Shifang Peng

Background: Sepsis remains a leading cause of mortality in intensive care units, with endotoxemia-induced hepatic inflammation being a major contributor to multiorgan failure. Bromodomain and extraterminal domain (BET) proteins are key epigenetic regulators of inflammation. While the BET inhibitor JQ1 shows protective effects in sepsis models, its toxicity limits translational application. NHWD-870, a next-generation BET inhibitor with improved potency and safety, has not been evaluated in septic liver injury. Methods: Eight-week-old male C57BL/6 mice were challenged with lipopolysaccharide (LPS) to induce endotoxemia and treated with NHWD-870 1 hour before LPS challenge. Survival rate was assessed and 0.5mg/kg was selected as the optimal dose. Liver injury and inflammation were assessed by histopathology, serum biochemistry, ELISA, RT-qPCR, and immunohistochemistry. In vitro, primary mouse hepatocytes (PMHs) and bone marrow-derived macrophages (BMDMs) were stimulated with LPS to examine NHWD-870 effects on cytokine production and macrophage polarization. Results: NHWD-870 improved survival in lethal LPS challenge and attenuated LPS induced ALT/AST elevations and hepatic inflammatory infiltration. Serum and hepatic IL6, TNFα, and MCP1 were reduced after NHWD-870; IL6 suppression at 12 h exceeded JQ1. NHWD-870 decreased hepatic macrophage and neutrophil infiltration and no obvious histopathological abnormalities were observed in the major organs at the active dose. In vitro, NHWD-870 dose dependently reduced LPS induced cytokine expression in PMH and BMDM and inhibited macrophage M1 polarization (CD86, iNOS). BRD4 and STAT1 expression increased after LPS; NHWD 870 suppressed both. Conclusion: NHWD-870 ameliorates endotoxemia-associated hepatic inflammation and improves survival in mice, at least in part via suppression of BRD4-STAT1 signaling and inhibition of macrophage M1 polarization. Further preclinical development is warranted.

Background: The prognosis of acute-on-chronic liver failure (ACLF) is impaired by etiology heterogeneity across regions. Currently, prognostic models incorporating nutrient anabolism–related indicators for patients with hepatitis B virus (HBV)–associated ACLF are lacking. Objectives: This study aimed to construct a nomogram that incorporates nutrition-related indexes alongside traditional predictors to estimate 12-week mortality in HBV-ACLF. Methods: We retrospectively analyzed adult patients with HBV-ACLF treated at our department between May 2020 and December 2021. A total of 242 HBV-ACLF patients were enrolled and categorized into survivor (n = 174) and progression (n = 68) groups. Independent prognostic factors were identified using logistic regression analysis and incorporated into a nomogram. Nomogram performance was evaluated in terms of discrimination, calibration, and clinical utility, with internal validation using bootstrap resampling. Results: Patients in the progression group were older, more prone to hepatorenal syndrome and spontaneous peritonitis, and had lower levels of prothrombin activity, L3 skeletal muscle index and ceruloplasmin (all p < 0.05). These six independent predictors were incorporated into the nomogram, which demonstrated superior discrimination ability, with an area under the receiver operating characteristic curve of 0.95, enabling accurate identification of patients at high risk of short-term mortality. The Hosmer–Lemeshow test confirmed excellent calibration, decision curve analysis confirmed the clinical benefit, and bootstrap validation confirmed the robustness. Conclusions: The developed nomogram, incorporating nutritional status, may provide complementary information to support short-term risk stratification and clinical decision-making in patients with HBV-ACLF awaiting liver transplantation.

Cholestasis is a complex pathophysiological syndrome characterized by impaired bile secretion and excretion. Extensive research has revealed that Ubiquitin D (UBD) plays a pivotal role in numerous types of malignancies and benign diseases. However, the underlying involvement of UBD in cholestasis remains unclear. The aim of this study was to analyze the role of UBD in Cholestasis. Transcriptome data from cholestasis patients (GSE61260, GSE159676 and GSE183754) were obtained from the Gene Expression Omnibus (GEO) database. These datasets, along with cholestatic mouse models and clinical specimens, were utilized to evaluate hepatic UBD expression. Subsequently, immunohistochemistry and immunofluorescence staining were applied to validate the expression and localization of hepatic UBD. Moreover, the association of hepatic UBD levels with clinical parameters was evaluated using Pearson's correlation analysis. In addition, Gene set enrichment analysis (GSEA) and xCell were employed to investigate the immune-related signatures and immune cell infiltration link to UBD in cholestasis, with findings further validated through immunofluorescence staining. Finally, the association of the hepatic UBD with T cell-related chemokine and chemokine receptor was explored using Pearson's correlation analysis. The results showed that UBD was significantly and consistently upregulated in the livers across cholestasis patient transcriptomic data, experimental mouse models, and clinical specimens. Hepatic UBD levels positively correlate with the severity of cholestasis and hepatocytes are identified as the primary source of UBD in cholestatic livers. Functional enrichment analysis indicated that immune-related pathway was significantly activated in the cholestatic liver with high expression of UBD group. Moreover, hepatic UBD expression was positively associated with the infiltration of T cells and with the expression of T cell-related chemokines and chemokine receptors in cholestasis. In conclusion, UBD is a key gene associated with disease severity and T cells infiltration in cholestasis. These findings provide new insight into the key biomarker of cholestasis and further highlight that UBD might be a promising novel therapeutic target for patients with cholestasis.

Metabolic dysfunction-associated steatohepatitis (MASH), a severe form of metabolic dysfunction-associated steatotic liver disease (MASLD), involves hepatic lipid accumulation, inflammation, and fibrosis. It can progress to cirrhosis or hepatocellular carcinoma without timely treatment. Current treatment options for MASH are limited. This study explores the therapeutic effects of fluorofenidone (AKF-PD), a novel small-molecule compound with antifibrotic and anti-inflammatory properties, on MASH in mouse model. Mice fed a choline-deficient, l-amino acid-defined, high-fat diet (CDAHFD) were treated with AKF-PD, resulting in reduced serum ALT, AST, hepatic lipid accumulation, liver inflammation, and fibrosis. Network pharmacology and RNA-sequencing analyses suggested that AKF-PD influenced multiple metabolic, inflammatory, and fibrosis-related pathways. Further experiments verified that AKF-PD activated hepatic AMPK signaling, leading to the inhibition of the downstream SREBF1/SCD1 pathway and the activation of autophagy. Additionally, AKF-PD suppressed the expression of various inflammatory factors, reduced macrophage infiltration, and inhibited NLRP3 inflammasome activation. Moreover, AKF-PD attenuated liver fibrosis by inhibiting TGFβ1/SMAD signaling. In conclusion, this study reveals that AKF-PD effectively decreases hepatic lipid accumulation, liver inflammation and fibrosis in a CDAHFD-induced MASH model, positioning AKF-PD as a promising candidate for the treatment of MASH.

This study aims to construct and validate noninvasive diagnosis models for evaluating significant liver fibrosis in patients with chronic hepatitis B (CHB). A cohort of 259 CHB patients were selected as research subjects. Through random grouping, 182 cases were included in the training set and 77 cases in the validation set. The nomogram was developed based on univariate analysis and multivariate regression analysis. Various machine learning models were employed to construct prediction models for significant liver fibrosis. The area under the ROC curve (AUC), sensitivity, specificity, NPV, PPV, and F1 score were used to evaluate the diagnostic performance. The new nomogram had excellent diagnostic efficiency (AUC 0.806, 95% CI: 0.740–0.872). Compared with other traditional noninvasive diagnostic models, the nomogram demonstrated higher AUC values and better prediction performance. Among six machine learning models, the random forest (RF) model achieved the highest AUC (AUC 0.819, 95% CI: 0.720–0.906). Finally, the importance of all variables in the RF model was ordered to illustrate the contribution of different variables, providing the clinical factors associated with the risk of significant liver fibrosis. This new nomogram may more reliably than other traditional models and the RF model demonstrated superior accuracy among six machine learning models.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent chronic liver disease that ranges from metabolic dysfunction-associated steatotic liver (MASL) to metabolic dysfunction-associated steatohepatitis (MASH), and may eventually progress to cirrhosis and hepatocellular carcinoma (HCC). The underlying mechanism of MASLD remains incompletely understood. This study aimed to identify key gene implicated in MASLD pathogenesis and validate its correlation with disease severity through an integration of bioinformatics and experimental approaches. Liver transcriptome data from MASLD patients were obtained from the Gene Expression Omnibus (GEO) database. A diet-induced MASLD mouse model was developed, and liver RNA-sequencing was performed. Liver specimens and clinical data from patients were collected for further analysis. A total of 120 differentially expressed genes (DEGs) were shared between datasets GSE89632 and GSE213621, with functional enrichment in inflammatory, metabolic, and cell cycle-related pathways. Protein-protein interaction (PPI) network analysis identified three modules associated with MASLD, with the cell cycle-related module being the most notable. EEF1A2 was identified as a novel hub gene and revealed to be elevated with MASLD progression through dataset analysis. EEF1A2 was confirmed to be highly expressed in the livers of both MASLD mouse models and patients. Moreover, the increased expression of EEF1A2 in MASH was positively correlated with higher serum alanine aminotransferase (ALT), alanine aminotransferase (AST), total cholesterol (TC), and body mass index (BMI). In conclusion, EEF1A2 is a novel hub gene significantly associated with MASLD severity and is a promising biomarker and therapeutic target for MASLD.

Background: Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease, with unclear pathogenesis. Although immune disorders, especially T cell infiltration, are thought to play a vital role in PSC, the specific pathogenesis mechanisms remain incompletely understood. This study evaluated the potential key gene associated with the PSC pathogenesis and analyzed the associations of the key gene with prognosis and immune cell infiltration by combining bioinformatics analysis and experimental verification. Methods: Transcriptome data of PSC and normal human liver tissues (GSE159676) were obtained from the gene expression omnibus database. Differentially expressed genes (DEGs) were identified, and differences in biological states were analyzed. A protein–protein interaction (PPI) network was constructed. Hub genes were identified, and their expression was verified using transcriptome data of mice fed 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) and Mdr2^−/− mice (GSE179993, GSE80776), as well as by immunohistochemistry staining on clinical samples. The correlations between the key gene and other factors were evaluated by Pearson’s correlation coefficient. Immune cell infiltration into human liver (GSE159676) was analyzed by xCell and verified by immunofluorescence staining on PSC liver samples. Results: Of the 185 DEGs identified, 113 were upregulated and 72 were downregulated genes in PSC. Genes associated with immune cell infiltration and fibrosis were significantly enriched in PSC. PPI network showed close interactions among DEGs. A module strongly associated with immune infiltration was identified, with annexin A1 (ANXA1) being the core gene. High expression of ANXA1 in PSC was confirmed in two public datasets and by immunohistochemistry staining on clinical samples. High ANXA1 expression was strongly associated with high-risk score for PSC. Also, ANXA1 expression was positively associated with chemokines and chemokine receptors and with the infiltration of immune cells, especially T cells, into liver with PSC. Immune infiltration, fibrosis, and cancer-related processes were markedly enriched in PSC with high expression of ANXA1. Conclusion: ANXA1 is a key gene associated with high risk and infiltration of immune cells, especially T cells, in PSC. These findings provide new insight into the key biomarker of PSC and suggest that targeting ANXA1 may be a valuable strategy for the treatment of PSC.

长期的乙型肝炎病毒(hepatitis B virus，HBV)感染可引起肝脏反复出现炎症坏死，进而发展为肝纤维化、肝硬化甚至肝癌。肝脏病理改变程度是指导慢性乙型肝炎(chronic hepatitis B，CHB)患者抗病毒治疗的重要依据之一。由于肝穿刺活检临床使用的局限性，有必要寻找到更好的无创性指标以评估CHB患者的肝脏病变程度，指导抗病毒治疗。本研究旨在分析CHB患者不同程度肝脏病变的临床特征，并探索谷丙转氨酶(alanine aminotransferase，ALT)正常的CHB患者肝脏炎症及纤维化程度的影响因素。 回顾性纳入完善肝脏病理活体组织检查(以下简称“活检”)的CHB患者310例。收集患者临床资料，以肝脏病理活检结果为金标准，分析总体样本相关临床指标与肝脏病理改变程度的关系。筛选ALT正常的CHB患者，探索影响其肝脏炎症及纤维化程度的独立危险因素。 310例CHB患者中，显著性肝脏炎症[肝脏炎症程度分级(liver inflammation grade，G)≥2]和显著性肝纤维化[肝纤维化程度分期(liver fibrosis stage，S)≥2]的患者分别为249例(80.3%)和119例(38.4%)。总体样本单因素分析结果显示：ALT、谷氨酰转肽酶、碱性磷酸酶水平以及HBV DNA定量与显著性肝脏病理改变(G/S≥2)相关(均P2 000 U/mL(OR=3.592，95% CI 1.534~8.409)；显著性肝纤维化的独立危险因素为碱性磷酸酶升高(OR=1.022，95% CI 1.002~1.043)、血小板计数降低(OR=0.990，95% CI 0.982~0.998)及乙型肝炎e抗原(hepatitis B e antigen，HBeAg)阳性(OR=14.845，95% CI 4.898~44.995)。根据多因素分析建立ALT正常的CHB患者显著性肝纤维化诊断模型，并绘制受试者操作特征曲线，曲线下面积为0.844(95% CI 0.779~0.910)。 肝脏的病理改变程度需综合不同临床指标进行评估。ALT正常的CHB患者中仍有相当一部分存在肝脏显著病理改变，需及时识别并进行抗病毒治疗。在这部分患者中，HBV DNA定量>2 000 U/mL提示存在显著性肝脏炎症；而基于碱性磷酸酶、血小板计数及HBeAg水平建立的显著性肝纤维化诊断模型有助于评估肝脏纤维化程度。