ETHNOPHARMACOLOGICAL RELEVANCE:Danggui-Shaoyao-San (DGSY) is a classic prescription in traditional Chinese medicine that has demonstrated therapeutic efficacy in treating fatty liver. However, its pharmacological mechanisms remain inadequately explored. AIM OF THE STUDY:This study aims to elucidate the molecular mechanisms through which DGSY may contribute to the modulation of MASH progression. MATERIALS AND METHODS:A MASH model was established in mice using a high-fat high-carbohydrate (HFHC) diet, followed by treatment with DGSY to assess its efficacy. RNA sequencing (RNA-Seq) analysis, western blotting, and immunofluorescence were employed to elucidate the potential mechanisms of DGSY in the treatment of MASH. The MASH mouse model and bone marrow-derived macrophages (BMDMs) stimulated with LPS + ATP were utilized to investigate the activation of the NLRP3 inflammasome in macrophages. Additionally, the mechanisms of DGSY were further validated through NLRP3 knockdown in vivo and knockout in vitro. UHPLC-Q-Orbitrap HRMS analysis was employed to identify the primary active components of DGSY. RESULTS:DGSY was effective in treating the HFHC diet-induced MASH mouse model, significantly alleviating liver inflammation. Mechanistically, RNA-Seq analysis indicated that the NOD-like receptor signaling pathway was significantly affected in DGSY-mediated effects. The protein expression levels of NLRP3, caspase1, and IL-1β were significantly elevated in liver macrophages from the MASH mouse model and in LPS + ATP-stimulated BMDMs, which were reversed by DGSY treatment. NLRP3 knockdown in vivo alleviated MASH but counteracted the therapeutic effects of DGSY. Both NLRP3 knockdown in vivo and knockout in vitro abolished its effect on inhibiting caspase1 activity and IL-1β release. In addition, UHPLC-Q-Orbitrap HRMS analysis identified four main active ingredients in DGSY, with atractenolide III and gallic acid notably inhibiting NLRP3 protein expression. CONCLUSIONS:DGSY exerts a protective effect against MASH, inhibiting hepatic macrophage NLRP3 inflammasome, suppressing caspase1 activity, and reducing IL-1β release. This study provides experimental evidence for the clinical application of DGSY in the treatment of MASH.

OBJECTIVE:CGA combination consisting of Cordyceps sinensis polysaccharide, gypenosides, and amygdalin, is derived from Fuzheng Huayu capsule (a traditional Chinese medicine approved for liver fibrosis) via previous Uniform Design Experimentation. The effect of CGA on metabolic dysfunction-associated steatohepatitis (MASH) and the potential mechanism based on bile acids (BAs) alternative biosynthesis pathway-farnesoid X receptor (FXR) axis are investigated here. METHODS:Alanine aminotransferase, hepatic triglyceride, malondialdehyde (MDA), BAs, fatty acids oxidation (FAO) activity, mRNA of inflammatory cytokines, α-smooth muscle actin (α-SMA), transforming growth factor-β, collagen type I, and FAO enzymes were detected. Liver sections underwent hematoxylin-eosin, Oil Red O, and Sirius red staining, and immunohistochemistry assay of F4/80 and α-SMA. Protein expression of peroxisome proliferator-activated receptor α (PPAR-α), FXR, BAs biosynthesis enzymes, small heterodimer partner (SHP), bile salt export pump (BSEP), adenosine triphosphate-binding cassette sub-family B member 4 (ABCB4) and nuclear factor-kappa B were detected. KEY FINDINGS:CGA ameliorated MASH, restoring FAO, PPAR-α, and FXR, accompanied by increased chenodeoxycholic acid proportion in the FXR agonist BAs pool, BAs alternative biosynthesis, and the FXR targets including SHP, BSEP, and ABCB4. CONCLUSION:CGA ameliorates MASH, promoting the hepatic BAs alternative biosynthesis pathway to activate FXR-PPARα restoring FAO.

To identify serum diagnostic biomarker for damp-heat (DH) pattern in chronic liver diseases using transcriptomics and metabolomics. Patients with chronic hepatitis B (CHB) or metabolic dysfunction-associated fatty liver disease (MAFLD) were categorized into DH and non-DH pattern groups. Serum RNA profiles were analyzed via RNA-seq/microarray, and metabolites were quantified by ultraperformance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). Biomarker screening and validation employed discovery (88 cases) and validation (85 cases) cohorts, utilizing Rank-in analysis and Least Absolute Shrinkage and Selection Operator (LASSO) regression for gene selection; reverse transcription-polymerase chain reaction (RT-PCR) and targeted UPLC-MS/MS for expression validation; random forest and receiver operating characteristic (ROC) curve analysis (with area under the curve, AUC) for diagnostic assessment. Compared with the non-DH pattern group, patients with DH pattern showed significantly elevated liver injury markers and reduced apolipoprotein A1 (P<0.05). Integrated transcriptome analysis (Rank-in) identified 315 dysregulated gene sets, primarily enriched in chemokine signaling. LASSO selected 27 genes for RT-PCR validation, confirming 5 differential genes. Metabolomics revealed 25 differential metabolites (discovery cohort), with 7 showing ⩾ 2-fold change; 6 maintained consistent trends in validation. A random forest model combining 4 genes (PTPN22, CTSD, TBX21, STAT4) and 2 metabolites (pyroglutamic acid and glutamic acid) achieved a validation AUC of 0.828 for DH diagnosis. A multi-omics diagnostic model incorporating 4 genes and 2 metabolites demonstrates promising diagnostic potential for DH pattern in patients with chronic liver diseases. (registration No. ChiCTR2000037248)

Compensated cirrhosis due to metabolic dysfunction-associated steatohepatitis (MASH) represents a major unmet need in hepatology, with no approved treatment to date. Compared with non-cirrhotic MASH, compensated cirrhosis is characterized by concurrent metabolic dysfunction, persistent inflammation, stabilized fibrosis and portal hypertension, making therapeutic development substantially more challenging. In recent years, several agents initially developed for non-cirrhotic disease have advanced into compensated MASH-related cirrhosis, including fibroblast growth factor 21 analogues, glucagon-like peptide-1 receptor/glucagon receptor dual agonists, thyroid hormone receptor-beta agonists and other antifibrotic approaches. Among these, fibroblast growth factor 21 analogues have shown the most encouraging efficacy signals. However, most candidates have not demonstrated clear histological or clinical benefit in this population. The predominance of negative trials suggests that limited progress reflects not only insufficient drug efficacy, but also disease complexity, marked patient heterogeneity and limitations of current endpoint frameworks. This review summarizes recent progress in drug development for compensated MASH-related cirrhosis, evaluates the efficacy and safety of major investigational agents, and discusses key barriers to development, including endpoint limitations, patient stratification and trial design. It also outlines future directions, including precision stratification, composite endpoints, noninvasive assessment tools, combination strategies and earlier screening and intervention.

INTRODUCTION:Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent chronic liver disease, and liver biopsy remains the gold standard for diagnosis. However, a subset of patients diagnosed with MASLD by imaging paradoxically exhibit steatosis grade S0 on liver biopsy, posing a diagnostic challenge. This study investigated the outcomes and causes of steatosis 0. METHODS:Steatosis 0 was defined as imaging-detected MASLD with a biopsy-defined steatosis grade S0. We used global cohorts to evaluate outcomes, histologic progression by paired biopsies, and possible causes. The cause analysis included MRI-proton density fat fraction (MRI-PDFF) for fat distribution, pathologist consistency testing, and comparison of serial biopsy sections. RESULTS:In a follow-up cohort of 3,273 biopsy individuals from 16 centers, 123 (3.8%) exhibited steatosis 0. Of these, 29.3% of them had advanced fibrosis (burnt-out MASLD) and demonstrated a risk of liver-related events, whereas those without advanced fibrosis had no such events. In the paired-biopsy cohort of 1,865 patients, 74 (4.0%) individuals initially showed steatosis grade S0; among them, 93.2% retained no or mild steatosis on follow-up biopsy. MRI-PDFF assessments in 42 MASLD patients revealed heterogeneous hepatic fat distribution, with some segments showing steatosis grade S0 despite elevated average liver fat. Interpathologist variability and discrepancies across consecutive biopsy sections contributed to misclassification of steatosis grade. DISCUSSION:Steatosis 0 represents a potential diagnostic gray zone in MASLD. It may be caused by burnt-out MASLD, uneven liver fat distribution, or variability in pathological assessment. Understanding the causes and implications of steatosis 0 is critical for diagnosis and management.

The emergence of glucagon-like peptide-1 receptor agonists (GLP-1RAs) represents a major breakthrough in the pharmacotherapy of obesity. However, their single-target mechanism has limitations. In recent years, combination strategies targeting multiple pathways and multiple targets have become a frontier in weight-loss drug development. This review systematically examines GLP-1RA-based combination weight-loss strategies, focusing on the scientific rationale and research progress in two main directions: single-agent multi-target agonists and multi-drug combination therapies. In single-agent multi-target strategies, glucagon-like peptide-1 receptor/glucose-dependent insulinotropic polypeptide receptor (GLP-1R/GIPR) dual agonists, glucagon-like peptide-1 receptor/glucagon receptor (GLP-1R/GCGR) dual agonists, glucagon-like peptide-1 receptor/amylin receptor (GLP-1R/AMYR) dual agonists, and triple agonists achieve weight loss beyond traditional single agents through synergistic regulation of appetite, energy metabolism, and lipolysis. In multi-drug combination therapy, free combinations offer flexibility, while fixed-dose combinations improve convenience and adherence. These strategies improve weight loss and metabolic outcomes, including hepatic fat and cardiometabolic parameters. In terms of safety, gastrointestinal events are the most common. However, as the number of targets increases and doses escalate, potential risks such as increased heart rate and hypoglycemia warrant attention, with particular consideration for thyroid and muscle safety during long-term use. Future research should focus on optimizing personalized treatment strategies, conducting head-to-head comparisons of agents with different mechanisms, developing oral formulations, and exploring long-term maintenance regimens to advance comprehensive obesity management.

Breast cancer-related fatigue is a persistent and subjective sense of tiredness caused by either the cancer itself or its treatments. This fatigue is disproportionate to recent physical activity and significantly interferes with daily life. The exact pathogenesis of Breast cancer-related fatigue remains unclear. Breast cancer cells induce metabolic abnormalities through reprogramming. This disruption in metabolic balance—particularly in various systems—may contribute to Breast cancer-related fatigue by compromising the body’s overall metabolic homeostasis, all while supporting the biosynthetic demands of cancer cell proliferation. Current pharmacological treatments for cancer-related fatigue have demonstrated limited and inconsistent efficacy. These treatments are often hindered by a narrow drug target profile, high rates of adverse effects, and the complex mechanisms underlying fatigue, particularly those related to metabolic changes. Traditional Chinese medicine, with its multi-target modulation and lower toxicity, holds significant potential as long-term adjunctive or alternative treatments for Breast cancer-related fatigue. This article reviews the current understanding of the metabolic causes of Breast cancer-related fatigue and explores the role of traditional Chinese medicine interventions, offering new insights into potential targeted therapies for this condition.

Dietary fat reshapes host-microbiota interactions, yet the upstream events that mediate overnutrition-driven microbiome alterations and metabolic dysfunction remain unclear. Here we compared mouse models of diet-induced and genetic obesity using multi-omics to identify the colonic mucus niche as an early, diet-sensitive driver of metabolic dysfunction. Excessive dietary lipids impaired glutamine metabolism and redox homeostasis in goblet cells, thinning the mucus layer and depleting the mucus-adapted symbiont Akkermansia muciniphila while expanding the bile-acid-transforming bacterium Clostridium scindens. Altered bile acid composition along the enterohepatic axis activates FXR-PLIN2 signalling in the small intestine and increases fat absorption. In parallel, enterocytes upregulate the PPARα-dependent uptake pathway that supports luminal lipid entry. Supplementation with glutamine restored goblet cell function and the gut microbiota-derived bile acid pool, thereby reducing intestinal FXR activation and lipid uptake. These findings reveal that dietary fat impairs colonic goblet cell function and reshapes microbial bile acid metabolism, influencing small-intestinal fat absorption.

Ubiquitin-specific protease 5 (USP5) is a key regulator in cancer, pain, and inflammatory disorders, yet its role in metabolic dysfunction-associated steatotic liver disease (MASLD) remains largely unexplored. This study aims to elucidate the functional significance and therapeutic potential of USP5 in MASLD. Hepatic USP5 expression was markedly reduced in liver specimens from MASLD patients, as well as in relevant in vivo and in vitro models. Hepatocyte-specific genetic ablation of USP5 exacerbated MASLD-associated hepatic steatosis and inflammation, whereas USP5 overexpression alleviated steatohepatitis. Mechanistically, co-immunoprecipitation coupled with mass spectrometry (Co-IP-MS), surface plasmon resonance (SPR), and ubiquitination assays revealed that USP5 directly interacts with carnitine palmitoyltransferase 1A (CPT-1A) and selectively cleaves K6-linked ubiquitin chains, thereby stabilizing CPT-1A via deubiquitination. In the context of MASLD, reduced USP5 expression disrupts this deubiquitination process, accelerating proteasomal degradation of CPT-1A. Consequently, USP5 deficiency impairs mitochondrial fatty acid oxidation (FAO) and promotes hepatic lipid accumulation. Notably, hepatocyte-specific restoration of CPT-1A substantially rescued the exacerbated lipid deposition induced by USP5 deficiency. Furthermore, docking-based virtual screening coupled with in vivo validation identified ligustroflavone (LF) as a potent USP5 activator. Animal studies clearly demonstrated that LF robustly enhanced USP5-mediated CPT-1A stabilization and significantly ameliorated lipid accumulation and inflammation inMASLD mice. Collectively, our findings uncover a previously unrecognized ubiquitination regulatory axis centered on USP5 and CPT-1A, and establish hepatic USP5 as an emerging therapeutic target for restoring mitochondrial FAO in MASLD.

AIMS:To examine obesity prevalence and reclassification under the new Lancet Commission on Obesity criteria among adults aged 18-59 years with metabolic dysfunction-associated steatotic liver disease (MASLD), including BMI-defined lean-MASLD. METHODS:We analysed NHANES 2017-2018 data from adults aged 18-59 years with MASLD, the age range available for dual-energy X-ray absorptiometry (DEXA) in adults of NHANES. Obesity was defined using the new Lancet Commission criteria and the traditional BMI-based criteria. The new criteria incorporated waist circumference, waist-to-hip ratio, waist-to-height ratio and body fat percentage measured by DEXA. Individuals were classified as having obesity if any of the following conditions were met: (1) BMI ≥ 40 kg/m2; (2) elevated BMI in combination with at least one elevated anthropometric measure; (3) at least two elevated anthropometric measures, irrespective of BMI; (4) excess body fat assessed by DEXA. Elevated BMI and traditional obesity was defined as BMI ≥ 27.5 kg/m2 for Asian and BMI ≥ 30 kg/m2 for other ethnicities. RESULTS:(1) The new criteria increased the estimated prevalence of obesity among patients with MASLD from 66.8% (95% CI, 60.6%-72.4%) to 98.3% (95% CI, 97.3%-99.0%). (2) Among MASLD patients classified as non-obese by traditional criteria, 94.9% were reclassified as obese under the new criteria. (3) Among BMI-defined lean-MASLD, 84.2% met the new obesity criteria. CONCLUSION:Among adults aged 18-59 years with MASLD, the new obesity criteria increased the estimated prevalence of obesity to 98.3%. Moreover, 94.9% of individuals considered non-obese by traditional BMI criteria and 84.2% of those with BMI-defined lean MASLD met the new obesity criteria.

BACKGROUND:Metabolic dysfunction and alcohol-related liver disease (MetALD) is characterised by the coexistence of metabolic dysfunction and moderate alcohol intake. In this population, the impact of specific drinking patterns and history, particularly persistent binge drinking (PBD) history, on disease severity and prognosis remains unclear. AIM:To evaluate the association between a history of PBD and the risks of advanced fibrosis and all-cause mortality in individuals with MetALD. METHODS:We analysed data from NHANES 1999-2016, including adults with MetALD. PBD is defined as having had a history of drinking ≥ 4 (women) or 5 (men) alcoholic beverages almost every day, based on ALQ150/151 questionnaires. Advanced fibrosis was defined as FIB-4 > 2.67, and mortality outcomes were obtained from the National Death Index. Multivariable logistic and Cox regression models were used to examine the associations between PBD and both liver fibrosis and all-cause mortality, adjusting for alcohol intake and other confounders. RESULTS:Among 865 individuals with MetALD, 326 (37.7%) reported a history of PBD. Compared to those without PBD, participants with PBD had a higher risk of advanced fibrosis (adjusted OR = 2.23, 95% CI: 1.12-4.45, p = 0.023). PBD was also associated with increased all-cause mortality (adjusted HR = 1.48, 95% CI: 1.03-2.13, p = 0.035). A higher risk of overnight hospitalisation in PBD supports these findings (adjusted OR = 1.88, 95% CI: 1.15-3.06, p = 0.012). CONCLUSION:PBD history is associated with increased risks of advanced fibrosis and mortality in MetALD. Clinical assessment should include an evaluation of drinking patterns and history, particularly persistent binge drinking.

OBJECTIVES:To investigate the causal relationship between gut microbiota, T-cell function, and the risk of colorectal cancer. METHODS:Gut microbiota data from the MiBioGen database and T-cell and colorectal cancer data from publicly available GWAS datasets were obtained for analyzing the causality between gut microbiota, T-cell subsets, and the risk of colorectal cancer with two-sample Mendelian randomization (MR) analyses, using inverse variance weighting as the primary analytical method supplemented with MR-Egger, weighted median, simple mode, and weighted mode methods. Horizontal pleiotropy was assessed using MR-PRESSO and MR-Egger regression. Cochran's Q test was used to evaluate heterogeneity, and sensitivity analysis was performed using the leave-one-out method. RESULTS:In the Forward MR analysis of gut microbiota and T cells, 11 gut microbiota species showed causal relationships. Six of these species exhibited positive correlations with T cells, including Prevotella7 (P=0.003), Ruminococcaceae UCG011 (P=0.033), Ruminococcaceae UCG004 (0.010), Ebacterium brachy group (P=0.005), Lachnospiraceae FCS020 group (P=0.028), and Coprobacter (P=0.033), and the remaining 5 species showed negative correlations with T cells. Forward MR analysis of T cells and colorectal cancer suggested that CD25++CD45RA-CD4+ non-regulatory T cells were negatively correlated with colorectal cancer risk (IVW: OR=0.935, 95% CI: 0.878-0.995; P=0.035). The analysis of gut microbiota and colorectal cancer suggested that 11 gut microbiota species were causally associated with colorectal cancer, and 6 of them (Eubacterium xylanophilum group, P=0.039; Selenomonadales, P=0.014; Negativicutes, P=0.014; Bifidobacteriaceae, P=0.048; Bifidobacteriales, P=0.048; and Coprococcus1, P=0.033) showed positive correlations and the remaining 5 showed negative correlations. CONCLUSIONS:Coprobacter spp. and Eubacterium xylanophilum group spp. are causally associated with both T cell activity and colorectal cancer risk, and the former bacteria induce inactivation of CD25++CD45RA-CD4+ non-regulatory T cells to promote colorectal cancer progression, whereas the latter bacteria promote CD25++CD45RA-CD4+ non-regulatory T cell activity to inhibit colorectal cancer development.

Metabolic dysfunction-associated steatotic liver disease (MASLD), as the hepatic manifestation of metabolic syndrome, is characterized by hepatic steatosis, inflammation, and fibrosis. Accumulating evidence suggests that metabolic reprogramming, particularly enhanced glycolysis, plays an important role in the pathogenesis of MASLD. Dysregulated glycolysis not only supplies energy and biosynthetic precursors for multiple hepatic cell types but also directly contributes to lipid accumulation, inflammatory activation, fibrogenesis, and even hepatocellular carcinogenesis. This review summarizes the mechanisms underlying glycolytic reprogramming in MASLD and examines the distinct roles of glycolytic alterations in various cell types, including hepatocytes, macrophages, hepatic stellate cells (HSCs), T cells, and bile duct epithelial cells (BECs). Furthermore, glycolysis mediates the crosstalk between different cell types and organ types in MASLD, modulating cytokine production and activating pro-inflammatory and pro-fibrotic signaling pathways. Finally, this article discusses the potential and challenges of targeting key glycolytic enzymes - such as hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase (PFKFB3) and Pyruvate kinase M2 (PKM2) - as therapeutic strategies, offering new perspectives for alleviating the disease burden of MASLD.

OBJECTIVE:To evaluate the effects of Sanren Decoction (SRD) on metabolic dysfunction-associated steatotic liver disease (MASLD) induced by high-fat diet (HFD) based on the protective effect of hepatocyte mitochondrial function. METHODS:Thirty-six male C57BL/6 mice were randomly assigned to 4 groups using stratified sampling based on body weight, including control, HFD, low-dose (10.09 g/kg) and high-dose (20.18 g/kg) SRD groups (n=9). MASLD model was induced in mice via a 16-week HFD. Liver histopathology was assessed using haematoxylin and eosin (HE) and Oil red O staining, while hepatic triglycerides (TG), serum alanine aminotransferase (ALT), fasting blood glucose (FBG), and fasting serum insulin levels were measured using commercial kits. Hepaticmetabolic profiling were analyzed and differential metabolite analysis was performed using partial least squares discriminant analysis and Kyoto Encyclopedia of Genes and Genomes pathways. Mitochondrial microstructure was assessed by electron microscopy. The protein expressions of respiratory chain complexes I-V were determined by Western blot analysis, while the activities of complexes I and II were measured using commercial kits. RESULTS:In the HFD-induced MASLD model, 4-week SRD treatment improved hepatic steatosis, inflammation, and hepatocyte ballooning (P<0.05). High-dose SRD treatment significantly reduced hepatic TG, ALT levels, and improved insulin sensitivity (both P<0.05). Low-dose SRD significantly reduced hepatic TG and FBG (P<0.05). Metabolomic analysis showed that differential liver metabolites were enriched in tricarboxylic acid cycle (TAC) pathway in mice of high-dose SRD and HFD groups. Electron microscopy showed that high-dose SRD improved mitochondrial morphology and enhanced adenosine triphosphate production and fatty acid oxidation activity (both P<0.05). Additionally, high-dose SRD significantly decreased the contents of hydrogen peroxide and malondialdehyde in liver tissue and increased the content of superoxide dismutase (both P<0.05). Treatment with high-dose SRD resulted in a significant increase in both protein expression and activity of mitochondrial complex II. Additionally, high-dose SRD enhanced the protein expression of mitochondrial complex I (both P<0.05). CONCLUSION:SRD exhibited hepatoprotective effects in MASLD, improving liver morphology, metabolism, and mitochondrial function, suggesting its potential as a therapeutic strategy for MASLD.