BACKGROUND:Hepatic ischemia-reperfusion injury (HIRI) is a serious clinical complication with limited effective interventions. Although mild hypothermic pretreatment (MHP) has demonstrated protective potential, the precise molecular mechanisms, particularly its role in modulating sterile inflammation and programmed cell death, remain poorly understood. METHODS:A murine model of 70% hepatic IR was established. Mice were pretreated with MHP (32 °C for 2 h) or normothermia. In vitro, AML-12 hepatocytes and isolated mouse neutrophils were subjected to oxygen-glucose deprivation (OGD) or stimulation with recombinant CIRP (rmCIRP). Inhibitors targeting CIRP (C23), TLR4 (TAK-242), and PAD4 (Cl-amidine) were used. Liver injury, inflammatory markers, NETosis markers (CitH3, PAD4), and pyroptosis (NLRP3, Caspase-1, GSDMD) were assessed via histology, ELISA, western blot, and immunofluorescence. RESULTS:MHP significantly attenuated liver damage, oxidative stress, and systemic inflammation following IR. It reduced the release of CIRP, neutrophil infiltration, and NETosis markers (PAD4 and CitH3). In vitro, CIRP released from injured hepatocytes directly induced NETosis via the TLR4/PAD4 pathway, which was inhibited by C23, TAK-242, or Cl-amidine (Cl). Furthermore, NETs promoted hepatocyte pyroptosis by upregulating NLRP3, Caspase-1, and GSDMD. MHP or inhibition of the CIRP-TLR4-PAD4 axis effectively suppressed both NETosis and pyroptosis. CONCLUSION:MHP protects against HIRI by inhibiting CIRP release, which subsequently reduces TLR4/PAD4-dependent NETosis and hepatocyte pyroptosis, thereby disrupting a self-amplifying inflammatory loop. These findings reveal a novel CIRP-NETs-pyroptosis axis as a key mechanism in HIRI and highlight potential therapeutic targets for liver I/R injury.

Abstract Background Hepatic ischaemia/reperfusion (I/R) injury poses a common clinical dilemma encountered during liver transplantation (LT), characterised by substantial cellular death and inflammation reactions. Ubc9, the sole E2 conjugating enzyme of SUMOylation, has long been recognised to regulate diverse biological and pathological processes. However, its impact on I/R‐induced liver damage is yet to be elucidated. Methods The expression levels of UBC9 in patients undergoing LT were analysed. Hepatocyte‐specific Ubc9‐deficient or transgenic mice were utilised in an in vivo model of hepatic I/R, alongside in vitro experiments that employed hypoxia/reoxygenation stimulation. The investigation focused on Ubc9's role in liver damage due to I/R and the underlying mechanisms through a range of phenotypic analyses and biological techniques. Results Herein, we found that hepatic tissues from patients with LT are featured by a significant downregulation of UBC9 expression. Studies in 68 donor hepatic biopsies further demonstrated a negative correlation between UBC9 expression and liver injury in patients with LT. Similarly, murine liver I/R was coupled with an obvious decrease in Ubc9 expression. Hepatocyte deficient in Ubc9 exacerbated liver injury in liver I/R, while Ubc9‐overexpression showed the opposite phenotype. Mechanistically, Ubc9‐mediated SUMOylation of Ninj1 at lysine K103 inhibited its membrane localisation and damage‐associated molecular patterns (DAMPs) release in hepatocytes, subsequently inhibited nuclear factor‐kappa B (NF‐κB) signalling in macrophages and curtailing inflammatory cytokines production. Conclusions These findings further suggest that Ubc9‐mediated SUMOylation of Ninj1 at lysine K103 may represent a potential therapeutic strategy for safeguarding the liver against I/R injury in clinical settings. Key points Ubc9 expression is downregulated in hepatocytes during hepatic ischaemia/reperfusion (I/R) injury. Higher UBC9 expression is associated with improved post‐liver transplantation (LT) liver function. Ubc9 ameliorates liver damage and inflammation responses in hepatic I/R injury. Ubc9‐mediated Ninj1 SUMOylation at K103 is essential for regulating the subcellular distribution of Ninj1. Ubc9 inhibits the release of hepatocyte‐derived damage‐associated molecular patterns (DAMPs) in a Ninj1 K103 SUMOylation‐dependent manner.

Lean metabolic dysfunction-associated steatotic liver disease (MASLD), defined as hepatic steatosis in individuals with normal BMI (<25 kg/m2 in non-Asians; <23 kg/m2 in Asians), is a clinically paradoxical phenotype in which patients face risks of progressive fibrosis, cardiovascular events, and liver-related mortality comparable to or exceeding obese counterparts. Visceral adiposity, dietary stressors, cholesterol overload, and genetic susceptibility involving PNPLA3 variants and PPARA polymorphisms synergistically disrupt hepatic lipid homeostasis. At the core of this dysregulation lies peroxisome proliferator-activated receptor alpha (PPARα), a master transcriptional regulator of hepatic fatty acid oxidation, ketogenesis, and anti-inflammatory signaling. PPARα suppression aries through convergent mechanisms, including aging-related RAGE signaling, fructose-mediated epigenetic silencing, gut‒liver axis disruption, and lipotoxicity, operating independently of systemic obesity. This review demonstrates that: (1) impaired PPARα activity underlies lean MASLD pathogenesis through transcriptional, epigenetic, and post-translational suppression; (2) synthetic PPARα agonists show limited applicability in this metabolically fragile population; and (3) natural product-derived modulators engage PPARα alongside FXR and LXR networks to restore metabolic balance. The PPARα-FGF21-ketogenesis axis and nuclear receptor crosstalk provide the mechanistic rationale for network-level therapeutic approaches. We advocate for spatially selective hepatic PPARα reactivation guided by lean-specific biomarker and genotype-stratified trial design.

Background and aimsAcute rejection (AR) is a common complication after liver transplantation. Current diagnostic modalities for acute rejection are either invasive or lack sufficient sensitivity. Therefore, the present study aimed to develop a novel and sensitive diagnostic tool for predicting AR after liver transplantation. Specifically, we investigated whether serum donor-derived cell-free DNA (dd-cfDNA) is closely associated with the occurrence of post-transplant AR.MethodsA prospective single-center diagnostic study enrolled 40 primary whole liver transplant recipients, divided into an indicative biopsy cohort (abnormal liver function requiring biopsy) and a protocol biopsy cohort (stable/mildly abnormal liver function without biopsy indication). The dd-cfDNA levels (absolute copy number and relative quantification) were dynamically monitored 14 days to 1 year post-transplant. Using pathological biopsy as the gold standard, receiver operating characteristic (ROC) curve analysis and logistic regression models compared the diagnostic efficacy of dd-cfDNA and liver function indices for AR.ResultsOur study demonstrated that circulating dd-cfDNA levels were significantly elevated in liver transplant recipients with acute rejection (AR) compared to those without. The diagnostic cutoffs were determined as dd-cfDNA% ≥10.39% (sensitivity: 95%, specificity: 90%) and dd-cfDNA ≥1928 copies/mL (sensitivity: 70%, specificity: 90%). The area under the curve (AUC) for dd-cfDNA% was 0.940, and for dd-cfDNA was 0.823, both outperforming routine liver function tests. After effective anti-rejection therapy, dd-cfDNA levels rapidly decreased, correlating with clinical improvement, as well as improvements in liver function and histopathology.ConclusionsAs a sensitive and non-invasive biomarker, dd-cfDNA can effectively predict the occurrence of AR after liver transplantation. Moreover, changes in dd-cfDNA levels facilitate the assessment of therapeutic efficacy, providing crucial references for optimizing immunosuppressive regimens and improving patient outcomes.

Donation after circulatory death (DCD) livers are subjected to severe ischemia-reperfusion injury (IRI) during transplantation. Hypothermic machine perfusion (HMP) offers superior mitochondria protection compared with conventional cold storage (CS), yet the underlying molecular mechanisms remain incompletely understood. In this study, we investigated the mechanistic basis of HMP protection using a rat DCD liver model combined with an in vitro hypoxia/reoxygenation model in human umbilical vein endothelial cells (HUVECs). Rats were subjected to 30 min of warm ischemia followed by CS, HMP and normothermic machine perfusion (NMP). HMP significantly ameliorated liver dysfunction, histopathological damage, oxidative stress and inflammation compared with CS. Mechanistically, HMP upregulated yes-associated protein 1 (YAP1) expression and promoted P53 nuclear translocation, thereby enhancing the expression of microautophagy-related proteins including mitochondria-eating protein (MIEAP), BCL2 interacting protein 3 (BNIP3) and BNIP3-like (BNIP3L), and facilitating the formation of mitochondrial-derived vesicles (MDVs). These effects were abrogated by the YAP1 inhibitor verteporfin in vivo and by YAP1 knockdown or MIEAP silencing in vitro. Collectively, our findings demonstrate that HMP reduced oxidative damage and inflammation of DCD liver through YAP1/P53-mediated micromitophagy.

Gastrointestinal malignancies pose a major public health challenge in China, where the majority of patients are diagnosed late and face poor survival outcomes. A clinlabomics data-based auxiliary screening tool could address this gap, yet previous studies lack the large-scale, multicenter validation necessary to support clinical translation. This study is a multicenter retrospective analysis involving 115,032 participants from four hospitals between January 1, 2019, and April 30, 2026. The cohort from Sichuan Cancer Hospital (SCH) was designated as the discovery set. After systematic comparison of various machine learning and deep learning models, glmnet was ultimately selected as the optimal predictive model. The final tool, Gastrointestinal Cancers Seek (GaSeek), was subsequently validated in three independent external cohorts, as well as in three real-world clinical cohorts. This GaSeek achieved an area under the receiver-operating characteristic curve (AUC) of 0.960 (95% CI: 0.952-0.968) in the internal validation cohort and an AUC of 0.852 (95% CI: 0.843-0.860) in the overall external validation cohort. Analysis of feature importance identified albumin (ALB) and hemoglobin (HGB) as the most influential predictors. Furthermore, the model demonstrated robust performance in detecting early-stage gastrointestinal cancers and was proficient in screening cases with high confidence for positive results. Using an appropriate threshold, GaSeek not only screened out gastrointestinal cancers but also identified other endoscopically detectable digestive tract tumors in real-world cohorts. We used “Screening Efficiency Gain (SEG)," defined as the ratio of the detection rate in the high-risk group to that in the overall cohort, to evaluate the improvement. In the SCH real-world cohort, whose non-cancer participants were healthy individuals, GaSeek delivered a high SEG of 15.70, showing excellent screening efficiency. The GaSeek was deployed as a publicly accessible web application (https://weiyanghe520.shinyapps.io/gastrointestinal-cancer-predictor/). It effectively identifies individuals at high risk for digestive tract tumors and could improve the efficiency of endoscopic screening for digestive tract tumors through risk stratification. This study has been registered with the Chinese Clinical Trial Registry, registration number: ChiCTR2500107671, registration date: August 15th, 2025, (https://www.chictr.org.cn/showproj.html?proj=276534).

The prognosis for patients diagnosed with hepatocellular carcinoma with bile duct tumor thrombus (HCC-BDTT) remains dismal, and there are presently no universally accepted treatment guidelines to address this complex condition. Long-term outcomes of liver transplantation (LT) for HCC-BDTT patients are unclear, and whether LT is a proper therapeutic option for HCC-BDTT patients remains to be determined. Therefore, we design a clinical trial to evaluate whether LT can improve recurrence-free survival (RFS) and overall survival (OS) in HCC-BDTT patients. This is an open-labeled, single-arm, prospective, multicenter and real-world study aiming to assess the survival outcomes of HCC-BDTT patients in LT. Patients will be enrolled based on histological confirmation of HCC with BDTT. The study is planned to take 4 years, 2 years for enrollment and 2 years for follow-up. We anticipate that LT confers beneficial survival outcomes for HCC-BDTT patients, specifically in terms of the pivotal parameters, such as RFS and quality of life. Upon successful completion of the trial, we will extend our monitoring over a longer follow-up time to accurately estimate important indicators such as OS. We expect that this study provides substantial evidence to refine treatment guidelines through thorough data analysis, ultimately contributing to better patient outcomes and advancing our understanding of the disease.Trial Register: Trial registered at www.clinicaltrials.gov (NCT06928415)

Donation after circulatory death (DCD) is a key source of liver grafts but it is associated with more severe ischemia‑reperfusion injury (IRI) and poorer transplant outcomes compared with donation after brain death. Hypothermic machine perfusion (HMP) effectively decreases DCD graft injury, but its protective molecular mechanisms remain unclear. Kruppel‑like factor 2 (KLF2) is an endothelial protective transcription factor induced by hemodynamic mechanical stimulation. However, the role of KLF2 in IRI during HMP in DCD livers is unclear. Rat livers undergoing DCD modeling followed by static cold storage (CS) or HMP were used to assess KLF2 expression and macrophage efferocytosis. Injury was assessed by serum alanine transferase/aspartate transferase levels, histology, TUNEL apoptosis assay and immunofluorescence (IF) for in situ efferocytosis. Protein markers were analyzed via western blotting, immunohistochemistry and IF. In vitro, HUVECs and macrophages were subjected to simulated CS/reperfusion. Macrophages efferocytosis was quantified using fluorescently labeled apoptotic Jurkat cells. Mechanisms were explored by RNA sequencing and co‑immunoprecipitation. Compared with the CS group, HMP decreased pathological injury, apoptosis and inflammation in DCD liver injury. KLF2 expression was upregulated. However, knockdown of KLF2 abrogated these endothelial protective effects in vitro. Furthermore, overexpression of KLF2 enhanced macrophage efferocytosis, whereas suppression of KLF2 impaired this. Moreover, enhanced efferocytosis contributed to inflammation resolution, ultimately improving overall graft injury and decreasing apoptosis. Mechanistically, KLF2 inhibited the NOD‑like receptor protein 3 (NLRP3) inflammasome to suppress pyroptosis, thereby indirectly enhancing efferocytosis. HMP alleviated IRI in DCD liver grafts by upregulating endothelial KLF2, which inhibited NLRP3 inflammasome‑mediated pyroptosis, thereby improving the inflammatory microenvironment and promoting macrophage efferocytosis.

The global burden of disease caused by multidrug-resistant (MDR) bacterial infections is escalating, necessitating the urgent development of novel antimicrobial materials that combine safety and high efficacy. Herein, a strategy combining bacterial capture with highly efficient broad-spectrum antimicrobial activity is employed to prepare a series of imidazolium salt chitosan (IMSCs) through a simple method. IMSCs-5 exhibits a capture-kill capability with a low propensity to induce bacterial resistance. It primarily induces bacterial death through intracellular reactive oxygen species generation and multiple synergistic mechanisms. Additionally, IMSCs-5 demonstrates strong therapeutic effects on wounds infected with methicillin-resistant Staphylococcus aureus, without causing notable systemic toxicity. These findings suggest that IMSCs provides new ideas for the rational design of novel antimicrobial materials, making it suitable for clinical applications in the treatment of MDR bacterial infections.

Non-compressible hemorrhage (NCH) caused by deep tissue injury remains a major cause of trauma-related death. Developing hemostatic materials with rapid expansion, effective coagulation promotion, good biocompatibility, and tissue regeneration is still urgently needed. This study reports a simple regioselective dual-oxidation (RDO) strategy for fabricating crosslinker-free hemostatic sponges from bacterial cellulose (BC) with high porosity. Carboxyl and aldehyde groups were both introduced into the BC scaffold after TEMPO-mediated oxidation and sodium periodate oxidation. The dual-oxidation BC (DOBC) sponges retained the 3D nanofibrous architecture and rapid shape recovery, associated with aldehyde-driven hemiacetal network formation and enhanced blood interactions. Meanwhile, the sponges exhibited favorable mechanical properties, excellent biocompatibility, high liquid absorption capacity, antibacterial activity, and rapid whole-blood coagulation. Notably, DOBC sponges effectively enrich red blood cells and platelets, further promoting rapid hemostasis. In vivo studies demonstrate that DOBC sponges effectively control bleeding in rat models of NCH and outperform gauze and commercial gelatin sponges. Moreover, they promoted cell infiltration, angiogenesis, and liver tissue regeneration. This study proposes a simple dual-oxidation, crosslinker-free strategy for preparing hemostatic sponges, thereby offering a promising solution for rapid hemostasis and subsequent tissue repair in non-compressible wounds.

Hemorrhage control remains a central unmet need in emergency medicine and surgery, as uncontrolled bleeding causes preventable mortality and is exacerbated by anticoagulation or coagulopathy, where many hemostats that depend on the coagulation cascade lose efficacy. Hierarchically structured, dual-crosslinked ultra-highly deacetylated chitosan (DC-UDCS) aerogels enable rapid, coagulation-independent hemostasis via a dual-crosslinked hierarchical capillary-elastic memory (DC-HCEM) mechanism. Directionally ice-templated pore architectures provide fast capillary infiltration that unlocks reversible hydrogen-bonded fixation, while covalent crosslinks store elastic energy to drive blood-triggered, ultrafast self-expansion and stable mechanical tamponade, physically occluding bleeding tracts and vessels. Concurrently, the cationic chitosan network promotes electrostatic erythrocyte aggregation and cellular adhesion, supporting sealing without requiring fibrin formation. In vitro and in vivo evaluations demonstrate rapid pressure generation and effective bleeding control in both normal and anticoagulated hemorrhage models, outperforming conventional gauze-type controls, while maintaining intrinsic antibacterial activity and favorable hemocompatibility/biocompatibility. This architecture-chemistry integrated aerogel platform provides a transferrable strategy for lifesaving hemostasis when coagulation is compromised.

Metabolic dysfunction-associated steatotic liver disease (MASLD) has emerged as a major global health burden, yet effective pharmacological interventions remain limited. Dysregulation of the gut microbiota-bile acid axis plays a pivotal role in MASLD pathogenesis; however, developing targeted therapies that address this complex interplay remains challenging. Here, we present a safe oral delivery system (QW) comprising in situ self-assembled nanofibrous porous microspheres derived from whey protein and quaternized chitosan, which integrates physical adsorption with bio-regulatory functions. In a murine MASLD model, oral QW administration reduced serum ALT (258.6 to 50.3 U/L) and AST (204.4 to 86.7 U/L) levels, and decreased hepatic triglyceride (0.54 to 0.18 mmol/g) and cholesterol (0.12 to 0.05 mmol/g) content. The NAS score decreased significantly, accompanied by marked histopathological improvements. Multi-omics analysis revealed that QW reshaped gut microbial composition, doubled the relative abundance of Bacteroidetes, modulated bile acid metabolism, and preserved intestinal barrier integrity through reactivation of the hepatic FXR-SHP signaling pathway. This biocompatible, multi-functional oral system offers a promising therapeutic strategy for MASLD and advances the paradigm of gut-liver axis-based interventions for metabolic liver diseases.

Hepatic ischemia-reperfusion injury (HIRI) is a common and inevitable pathological event during liver transplantation and hepatectomy, which significantly impairs postoperative liver function recovery and patient prognosis. However, the molecular and cellular mechanisms of HIRI have not been fully elucidated and further research is urgently needed. In recent years, the rapid development of bioinformatics analysis technology and the research method combining multi-dimensional data mining with experimental verification have become important strategies for exploring the mechanisms of complex diseases. Building on this, this study aims to screen and analyze the potential roles and mechanisms of key regulatory factors in the process of HIRI through systematic bioinformatics analysis and experimental verification, providing a basis for clarifying its pathogenesis and finding potential therapeutic targets. In this study, two transcriptome microarray datasets (GSE14951 and GSE7706) of human liver tissue were systematically analyzed, and candidate genes related to IRI were initially screened through differential expression analysis. Combined with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, protein-protein interaction (PPI) network construction and modular analysis were performed to identify potential key regulatory factors, ultimately highlighting HSPH1 and DNAJB1. Subsequently, based on a mouse liver IRI model, paired reperfusion tissue samples from clinical liver transplant patients, and an AML12 cell hypoxia/reoxygenation (H/R) model, the two genes were experimentally validated from multiple perspectives, including transcriptional expression, protein levels, and subcellular localization. A combination of quantitative PCR (qPCR), Western blotting, immunohistochemistry (IHC), immunofluorescence (IF), and co-immunoprecipitation (Co-IP) was employed to comprehensively evaluate their expression dynamics, subcellular distribution, and protein-protein interaction characteristics. Differential expression analysis identified 154 genes with consistent expression trends across the two datasets, which were significantly enriched in metabolic, stress response, inflammatory, and protein folding pathways. PPI network construction and module analysis further identified HSPH1 and DNAJB1 as core components of a heat shock protein interaction cluster. Validation using a mouse IRI model, paired reperfusion tissue samples from clinical liver transplant patients, and an AML12 cell hypoxia/reoxygenation (H/R) model demonstrated that both genes were significantly upregulated under IRI conditions, localized in the cytosol, and exhibited co-localization and physical interaction. Transcription factor prediction analysis suggested that STAT3 and NR1I2 might be involved in their transcriptional regulation. In conclusion, HSPH1 and DNAJB1 are co-expressed and physically interact in hepatic IRI, suggesting that they may be involved in the regulation of protein homeostasis and cellular stress responses related to liver IRI, providing important experimental evidence for a deeper understanding of the molecular characteristics of liver IRI.

Hepatic ischemia-reperfusion injury (IRI), driven primarily by excessive mitochondrial reactive oxygen species (ROS) generation, is a major cause of liver dysfunction, graft failure, and postoperative complications. However, no pharmacological agents have been clinically approved for its prevention or treatment, and there is an urgent need for effective therapeutic strategies. In this study, we established a nanoplatform composed of PEGylated polydopamine nanoparticles modified with the mitochondrial-targeting peptide SS-31 (PPS NPs). SS-31 peptide modification confers PPS NPs with efficient mitochondrial-targeting capability, thereby restoring mitochondrial membrane potential and reducing ROS accumulation in the hypoxia/reoxygenation model. Furthermore, treatment with PPS NPs significantly mitigates liver injury, decreases inflammatory factor levels, and inhibits neutrophil recruitment in mice subjected to IRI. Transcriptome sequencing and metabolomics analyses indicate that PPS NPs can protect the liver from ischemia-reperfusion injury by preserving mitochondrial integrity, reducing ROS generation, and regulating arachidonic acid and glutathione metabolism. By preserving mitochondrial function, maintaining cellular redox homeostasis, and suppressing inflammatory cascades, PPS NPs ultimately inhibit mitochondria-dependent apoptosis and confer protection against liver IRI, providing a practical therapeutic strategy for hepatic IRI clinical management.

>To the Editor: Chylothorax is a serious disease characterized by rupture of the thoracic tube and milky exudation from the pleural cavity,which can lead to a variety of pathological symptoms and is life threatening[1].Chylothorax is common after thoracic surgery or trauma.The non-traumatic chylothorax is rare in the clinical practice,and the etiology is complex and often associated with the primary disease[2].Chylothorax is a rare complication of patients with advanced cirrhosis[3],most of which are manifested as dyspnea,cough,chest pain,and the medical treatment effect is relatively poor.We performed orthotopic liver transplantation (OLT) in a patient with advanced cirrhosis combined with massive chylothorax and chyloperitoneum.The 2-year follow-up showed that the patient’s liver function was stable and no recurrence of chylothorax.