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.

The shortage of donor organs is the primary factor limiting the availability of liver transplantation (LT) and is a leading cause of death among patients on the waiting list. The reuse of liver allografts, while rare, represents a significant and unconventional donor resource, offering a promising strategy to expand the donor pool. This approach has been documented in international literature, demonstrating favorable surgical outcomes and long-term follow-up results. Here, we report the first case of liver allograft reuse in the Liver Transplantation Center, Department of General Surgery, Huashan Hospital, Fudan University. In this case, the first recipient underwent orthotopic LT for acute liver failure and hepatic encephalopathy. However, their condition deteriorated on the seventh postoperative day, culminating in brain death. Following evaluation and maintenance, the liver allograft was successfully re-transplanted into a second recipient, who had undergone LT six days earlier but experienced acute hepatic artery embolism leading to rapid liver function deterioration. The second recipient's liver function recovered smoothly after surgery, and they were discharged on the 28th postoperative day. This case highlights the significant value of liver allograft reuse in expanding the donor pool and providing life-saving options for critically ill patients requiring urgent LT.

The shortage of donor organs is the primary factor limiting the availability of liver transplantation (LT) and is a leading cause of death among patients on the waiting list. The reuse of liver allografts, while rare, represents a significant and unconventional donor resource, offering a promising strategy to expand the donor pool. This approach has been documented in international literature, demonstrating favorable surgical outcomes and long-term follow-up results. Here, we report the first case of liver allograft reuse in the Liver Transplantation Center, Department of General Surgery, Huashan Hospital, Fudan University. In this case, the first recipient underwent orthotopic LT for acute liver failure and hepatic encephalopathy. However, their condition deteriorated on the seventh postoperative day, culminating in brain death. Following evaluation and maintenance, the liver allograft was successfully re-transplanted into a second recipient, who had undergone LT six days earlier but experienced acute hepatic artery embolism leading to rapid liver function deterioration. The second recipient's liver function recovered smoothly after surgery, and they were discharged on the 28th postoperative day. This case highlights the significant value of liver allograft reuse in expanding the donor pool and providing life-saving options for critically ill patients requiring urgent LT.

BACKGROUND:Glucocorticoids can inhibit the expression of hub genes in the parathyroid hormone type Ⅰ receptor(PTH1R)/protein kinase A(PKA)signaling axis and interfere with the osteogenic and angiogenic differentiation of bone marrow mesenchymal stem cells,leading to the disruption of blood supply in bone and bone tissue structures.Previous studies of the research team showed that Gubitongxiao granules can induce blood vessel formation and inhibit osteoblast apoptosis,which has a certain effect on the prevention and treatment of steroid-induced femoral head necrosis. OBJECTIVE:To observe the therapeutic effect of Gubitongxiao granules in a mouse model of steroid-induced femoral head necrosis,and to explore its mechanism from the PTH1R/PKA signaling axis. METHODS:An animal model of steroid-induced necrosis of the femoral head was established by intraperitoneal injection of lipopolysaccharide and gluteal muscle injection of prednisolone acetate.After identification by nuclear magnetic resonance method,60 mice that were successfully modeled were divided into model group,Gubitongxiao granule group and Tongluo Shenggu capsule group,with 20 mice in each group.Another 12 normal mice were used as control group.The corresponding groups were intragastrically given the corresponding drugs for 12 weeks,and then the samples were taken under anesthesia.Histomorphology of femoral head samples was observed by hematoxylin-eosin staining.Enzyme-linked immunosorbent assay was used to detect the serum levels of bone alkaline phosphatase,type Ⅰ amino-terminal extension peptide,parathyroid hormone,osteocalcin and alkaline phosphatase.Western blot and RT-qPCR were used to detect PTH1R,PKA,myocyte enhancer factor 2,sclerostin and guanylate-binding protein activity-stimulating peptide at protein and gene expression levels,respectively. RESULTS AND CONCLUSION:Gubitongxiao granules may reduce the serum PTH level in mice,inhibit the activation of the PTH1R/PKA signal axis,further up-regulate the protein expressions of sclerostin and myocyte enhancer factor 2,and increase the levels of bone alkaline phosphatase,type Ⅰ amino-terminal extension peptide,osteocalcin and alkaline phosphatase in mice,thus improving femoral head necrosis,which is comparable to the intervention effect of Tongluo Shenggu capsules.It is speculated that Gubitongxiao granules may prevent and treating hormonal femoral head necrosis by regulating the PTH1R/PKA signaling axis.

Objective To explore the expression and significance of neutrophil extracellular traps(NETs)in peripheral blood of primary Sj?gren's syndrome(pSS)patients across different disease activity levels,and the predictive value of NETs and routine laboratory markers antithrombin Ⅲ(AT Ⅲ),alkaline phosphatase(ALP)and carbohydrate antigen 125(CA125)for pSS disease activity.Methods A total of 94 newly diagnosed pSS patients at the Affiliated Lianyungang Hospital of Xuzhou Medical University from October 2021 to December 2023 were categorized into active(n=49)and non-active(n=45)groups based on the European League Against Rheumatism(EULAR)Sj?gren's Syndrome disease activity index(ESSDAI).The levels of NETs biomarkers,namely serum myeloperoxidase(MPO)-DNA,were measured using ELISA.Laboratory routine indicators and MPO-DNA were integrated into multivariate Logistic regression to screen for independent influencing factors of pSS disease activity.Pearson's correlation was used to evaluate the relationship between MPO-DNA levels and ESSDAI scores.The efficacy of MPO-DNA alone or in combination with AT Ⅲ,ALP and CA125,for predictors of disease activity was evaluated using ROC curve.Results Serum MPO-DNA(23.884±3.494 μg/L),ALP(80.159±34.318 U/L)and CA125(20.300±16.560 U/ml)levels of active pSS patients were higher than those in the non-active patients(19.024±3.324 μ g/L,67.500±21.166U/L,13.200±6.340 U/ml),while AT Ⅲ(89.180±15.040 ng/ml)was lower than that in non-active patients(94.650±11.620 ng/ml),with significant differences(t=-7.921,-2.426,-2.925,2.094,all P＜0.05).Multivariate Logistic regression analysis showed laboratory indicator MPO-DNA,ALP and CA125 were independent risk factors,while AT Ⅲ was an independent protective factor(all P＜0.05).MPO-DNA was positively correlated with ESSDAI scores(r=0.602,P＜0.01).The AUC(95％CI)of the combination of ALP,CA125 and AT Ⅲ for predicting disease activity in pSS was 0.711(0.612～0.810).The AUC(95％CI)of MPO-DNA alone for predicting disease activity in pSS was 0.837(0.758～0.915),and the AUC(95％CI)of combination of MPO-DNA,ALP,CA125 and AT Ⅲ for predicting disease activity in pSS was 0.866(0.797～0.935),showing an improving in the predictive value.Conclusion The involvement of NETs in the occurrence and expression levels of pSS is related to its disease activity.NETs combined with ALP,CA125 and AT Ⅲ have effective diagnostic performance for the disease activity of pSS.This tool can serve as a biological indicator for predicting the disease activity of pSS.

Objective To investigate the protective effect of Gubi Tongxiao granules on steroid-induced necrosis of femoral head(SANFH).Methods Femoral head and bone marrow tissues of patients with SANFH and femoral neck fracture(FNF)undergoing hip replacement were collected and studied.The expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),FoxOl and cholesterol regulatory binding protein(SREBP)were detected in bone tissue.Human bone marrow mesenchymal stem cells(hBMSCs)were isolated and cultured.The effect of the drug-containing serum of Gubitongxiao granule on the proliferation of hBMSCs was detected by CCK-8 method.Oil red O and alizarin red staining were used to observe the adipogenic and osteogenic differentiation of cells.The expression of peroxisome proliferator-activated receptor γ(PPARy),CCATT enhancer binding protein α(C/EBPα),Runt-associated transcription factor 2(Runx2),alkaline phosphatase(ALP)mRNA and pathway protein were analyzed by RT-qPCR and Western blot.Results Western blot results showed that the expression of PI3K,AKT and SREBP protein in femoral head tissue of SANFH patients was significantly increased compared with that of normal group,while FoxOl expression was significantly decreased(P＜0.05);Different volume fractions of Gubi Tongxiao granules could promote the proliferation of hBMSCs,especially in 5％group after 48h.Compared with the model group,5％drug-containing serum could significantly reduce the lipid droplet coloring in the cytoplasm and increase the orange-red mineralized nodules in the calcium salt deposit,down-regulate the mRNA expression levels of PPARγ and C/EBPα,and up-regulate the mRNA expression levels of Runx2 and ALP(P＜0.05).In addition,after the intervention of drug-containing serum,the expression of FoxO1 protein in cells was significantly increased,and the expression of AKT,p-FoxO1 and SREBP protein was decreased(P＜0.05).Conclusion After the intervention of Gubi Tongxiao granules,the lipid differentiation of hBMSCs decreased and the osteogenic differentiation increased,which can effectively improve the lipid deposition in glucocorticoid environment,and the mechanism may be related to the regulation of PI3K/AKT/FoxO1 signaling pathway.

Objective To investigate the protective effect of Gubi Tongxiao granules on steroid-induced necrosis of femoral head(SANFH).Methods Femoral head and bone marrow tissues of patients with SANFH and femoral neck fracture(FNF)undergoing hip replacement were collected and studied.The expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),FoxOl and cholesterol regulatory binding protein(SREBP)were detected in bone tissue.Human bone marrow mesenchymal stem cells(hBMSCs)were isolated and cultured.The effect of the drug-containing serum of Gubitongxiao granule on the proliferation of hBMSCs was detected by CCK-8 method.Oil red O and alizarin red staining were used to observe the adipogenic and osteogenic differentiation of cells.The expression of peroxisome proliferator-activated receptor γ(PPARy),CCATT enhancer binding protein α(C/EBPα),Runt-associated transcription factor 2(Runx2),alkaline phosphatase(ALP)mRNA and pathway protein were analyzed by RT-qPCR and Western blot.Results Western blot results showed that the expression of PI3K,AKT and SREBP protein in femoral head tissue of SANFH patients was significantly increased compared with that of normal group,while FoxOl expression was significantly decreased(P＜0.05);Different volume fractions of Gubi Tongxiao granules could promote the proliferation of hBMSCs,especially in 5％group after 48h.Compared with the model group,5％drug-containing serum could significantly reduce the lipid droplet coloring in the cytoplasm and increase the orange-red mineralized nodules in the calcium salt deposit,down-regulate the mRNA expression levels of PPARγ and C/EBPα,and up-regulate the mRNA expression levels of Runx2 and ALP(P＜0.05).In addition,after the intervention of drug-containing serum,the expression of FoxO1 protein in cells was significantly increased,and the expression of AKT,p-FoxO1 and SREBP protein was decreased(P＜0.05).Conclusion After the intervention of Gubi Tongxiao granules,the lipid differentiation of hBMSCs decreased and the osteogenic differentiation increased,which can effectively improve the lipid deposition in glucocorticoid environment,and the mechanism may be related to the regulation of PI3K/AKT/FoxO1 signaling pathway.

【Objective】 To compare the similarities and differences of polymerized human cord hemoglobin (PolyCHb) and free hemoglobin (FHb) on partial coagulation indexes in vitro, so as to analyze the effect of PolyCHb on coagulation dysfunction. 【Methods】 Using normal saline, two concentrations of FHb and PolyCHb and 36% methemoglobin-containing PolyCHb to mix with fresh whole blood or plasma-rich plasma (PRP) in equal proportions, and incubate at 37°C for 30 minutes to detect prothrombin time (PT), activated partial thromboplastin time (APTT), coagulation factor Ⅱactivity (FⅡ∶C), coagulation factor Ⅴactivity (FⅤ∶C), coagulation factor Ⅷactivity (FⅧ∶C), coagulation factor Ⅸactivity (FⅨ∶C), von Willebrand factor (vWF) and platelet P-selectin (CD62P). 【Results】 1) NaCl group: PT(22.68±1.76) s; APTT(59.58±7.52) s; FⅡ∶C(45.91±3.27) %; FⅤ∶C(30.86±4.43) %; FⅧ∶C(41.32±12.94) %; FⅨ∶C(23.96±5.10) %; vWF (2.14±0.54) mg/L; CD62P(7.44±4.47) %. This group kept as a diluted control. 2) 2% FHb group compared with 7% FHb group: FⅧ∶C (42.16±12.31) %vs (56.64±12.22 ) % (P<0.05). No significant difference was found in other indexes (P>0.05). 3) There is no significant difference between 2% PolyCHb group and 7% PolyCHb group (P>0.05). 4) There is no significant difference between 2% FHb group and 2% PolyCHb group (P>0.05). 5) 7% FHb group compared with 7% PolyCHb group: PT(23.31±1.34)s vs (21.97±1.56)s (P<0.05); APTT(50.12±5.72)s vs (55.43±5.43)s (P<0.05); FⅧ∶C (56.64±12.22) %vs (42.37±13.00)% (P<0.05); vWF (1.41±0.30) mg/L vs (2.25±0.41) mg/L (P< 0.05). No significant difference was found in other indexes (P>0.05). 6) 7% PolyCHb group Compared with Met-PolyCHb group: APTT(55.43±5.43) s vs (46.33±4.86)s (P<0.05); FⅧ∶C (42.37±13) %vs (60.51±10.16) % (P<0.05). No significant difference was found in other indexes (P>0.05). 【Conclusion】 The effect of PolyCHb on coagulation markers is different from FHb. At the concentration of this study, PolyCHb will not cause coagulation disorders. However, if the methemoglobin (MetHb) content is too high, it will activate the intrinsic coagulation pathway.

Objective To explore the postoperative psychological status of donors during pediatric living donor liver transplantation (LDLT) to elucidate the correlation between resilience,anxiety and depression.Methods Random sampling was employed for selecting 60 pediatric LDLT donors undergoing LDLT from September 2014 to February 2019.They were requested to answer a questionnaire.The questionnaire concluded general information,self-rating anxiety scale (SAS),selfrating depression scale (SDS) and Chinese version of Resilience Scale.Results The score of anxiety was (46.06 ± 10.06) and depression was (50.32 ± 11.49).Both values were higher than those of Chinese norm.The score of resilience was (59.55 ± 14.62).And the total score of resilience and the score of each dimension were negatively correlated with anxiety and depression (P＜0.01).Conclusions The postoperative anxiety and depression level of donors during LDLT are higher than the ordinary.Resilience is negatively correlated with the level of anxiety and depression.The lower level of resilience,the higher anxiety and depression of donors during LDLT.For clinicians,appropriate intervention measures should be taken for improving the resilience,reducing negative emotions and boosting the quality-of-life of donors during LDLT.

Objective To observe the relationship between the peripheral blood IFN-γ secretion level and prognosis in the patients with early stage of acute cerebral hemorrhage.Methods Sixty-six inpatients with acute intracerebral hemorhage in the emergency department of this hospital from July 2015 to August 2016 were selected as the experimental group,and 60 healthy people in the physical examination center served as the blank control group.The peripheral blood mononuclear cells(PBMCs) and plasma were separated.The helper T cells(Th)1/Th2 eytokines(IFN-γ,IL-2,IL-4 and IL-10) were detected by using ELISA.The flow cytometry was used to detect intracellular cytokines;the disease condition change from 1 d of onset to 30 d in the experiment group was analyzed by using NIHSS.The relationship between the cytokines and prognosis on 30 d evaluated by NIHSS score was analyzed.Results The peripheral blood IFN-γ level had statistically significant difference between the two groups(P＜0.05),while the levels of IL-2,IL-4 and IL-10 had no statistical difference(P＞0.05).The peripheral blood IFN-γ secretion level during early stage in the experimental group was positively correlated with the prognosis(P＜0.05).Conclusion The increase of peripheral blood IFN-γ during early stage is correlated with the poor prognosis in the patients with acute intracerebral hemorrhage.