The role of folic acid in male reproductive health remains incompletely understood, with existing findings presenting conflicting results. Reproductive tract inflammation, a key factor influencing semen quality, may modulate the effects of folic acid on semen parameters. However, this hypothesis remains unverified in a systematic manner. This retrospective study enrolled 98 men. Red blood cell folic acid levels, seminal plasma elastase, and semen parameters were measured. Polynomial regression models were employed to analyze nonlinear relationships and interaction effects, while machine learning methods were applied for model validation. The research found that the impact of red blood cell folate on semen quality is parameter-specific: it shows a significant U-shaped relationship with the sperm DNA fragmentation index (quadratic term β = 2.560 × 10⁻⁴, p = 0.015), and this relationship is significantly modulated by the inflammatory state (interaction term β = 2.825 × 10⁻⁷, p = 0.035), while it shows a stable linear positive correlation with the sperm forward motility rate (β = 0.024, p = 0.015), and is affected by age. Machine learning analysis further verified the core position of folate and inflammatory indicators in predicting semen quality. The effects of folic acid on semen quality are multifaceted. Its U-shaped influence on sperm DNA integrity is finely modulated by the inflammatory microenvironment, while simultaneously enhancing sperm motility. These findings suggest that clinical folic acid intervention strategies should consider the distinct responses of specific semen parameters and individual inflammatory states, offering novel insights into the precise prevention and treatment of male infertility.

BACKGROUND:Paclitaxel produces a chronic, glove-stocking neuropathy that is frequently accompanied by anxiety and depression. However, the neural circuits and molecular signals driving this pain-affect comorbidity remain unclear. METHODS:Chemotherapy-induced neuropathic pain (CINP) was established by paclitaxel injections in male rats. Mechanical and thermal sensitivity and negative emotional behaviors were assessed using behavioral tests. Parabrachial nucleus (PBN)-central amygdala (CeA) projections were traced with viral vectors, and optogenetics was used to manipulate PBN terminals. Prodynorphin (PDYN) and κ-opioid receptor (KOR) mRNA levels were quantified by reverse transcription quantitative polymerase chain reaction (RT-qPCR). RESULTS:CINP rats developed persistent pain hypersensitivity and negative emotional behaviors, accompanied by increased CeA c-Fos and PDYN expression without changes in KOR mRNA. Intra-CeA administration of the KOR antagonist Navacaprant alleviated both pain and negative emotional behaviors. Optogenetic inhibition of the PBN-CeA pathway reduced CeA activation, attenuated pain hypersensitivity, and improved negative emotional behaviors, whereas pathway activation elicited both changes in naïve rats. Chronic inhibition of this pathway lowered CeA PDYN mRNA levels in CINP rats. CONCLUSION:The PBN-CeA pathway contributes to paclitaxel-induced neuropathic pain and anxiety-like behavior, and is associated with increased dynorphin expression in the CeA. Targeting this pathway or its downstream KOR may provide a therapeutic strategy for chemotherapy-related sensory and affective disturbances.

Oxytocin has been found to modulate and improve pain in humans, but the mechanisms underlying these antinociceptive properties, especially in visceral hypersensitivity, are still unclear. Irritable bowel syndrome (IBS) models were established by colorectal distention in newborn rats aged 8 to 14 days, and visceral hypersensitivity was assessed using electromyogram (EMG). Oxytocin or saclofen was administered intrathecally to evaluate visceral hypersensitivity in the rats. The protein expressions of oxytocin receptor (OTR), γ-aminobutyric acid type B1 receptor (GABAB1), and transient receptor potential vanilloid 1 (TRPV1) in the lumbosacral spinal cord regions were measured. IBS rats exhibited a unique spinal cord molecular signature comprising decreased OTR/GABAB1 and increased TRPV1 expression. Intrathecal oxytocin treatment not only normalized these molecular alterations (increasing GABAB1 while decreasing TRPV1) but also ameliorated visceral pain behaviors. Crucially, this therapeutic effect was fully reversed by GABAB1 inhibition, establishing the necessity of intact GABAergic signaling for oxytocin-mediated analgesia. Collectively, these findings indicate that oxytocin relieves visceral hypersensitivity through the regulation of GABAB1 and TRPV1 in the spinal cord of IBS rats.

QuestionIs anti-programmed cell death 1 therapy effective and safe as first-line therapy in patients with advanced melanoma predominantly of acral subtype?FindingsIn this phase 3 randomized clinical trial of 256 patients with advanced melanoma (160 acral subtype [62.7%]), toripalimab as first-line therapy significantly improved blinded independent central review-assessed progression-free survival compared with dacarbazine with an acceptable safety profile.MeaningToripalimab could be used as a new first-line treatment option for advanced melanoma, particularly where acral cutaneous melanoma is the predominant subtype. ImportanceProgrammed cell death 1 (PD-1) inhibitors have been the standard first-line treatment for advanced melanoma; however, their clinical benefit in advanced melanoma predominantly of acral subtype remains unclear.ObjectiveTo evaluate the efficacy and safety of toripalimab (a PD-1 inhibitor) vs dacarbazine as the first-line treatment in advanced melanoma predominantly of acral subtype.Design, Setting, and ParticipantsThis multicenter, open-label, positive-control phase 3 randomized clinical trial enrolled patients with advanced melanoma from January 22, 2018, to July 12, 2023. Eligible patients had histologically confirmed stage III or IV melanoma and no prior systemic therapy for advanced melanoma. Data were analyzed from September 23 to November 27, 2023.InterventionPatients were randomized (1:1) to receive toripalimab, 240 mg, every 2 weeks for up to 2 years, or dacarbazine, 1000 mg/m2, every 3 weeks until disease progression or intolerable toxic effects. Patients in the dacarbazine group were allowed to receive toripalimab after radiographic disease progression.Main Outcomes and MeasuresProgression-free survival (PFS) assessed by blinded independent central review (BICR) per Response Evaluation Criteria in Solid Tumors, version 1.1.ResultsThe analysis included 256 patients (median [range] age, 58 [18-85] years; 113 females [44.3%] and 142 males [55.7%]), among whom 160 participants (62.7%) had acral melanoma; 127 were randomized to receive toripalimab and 128 to dacarbazine (1 patient was excluded for Good Clinical Practice violation). The median (range) follow-up period was 11.8 (0.1-62.5) months. Toripalimab significantly reduced the risk of disease progression or death by 29.2% (hazard ratio, 0.71; 95% CI, 0.53-0.95; P = .02) compared with dacarbazine as assessed by BICR . Consistent PFS benefit was observed in most predefined subgroups, including patients with acral subtype. The BICR-assessed objective response rate of 11.0% (95% CI, 6.2%-17.8%) vs 8.6% (95% CI, 4.4%-14.9%), and the median duration of response of 13.8 (95% CI, 5.9-not evaluable) months vs 6.9 (95% CI, 3.8-not evaluable) months, respectively, also favored toripalimab over dacarbazine. Grade 3 or worse treatment-related adverse events occurred in 36 patients (28.3%) receiving toripalimab, with the most common (>= 3%) being increased lipase (11 patients [8.7%]), anemia (5 patients [3.9%]), increased gamma-glutamyltransferase (4 patients [3.1%]), hyponatremia (4 patients [3.1%]), and increased blood triglycerides (4 patients [3.1%]).Conclusions and RelevanceThis phase 3 randomized clinical trial found that as a first-line treatment for advanced melanoma predominantly of acral subtype, toripalimab showed a significant PFS benefit over dacarbazine and an acceptable safety profile.Trial RegistrationClinicalTrials.gov Identifier: NCT03430297 This randomized clinical trial evaluates the efficacy and safety of toripalimab vs dacarbazine as the first-line treatment in advanced melanoma predominantly of acral subtype.

SHR-A1811, an antibody‒drug conjugate consisting of the anti-HER2 antibody trastuzumab conjugated via a cleavable linker to a topoisomerase I inhibitor payload, demonstrated substantial antitumor activity in patients with heavily treated HER2-expressing or mutated advanced solid tumors. The main analysis was reported, and this is a long-term follow-up of the HORIZON-X trial (NCT04446260). This global, multicenter, first-in-human, phase 1 trial enrolled patients aged ≥ 18 years with unresectable, advanced, or metastatic HER2-expressing or mutated solid tumors refractory or intolerant to standard therapies across 38 hospitals. SHR-A1811 was administered intravenously at doses ranging from 1.0 to 8.0 mg/kg every three weeks. The primary endpoints included dose-limiting toxicity, safety, and the recommended phase 2 dose. From September 7, 2020, to June 4, 2024, 396 patients with a median of three prior treatment regimens (IQR 2-5) received SHR-A1811. As of March 12, 2025, the median follow-up was 17.1 months for HER2-positive breast cancer, 10.6 months for HER2-low expressing breast cancer, and 4.3 to 8.2 months in non-breast cancers. The safety profile remained consistent with that of previous reports. Grade 3 or higher treatment-related adverse events occurred in 261 patients (65.9%), and any grade interstitial lung disease was observed in 10 patients (2.5%). The median progression-free survival was 25.0 months (95% CI 17.2-33.6) for HER2-positive breast cancer, 11.0 months (95% CI 8.2-13.8) for HER2-low expressing breast cancer, and 3.5 to 17.2 months for non-breast tumors. This final analysis further confirmed the long-term efficacy and favorable safety profile of SHR-A1811 among heavily prior-treated advanced solid tumors, reinforcing its potential as an effective HER2-targeted therapy.

BackgroundGastric cancer (GC) is often associated with a poor prognosis, and the precise molecular mechanisms driving its pathogenesis are not yet fully characterized. Stress granules (SGs) are now understood to play a crucial role in tumor progression, yet the prognostic value of SG-related markers in GC remains unclear. This study aimed to identify SG-related prognostic genes, clarify their clinical and biological significance in GC, and validate their potential as predictive indicators for patient overall survival (OS).MethodsSingle-cell and transcriptomic data for gastric cancer, along with genes related to stress granules (SGRGs), were acquired from public databases and literature. Candidate genes were identified by intersecting differentially expressed genes (DEGs) with SGRGs. Prognostic genes were identified through univariate Cox regression, and a risk score model was constructed. The model’s performance was validated in an independent cohort. Based on risk stratification, functional enrichment analysis, immune cell infiltration pattern assessment, and chemotherapy drug sensitivity analysis were conducted. Cell types expressing the prognostic genes were identified using single-cell RNA sequencing (scRNA-seq), and the related key cell clusters were identified.ResultsSERPINE1, CD36, MMRN1, and GRP were identified as prognostic genes. The risk model demonstrated good performance in predicting the survival status of GC patients. GSEA revealed that significantly enriched pathways included neuroactive ligand-receptor interaction and extracellular matrix (ECM)-receptor interaction pathways. CD36, MMRN1, and SERPINE1 demonstrated significant positive correlations with mast cells (correlation coefficients (r) > 0.3, P < 0.001). Chemotherapy drugs exhibited greater efficacy in high-risk GC patients. Moreover, endothelial cells were considered key cells and played a critical role in GC. Finally, SERPINE1 expression was associated with clinical features and prognosis in GC.ConclusionIn summary, we identified a four-gene SG-related signature strongly associated with prognosis in GC and constructed a predictive model with clinical potential. Our integrated analysis identified endothelial cells as a candidate population linked to the expression of these genes. These findings provide associative evidence linking SGs to GC outcomes and highlight potential targets for future mechanistic and therapeutic exploration.

Importance:Programmed cell death 1 (PD-1) inhibitors have been the standard first-line treatment for advanced melanoma; however, their clinical benefit in advanced melanoma predominantly of acral subtype remains unclear. Objective:To evaluate the efficacy and safety of toripalimab (a PD-1 inhibitor) vs dacarbazine as the first-line treatment in advanced melanoma predominantly of acral subtype. Design, Setting, and Participants:This multicenter, open-label, positive-control phase 3 randomized clinical trial enrolled patients with advanced melanoma from January 22, 2018, to July 12, 2023. Eligible patients had histologically confirmed stage III or IV melanoma and no prior systemic therapy for advanced melanoma. Data were analyzed from September 23 to November 27, 2023. Intervention:Patients were randomized (1:1) to receive toripalimab, 240 mg, every 2 weeks for up to 2 years, or dacarbazine, 1000 mg/m2, every 3 weeks until disease progression or intolerable toxic effects. Patients in the dacarbazine group were allowed to receive toripalimab after radiographic disease progression. Main Outcomes and Measures:Progression-free survival (PFS) assessed by blinded independent central review (BICR) per Response Evaluation Criteria in Solid Tumors, version 1.1. Results:The analysis included 256 patients (median [range] age, 58 [18-85] years; 113 females [44.3%] and 142 males [55.7%]), among whom 160 participants (62.7%) had acral melanoma; 127 were randomized to receive toripalimab and 128 to dacarbazine (1 patient was excluded for Good Clinical Practice violation). The median (range) follow-up period was 11.8 (0.1-62.5) months. Toripalimab significantly reduced the risk of disease progression or death by 29.2% (hazard ratio, 0.71; 95% CI, 0.53-0.95; P = .02) compared with dacarbazine as assessed by BICR . Consistent PFS benefit was observed in most predefined subgroups, including patients with acral subtype. The BICR-assessed objective response rate of 11.0% (95% CI, 6.2%-17.8%) vs 8.6% (95% CI, 4.4%-14.9%), and the median duration of response of 13.8 (95% CI, 5.9-not evaluable) months vs 6.9 (95% CI, 3.8-not evaluable) months, respectively, also favored toripalimab over dacarbazine. Grade 3 or worse treatment-related adverse events occurred in 36 patients (28.3%) receiving toripalimab, with the most common (≥3%) being increased lipase (11 patients [8.7%]), anemia (5 patients [3.9%]), increased γ-glutamyltransferase (4 patients [3.1%]), hyponatremia (4 patients [3.1%]), and increased blood triglycerides (4 patients [3.1%]). Conclusions and Relevance:This phase 3 randomized clinical trial found that as a first-line treatment for advanced melanoma predominantly of acral subtype, toripalimab showed a significant PFS benefit over dacarbazine and an acceptable safety profile. Trial Registration:ClinicalTrials.gov Identifier: NCT03430297.

Ulcerative colitis (UC) is characterized by the dysregulation of inflammatory pathways, leading to extensive tissue damage and chronic inflammation. NLRP3 inflammasome promotes inflammatory cytokine maturation and pyroptosis. Meteorin-like protein (Metrnl) has anti-inflammatory effects; however, its specific mechanisms in regulating these pathways remain unclear. This study investigated the effect of Metrnl on UC via regulation of the NLRP3 inflammasome through regulation of its key component, apoptosis-associated speck-like protein containing CARD (ASC). NCM460 and FHC cells were treated with TNF-α to establish a cell validation model. Cells with Metrnl interference or overexpression were intervened with three inflammasome activators (ATP, nigericin, or alum), and a caspase-1 inhibitor (belnacasan). The inflammatory factor levels, ASC mRNA and protein expressions, pyroptosis-related proteins, and inflammasome were detected. Interactions between ASC, Metrnl and inflammasome were determined by protein immunoprecipitation. UC models were constructed using wild-type and Metrnl knockout mice. Histopathological changes and differences in pyroptosis-related protein and inflammasome expressions were examined. Metrnl expression was significantly decreased in colitis, and the inflammatory factors IL-1β and LDH expressions increased at the cellular and animal levels, promoting ASC expression and pyroptosis, and activating the inflammasome. Metrnl interference accelerated inflammation and pyroptosis, which were reversed by Metrnl overexpression. Interactive relationships between Metrnl and ASC were noted. Metrnl may mediate ASC protein expression, and reduce IL-1β expression and pyroptosis in colon epithelial cells through the inhibition of ASC-NLRP3 binding. In vivo studies with DSS-induced colitis models confirmed the significance of these findings. After Metrnl knockout, the colon of normal mice was shorter, and Metrnl-/- UC mice showed more severe colon tissue damage, reduced goblet cells, increased inflammation, and accumulation of pyroptosis protein expression, indicating that Metrnl knockout increases susceptibility to UC. Metrnl interferes with key inflammatory signaling pathways in UC by inhibiting the interaction between ASC and NLRP3, suggesting its potential in UC treatment.

Tropomyosin receptor kinases (TRK) tyrosine kinase inhibitors (TKIs) have demonstrated marked efficacy in neurotrophic receptor tyrosine kinase (NTRK) fusion positive tumors. However, resistance inevitably develops. Eratrectinib (VC004) is a next-generation selective TRK TKI which can overcome drug resistance. In this phase 1 study, patients with locally advanced or metastatic solid tumors were enrolled in the dose-escalation part. A standard 3 + 3 design was adopted to sequentially assign patients at 25 mg twice daily (BID), 50 mg BID, 100 mg BID, and 200 mg BID dose levels of oral eratrectinib. After each dose had been established, we expanded specific dose cohorts in patients with NTRK fusion positive locally advanced or metastatic solid tumors to obtain adequate data for safety, pharmacokinetics (PK), and efficacy. The primary endpoints of this study were safety, maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D). Between December 4, 2020 and November 4, 2021, 16 patients were enrolled in the dose-escalation part and a dose of 100 mg BID was identified as MTD. Between May 8, 2021 and March 19, 2025, 75 patients were enrolled in the dose-expansion part at doses of eratrectinib 25 mg BID, 50 mg BID, and 75 mg BID. Treatment-emergent adverse events (TEAEs) occurred in 94.7

BackgroundInflammatory bowel disease (IBD) is a complex chronic intestinal inflammatory disorder. Oxidative stress (OS) is crucial in the pathogenesis of IBD by promoting inflammation and disrupting the intestinal epithelial barrier.MethodsWeighted gene co-expression network analysis and machine learning were applied to publicly available gene expression data to identify OS-related biomarkers for IBD. Receiver operating characteristic curve, immune infiltration analysis, and single-cell analysis were used to evaluate diagnostic performance and cellular localization. RT-qPCR and Western blotting were conducted to validate gene expression.ResultsThree hub genes (LCN2, APP, and TNFSF4) demonstrated strong diagnostic accuracy (AUC: 0.781, 0.805, and 0.754, respectively) in the discovery cohort (GSE3365). Furthermore, external validation in three independent cohorts also confirmed the robust diagnostic performance of the three hub genes. These genes were enriched in OS and inflammatory pathways, strongly correlated with infiltration of macrophages M0, monocytes, and neutrophils (p<0.01), and were highly expressed in goblet cells, stem cells, and T cells. DSS model validation confirmed significant upregulation at mRNA and protein levels.ConclusionThese OS-associated genes represent novel molecular targets for precise diagnosis and personalized treatment of IBD. This finding highlights the interplay among OS, immune dysregulation, and inflammation in IBD pathogenesis.

2527 Background: QLF31907, a bispecific antibody that simultaneously block PD-1/L1 immunosuppressive pathway on cancer cells and conditionally activate 4-1BB co-stimulatory pathway on tumor-specific T cells, was designed to restrict 4-1BB agonism to the tumor microenvironment, which might overcome resistance to PD-(L)1 inhibitor and reduce hepatoxicity as traditional 4-1BB monoclonal antibodies reported. Although immunotherapy (IO) has revolutionized the treatment of melanoma, a significant proportion of patients (pts), particularly those with mucosal and acral subtypes, either fail to respond initially or experience disease relapse after treatment. Here, we present results of QLF31907 in previously-treated pts with advanced melanoma, including IO-exposed. Methods: This phase 2 trial was comprised of safety observation stage and efficacy expansion stage. Pts with unresectable locally advanced or metastatic melanoma who failed, were intolerable to, or refused standard treatment were recruited and administered QLF31907 via intravenous infusion from 5 mg/kg to 20 mg/kg every 2 weeks (Q2W) or 3 weeks (Q3W). The primary endpoints were dose-limiting toxicity (DLT) and safety in safety observation stage, and was objective response rate (ORR) per RECIST v1.1 assessed by investigator in efficacy expansion stage. Results: As of Dec 31, 2025, 59 pts were enrolled (median age: 57.0 years; male: 47.5%; ECOG PS of 1: 42.4%; stage IV: 93.2%). The mucosal subtype accounted for the most (40.7%), followed by acral (33.9%), cutaneous (non-acral; 18.6%) and primary unknown (6.8%). Median prior lines of therapies were 2.0 (range, 1–5). Fifty-five (93.2%) pts received prior immunotherapy, including 50 (87.7%) pts received prior anti-PD-1/PD-L1 agents. No DLT occurred. Grade ≥3 treatment-emergent adverse events (TEAEs) occurred in 34 (57.6%) pts. The most common grade ≥3 TEAEs (≥10%) were liver injury (18.6%), neutrophil count decreased (15.3%), anemia (13.6%), white blood cell count decreased (11.9%). In 57 efficacy-evaluable pts, seven had partial response. The ORR and disease control rate (DCR) was 12.3% (95% confidence interval [CI], 5.1%-23.7%) and 56.1% (95% CI, 42.4%-69.3%), respectively. The median progression-free survival, duration of response, and overall survival was 2.6 months (95% CI, 1.6-3.7), 5.8 months (95% CI, 2.3-not evaluable [NE]), and 15.3 months (95% CI, 11.6-NE), respectively. Conclusions: QLF31907 showed potential anti-tumor activity and acceptable safety profile in heavily-treated pts with advanced melanoma, including IO-exposed pts. These results warrant validation in further clinical trials. Clinical trial information: NCT05823246 .

Cutaneous melanoma (CM) is an aggressive cancer where early intervention is crucial, but the prognostic role and mechanisms of ubiquitination-related genes (URGs) in immune regulation remain unclear. This study aimed to develop a URG-based prognostic signature and explore its relationship with immune modulation in CM. We integrated single-cell RNA sequencing (scRNA-seq) and bulk RNA-seq data, identifying prognostic URGs through univariate and multivariate Cox regression. A six-gene signature (UBE2L6, SPSB1, PSMB9, PSMB10, RNF213 and ATXN3) was established and validated. The signature effectively stratified patients into high- and low-risk groups, with significant survival differences. Pathway analysis revealed immune-related processes, such as 'cytokine-cytokine receptor interaction' and 'antigen processing and presentation', enriched in the low-risk group. Immune cell infiltration analysis demonstrated significant differences in the abundance of 12 immune cell types between risk groups. Notably, PSMB9 expression was positively correlated with CD8+ T cell abundance (r = 0.64, p < 0.05). scRNA-seq analysis highlighted T cells as a key cell type, with all six prognostic genes showing dynamic expression changes during T cell differentiation. Our findings suggest that URGs influence CM prognosis by modulating the immune microenvironment, offering new insights for immunotherapeutic strategies.

As an emerging biomarker, tumor mutational burden (TMB) has attracted increasing attention from clinicians in predicting the efficacy of tumor immunotherapy. Currently, TMB is detected primarily by whole-exome sequencing or targeted panel sequencing on high-throughput sequencing platforms. However, the lack of uniformity in detection methods, threshold settings, and reporting formats, as well as the significant differences in TMB values among different cancer types, have hindered the standardized application of this biomarker in clinical practice. This consensus focuses on the definition, standardization of detection, clinical significance, and limitations of TMB, and provides consensus recommendations for the clinical application of TMB in real-world practice in China. This consensus is aimed at helping clinicians and laboratory personnel understand the clinical significance and testing standards of TMB, promoting more accurate interpretation of test results, and improving patient care.

Malignant peritoneal mesothelioma (MPeM) is a malignant tumor originating from the peritoneum. In recent years, the incidence of MPeM has been increasing. Because MPeM is insidious in onset and of strong local invasiveness, and most patients are found in advanced stage, early screening and treatment of the population at the high risk of malignant mesothelioma are of vital importance. The main treatment methods for MPeM include cytoreductive surgery, hyperthermic intraperitoneal chemotherapy, systemic chemotherapy and immunotherapy. To promote the homogenization and normalization of the diagnosis and treatment of malignant peritoneal mesothelioma, based on existing clinical research evidence, the Chinese Alliance of Research for Mesothelioma (ChARM) formulated the National Expert Consensus on the Clinical Diagnosis and Treatment of Malignant Peritoneal Mesothelioma in combination with expert opinions nationwide. The contents of expert consensus cover epidemiology, diagnosis, treatment and prognosis follow-up.

Immune checkpoint inhibitors (ICIs) have transformed cancer treatment, yet immune-related adverse events (irAEs) including immunotherapy-related gastritis (IRAEG) pose significant clinical challenges—often necessitating treatment interruption that may compromise antitumor efficacy. IRAEG presents with atypical symptoms, lacks specific biomarkers, and shows histopathological overlap with other forms of gastritis, complicating diagnosis and management. Despite increasing clinical recognition, a systematic understanding of spatial molecular alterations across the full disease course remains limited. Here, we used spatial proteomics to map the molecular landscape of IRAEG during disease progression and to define stage-specific patterns of molecular evolution relevant to cancer immunotherapy management. We analyzed tissue samples from seven patients, including four non-immunotherapy-related gastritis controls and three cancer patients who developed IRAEG following ICI therapy for solid tumors, sampled longitudinally across four disease stages: baseline (G1), acute severe inflammation (G2), early recovery (G3), and complete recovery (G4). Using laser capture microdissection coupled with data-independent acquisition mass spectrometry, we profiled 177 spatially resolved gastric tissue regions. Multiplex immunohistochemistry and immunofluorescence characterized features of the immune microenvironment, while Gene Ontology, KEGG pathway analysis, Gene Set Variation Analysis, and xCell inference enabled functional, metabolic, and immune profiling. Key immune and NET-related findings were further validated by multiplex immunofluorescence in an independent, expanded cohort of IRAEG and non-immunotherapy-related gastritis samples. IRAEG was characterized by widespread HLA molecule activation and enhanced antigen processing, resembling the immune phenotype observed in organ transplant rejection. The acute G2 stage exhibited excessive neutrophil extracellular trap formation, profound metabolic suppression, and collapse of immune homeostasis—features that may inform early intervention strategies to preserve ICI treatment continuity. During early recovery (G3), inflammatory injury transitioned toward repair, marked by activation of fatty acid metabolism and PPAR signaling. Notably, even at complete clinical recovery (G4), more than 1,000 proteins remained differentially expressed, reflecting sustained enhancement of metabolic and immune functions and establishing a distinct molecular "memory" state with implications for ICI rechallenge decisions. These findings define four molecularly distinct stages of IRAEG progression and recovery. The stage-specific signatures identified here serve as candidate biomarkers for diagnosis, disease staging, and therapeutic response assessment, and may guide clinical decisions regarding irAE management, treatment modification, and safe ICI rechallenge to support continued antitumor therapy.

Irritable bowel syndrome (IBS) is characterized by chronic visceral pain and anxiety comorbidity, yet the central mechanisms linking gut signals to persistent pain–emotion dysregulation remain unclear. This study investigated whether hippocampal Lipocalin-2 (LCN2) drives neuronal ferroptosis to mediate visceral hypersensitivity and anxiety-like behaviors in a neonatal rat IBS model. Using a colorectal distension model, we conducted behavioral tests, molecular assays, and electron microscopy. IBS-like rats exhibited upregulated hippocampal LCN2 expression, predominantly in neurons, alongside ferroptosis hallmarks including increased ACSL4, decreased GPX4, elevated Fe2⁺ and MDA levels, and mitochondrial damage. Inhibition of ferroptosis reversed visceral pain and anxiety-like behaviors. LCN2 knockdown in the hippocampal CA1 region alleviated IBS-related phenotypes, while LCN2 overexpression in normal rats induced similar behavioral and ferroptotic changes. These findings identify a novel LCN2–ferroptosis axis in hippocampal neurons that critically mediates gut–brain dysregulation in IBS. Our study provides the first causal evidence linking LCN2-driven ferroptosis to visceral pain–anxiety comorbidity, offering new mechanistic insights and potential therapeutic targets for IBS and related gut–brain disorders.

Visceral hypersensitivity is a hallmark feature of irritable bowel syndrome (IBS), yet its underlying mechanisms remain incompletely understood. In the present study, we found that miRNA-let7b5p was downregulated in the spinal cord of IBS model rats induced by neonatal colorectal distension. Concurrently, microglia exhibited a shift toward a pro-inflammatory M1 phenotype and selectively engulfed inhibitory synapses, resulting in impaired GABAergic neuronal function and disruption of the excitatory/inhibitory balance. Intrathecal administration of a miRNA-let7b5p agomir suppressed M1-type microglial activation in the spinal cord, reduced pro-inflammatory cytokine levels, and alleviated visceral hypersensitivity, whereas antagomir treatment induced visceral hypersensitivity in control rats. Mechanistically, MAP3K3 was identified as a direct target of miRNA-let7b5p, and its knockdown recapitulated the protective effects conferred by miRNA upregulation. Collectively, these findings demonstrate that miRNA-let7b5p attenuates IBS-associated visceral hypersensitivity by downregulating MAP3K3, thereby inhibiting spinal microglial activation and restoring GABAergic neuronal function. This study provides novel insights into the pathogenesis of IBS-related visceral hypersensitivity and highlights a potential therapeutic target for drug development.

Background: Programmed cell death protein (PD-1) inhibitors constitute the standard adjuvant therapy for cutaneous melanoma (CM), but well-established strategies for acral melanoma (AM) remain limited. Objective: To compare the efficacy of adjuvant anti-PD-1 immunotherapy versus high-dose interferon α-2b (HDI) in patients with stage IIB-IV AM and CM. Design: This multicenter, retrospective study enrolled 511 patients with resected stage IIB-IV AM and CM between January 2017 and December 2023. Methods: Patients were divided into four groups by subtype and treatment: patients with CM and treated with anti-PD-1 (CM-PD-1), patients with CM and treated with HDI (CM-HDI), patients with AM and treated with anti-PD-1 (AM-PD-1), and patients with AM and treated with HDI (AM-HDI). Recurrence-free survival (RFS), overall survival (OS), and patient safety were evaluated. Results: This study comprised 362 AM and 149 CM cases. AM cases presented with thicker primary lesions, higher ulceration rates, and fewer BRAF mutations. Median follow-up was 49 months. Among patients with stage IIB/C, median RFS was not reached in any groups. Among stage III/IV patients, median RFS was 14.6 (CM-PD-1), 13.7 (CM-HDI), 13.3 (AM-PD-1), and 11.7 months (AM-HDI), and median OS was 61.6, 40.7, 42.4, and 53.4 months, respectively, without significant intergroup differences. Anti-PD-1 significantly improved RFS in patients with stage III/IV AM with KIT mutations (9.1 vs 5.0 months, p = 0.048) and ⩾4 lymph node metastases (10.5 vs 6.6 months, p = 0.036). Anti-PD-1 had significantly fewer adverse effects than HDI (60.4% vs 88.6%, p < 0.001), including fewer grades 3–4 events (4.6% vs 30.7%, p < 0.001). Conclusion: Adjuvant anti-PD-1 therapy provides RFS comparable to that of HDI in AM and CM, with a superior safety profile. Patients with AM harboring KIT mutations derive greater benefits from adjuvant anti-PD-1 therapy.