperative pulmonary complications, 30-day readmission, and 30-day mortality. Sensitivity analyses using inverse probability of treatment weighting (IPTW) and overlap weighting were performed to validate the robustness of findings. Results: Among 3579 patients, 16.6% ( n = 594) had mildly elevated PASP. After PSM (491 pairs), patients with mildly elevated PASP experienced significantly higher rates of intraoperative hypotension [46.4 vs. 35.2%; odds ratio (OR) = 1.525; 95% CI: 1.190–1.957; P < 0.001]. Both short-duration (P < 0.001). These associations remained consistent across IPTW and overlap weighting analyses. Independent predictors of elevated PASP included smoking history (OR = 2.817), COPD (OR = 2.199), higher BMI (OR = 1.168 per unit), and older age (OR = 1.095 per year). Conclusions: Mildly elevated PASP significantly increases the risk of intraoperative hypotension and myocardial injury during thoracoscopic lobectomy. These findings identify patients with mildly elevated PASP as a high-risk population requiring careful hemodynamic monitoring and management. Preoperative assessment of PASP may enhance risk stratification for patients undergoing thoracoscopic lobectomy.

In sepsis, acute lung injury (ALI) is a severe complication and a leading cause of death, involving complex mechanisms that include cellular and molecular interactions between immune and lung parenchymal cells. In recent decades, the role of Toll-like receptor 4 (TLR4) in mediating infection-induced inflammation has been extensively studied. However, how TLR4 facilitates interactions between innate immune cells and lung parenchymal cells in sepsis remains to be fully understood. This study aims to explore the role of TLR4 in regulating macrophage immunity and metabolism in greater depth. It also seeks to reveal how changes in these processes affect the interaction between macrophages and both pulmonary endothelial cells (ECs) and lymphatic endothelial cells (LECs). Using TLR4 knockout mice and the combined approaches of single-cell RNA sequencing and experimental validation, we demonstrate that in sepsis, TLR4-deficient macrophages upregulate Abca1, enhance cholesterol efflux, and reduce glycolysis, promoting M2 polarization and attenuating inflammation. These metabolic and phenotypic shifts significantly affect their interactions with pulmonary ECs and LECs. Mechanistically, we uncovered that TLR4 operates through multiple pathways in endothelial dysfunction: macrophage TLR4 mediates inflammatory damage to ECs/LECs, while endothelial TLR4 both directly sensitizes cells to lipopolysaccharide-induced injury and determines their susceptibility to macrophage-derived inflammatory signals. These findings reveal the complex role of TLR4 in orchestrating both immune-mediated and direct endothelial responses during sepsis-induced ALI, supporting that targeting TLR4 on multiple cell populations may present an effective therapeutic strategy.

Sepsis-induced acute lung injury (ALI), characterized by severe hypoxemia and pulmonary leakage, remains a leading cause of mortality in intensive care units. The exacerbation of ALI during sepsis is largely attributed to uncontrolled inflammatory responses and endothelial dysfunction. Emerging evidence suggests an important role of Z-DNA binding protein 1 (ZBP1) as a sensor in innate immune to drive inflammatory signaling and cell death during infections. However, the role of ZBP1 in sepsis-induced ALI has yet to be defined. We utilized ZBP1 knockout mice and combined single-cell RNA sequencing with experimental validation to investigate ZBP1’s roles in the regulation of macrophages and lung endothelial cells during sepsis. We demonstrate that in sepsis, ZBP1 deficiency in macrophages reduces mitochondrial damage and inhibits glycolysis, thereby altering the metabolic status of macrophages. Consequently, this metabolic shift leads to a reduction in the differentiation of macrophages into pro-inflammatory states and decreases macrophage pyroptosis triggered by activation of the NLRP3 inflammasome. These changes significantly weaken the inflammatory signaling pathways between macrophages and endothelial cells and alleviate endothelial dysfunction and cellular damage. These findings reveal important roles for ZBP1 in mediating multiple pathological processes involved in sepsis-induced ALI by modulating the functional states of macrophages and endothelial cells, thereby highlighting its potential as a promising therapeutic target.

Purpose: Ischemia reperfusion injury (IRI) unavoidably occurs during lung transplantation, further contributing to primary graft dysfunction (PGD). Neutrophils are the end effectors of IRI and activated neutrophils release neutrophil extracellular traps (NETs) to further amplify damage. Nevertheless, potential contributions of NETs in IRI remain incompletely understood. This study aimed to explore NET-related gene biomarkers in IRI during lung transplantation. Methods: Differential expression analysis was applied to identify differentially expressed genes (DEGs) for IRI during lung transplantation based on matrix data (GSE145989, 127003) downloaded from GEO database. The CIBERSORT and weighted gene co-expression network analysis (WGCNA) algorithms were utilized to identify key modules associated with neutrophil infiltration. Moreover, the least absolute shrinkage and selection operator regression and random forest were applied to identify potential NET-associated hub genes. Subsequently, the screened hub genes underwent further validation of an external dataset (GSE18995) and nomogram model. Based on clinical peripheral blood samples, immunofluorescence staining and dsDNA quantification were used to assess NET formation, and ELISA was applied to validate the expression of hub genes. Results: Thirty-eight genes resulted from the intersection between 586 DEGs and 75 brown module genes, primarily enriched in leukocyte migration and NETs formation. Subsequently, four candidate hub genes (FCAR, MMP9, PADI4, and S100A12) were screened out via machine learning algorithms. Validation using an external dataset and nomogram model achieved better predictive value. Substantial NETs formation was demonstrated in IRI, with more pronounced NETs observed in patients with PGD ≥ 2. PADI4, S100A12, and MMP9 were all confirmed to be up-regulated after reperfusion through ELISA, with higher levels of S100A12 in PGD ≥ 2 patients compared with non-PGD patients. Conclusion: We identified three potential NET-related biomarkers for IRI that provide new insights into early detection and potential therapeutic targets of IRI and PGD after lung transplantation.

Introduction Echocardiography provides a non-invasive estimation of pulmonary artery systolic pressure (PASP) and is the first diagnostic test for pulmonary hypertension. Recent studies have demonstrated that PASP of more than 30 mm Hg related to increased mortality and morbidity. However, perioperative risks and management for patients with mildly elevated PASP are not well established. This study aims to explore the association between mildly elevated PASP and perioperative adverse outcomes. Methods and analysis This will be a retrospective cohort study conducted at Shanghai Pulmonary Hospital in Shanghai, China. Eligible patients are adults (≥18 years) who performed preoperative echocardiography and followed thoracoscopic lobectomy. Our primary objective is to determine the effect of preoperative mildly elevated PASP on the incidence of hypotension during surgery. Whether mildly elevated PASP is related to other perioperative adverse events (including hypoxaemia, myocardial injury, new-onset atrial fibrillation, postoperative pulmonary complications, 30-day readmission and 30-day mortality) will be also analysed. An estimated 2300 patients will be included. Ethics and dissemination The study has been approved by the institutional review board of Shanghai Pulmonary Hospital (approval No: 2022LY1143). The research findings intend to be published in peer-reviewed scientific publications. Trial registration number Chinese Clinical Trial Registry (ChiCTR2200066679).

Introduction Patients with a mediastinal mass are at risk of pulmonary complications in the perioperative period. Preoperative spirometry tests are recommended in patients scheduled for thoracic surgery. Our objective is to investigate the association between preoperative spirometry results and the incidence of postoperative pulmonary complications in patients following mediastinal mass resection, which may determine the usefulness of spirometry tests in the prediction of the perioperative respiratory risk. Methods and analysis This protocol describes a retrospective cohort study of patients with mediastinal masses in Shanghai Pulmonary Hospital between 1 September 2021 and 1 September 2022, with a planned sample size of 660 patients. The primary aim of this study is to explore the association between preoperative spirometry results and the occurrence of postoperative pulmonary complications after mediastinal mass resection. Logistic regression analysis will be used to calculate the adjusted incidence rate difference and incidence rate ratios (with 95% CIs). Ethics and dissemination The study was approved by the ethics committee of Shanghai Pulmonary Hospital (K21-372Y). The results of the study will be submitted to a peer-reviewed biomedical journal for publication and presented at relevant conferences.

Purpose: Simultaneous bilateral pulmonary resection via uniportal video-assisted thoracoscopic surgery (UVATS) was safe and feasible for the treatment of bilateral multiple pulmonary nodules. But, it should be noted that considerable postoperative pain at the bilateral surgical site was a crucial issue. The safety and efficacy of bilateral thoracic paravertebral block (TPVB) have been reported for postoperative analgesia. But, whether bilateral sequential TPVB can be safely and effectively used in simultaneous bilateral UVATS remains unknown. Therefore, this study aimed to determine the analgesic efficacy and safety of bilateral sequential TPVB after simultaneous bilateral UVATS. Study Design and Methods: In this study, 80 participants scheduled for UVATS will be randomly allocated to the bilateral sequential TPVB group (G₂) and the control group (G₁). The patient of G₂ will be performed bilateral TPVB at 2 time-points: before the start of the first side of pulmonary resection and before the start of the contralateral pulmonary resection. G₁ will only receive standard analgesia protocol. The primary outcome is the numeric rating scale score during coughing at 24 h postoperatively. The secondary outcomes include the Prince Henry Pain Score scores, sufentanil consumption, postoperative nausea and vomiting, levels of inflammatory factors, and the Quality of Recovery-40 scores at different time points, as well as chronic pain at postoperative day (POD) 90. Discussion: This is the first prospective trial to determine the safety and effectiveness of ultrasound-guided bilateral sequential TPVB for postoperative analgesia following simultaneous bilateral UVATS. This study also intended to evaluate the effect of this intervention on postoperative quality of recovery and inflammation levels. The final results will provide clinical evidence related to bilateral sequential TPVB, and promote the application of that acting as a more appropriate analgesic method for simultaneous bilateral UVATS.