BackgroundThe gut microbiome has emerged as a potential modulator of transplant rejection. However, the relevance of anaerobe-associated gut microbial communities to transplant rejection responses remains unclear.MethodsIn a murine islet transplantation model, we assessed the effects of metronidazole pretreatment on allograft survival and function. 16S rRNA sequencing was performed to characterize treatment-associated changes in gut microbial composition. Separately, an exploratory two-sample MR analysis using human GWAS data was performed to prioritize gut microbial taxa associated with a composite outcome of transplant failure or rejection. Biological annotation was subsequently used to prioritize candidate host pathways. Immunohistochemistry and flow cytometry were used to evaluate post-transplant immune responses after metronidazole pretreatment.ResultsMetronidazole treatment remodeled gut microbiota composition and significantly delayed islet allograft rejection. MR analyses identified several anaerobe-associated gut microbial taxa potentially relevant to transplant rejection risk, including the family Defluviitaleaceae and the genera Intestinibacter, Bilophila, Ruminococcus and Eubacterium fissicatena. Integrative biological annotation highlighted glycosylation-related pathways and prioritized ST3GAL4 for subsequent expression assessment in the murine transplant model. Furthermore, metronidazole treatment reduced CD4+ T-cell infiltration around the allograft and Th17-related inflammatory responses.ConclusionAltogether, our findings document gut microbial remodeling and delayed allograft rejection following metronidazole pretreatment. These findings are hypothesis-generating and warrant further mechanistic investigation.

PurposeTo elucidate the role of protein lactylation in gastric adenocarcinoma progression, focusing on lactate dehydrogenase B (LDHB) delactylation at K58 and its effects on metastasis, glutathione (GSH) metabolism, and ferroptosis resistance.MethodsQuantitative proteomics profiled lactylation in gastric adenocarcinoma versus adjacent normal tissues. Bioinformatics and Kaplan-Meier analyses assessed differentially lactylated genes and survival correlations. LDHB lactylation sites were validated via immunoprecipitation and Western blotting in lactate-stimulated cells. Functional assays evaluated LDHB-K58R (delactylation mimic) effects on proliferation, invasion, GSH levels, cystine uptake, STAT1/SLC7A11/GPX4 expression, and RSL3-induced ferroptosis in vitro. EMT markers were examined by immunofluorescence and Western blotting. In vivo lung metastasis was assessed in nude mice injected with modified AGS cells, treated with RSL3 or DMSO.ResultsProteomics identified 121 upregulated and 53 downregulated lactylation sites in tumors, with LDHB as a key differentially lactylated protein. High LDHB expression correlated with poor progression-free and disease-specific survival. LDHB was primarily lactylated at K58; K58R delactylation enhanced proliferation, colony formation, invasion, cystine uptake, and SLC7A11-dependent GSH synthesis by suppressing STAT1 and downregulating GPX4, without affecting EMT or lactate release. K58R conferred robust resistance to RSL3-induced ferroptosis, reducing lipid ROS and preserving malignant phenotypes in vitro. In vivo, K58R promoted lung metastasis and suppressed RSL3-induced ferroptosis and lipid peroxidation (4-HNE).ConclusionLDHB-K58 delactylation drives gastric cancer metastasis via SLC7A11-mediated GSH synthesis and ferroptosis resistance, suggesting therapeutic targeting of this axis for overcoming therapy resistance.

Background The optimal surgical strategy for adenocarcinoma of oesophagogastric junction (AEG) remains debated, particularly regarding lymphadenectomy extent, gastrectomy type and surgical approach, with real-world prospective evidence being scarce. Objective To map lymph node metastasis (LNM) patterns and assess surgical outcomes in a large multicentre cohort of patients with AEG undergoing radical resection. Design The Chinese League of Adenocarcinoma of Esophagogastric Junction (CLAEG) registry, initiated in 2022 across 44 high-volume Chinese centres, prospectively enrolled AEG patients. This analysis included 2044 radical resections, with LNM assessed by station, stratified by Siewert type and neoadjuvant therapy. Surgical outcomes were compared between total versus proximal gastrectomy and laparoscopic versus open resection. Results Most tumours were Siewert type II (64.6%) or III (33.4%). LNM was substantially higher in abdominal than mediastinal stations; category-1 nodes (metastasis, >10%) comprised stations 1, 2, 3, 4, 7, 8a, 9 and 11p. The LNM rates for mediastinal stations were 2.77% (No. 110), 0.71% (No. 111) and 0.68% (No. 112). Patients who received neoadjuvant therapy had lower LNM rates, indicating nodal downstaging. Among those undergoing gastrectomy, patients who underwent total gastrectomy had a lower postoperative complication rate than those who underwent proximal gastrectomy (14.8% vs 21.0%; p=0.001) and achieved more extensive lymphadenectomy. Compared with open surgery, patients who underwent laparoscopic resection experienced faster postoperative recovery without higher complication rates (16.5% vs 17.3%). No perioperative mortality occurred. Conclusion The CLAEG study shows that abdominal lymphadenectomy should be prioritised in AEG, with neoadjuvant therapy, total gastrectomy and laparoscopy associated with favourable short-term outcomes.

Colorectal cancer peritoneal metastasis (CRC-PM) often has poor prognosis after traditional hyperthermic intraperitoneal chemotherapy (HIPEC) due to constraints of respiratory movements, intestinal peristalsis and peritoneum plasma barrier. Herein, cisplatin and 17AAG were loaded into a thermoresponsive biodegradable polyurethane (PU) hydrogel, forming the CDDP/17AAG@PU system. This system showed favorable in vitro sustained release properties, controllable thermosensitive gelation at 43°C and excellent biocompatibility. Benefiting from the hydrogel's sustained release at safe doses, compared with the free drug group, this system significantly reduced serum biochemical abnormalities, as indicated by lower serum ALT/CREA levels and less apoptosis of normal intestinal cells. Moreover, its thermosensitive treatment enhanced antitumor efficacy by promoting DNA cross-linking and inhibiting oncogenic signaling-related proteins (p-PI3K, p-AKT, MMP2). In vivo experiments showed it more effectively inhibited CRC-PM compared to single-drug treatment and the control group. In conclusion, the safe, degradable CDDP/17AAG@PU system spreads to cover CRC-PM tumors at 37°C and gels rapidly at 43°C (clinical HIPEC temperature). It reduces antitumor drug induced hepatorenal toxicity, exerts good efficacy against CRC-PM and thus hold considerable potential for clinical translation.

Regulatory T cells (Tregs) have been shown to be involved in the induction of transplantation tolerance in numerous models. Our previous work demonstrated that methyltransferase-like 14 (METTL14) loss impaired Treg function and hindered the establishment of transplantation tolerance. However, the underlying mechanisms remain unclear. In this study, we found that METTL14 knockdown in Tregs significantly impaired their regulatory function, leading to poor allograft function and accelerated transplant rejection. Using methylated RNA immunoprecipitation- and mRNA-sequencing approaches, we discovered that METTL14 deficiency fostered the expression of semaphorin 4D (Sema4D) mRNA, a key semaphorin family member with immunoregulatory activity. Methylation of target adenosines reduced Sema4D mRNA degradation, a process mediated by the METTL14-YTH N6-methyladenosine RNA binding protein 2 axis. Inhibition of Sema4D suppressed its interaction with its receptor, thereby preserving Treg immunoregulation capability and prolonging allograft survival through the p21-activated kinase-signal transducer and activator of transcription 5signaling pathway. Importantly, Sema4D expression in kidney transplant biopsies were negatively correlated with renal allograft survival. In summary, our findings suggest that METTL14 deficiency in Tregs leads to transplant rejection and reveal for the first time that Sema4D may serve as a potential therapeutic target to enhance Treg function in transplantation.

Radiotherapy (RT) is a commonly employed therapeutic approach for malignant tumors located in the abdomen. However, radiation proctopathy (RP) is a significant side effect that occurs during RT, and there are limited pharmaceutical interventions available. This study aimed to investigate the effectiveness of Liriodendron, a primary extract derived from the traditional Chinese medicine Fibraureae Caulis, in the prevention and treatment of radiation proctopathy. We established a mouse model of radiation proctopathy through local irradiation and analyzed various clinicopathological measures over the subsequent eight weeks. Our findings indicate that both the Liriodendron gavage group and the Liriodendron enema group exhibited a reduction in the production of pro-inflammatory cytokines and an increase in the secretion of anti-inflammatory cytokines, as determined by transcriptome sequencing analysis and ELISA examination. Furthermore, Liriodendron administration led to a decrease in the levels of reactive oxygen species (ROS) and apoptotic cells in the intestinal tissue of mice. MASSON staining and immunohistochemistry demonstrated that Liriodendron could reduce the expression of α-SMA in rectal tissue and the extent of intestinal fibrosis in mice with radiation proctopathy. In conclusion, this study suggests that Liriodendron holds potential as a therapeutic agent for the treatment of RP.

Murine renal subcapsular islet transplantation presents a promising technique for diabetes treatment by addressing challenges such as immune rejection and reliance on immunosuppressive drugs. Here, we present a protocol for the isolation, purification, and transplantation of mouse pancreatic islets that overcomes these challenges. Specifically, we describe steps for inducing diabetes with streptozotocin, pancreatic perfusion and isolation, and islet cell purification. We then detail procedures for renal subcapsular islet transplantation, dual antibody therapy, and immune cell and graft monitoring.For complete details on the use and execution of this protocol, please refer to Liu et al.1

INTRODUCTION:Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide, with most Chinese patients diagnosed at a locally advanced stage. Neoadjuvant chemotherapy (NAC) is increasingly used to improve resectability and survival. Laparoscopy-assisted distal gastrectomy (LADG) provides short-term recovery benefits compared with open distal gastrectomy (ODG), but its safety and oncologic efficacy following NAC remain uncertain. This trial aims to determine whether LADG is non-inferior to ODG in terms of long-term survival outcomes in patients with locally advanced distal gastric cancer (LAGC) after NAC. METHODS AND ANALYSIS:This is a multicentre, randomised, controlled, non-inferiority trial conducted at high-volume GC centres in China. Eligible patients (aged 18-75 years; cT3-4a, N0/+, M0) with histologically confirmed distal gastric adenocarcinoma who have completed standard NAC will be randomised 1:1 to LADG or ODG with D2 lymphadenectomy. Surgical quality will be standardised through operative manuals, intraoperative video recording and central auditing. The primary endpoint is 3-year disease-free survival. Secondary endpoints are 3- and 5-year overall survival. A total of 998 patients (499 per arm) will be enrolled, providing 80% power to test non-inferiority with an absolute 8% margin, accounting for 15% attrition. Analyses will follow the intention-to-treat principle, with Cox models used for survival comparisons and subgroup analyses according to nodal status, tumour size and pathological response. ETHICS AND DISSEMINATION:This trial has been reviewed and approved by the Biomedical Ethics Committee of West China Hospital, Sichuan University (Approval No. 2025 (865), 16 July 2025). Written informed consent will be obtained from all participants. The results will be disseminated through peer-reviewed journals and international conferences, providing high-level evidence to guide the surgical management of LAGC after NAC. TRIAL REGISTRATION NUMBER:Chinese Clinical Trial Registry, ChiCTR2500109677; registered on 23 September 2025. Protocol V.2.1, dated 29 June 2025.

Achieving immune tolerance is a key goal in organ transplantation, as it eliminates the need for long-term immunosuppression. Regulatory B cells (Bregs) present a promising strategy for inducing tolerance. Our previous findings demonstrated that the adoptive transfer of ex vivo-expanded murine splenic B regulatory cells, referred to as Toll-like receptor-Bregs (TLR-Bregs) (Toll-like receptors 9 and 4 stimulated), induces tolerance to allografts. Here, we identified that circulating T cells increase programmed cell death protein 1 (PD-1) expression after TLR-Breg cellular therapy. To investigate whether programmed death-ligand 1 (PD-L1)/PD-1 signaling is involved in tolerance induction by TLR-Bregs, we generated TLR-Bregs from wild-type or PD-L1-deficient B6 splenocytes and assessed their regulatory functions. Our findings revealed that TLR-Bregs express high levels of PD-L1 and extend graft survival via a PD-L1-dependent mechanism. Mechanistically, PD-L1 enhances the activation and survival of TLR-Bregs; PD-L1 is required for TLR-Bregs to induce PD-1 expression and an optimal inhibition of effector CD4+ T cells while promoting the forma-tion of functional Foxp3hiCCR7hiCTLA-4hi regulatory T cells. This study highlights the foundational role of PD-L1 in inducing tolerance via ex vivo-expanded TLR-Bregs.

Purpose:The objective of this study was to ascertain the safety of laparoscopic distal D2 radical gastrectomy in treating gastric cancer patients after NAC with locally advanced disease (cT3-4a, N0/ +, M0) by evaluating postoperative complications. Study Design:A prospective, multicenter, single-arm clinical trial. Methods:This clinical trial was conducted at 14 hospital centers in China. Adults aged 18-75 years with histologically confirmed LAGC (cT3-4a, N0/ +, M0) were enrolled in the study. Participants received three cycles of administration of intravenous oxaliplatin (130 mg/m2 on day 1 of each cycle) plus oral capecitabine (1000 mg/m2 twice daily on days 1 to 14 of each cycle), repeated every three weeks prior to undergoing laparoscopic distal gastrectomy. The primary endpoint was the postoperative overall morbidity rate. Secondary endpoints included postoperative mortality rate, surgery-related complications, R0 resection rate, and the rate of conversion to laparotomy, response of neoadjuvant chemotherapy (NAC), adverse event rate of NAC, operation time, blood loss, postoperative severe morbidity rate, and postoperative recovery course. Results:A total of 153 patients who underwent NAC prior to laparoscopic D2 distal gastrectomy were included in the final analysis. The study reported a postoperative overall morbidity rate of 20.9% (95%CI: 15.2%-28.0%), with a postoperative mortality rate of 0%. Pneumonia is the most common complication (9.2%). Ten patients exhibited elevated levels of body fluid amylase without presenting any clinical symptoms or undergoing additional clinical intervention. The R0 resection rate was achieved at 100%. The rate of conversion to laparotomy was 1.3%. 9.2% of patients achieved pathological complete response (pCR) following NAC. The overall incidence of adverse effects after NAC was 20.3% (95%CI: 14.7%-27.3%). The most common grade 3-4 treatment-related adverse events during neoadjuvant treatment were a decrease in platelet count and vomiting, each occurring in 0.7% of patients. Conclusion:The laparoscopic distal D2 radical gastrectomy demonstrated a favorable safety profile in the treatment of gastric cancer patients with advanced disease (cT3-4a, N0/ +, M0) following NAC.