BACKGROUND:Chronic rhinosinusitis with nasal polyps involves mixed type 2-type 3 inflammation, associated with disease severity and treatment resistance, yet mechanisms remain unclear. OBJECTIVE:We sought to investigate the role of IL-2 and its interaction with type 3 inducers (TNF-α, IL-1β, and IL-23) in driving mixed type 2-type 3 inflammation. METHODS:IL-2 and receptor expression in nasal polyps was analyzed using tissue homogenates and public RNA-sequencing data. Dispersed nasal polyp cells were treated with IL-2/type 3 inducers, with cytokine production, proliferation, and gene expression analyzed via immunoassays, flow cytometry, and RNA sequencing. IL-2 receptor/Janus kinase (JAK) blockade studies were conducted. Furthermore, CD4+ and CD8+ T cells were magnetically isolated from nasal polyps to evaluate their response to cytokine stimulation. RESULTS:IL-2 levels were increased in chronic rhinosinusitis with nasal polyps, particularly in type 3-dominant and mixed type 2-type 3 subgroups, and correlated with type 3 cytokines. RNA sequencing supported upregulated IL-2 receptors and their coexpression with type 3 genes. IL-2 synergized with type 3 inducers to enhance both type 2 and type 3 cytokine production in dispersed nasal polyp cells. Direct functional evidence from isolated CD4+ and CD8+ T cells confirmed this synergy, with CD8+ T cells emerging as a novel source of IL-13. Flow cytometry further supported these findings, showing synergistic cytokine production across diverse cell populations, including T-cell subsets and natural killer cells. Memory T cells mediated T-cell receptor-independent cytokine production. Transcriptomic analysis identified activated type 3, type 2, and JAK-signal transducer and activator of transcription signaling pathways. IL-2 receptor blockade, particularly JAK inhibition, attenuated IL-2/type 3 inducer-mediated synergistic inflammation. CONCLUSIONS:A novel mechanism was identified whereby IL-2 synergizes with proinflammatory type 3 inducers to amplify mixed type 2-type 3 inflammation via innate-like T-cell activation, and targeted JAK inhibition was validated as a potential therapy.

Observational studies have long suggested an association between gut microbiota dysbiosis and gastric cancer, yet the causal relevance and underlying biological pathways remain poorly defined. Here, we applied a comprehensive Mendelian randomization (MR) framework to gut microbiota and gastric cancer GWAS summary statistics to identify 14 microbial taxa significantly associated with gastric cancer risk. Sensitivity analyses, including tests for horizontal pleiotropy, heterogeneity, and genetic co-localization, revealed no strong evidence of shared causal variants (H4 < 2%); however, convergent results across five complementary MR models support overall causal inference. Mediation analysis further identified a partial pathway from the taxon ebi-a-GCST90027718 to gastric cancer via the metabolite met-c-926 (sdLDL), accounting for 4.31% of the total effect. Preliminary 16S rRNA sequencing in a pilot cohort (n = 5 per group) revealed divergent shifts in Ruminococcus subgroups without disrupting overall community stability, though these observations require replication. Collectively, this study establishes a genetically anchored evidence chain linking host genetics, gut microbiota, and gastric cancer pathogenesis, providing insights for microbiota-targeted prevention strategies.

Chronic obstructive pulmonary disease (COPD) is a highly heterogeneous disease with complex pathogenesis. Identifying high-risk populations and implementing timely prevention strategies are critical to reducing the disease burden. Single-cell RNA sequencing of lung tissue from control never-smokers, patients with pre-COPD, and COPD patients revealed a novel T cell subset characterized by high expression of metallothionein (MT) genes, designated MT-high T cells. These cells were progressively depleted in the lungs with disease progression. A similar decline was observed in the peripheral blood using flow cytometry, highlighting the potential of these cells to serve as an accessible biomarker of disease progression. Functional analysis indicated that MT-high T cells suppress CD8+ T cell cytotoxic activity, suggesting a key immunoregulatory role in disease pathogenesis. Receiver operating characteristic curve analysis demonstrated the excellent potential of MT-high T cell frequency to predict susceptibility to COPD. These findings establish MT-high T cells as promising biomarkers for identifying individuals at risk for COPD and as novel targets for future therapeutic and prophylactic strategies.

Background:When systemic lupus erythematosus (SLE) affects the lymphatic system, it disrupts the normal physiological function of the lymphatic circulation, leading to clinical chylous effusion and lymphedema. Currently, the computed tomography lymphangiography (CTL) or technetium-99m dextran (99Tcm-DX) lymphoscintigraphy features of patients with SLE complicated by chylous effusion and lymphedema have not been published, and studies on the imaging features of different subtypes are lacking. This study aimed to analyze the imaging characteristics of CTL and 99Tcm-DX lymphoscintigraphy in 23 patients with SLE complicated with chylous effusion and/or lymphedema, and describe the differences in imaging findings among different subtypes, so as to provide a reference for the clinical understanding of this disease. Methods:A retrospective analysis was conducted on the clinical and imaging data of 23 patients with SLE complicated by chylous effusion and/or lymphedema. All patients underwent CTL and 99Tcm-DX lymphoscintigraphy. The patients were divided into three subgroups: chylous effusion (n=19), chylous effusion and lymphedema (n=3), and lymphedema (n=3). The signs of lymphatic abnormalities detected by the two examinations were compared using the McNemar test (with the exact probability method), and diagnostic consistency was evaluated using the kappa test (P<0.05 was considered statistically significant), and the differences in imaging features among the subgroups were described. Results:No significant differences were found between CTL and 99TCm-DX lymphoscintigraphy in depicting thoracic duct outlet obstruction, thoracic segment tortuosity or dilation of the thoracic duct, left subclavian trunk visualization, abnormal perineal distribution, chylothorax, or chylous ascites (P>0.05), with kappa values of 0.623, 0.862, 0.511, 0.646, 0.425, and 0.777, indicating diagnostic agreement levels of good, almost perfect, good, good, moderate, and good, respectively. However, the two methods differed significantly in showing combined chylothorax and chylous ascites (P<0.05), with minor diagnostic agreement (kappa =0.157). CTL additionally identified contrast extravasation in the left cervical trunk, mediastinum, lung, perirenal, and peritoneal areas, as well as iliac and lumbar trunk dilation, which were not detected by lymphoscintigraphy. Subgroup analysis showed that the chylous effusion group mainly exhibited lesions of the thoracic duct, iliac lymphatic vessels, and lumbar trunks; the chylous effusion and lymphedema group displayed multi-region involvement with abnormal lower-extremity lymphatic reflux; and the lymphedema group had thoracic duct outlet obstruction and distal lymphatic occlusion in the affected limb. Conclusions:CTL demonstrates subtle morphological abnormalities of lymphatic vessels and provides precise anatomical localization. 99TCm-DX lymphoscintigraphy is useful for dynamically evaluating lymphatic drainage and detecting chylous effusion. The two modalities are complementary and provide comprehensive information for SLE complicated by chylous effusion and/or lymphedema. Distinct imaging patterns among the three subgroups may support clinical subtyping of this condition.

Abstract Background This study aimed to elucidate B cell subset pathology in COPD, a poorly characterized area, with a focus on its similarities to and differences from classical autoimmune disorders. Methods The single-cell RNA-sequencing (scRNA-seq) data from COPD and autoimmune diseases were obtained from Gene Expression Omnibus (GEO) for comparative analyses of B cell subsets and functions via differentially expressed genes (DEGs), KEGG, protein-protein interaction (PPI), and cell–cell communication analyses. Serum IgG4 was measured by ELISA and correlated with clinical parameters. The peripheral blood B cells were sorted by flow cytometry for single-cell B cell receptor (BCR) sequencing. A v-Abl/Bcl2 pro-B cell line was stimulated with cigarette smoke extract (CSE) to assess abnormal development in vitro . Results In lung tissue, IgG4 + plasma cells were enriched and expressed BCR activation/inflammatory genes and TNF/NF-κB/MAPK pathways. Serum IgG4 concentrations correlated negatively with pre-and post-bronchodilator FEV 1 /FVC. B cell interacted with monocytes, macrophages, fibroblasts and endothelial cells via IL-1β/IL-6, integrin and chemokine signalling, contributing to chronic inflammation and remodelling. In peripheral blood, transitional T1 B cells were increased, accompanied by λ-chain enrichment and increased IGLV1-47 usage, as well as enrichment of autoimmune pathways. In the bone marrow, the numbers of pre-B I cells were increased while those of small pre-B III cells were reduced, with altered expression of BCR development genes. CSE stimulation of the pro-B cell line reduced λ5 expression in a concentration-dependent manner. Conclusions The autoimmune abnormalities in COPD appear more restricted, although IgG4 antibody generation may contribute to immune-mediated lung damage.

Acorus tatarinowii, a traditional Chinese medicinal herb, is used clinically to treat neurological and respiratory disorders. Our previous work showed that α-asaronol (α-AOL), a low-toxicity natural constituent of A. tatarinowii, exerts robust antiepileptic effects through γ-aminobutyric acid (GABA)-ergic modulation. Given the critical role of the peripheral GABAergic system in asthma pathogenesis and inflammation regulation, the therapeutic potential of α-AOL for asthma warrants investigation. This study evaluated the efficacy, safety, and pulmonary targeting of both nebulized and oral α-AOL. Murine asthma models were established using ovalbumin (OVA) and house dust mite (HDM). Both nebulized α-AOL (5, 10, 20 mg/kg) and oral α-AOL were administered, with α-asarone, and albuterol as references. Histological, biochemical, and functional assessments were performed to evaluate lung inflammation, mucus secretion, cytokine levels (IL-4, IL-5, IL-13, TNF-α), airway smooth muscle (ASM) relaxation, and airway hyperresponsiveness (AHR). Pharmacokinetic and safety profiles were determined via HPLC tissue distribution analysis, human ether-à-go-go related gene (hERG) assays, and monitoring of respiratory and cardiovascular function. Nebulized α-AOL dose-dependently alleviated OVA/HDM-induced airway inflammation, characterized by reduced inflammatory cell infiltration, cytokine release, mucus hypersecretion, and AHR. It also induced concentration-dependent ASM relaxation and demonstrated dual therapeutic activity by both preventing and reversing methacholine (Mch)-induced acute bronchospasm. Nebulization provided sustained lung targeting (≥60 min retention). α-AOL exhibited negligible hERG inhibition (IC50 > 100 μM), no adverse respiratory effects, and no significant impact on blood pressure. In conclusion, nebulized and oral α-AOL possesses potent anti-inflammatory and bronchodilatory activities; the nebulized formulation shows favorable lung-targeted delivery and an excellent safety profile, positioning it as a promising inhaled candidate for asthma therapy, particularly in pediatric populations.

Pulmonary hypertension (PH) is a severe disease characterized by pulmonary vascular remodeling in which various immune cells play a critical role in vascular remodeling, although the details are still vague. Furthermore, current clinical treatments primarily focus on pulmonary vasodilation, but do not fundamentally address vascular remodeling itself. Here, first significant changes in neutrophils during the development of PH are demonstrated and show that neutrophil depletion can effectively attenuate disease progression. Moreover, the data show that neutrophil-derived S100A9 is the key mediator to promote vascular remodeling, while both knockout and inhibition of S100A9 can prevent PH. In a co-culture system of neutrophils and endothelial cells (ECs), hypoxic stimulation leads to increased S100A9 secretion by neutrophils, which activates the RAGE/PI3K/AKT pathway and causes dysfunction of ECs. These findings suggest that neutrophil-derived S100A9 mediated neutrophil-EC crosstalk plays an important role in pulmonary vascular remodeling, providing a promising strategy for treatment of PH.

ABSTRACT Cancer is more than just a collection of tumor cells. The complex tumor system, including the tumor immune microenvironment (TIME), is continually changing. Tumor cells are in constant communication with all stromal elements (e.g., fibroblasts, endothelial cells, and extracellular matrix) and immune effector cells (e.g., T cells, B cells, natural killer cells, dendritic cells, macrophages, and myeloid‐derived suppressor cells). Together, these intricate interactions among cell and molecular signaling pathways collectively drive tumor growth, tumor invasion, and metastasis and significantly affect the efficacy of cancer treatments. Recent investigations, from a tumor‐centric research paradigm to a complete evaluation of the local tumor microenvironment, have revealed the importance of the TIME. Although reviews in these fields typically focus on cellular/molecular breakdowns of the TIME and evasion of the immune system, a systematic study of its dynamic evolution is lacking. This review comprehensively discusses the major regulators and networks involved in the dynamic evolution of the TIME, the spatiotemporal dynamics of TIME components, metabolic reprogramming as an engine of TIME evolution, the targeting of metabolic regulators, and niches for TIME modulation, clinical and translational challenges, and future prospects. This information could help researchers explore the TIME and generate new therapeutic strategies.

To the Editor: Common respiratory diseases such as chronic obstructive pulmonary disease (COPD) and asthma induce significant health burdens worldwide.[1,2] Airway/lung microbiome and microbial metabolites have been proven to be closely related to multiple respiratory diseases.[3] However, few relevant studies show characteristics and horizontal comparisons of metabolites of dominant bacteria in respiratory diseases. Additionally, most results are based on association analyses of microbiome sequences, especially those of the gut microbiome, and untargeted metabolomics, without evidence that the metabolites are directly produced and secreted by certain bacteria.[4] In this context, our study aimed to analyze and compare the key metabolites and metabolic pathways of common bacteria involved in respiratory diseases. Based on the culture results of bacteria in clinical samples of patients with respiratory diseases and the reports of cohort studies,[5] we analyzed metabolic changes of five common bacteria, namely, Streptococcus pneumoniae (Sp), Staphylococcus aureus (Sa), Moraxella catarrhal (Mc), Pseudomonas aeruginosa (Pa), and Haemophilus influenzae (Hin). Firstly, we cultivated type strains of the five bacteria until the optical density at 600 nm (OD600) reached 1.0, collected their supernatants [Supplementary Materials, https://links.lww.com/CM9/C341], and then conducted untargeted metabolomics analysis of supernatants to investigate the metabolic profile under the same conditions, attempting to minimize the influence of variabilities and unknown factors. The metabolites with variable influence in projection (VIP) >1 and P <0.05 and fold change (FC) ≥2 or FC ≤0.5 were considered differentially abundant metabolites. Then, we compared the similarity and specificity of differentially abundant metabolites among these bacteria to identify possible key metabolites and related pathways, which might provide a shortcut for further exploration of the role of bacterial metabolites in respiratory diseases. Metabolites annotated by each secondary classification under the primary classification of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were shown in the positive and negative ion modes. In the positive ion mode, most metabolites were annotated to biological pathways such as metabolism. The top 5 secondary classifications included global and overview maps (100 metabolites), amino acid metabolism (47 metabolites), metabolism of cofactors and vitamins (20 metabolites), nucleotide metabolism (17 metabolites), and metabolism of other amino acids (16 metabolites), accounting for 81.97% (200/244) of the metabolites annotated to the metabolism pathway. In the negative ion mode, most metabolites were annotated to biological pathways such as metabolism again. The top 5 secondary classifications included global and overview maps (107 metabolites), amino acid metabolism (38 metabolites), carbohydrate metabolism (29 metabolites), nucleotide metabolism (27 metabolites), and lipid metabolism (22 metabolites), accounting for 76.90% (223/290) of the metabolites annotated to the metabolism pathway. The metabolites annotated by superclass in the human metabolome database (HMDB) in the positive ion mode were as follows: hydrocarbon derivatives (1 metabolite), alkaloids and derivatives (6 metabolites), organic nitrogen compounds (11 metabolites), phenylpropanoids and polyketides (14 metabolites), nucleosides, nucleotides, and analogues (20 metabolites), organic oxygen compounds (20 metabolites), benzenoids (34 metabolites), lipids and lipid-like molecules (58 metabolites), organoheterocyclic compounds (65 metabolites), and organic acids and derivatives (98 metabolites). In the negative ion mode, the metabolites annotated by the superclass in the HMDB were as follows: alkaloids and derivatives (3 metabolites), organic nitrogen compounds (5 metabolites), phenylpropanoids and polyketides (10 metabolites), benzenoids (19 metabolites), organic oxygen compounds (26 metabolites), nucleosides, nucleotides, and analogues (29 metabolites), organoheterocyclic compounds (32 metabolites), organic acids and derivatives (64 metabolites), lipids and lipid-like molecules (68 metabolites). The results of screening of differentially abundant metabolites revealed that between the supernatants of the Sp group and the control medium group, 432 differentially abundant metabolites were identified in the positive ion mode, including 176 upregulated and 256 downregulated metabolites. In the negative ion mode, 249 differentially abundant metabolites were identified, including 137 upregulated and 112 downregulated metabolites. Among the supernatants of the Sa group and the control medium group, 103 differentially abundant metabolites were identified in the positive ion mode, including 38 upregulated and 65 downregulated metabolites. In the negative ion mode, 56 differentially abundant metabolites were identified, including 16 upregulated and 40 downregulated metabolites. Between the supernatants of Mc group and the control medium group, 137 differentially abundant metabolites were identified in the positive ion mode, including 71 upregulated and 66 downregulated metabolites. In the negative ion mode, 76 differentially abundant metabolites were identified, including 33 upregulated and 43 downregulated metabolites. Among the supernatants of the Pa group and the control medium group, 407 differentially abundant metabolites were identified in the positive ion mode, including 178 upregulated and 229 downregulated metabolites. In the negative ion mode, 253 differentially abundant metabolites were identified, including 123 upregulated and 130 downregulated metabolites. Among the supernatants of the Hin group and the control medium group, 133 differentially abundant metabolites were identified in the positive ion mode, including 73 upregulated and 60 downregulated metabolites. In the negative ion mode, 81 differentially abundant metabolites were identified, including 31 upregulated and 50 downregulated metabolites. We then compared overlapping and unique differentially abundant metabolites among different bacteria [Figure 1]. In the positive ion mode, there were no common metabolites among the five kinds of bacteria. However, 114, 18, 33, 76, and 46 metabolites were found to be specific for Sp, Sa, Mc, Pa, and Hin, respectively. The top 5 upregulated metabolites in the supernatant of Sp were thioguanine, bicyclo prostaglandin E2, triacanthine, 4-hydroxyretinoic acid, and xanthurenic acid, whereas the top 5 downregulated metabolites were alanine-leucine (ala-leu), 3′-hydroxy-stanozolol, maltotetrose, 6-hydroxymelatonin, and S-adenosylhomocysteine. The top 5 upregulated metabolites in the supernatant of Sa were dihydroceramide (d16:0/16:0), 8-hydroxy-eicosapentaenoic acid, digalactosyl monoacylglycerol (18:2), tetramethylpyrazine, and norfentanyl, whereas the top 5 downregulated metabolites were ergothioneine, cafestol, trans-2-butene-1,4-dicarboxylic acid, 4-hydroxy estrone 1-N3-adenine, and prostaglandin A3. The top 5 upregulated metabolites in the supernatant of Mc were L-tyrosine methyl ester, urocanic acid, 6-aminonicotinamide, T-2 triol, and haloperidol-d4, whereas the top 5 downregulated metabolites were pantethine, ingenol-3-angelate, UR-144 N-(2-hydroxypentyl) metabolite, all-trans-retinal and gatifloxacin. The top 5 upregulated metabolites in the supernatant of the Pa were sterigmatocystin, astragaloside IV, 6β-naltrexol-d3, norbutorphanol, and milbemycin A4 oxime, whereas the top 5 downregulated metabolites were stearamide, N-acetyl-L-tyrosine, L-asparagine, L-adrenaline, and S-adenosyl-L-methionine. The top 5 upregulated metabolites in the supernatant of Hin belonged to phosphatidyl ethanolamine, including PE (12:0/12:0), PE (8:0/8:0), PE (9:0/9:0), PE (11:0/11:0) and PE (11:0/13:1), whereas the top 5 downregulated metabolites were spermidine, cytosine, cytidine 5′-monophosphate (hydrate), N4-acetylcytidine, and gedunin.Figure 1: Common and specific differentially abundant metabolites among supernatants of different bacteria in the positive (A) and negative (B) ion modes. BHI: Brain-heart infusion medium; Hin: Haemophilus influenzae; Mc: Moraxella catarrhal; Pa: Pseudomonas aeruginosa; Sa: Staphylococcus aureus; Sp: Streptococcus pneumoniae.In the negative ion mode, docosapentaenoic acid and 8,11,14-eicosatrienoic acid were two common metabolites among the five kinds of bacteria. In addition, there were 75, 5, 14, 53, and 37 metabolites specific for Sp, Sa, Mc, Pa, and Hin, respectively. The top 5 upregulated metabolites in the supernatant of Sp were deoxyribose 5-phosphate, d-mannose 6-phosphate, d-glucose 6-phosphate, chaetocin, and piceatannol, whereas the top 5 downregulated metabolites were stachyose, methylmalonate, pantothenic acid, serotonin, and xanthosine. The upregulated metabolites in the supernatant of Sa were N-lauroylsarcosine, thromboxane B1, inosine-5′-monophosphate and N-acetyl-L-histidine, whereas the only one downregulated metabolite was 5′-adenylic acid. The top 5 upregulated metabolites in the supernatant of Mc were guanosine 5′-diphosphate (GDP), 18-hydroxy-eicosapentaenoic acid, nicotinamide N-oxide, 19-nortestosterone and geranylgeranyl pyrophosphate, whereas the top downregulated metabolites were oleoyl ethanolamide, bilirubin, stearic acid and cymarin. The top 5 upregulated metabolites in the supernatant of Pa were 3-hydroxydecanoic acid, 12-hydroxydodecanoic acid, 3-hydroxypicolinic acid, cholic acid, and protoporphyrin IX, whereas the top 5 downregulated metabolites were adenylosuccinic acid, adenine, quinic acid, 3-(3-methoxyphenyl) propionic acid and 7-hydroxy-3,4-dihydrocarbostyril. The top 5 upregulated metabolites in the supernatant of Hin belonged to phosphatidyl ethanolamine class, including PE (12:0/12:0), PE (8:0/8:0), PE (9:0/9:0), PE (11:0/11:0), and PE (11:0/13:1), and top 5 downregulated metabolites were uridine monophosphate (UMP), guanosine-3′,5′-cyclic monophosphate, adenosine-3′,5′-cyclic monophosphate, biopterin and ribulose-5-phosphate. Metabolic pathway enrichment analysis revealed that in the positive ion mode, there were no significantly enriched metabolic pathways of differentially abundant metabolites in supernatants of Sp and Pa. The most significantly enriched pathways were purine metabolism in the supernatant of Sa and arachidonic acid metabolism in the supernatant of Mc. The significantly enriched pathways of the differentially abundant metabolites in the supernatant of Hin were purine metabolism and biosynthesis of unsaturated fatty acids, respectively. In the negative ion mode, there were no significantly enriched metabolic pathways of differentially abundant metabolites in the supernatants of Sp and Pa. The most enriched significant pathway was purine metabolism in the supernatant of Sa. The most enriched pathways in the supernatant of Mc were biosynthesis of unsaturated fatty acids, nicotinate and nicotinamide metabolism, fatty acid biosynthesis, and carbon fixation in photosynthetic organisms, respectively. The significantly enriched pathways of differentially abundant metabolites in the supernatant of Hin were also identified, including pathways such as alanine, aspartate and glutamate metabolism, the citrate cycle, carbon fixation pathways in prokaryotes, β-alanine metabolism, furfural degradation, nicotinate and nicotinamide metabolism, histidine metabolism, amino sugar and nucleotide sugar metabolism, glyoxylate and dicarboxylate metabolism, and the biosynthesis of unsaturated fatty acids. In summary, we conducted untargeted metabolomics on common bacteria detected clinically in respiratory diseases and described the features of their metabolic profiles. We also screened a series of common and specific differentially abundant metabolites of each bacterium. We hope these data will provide the basis for further exploring the role of microbiome metabolites in the pathogenesis, diagnosis, and prevention of respiratory diseases. Acknowledgments We thank Robin James Storer, PhD, from Liwen Bianji (Edanz) (www.liwenbianji.cn) for editing the English text of a draft of this manuscript. Funding This study was supported by grants from the National Natural Science Foundation of China (Nos. 82090013, 82071805, and 82241011) and Beijing Natural Science Foundation (Nos. 7212001, 7242010, and 7242002). Conflicts of interest None.

Group 2 innate lymphoid cells (ILC2s) directly contribute to local inflammation in type 2 inflammatory airway diseases. Here, we identify ILC2 subsets by single cell RNA sequencing in chronic rhinosinusitis with nasal polyps (CRSwNP) and in a memory inflammatory mouse model. We find that toll-like receptor 4 (TLR4)+ILC2s, with similar markers to their human counterparts, expresse memory cell markers, persist over time, and respond more vigorously to a secondary unrelated antigen challenge in the mouse model. Genetic ablation of TLR4 or blockade by anti-TLR4 antibodies leads to the reduction of IL-13 expression from ILC2s and mucus production in mice. The assay for transposase-accessible chromatin sequencing further confirms the importance of accessible TLR4 gene loci and its down-stream signaling pathway in maintaining trained immunity of TLR4+ILC2s after repeated stimulation by HDM. Taken together, TLR4 has a function in trained immunity maintenance within ILC2s, which may contribute to disease chronicity through a non-specific immunological memory.

Background: Primary thoracic lymphangioma is a rare disease. Most of the previous studies are comprised of individual case reports, with a very limited number of patients included. Objective: This study aims to investigate the chest computed tomography (CT) imaging features and clinical manifestations of thoracic lymphangioma, thereby enhancing our understanding of the condition. Methods: A retrospective analysis was conducted on 62 patients diagnosed with thoracic lymphangioma, comprising 32 males and 30 females. The study focused on analyzing the chest CT imaging features and the clinical manifestations observed in these patients. Results: The incidence rates of thoracic lymphangioma did not differ significantly between males and females; however, it was more frequently observed in children and adolescents. The most common clinical symptoms included cough, fever, chylothorax, chylous pericardium, and lymphedema. The mediastinum (82.3%) emerged as the most frequent location for thoracic lymphangioma, followed by the chest wall (62.9%), bone (40.3%), and pleura (32.3%). Pulmonary lymphangioma, the least prevalent subtype (19.4%), exhibited a propensity to induce respiratory symptoms, frequently manifesting as a generalized lymphatic anomaly (GLA). Furthermore, elevated levels of D-dimer were detected in 34 patients (54.8%) with thoracic lymphangioma. Conclusions: Imaging examinations play a crucial role in assisting clinicians in making more accurate early diagnoses of thoracic lymphangioma. They are also helpful for assessing the extent of systemic infiltration and enhancing diagnostic precision. With radiological assessment, clinicians could more readily select appropriate therapeutic treatments and monitor the progression of follow-up care.

B-1 cells are derived from a subpopulation of B lymphocytes which have a specific developmental process, unique phenotype and location, and distinct functions in comparison with conventional B-2 cells. The origin of B-1 cells is not completely clear, with two existing hypotheses concerning their lineage and differentiation pathways. B-1 cells are located principally in the peritoneal and pleural cavities, but are also distributed in secondary lymphoid tissues, at mucosal sites and in the blood and bone marrow. B-1 cells regulate immune responses and maintain homeostasis by secretion of natural antibodies (nAbs), and participate in the adaptive immune response through phagocytosis and presentation of antigens to T cells. B-1 cells are associated with many diseases including autoimmune, infectious and inflammatory diseases. This review focuses on the origin and biological functions of B-1 cells as well as their involvement in human disease, and discusses advances in the understanding of the heterogeneity of B-1 cells under specific pathophysiological features, as partly clarified by single-cell sequencing analysis.

The chronic autoimmune disease multiple sclerosis (MS) now remains incurable. Paeoniflorin (PF), which is a monoterpene glucoside obtained from Paeonia lactiflora Pall, is recognized for neuroprotective and anti-inflammatory properties. However, the precise mechanism by which PF regulates MS is unclear. This work aims to elucidate the underlying mechanisms of PF in EAE, a well established animal model of MS, and to discover the target proteins that PF directly acts on. Our results revealed that PF administration can significantly attenuate the clinical symptoms of EAE and alleviate the central nervous system (CNS) inflammatory environment by inhibiting M1-type microglia/macrophages. Mechanistically, PF was found to directly interact with the glycolytic enzyme α-enolase (ENO1), inhibiting its enzymatic activity and expression to impair glucose metabolism, thereby suppressing microglia/macrophage M1 polarization and ameliorating CNS inflammation. Significantly, Eno1 knockdown in microglia/macrophages diminished their pro-inflammatory phenotype, while treatment with ENOBlock or the specific knockout of Eno1 in microglia led to EAE remission, underscoring the critical role of ENO1 in EAE progression. This study uncovers the molecular mechanism of PF in treating EAE, linking the anti-inflammatory property of PF to the glucose metabolism process, which will broaden the prospective applications of PF.

Myocardial fibrosis (MF) is a common pathological hallmark of cardiovascular diseases, reflecting shared mechanisms in their progression. However, the lack of reliable MF models that accurately mimic its pathogenesis has hindered drug discovery, highlighting the urgent need for more effective therapeutic agents. Herein, a novel contractile three-dimensional (3D) myocardial tissue model integrating cardiomyocytes, cardiac-fibroblasts, and bone marrow-derived macrophages in collagen hydrogel was developed to simulate the fibrotic changes of cardiovascular disease, and facilitate the screening of anti-MF compounds. The 3D myocardial tissue model exhibited precise, visualizable, and quantifiable contractile characteristics under hypoxia and drug interventions. 76 compounds extracted from the resins of Toxicodendron vernicifluum, a traditional Chinese medicine with clear clinical benefits for fibrotic diseases, were screened for anti-fibrotic activity. Using an in vitro 3D oxygen-glucose deprivation (OGD)-treated myocardial tissue model instead of a two-dimensional transforming growth factor-β treated cardiac-fibroblasts model, two candidates including LQ-40 and SQ-3 exert impressive anti-MF activity, which was further validated in left anterior descending coronary artery ligation-induced MF mouse model. The current results demonstrate the feasibility and advantage of the novel contractile 3D tissue model with multi-cell types in discovering candidates for MF, further stressing the great potential of regulating macrophages in the treatment of MF.

The adenoids and tonsils are important immune organs of the nasopharynx that often become hypertrophic in childhood because of recurrent pathogen infection. However, the differences in the immune microenvironment of adenoid hypertrophy (AH) and tonsil hypertrophy (TH) are unclear. Here, we show the epidemiological characteristics and peripheral blood cell indices of 1209 pediatric patients (1–15 years old) diagnosed with AH, and find that AH is often accompanied by TH and characterized by specific changes in immune cell types. Single-cell RNA sequencing analysis show that 12 paired AH and TH samples contain large numbers of B, T cells and some exhausted effector memory CD4 + T cells. Compared with matched TH, AH have more naïve B cells and regulatory CD4 + T cells and less plasma B cells. Weaker antigen presentation and more significant immunosuppression are also observed in AH. In contrast, the number and cytotoxicity of cytotoxic CD8 + T cells decrease with AH grade. These findings will help our understanding of the immune response to nasopharyngeal infection.

Background:Type 2 (T2) inflammatory respiratory diseases encompass a range of conditions characterized by inflammation affecting the airways and lung parenchyma, with their pathogenesis rooted in T2 inflammation. Biological treatments that mitigate T2 inflammation revolutionize the therapeutic landscape for these respiratory diseases. However, there are decision-making difficulties in terms of the target population, timing of initiation, and type selection for biological targeted therapy. Methods:Search strategies were focused on relevant issues related to T2 inflammatory respiratory diseases from PubMed with search date from 2014 to 2024. The quality of evidence and grading recommendations were assessed with the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system. Consensus was achieved through two rounds of anonymous voting with a strong recommendation demanding at least 70% approval from the participants. Results:A total of 370 basic research results and clinical evidence-based medical data were collected and reviewed. The latest research advances, clinical evidence, and expert insights relating to the use of biological treatments aiming at T2 inflammation in respiratory diseases and their co-morbidities were discussed rigorously and iteratively by an expert panel, and a consensus report with recommendations is presented. Conclusions:This consensus outlines the pathogenesis, assessment of T2 inflammation, biological therapies targeted at T2 inflammation, and management strategies for T2 inflammatory respiratory diseases and their comorbidities. It will serve as a valuable guide for clinicians in China, empowering them to diagnose and manage these conditions more effectively.

Chyloptysis indicates the presence of a lymphangio-bronchial fistula. This study compared the chest CT imaging features and clinical manifestations between CLA (complex lymphatic anomalies) patients with and without chyloptysis. A retrospective analysis was performed on 291 thoracic CLA patients (142 males, 149 females) admitted from December 2008 to February 2024. Divided into chyloptysis (152) and non—chyloptysis (139) groups, we compared their clinical data and chest CT features to find differences related to chyloptysis in CLA patients. The chyloptysis group had a significantly higher incidence of chyle pericardium, as well as more cystic lesions at the right lymphatic duct, and of the thoracic duct itself. In contrast, the non-chyloptysis group showed more cystic lesions of the liver, spleen, abdominal cavity, neck, and skeleton. All these differences were statistically significant (P < 0.05).The incidence of lymphedema in the non-chyloptysis group was higher than that in the chyloptysis group (P < 0.05), among which there was a significant statistical correlation between lymphedema of the lower limbs, lymphedema of the scrotum, lymphedema of the face and lymphedema of the upper limbs and chyloptysis (P < 0.05). The severity of chylopulmonary disease was closely related to the appearance of chyloptysis (P < 0.001). In terms of the characteristics of chest CT findings, the incidence of GGO, consolidation, grid shadow, vacuolar sign, nodular pleural thickening, solid nodules, perifascicular interstitial thickening, hilar opacity or swelling, and parapericardial effusion were higher in the chyloptysis group than in the non-chyloptysis group (P < 0.05). The analysis of these differences is of great clinical significance for understanding the distribution of chyloptysis in CLA patients with different CT signs, and it is convenient for clinicians to strengthen the risk prediction and management of chyloptysis in CLA patients.

Respiratory syncytial virus (RSV) infection has been associated with disruption of the airway epithelial barrier, potentially increasing the risk of asthma development. However, whether and how RSV and RSV-induced IL-33 contribute to this process are still unclear. In vivo, 7-day-old C57BL/6 mice were infected perinasally with RSV, then viral replication, lung inflammation and barrier integrity were evaluated at various time points postinfection. In vitro, human epithelial cells were infected with RSV in the presence or absence of IL-33, and the expression and localization of apical junction complex proteins (AJC) were assessed by western blot analysis and immunofluorescence staining. The involvement of components of the IL-33/ST2/MyD88 axis was further verified through blockade of endogenous IL-33 signaling and pharmacological inhibition of MyD88. Exposure to RSV infection resulted in impairment of the airway epithelial barrier, as indicated by reduced expression of tight junction proteins (ZO-1, Occludin) and adherents junction protein (E-cadherin) in the lung tissues. These effects on epithelial barrier disruption were significantly attenuated in St2-/- mice compared with wild-type controls. In vitro, the RSV-induced epithelial barrier disruption was exacerbated by topical application of exogenous IL-33, partially through activation of MyD88-mediated NF-κB signaling. Notably, knockdown of St2 by siRNA transfection or pharmacological inhibition of MyD88 partially restored the expression of E-cadherin, ZO-1 and Occludin in RSV-infected epithelial cells. RSV infection triggers robust IL-33 release from airway epithelial cells, leading to disrupted expression of AJC protein via activation of the MyD88-dependent NF-κB signaling pathway. These findings highlight the IL-33/ST2/MyD88 axis as a critical mediator of epithelial barrier dysfunction, which may represent a potential target for therapeutic intervention in RSV-mediated lung diseases.

Oral fungal microbiota plays an important role in many diseases, however, the role of oral fungal microorganisms in the development of patients infected with Omicron has not been reported. A total of 963 tongue coating samples were prospectively included in this study, and finally 336 samples from patients infected Omicron variant (PIOV), 234 samples from recovered patients infected with Omicron (RP), 71 samples from patients infected original strain of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) (PIOS), 299 samples from healthy controls (HC) completed internal transcribed spacer (ITS) sequencing after screening and quality control. By comparing the difference of oral fungal microorganisms between PIOV, RP and HC, we found that with the recovery of PIOV, their oral fungal microecological diversity increased gradually. Besides, at the species level, there were 24 oral fungal species such as Zanclospora_jonesii increased gradually, while there were 24 oral fungal species such as Saccharomyces_cerevisiae decreased gradually. In addition, by comparing PIOS and PIOV, we found that the alpha diversity of oral fungal microorganisms in PIOV was significantly lower than PIOS and the main species of the two groups were different. At the same time, we randomly divided PIOV and HC into training and validation set. Based on random forest model and five-fold cross-validation, we identified three optimal microbial markers of oral fungi and constructed a diagnostic model of PIOV. The area under the curve (AUC) value of PIOV group was 99.01% in discovery phase and 97.84% in verification phase. In summary, based on large-scale samples, this study is the first to elucidate the characteristics of oral fungal microbiota changes during PIOV recovery and establish a supplemental non-invasive diagnostic model for PIOV based on the oral fungal microbiome.