Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

Objective:To investigate the effect of donor blood lipid levels and the degree of fat deposition on the pancreatic surface on the outcome of islet isolation.Method:A retrospective analysis was conducted on 171 cases of islet isolation data from organ donors between May 2015 and December 2024. According to the percentage of fat deposition area on the surface of the pancreatic capsule after trimming, the samples were divided into three groups: mild surface fat group (<30%, 60 cases) , moderate surface fat group (30%–70%, 55 cases) , and severe surface fat group (>70%, 56 cases). The modified Ricordi method was used to digest pancreatic tissue, and islets were purified by continuous density gradient centrifugation. The digestion efficiency, digestion time, islet yield (islet equivalent/quantity) , purity, score, and size were analyzed and compared among groups. One-way ANOVA was used for inter-group comparisons, and Pearson correlation analysis and multiple linear regression analysis were used to explore the relationship between blood lipid levels and islet isolation parameters.Result:The severe surface fat group had significantly higher pre-purification and post-purification islet equivalents, islet number, amount of digested pancreatic tissue, donor triglyceride levels, and low density lipoprotein (LDL) levels than the other groups (all P<0. 05) . Correlation analysis showed that LDL level was positively correlated with pre-purification islet equivalents (62 cases, r=0. 298, P=0. 019) and islet number (58 cases, r=0. 285, P=0. 030) . Donor high density lipoprotein (HDL) level was negatively correlated with post-purification islet equivalents (54 cases, r= – 0. 282, P=0. 039) ; donor triglyceride level was positively correlated with the amount of digested tissue (56 cases, r=0. 268, P=0. 046) and negatively correlated with purity (51 cases, r= - 0. 297, P=0. 035) ; donor very low density lipoprotein (VLDL) level was positively correlated with the amount of digested tissue (67 cases, r=0. 337, P=0. 005) and negatively correlated with purity (61 cases, r=- 0. 348 , P=0. 006) ; donor total cholesterol level was negatively correlated with pancreatic digestion efficiency (34 cases, r= - 0. 370, P=0. 032) , and the above differences were all statistically significant. Conclusion:Pancreata with heavier surface fat deposition can yield a higher number of islets. Meanwhile, donor blood lipid levels are correlated with islet isolation outcomes and can serve as important indicators for donor pancreas selection.

Liver transplantation (LT) has become a standard treatment for end-stage liver diseases, and graft injury is intricately associated with poor prognosis. Granzyme B (GZMB) plays a vital role in natural killer (NK) cell biology, but whether NK-derived GZMB affects graft injury remains elusive. Through the analysis of single-cell RNA-sequencing data obtained from human LT grafts and the isolation of lymphocytes from mouse livers following ischemia-reperfusion injury (IRI), we demonstrated that 2NK cells with high expression of GZMB are enriched in patients and mice. Both systemically and liver-targeted depletion of NK cells led to a notable reduction in GZMB⁺ cell infiltration, subsequently resulting in diminished graft injury. Notably, the reconstitution of Il2rg ^-/- Rag2 ^-/- mice with purified Gzmb-KO NK cells demonstrated superior outcomes compared to those with wild-type NK cells. Crucially, global knockout of GZMB and pharmacological inhibition exhibited remarkable improvements in liver function in both mouse IRI and rat LT models. Moreover, a phosphorylated derivative of FDA-approved vidarabine was identified as an effective inhibitor of mouse GZMB activity by molecular dynamics, which could provide a potential avenue for therapeutic intervention. Therefore, targeting NK cell-derived GZMB during the LT process suggests potential therapeutic strategies to improve post-transplant outcomes.

Objective:To investigate the molecular mechanism of the 5-methylcytosine(m5C)reader Y-box binding protein 1(YBX1)in participating in the development and progression of colorectal cancer(CRC)by regulating the stability of the ferroptosis inhibitor membrane-spanning 4-domains subfamily A 15(MS4A15).Methods:Bioinformatics databases were used to investigate the mRNA and protein expression levels of YBX1 in CRC.RT-qPCR was used to measure the expression level of YBX1 in CRC cells.LC-MS was used to measure the level of m5C modification in CRC cells and nor-mal colorectal mucosal cells.CCK-8 assay was used to observe the effect of YBX1 on the proliferation of CRC cells,Transwell assay was used to observe its effect on the migration ability of CRC cells,and flow cytometry was sued to observe its effect on the apoptosis of CRC cells.Bioinformatics methods were used to identify the ferrop-tosis inhibitors that can interact with YBX1 and potential m5C modification sites.GEPIA2 was used to analyze the association between the expression of YBX1 and MS4A15.The expression of YBX1 was inhibited,and then the mRNA expression level and m5C modifica-tion level of MS4A15 were analyzed.The catRAPID database was used to find the binding sites between YBX1 protein and MS4A15 mRNA.CRC cells were treated with actinomycin D after inhibition of YBX1 expression,and RT-qPCR was used to measure the stabil-ity of MS4A15 mRNA.The expression of MS4A15 was inhibited,and then the proliferative activity,migration ability,and apoptosis rate of cells were measured,as well as the expression levels of the key indicators for ferroptosis,including MDA,ROS,and Fe2+.Results:High mRNA and protein expression levels of YBX1 were observed in CRC,and YBX1 was highly expressed in CRC cells.The m5C modification level of CRC cells was significantly higher than that of normal colorectal mucosal cells.YBX1 could promote the prolifera-tion and migration of CRC cells and inhibit the apoptosis of CRC cells.The bioinformatics analysis showed that YBX1 was positively correlated with the expression level of the ferroptosis inhibitor MS4A15,and there were multiple m5C modification sites on MS4A15.Inhibition of YBX1 expression could reduce the mRNA expression level and m5C modification level of MS4A15 and the stability of MS4A15 mRNA.There were significant reductions in the proliferative activity and migration ability of CRC cells and a significant in-crease in the apoptosis rate of CRC cells after inhibition of MS4A15 expression,with significant increases in the content of MDA,ROS,and Fe2+.Conclusion:These results show that YBX1 promotes the development and progression of CRC by stabilizing MS4A15 via m5C modification,which provides a promising targeted therapeutic strategy for CRC patients.

Objective:To investigate the effect of donor blood lipid levels and the degree of fat deposition on the pancreatic surface on the outcome of islet isolation.Method:A retrospective analysis was conducted on 171 cases of islet isolation data from organ donors between May 2015 and December 2024. According to the percentage of fat deposition area on the surface of the pancreatic capsule after trimming, the samples were divided into three groups: mild surface fat group (<30%, 60 cases) , moderate surface fat group (30%–70%, 55 cases) , and severe surface fat group (>70%, 56 cases). The modified Ricordi method was used to digest pancreatic tissue, and islets were purified by continuous density gradient centrifugation. The digestion efficiency, digestion time, islet yield (islet equivalent/quantity) , purity, score, and size were analyzed and compared among groups. One-way ANOVA was used for inter-group comparisons, and Pearson correlation analysis and multiple linear regression analysis were used to explore the relationship between blood lipid levels and islet isolation parameters.Result:The severe surface fat group had significantly higher pre-purification and post-purification islet equivalents, islet number, amount of digested pancreatic tissue, donor triglyceride levels, and low density lipoprotein (LDL) levels than the other groups (all P<0. 05) . Correlation analysis showed that LDL level was positively correlated with pre-purification islet equivalents (62 cases, r=0. 298, P=0. 019) and islet number (58 cases, r=0. 285, P=0. 030) . Donor high density lipoprotein (HDL) level was negatively correlated with post-purification islet equivalents (54 cases, r= – 0. 282, P=0. 039) ; donor triglyceride level was positively correlated with the amount of digested tissue (56 cases, r=0. 268, P=0. 046) and negatively correlated with purity (51 cases, r= - 0. 297, P=0. 035) ; donor very low density lipoprotein (VLDL) level was positively correlated with the amount of digested tissue (67 cases, r=0. 337, P=0. 005) and negatively correlated with purity (61 cases, r=- 0. 348 , P=0. 006) ; donor total cholesterol level was negatively correlated with pancreatic digestion efficiency (34 cases, r= - 0. 370, P=0. 032) , and the above differences were all statistically significant. Conclusion:Pancreata with heavier surface fat deposition can yield a higher number of islets. Meanwhile, donor blood lipid levels are correlated with islet isolation outcomes and can serve as important indicators for donor pancreas selection.

Objective:To investigate the changes of β-cell dedifferentiation in type 2 diabetes mellitus(T2DM) and aging subjects.Methods:Using pancreatic samples from 12 prediabetic individuals, 21 type 2 diabetic patients, and 27 control, the level of β-cell dedifferentiation was assessed by immunofluorescence staining for FOXO1 and insulin expression. Correlation analyses were performed between β-cell dedifferentiation levels and hemoglobin A 1C(HbA 1C) level in 60 human pancreatic samples. Correlation analyses were performed between β-cell dedifferentiation levels and age in non-diabetic and T2DM. Results:FOXO1 was mainly expressed in the cytoplasm of human islet β-cells. The proportion of FOXO1 -INS + /INS + cells in T2DM significantly increased compared with control and pre-diabetes, and positively correlated with HbA 1C level( r=0.623, P<0.001). The proportion of FOXO1 -INS + /INS + cells in the young group of T2DM was significantly higher than that in the non-diabetic young and elderly groups, and further significantly increased in elderly group. In T2DM, the proportion of FOXO1 -INS + /INS + cells was positively correlated with age( r=-0.53, P<0.05). Conclusion:Hyperglycemia and aging lead to an increased level of β-cell dedifferentiation in T2DM.

Idiopathic pulmonary fibrosis(IPF)is a common interstitial fibrosis disease with poor prognosis and lack of effec-tive treatment to prolong the survival time of patients.Cell senescence can induce tissue inflammation and accelerate the patho-logical process of fibrosis.Excessive oxidative stress,nuclear DNA damage and mitochondrial dysfunction in the pathological en-vironment of IPF are the main causes of cell senescence.Nicotinamide adenine dinucleotide(NAD+)is an important coenzyme in redox reaction,which can regulate a series of biological functions by blocking cell senescence.Its active expression is closely re-lated to the occurrence and development of IPF.Therefore,to explore the mechanism of NAD+regulating cell senescence under pathological condition is of great significance for the prevention and treatment of IPF.

Idiopathic pulmonary fibrosis(IPF)is a chronic progressive interstitial lung disease of unknown etiology,with a rapid disease course,poor prognosis,and the absence of effective therapeutic drugs.Mitochondrial dys-function is one of the crucial causes of inducing IPF.Silent information regulator 3(SIRT3)can restore mitochondrial ho-meostasis by inhibiting mitochondrial oxidative stress,repairing mitochondrial DNA damage,and ameliorating abnormal mitochondrial lipid metabolism.This paper summarizes the role and mechanism of SIRT3 in attenuating mitochondrial dys-function based on delineating the relationship between mitochondrial dysfunction and IPF,aiming to provide references for finding effective treatment methods for IPF.

Hepatocellular carcinoma (HCC) is one of the most common malignancies and is a major cause of cancer-related mortalities worldwide (Forner et al., 2018; He et al., 2023). Sarcopenia is a syndrome characterized by an accelerated loss of skeletal muscle (SM) mass that may be age-related or the result of malnutrition in cancer patients (Cruz-Jentoft and Sayer, 2019). Preoperative sarcopenia in HCC patients treated with hepatectomy or liver transplantation is an independent risk factor for poor survival (Voron et al., 2015; van Vugt et al., 2016). Previous studies have used various criteria to define sarcopenia, including muscle area and density. However, the lack of standardized diagnostic methods for sarcopenia limits their clinical use. In 2018, the European Working Group on Sarcopenia in Older People (EWGSOP) renewed a consensus on the definition of sarcopenia: low muscle strength, loss of muscle quantity, and poor physical performance (Cruz-Jentoft et al., 2019). Radiological imaging-based measurement of muscle quantity or mass is most commonly used to evaluate the degree of sarcopenia. The gold standard is to measure the SM and/or psoas muscle (PM) area using abdominal computed tomography (CT) at the third lumbar vertebra (L3), as it is linearly correlated to whole-body SM mass (van Vugt et al., 2016). According to a "North American Expert Opinion Statement on Sarcopenia," SM index (SMI) is the preferred measure of sarcopenia (Carey et al., 2019). The variability between morphometric muscle indexes revealed that they have different clinical relevance and are generally not applicable to broader populations (Esser et al., 2019).
Humans ; Aged ; Sarcopenia/diagnostic imaging* ; Carcinoma, Hepatocellular/diagnostic imaging* ; Muscle, Skeletal/diagnostic imaging* ; Deep Learning ; Prognosis ; Radiomics ; Liver Neoplasms/diagnostic imaging* ; Retrospective Studies

To construct monoclonal antibodies against Haemonchus contortus native H11 protein.In this study,five 4-6 weeks female BALB/c mice were immunized with native H11 protein extrac-ted from adult worms by Concanavalin A lectin.Spleen cells were isolated and fused with SP2/0 cells after 3 times of immunization.Two hybridoma cell lines,named A1E3 and A10E1,which could stably secrete monoclonal antibodies against H11 protein were obtained.The subtype identi-fication and immunological analysis showed that the heavy chain of the two monoclonal antibodies belonged to IgG1 and the light chain was κ type,and both monoclonal antibodies recognized the natural antigen H11.Immunohistochemical localization and larval developmental inhibition test in vitro showed that the mAb A1E3 could be localized to the intestinal microvilli of the adult worm,and that the antibody can inhibit the growth of the fourth-stage larvae.The successful production of two monoclonal antibodies not only lays the foundation for the study of protective antigenic epitopes of the H11 protein and the development of epitope vaccines,but also provides a potential application of the monoclonal antibody for the treatment of haemonchusis in animals.