BACKGROUND Chronic unconjugated hyperbilirubinemia requires careful differentiation between disorders caused by uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene mutation (Gilbert's and Crigler-Najjar's syndromes) and those caused by hemolytic diseases, as treatment goals and prognoses vary significantly. Due to the clinical overlap and the complexity of current specialized testing, accurately distinguishing these two etiologies presents a major clinical challenge. AIM To establish a convenient nomogram model to distinguish chronic unconjugated hyperbilirubinemia associated with UGT1A1 gene mutation from hemolytic diseases. METHODS In this retrospective study, patients diagnosed with chronic unconjugated hyperbilirubinemia at Beijing YouAn Hospital from January 2022 to May 2025 were enrolled and categorized into the UGT1A1 mutation-associated group and the hemolytic disease-associated group. To create a nomogram, least absolute shrinkage and selection operator regression and multivariate logistic regression were used to screen for differential diagnosis factors. The performance of the nomogram was evaluated by the receiver operating characteristic curve, precision-recall curve, calibration curve, and decision curve analysis. RESULTS A total of 429 patients (357 with UGT1A1 mutation-associated group, 72 with hemolytic disease-associated group) were enrolled. Patients diagnosed from January 2022 to December 2024 were randomly divided into training (n = 265) and internal validation (n = 114) cohorts. Patients diagnosed from January 2025 to May 2025 (n = 50) were used for external validation. Four key variables - abnormality of peripheral blood smear, hematocrit, red cell distribution width standard deviation, and reticulocyte percentage - were selected to construct the nomogram. External validation yielded an area under the receiver operating characteristic curve of 0.986, sensitivity of 100%, specificity of 90%, area under the precision-recall curve of 0.938, and F1-score of 0.833. Calibration curves showed good agreement between predicted and actual outcomes. Decision curve analysis confirmed the clinical utility of this nomogram. CONCLUSION We developed an effective nomogram model for the differential diagnosis of UGT1A1 gene mutation-associated and hemolytic disease-associated unconjugated hyperbilirubinemia, which improves clinical preliminary screening.

This multicenter study evaluated serum HBV RNA as a prognostic biomarker for nucleos(t)ide analogue (NA) therapy in chronic hepatitis B (CHB), focusing on correlations with HBV DNA/HBsAg and predictive value for virological response (VR) and HBeAg seroconversion. From 2020 to 2024, 427 treatment-naive HBeAg-positive CHB patients across nine Chinese hospitals were enrolled. Longitudinal HBV RNA-DNA-HBsAg correlations were analyzed during 24-month NA therapy. Predictive performance for VR at 12 months and HBeAg seroconversion at 24 months was assessed. Non-VR patients had higher baseline HBV DNA (median 7.42 vs. 5.38 log10 IU/mL, P = 0.001) and HBV RNA (6.63 vs. 4.60 log10 copies/mL, P = 0.001). Pretreatment HBV RNA correlated with HBV DNA (r = 0.638, P = 0.001), shifting to HBsAg post-24-month treatment (r = 0.917, P = 0.0005). HBV DNA and RNA levels at 6 months independently predicted therapeutic outcomes. For VR, HBV DNA demonstrated a hazard ratio (HR) of 0.16 (P < 0.001) with an area under the ROC curve (AUROC) of 0.889, while HBV RNA showed an HR of 0.77 (P = 0.03) and AUROC of 0.754. For HBeAg seroconversion, HBV DNA yielded an HR of 0.01 (P = 0.015) with AUROC 0.900, and HBV RNA exhibited an HR of 0.72 (P = 0.03) and AUROC 0.703, underscoring their consistent prognostic value at this critical timepoint. HBV RNA dynamics transition from DNA- to HBsAg-associated correlations during NA therapy. Early declines in HBV DNA/RNA at 6 months predict VR and HBeAg seroconversion, establishing this time as a a robust and clinically practical early prediction point. HBV RNA complements existing biomarkers for optimizing CHB management.

Background Wilson disease (WD) is a rare inherited copper metabolism disorder with variable prevalence worldwide. Many regions lack reliable data, leading to underestimation of burden. WD presents with hepatic, neurological, and psychiatric symptoms, but mental health impacts and economic costs are often overlooked. Methods We systematically searched PubMed up to July 2025 for studies reporting WD prevalence, clinical features, quality of life, or economic burden. Disability-adjusted life years (DALYs) were estimated, and temporal trends were analyzed. Meta-analyses pooled epidemiological and clinical data. Forecast models predicted future prevalence and costs. Results A total of 136 studies were included. Prevalence varied, being higher in Southern Europe and parts of Asia, but lower in Northern/Eastern Europe. Rising prevalence was observed in Morocco, South Korea, Spain, and Germany, likely due to improved recognition. Forecasts suggest a gradual global increase through 2030. WD mainly affects young individuals (mean age 19.9 years), with hepatic involvement in 72% and acute liver failure in 31%. Overall mortality was 14%. Mental health complications were frequent, with depression in 47.7% and anxiety in 40%. DALYs have risen globally since the 1970s, with China, the U.S., and Poland projected to carry the heaviest future burden. Economic costs are substantial, particularly in the U.S., where annual hospitalization per patient may exceed USD 60,000. Conclusion WD imposes significant clinical, psychological, and economic burdens worldwide. Improved early diagnosis, standardized treatment, and comprehensive care are needed to reduce morbidity, enhance quality of life, and lower healthcare costs.

SARS-CoV-2 infection remains a global health threat, yet its pathogenic mechanisms are incompletely understood. Here, we show that viral nucleocapsid protein (NP) is detectable in the serum of patients with SARS-CoV-2 infection independently of viral RNA. Using virus-like particle (VLP) systems and in vitro infection models, we demonstrate that NP is secreted in a vesicle-free form via type I unconventional protein secretion (UPS) pathway. This process is regulated by NP phosphorylation and oligomerization, coordinated by viral structural proteins, and dependent on membrane components including heparan sulfate proteoglycans (HSPGs) and phosphatidylinositol 4,5-bisphosphate (PI (4,5) P2). Secreted NP is preferentially taken up by granulocytes and induces the release of inflammatory cytokines, including IL-6 and TNF-α. Our findings shed a light on the secretion mechanism of this pro-inflammatory NP, highlighting it as a potential therapeutic target for COVID-related systemic inflammation.

BACKGROUND:Sphingolipids play an important role in the development of multiple metabolic disease. However, no study explored the impact of bilirubin on sphingolipid metabolism in human. This study aims to investigate the plasma sphingolipid profiles and their association with blood lipids in mild hyperbilirubinemia (Gilbert's syndrome; GS) individuals. METHODS:This cross-sectional study enrolled 224 participants including 112 individuals with GS and 112 age- and gender-matched healthy controls. Liquid chromatography-mass spectrometry was employed to quantify 53 plasma sphingolipid metabolites in a subset of 55 GS and 55 age- and gender-matched healthy controls. OPLS-DA model was constructed using SIMCA 14.1 software to identify distinct plasma sphingolipid metabolites between two groups. RESULTS:The median age of 224 subjects was 35 years, with males comprising 29.5%. The GS group exhibited higher levels of total bilirubin and high-density lipoprotein cholesterol, along with lower levels of triglycerides (all P < 0.05). Analysis of plasma sphingolipid revealed significant differences in 14 sphingolipids between two groups. Compared to the healthy control group, the GS group had lower levels of Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/25:0), Cer(d18:1/26:0), Cer(d18:2/16:0), CerP(d18:1/22:0), HexCer(d18:1/22:0), HexCer(d18:1/24:1), HexCer(d18:1/24:0), LacCer(d18:1/16:0), as well as higher levels of CerP(d18:1/12:0), LacCer(d18:1/24:0), sphingosine-1-phosphate, and sphinganine (all P < 0.05). Correlation analysis of 110 subjects indicated that Cer(d18:1/25:0), Cer(d18:1/26:0), Cer(d18:2/16:0), CerP(d18:1/22:0), HexCer(d18:1/22:0), HexCer(d18:1/24:0) were positively correlated with total cholesterol. Additionally, sphinganine was positively correlated with high-density lipoprotein cholesterol and sphingosine 1-phosphate was negatively correlated with triglycerides (all P < 0.05). CONCLUSION:This study demonstrated that plasma ceramide levels decreased in Gilbert's syndrome, which correlates with a favorable blood lipid profile.

Despite achieving functional cure with pegylated interferon-based therapy, a significant proportion of chronic hepatitis B (CHB) patients experience recurrence, even in the presence of high anti-HBs titers. Robust predictors to identify these at-risk individuals are lacking. We aimed to develop and validate a novel model integrating viral (HBcrAg) and host immune (anti-HBc, anti-HBs) biomarkers for personalized recurrence prediction. In this ambispective cohort, 84 CHB patients with functional cure (28 recurrence [R], 56 non-recurrence [NR]) were rigorously selected via 1:2 propensity score matching and followed for ≥ 96 weeks. Serum levels of HBcrAg, anti-HBc, and anti-HBs at end-of-treatment (EOT) were quantified. Independent predictors were identified by logistic regression, and diagnostic performance was evaluated by ROC analysis. At EOT, the R group had significantly higher HBcrAg levels than the NR group (3.30 ± 1.13 vs. 2.26 ± 1.04 log10 U/mL; p < 0.001), along with lower anti-HBc levels (2.53 ± 0.77 vs. 3.22 ± 0.53 log10 IU/mL; p < 0.001) and lower anti-HBs levels (1.89 ± 0.82 vs. 2.26 ± 0.56 log10 IU/mL; p = 0.014). Multivariate analysis confirmed HBcrAg (OR = 3.48, p = 0.001) and anti-HBc (OR = 0.10, p = 0.001) as independent predictors, while anti-HBs lost significance. The combined three-marker model outperformed any single biomarker, achieving an AUC of 0.88 (95

BackgroundWhile functional cure (FC) is the elusive endpoint of chronic hepatitis B (CHB) therapy, a clear consensus on its practical expectations is lacking. To inform the development of new treatments and clinical guidelines, we conducted a nationwide survey to quantify Chinese physicians' perceptions of FC and their benchmarks for successful novel therapies.MethodsIn this cross-sectional study, we administered a detailed online questionnaire to 151 attending physicians and above with extensive experience treating CHB. A quota sampling method was employed to ensure a geographically balanced cohort representative of practice patterns across China.ResultsA total of 151 physicians were surveyed, with the vast majority (70.8%) endorsing functional cure (FC) as the ultimate treatment goal. The most valued clinical benefit of FC was the reduction in liver cirrhosis and hepatocellular carcinoma (mean score 9.6/10). Combination therapy containing Peg-IFNα was favored by 76.1% of respondents as the preferred strategy to achieve FC. Post-treatment, physicians strongly recommended a minimum of one year of follow-up and adjunct consolidation therapy to mitigate relapse risk. Critically, a consensus emerged on key benchmarks for novel therapies: a minimal acceptable FC rate of 30% and a strong preference for regimens based on Nucleos(t)ide Analogs (NAs) and/or Peg-IFNα.ConclusionsOur findings translate the collective expertise of Chinese physicians into actionable benchmarks for HBV functional cure. The consensus on a minimal 30% cure rate and a preference for combination therapy provide crucial guidance for clinical trial design and the development of novel antiviral strategies.

Nucleos(t)ide analog (NAs) treatment achieves limited HBsAg loss rates; switching to or adding Peginterferon (PegIFN) therapy may improve outcomes. This study aimed to evaluate the efficacy of adding PegIFNα-2b therapy in chronic hepatitis B (CHB) patients who are virally suppressed by NAs. 250 CHB patients on NAs treatment with HBsAg < 1500 IU/mL, serum HBV DNA < 20 IU/mL, and ALT ≤ 1.5 × ULN were enrolled. Patients continued their NAs regimen and initiated PegIFNα-2b (180 µg/week) for 48 weeks, with follow-up until week 72. In addition to routine biochemistry, HBsAg and HBV RNA levels were measured at each visit; HBcrAg was assessed at baseline, week 12, and week 24. 214 patients completed 48 weeks of PegIFNα-2b therapy, and 34.6

Background and aims: The impact of metabolic dysfunction-associated steatotic liver disease (MASLD) on pegylated interferon-alpha (PEG-IFNα)-induced HBsAg clearance remains uncertain. We investigated whether MASLD severity and cumulative metabolic burden were associated with HBsAg clearance in HBeAg-negative chronic hepatitis B (CHB) patients with low baseline HBsAg. Methods This multicenter cohort study included HBeAg-negative CHB adults with low baseline HBsAg (≤ 1500 IU/mL) and suppressed/low-level HBV replication completing 48 weeks of PEG-IFNα therapy. Patients were classified as having CHB without steatosis or with MASLD (mild, n = 68; moderate, n = 40; severe, n = 99) based on controlled attenuation parameter, metabolic risk factors, and alcohol exclusion. The primary endpoint was week-48 HBsAg clearance. Results Among 526 patients, 206 (39.2%) achieved HBsAg clearance by week 48. While overall clearance did not differ between non-steatosis and MASLD groups (40.8% vs. 36.7%, P = 0.354). However, severe MASLD was associated with a lower HBsAg clearance rate than no hepatic steatosis (24.2% vs. 40.8%, adjusted P = 0.018), whereas clearance rates were 44.1% and 55.0% in patients with mild and moderate MASLD, respectively. After inverse probability of treatment weighting and multivariable Cox adjustment, severe MASLD remained independently associated with reduced HBsAg clearance (adjusted HR = 0.539, 95% CI: 0.323–0.899, P = 0.018). Moreover, ≥ 3 cardiometabolic risk factors predicted lower clearance (adjusted HR = 0.382, 95% CI: 0.164–0.888, P = 0.025). Conclusions Severe MASLD and high cumulative metabolic burden, rather than MASLD per se, were associated with impaired PEG-IFNα-induced HBsAg clearance in HBeAg-negative CHB patients with low baseline HBsAg, aiding selection for interferon-based functional cure strategies.

Aim: To investigate the clinical relevance of autoimmune phenomena (AP), including autoantibody positivity and elevated serum immunoglobulins, in patients with Wilson's disease (WD), particularly with respect to disease severity and prognosis. Methods: We retrospectively enrolled treatment-na & iuml;ve WD patients (Leipzig score >= 4) and classified them as WD with AP (AP-WD) or WD without AP (NAP-WD) based on autoantibody positivity (titer >= 1:100) and/or immunoglobulin G (IgG) above the upper limit of normal. Baseline laboratory data, clinical complications, and liver histopathology were compared. Patients received standard anti-copper therapy and were followed longitudinally to evaluate the impact of AP on liver-related outcomes. Results: Eighty-six treatment-na & iuml;ve WD patients were included (48 AP-WD, 38 NAP-WD). At baseline, AP-WD patients exhibited more severe laboratory and clinical features, including lower platelet counts, reduced albumin, prolonged international normalized ratio, higher aspartate aminotransferase-to-platelet ratio index (APRI), Model for End-Stage Liver Disease (MELD)/Pediatric End-Stage Liver Disease (PELD), and Child-Pugh scores, and greater prevalence of ascites (all P < 0.05). Histopathological analysis demonstrated increased plasma cell infiltration and heightened portal inflammatory activity in the AP-WD group (P < 0.05). Longitudinal follow-up revealed that the presence of AP was independently associated with an increased risk of adverse liver-related events, including liver transplantation, or death. Conclusion: AP is common in WD and correlates with more severe hepatic dysfunction and poorer long-term outcomes. Screening for autoantibodies and IgG levels in newly diagnosed WD patients may provide important prognostic insight and facilitate early risk stratification, guiding tailored monitoring and management strategies.

Introduction:Low-level viremia (LLV) is associated with the progression of liver fibrosis and a high risk of hepatocellular carcinoma in patients with chronic hepatitis B (CHB). The present study aimed to compare the efficacy between nucleos(t)ide analogs (NAs) therapy and combination therapy of NAs and pegylated interferon-α (pegIFN-α) in entecavir (ETV)-treated CHB patients with LLV. Methods:This was a retrospective cohort study. ETV-treated CHB patients with LLV were included and divided into the NA group and the NA+IFN group. The NA group comprised patients switching to tenofovir alafenamide fumarate, whereas the NA+IFN group comprised those adding on pegIFN-α additionally. We compared changes in HBV markers and complete virological response (CVR) between the two groups. Results:A total of 127 patients were enrolled, including 51 in NA+IFN group and 76 in NA group. In the NA+IFN group, the decline in HBsAg level from baseline (△ HBsAg) was significantly greater (-0.17 log10IU/mL vs. -0.06 log10IU/mL, P = 0.011) at week 24, and HBsAg clearance rate and △ HBsAg were significantly higher (8.9% vs. 0%, P = 0.017; -0.27 log10IU/mL vs. -0.11 log10IU/mL, P = 0.023) at week 48. The 48-week CVR rate in the NA+IFN group was 66.7% (34/51), which was comparable to 68.4% (52/76) in the NA group (P = 0.836). Conclusion:In ETV-treated patients with LLV, receiving NAs plus pegIFN-α tends to increase the effect of HBsAg clearance.

Background and Aims:Inherited metabolic liver diseases (IMLDs) have complex etiologies and vary widely in clinical presentation, with a significant overall incidence. With the advancements in diagnostic and treatment technologies, an increasing number of children with inherited metabolic diseases are surviving into adolescence and adulthood. These advancements have improved our understanding of the IMLD disease spectrum and clinical outcomes. This study aimed to analyze changes in the disease spectrum and epidemiological characteristics of inherited metabolic liver diseases (IMLD) over the past 20 years in two specialized liver disease hospitals in northern China. Methods:A retrospective analysis was conducted on IMLD cases diagnosed between January 1, 2002, and December 31, 2023, at two liver disease specialty hospitals in Beijing. Data were obtained from inpatient and outpatient hospital information systems, with diagnoses based on national and international IMLD diagnosis and treatment guidelines. Results:A total of 2,103 IMLD patients were analyzed, including 1,213 adults and 890 children. IMLD accounted for 4.58‰ of hospitalized liver disease patients during this period. The most common IMLD was Wilson's disease, comprising 68% of all IMLD cases. The number of diagnosed IMLD types increased from 15 to 32 across two 11-year periods (2002-2012 and 2013-2023). Among pediatric patients, glycogen storage disease and Alagille syndrome were more prevalent in those under one year of age, while Wilson's disease was prevalent across all age groups. In adult IMLD patients, Wilson's disease, polycystic liver disease, and hereditary hyperbilirubinemia were more frequently observed. Conclusions:Over the past 20 years, both the number of diagnosed IMLD cases and disease diversity have significantly increased, with Wilson's disease remaining the most prevalent IMLD. These findings provide valuable insights for the long-term management of IMLD patients and the allocation of healthcare resources.

Objectives: GLS4 is a first-in-class hepatitis B virus (HBV) capsid assembly modulator that inhibits HBV replication by interfering with assembly and disassembly of the virus nucleocapsid, this prospective, open-label, comparative, phase 2b trial evaluated the antiviral activity and safety of GLS4/ritonavir (RTV) combined with entecavir in hepatitis B e antigen-positive patients. Methods: 250 CHB patients were enrolled, including treatment-na & iuml;ve patients and those interrupted anti-HBV drugs for >= 6 months (Part A, n=125), and patients who had taken ETV for >= 1 year and had achieved viral suppression (Part B, n=125). Patients were randomly allocated to receive 120 mg GLS4/100 mg RTV plus 0.5 mg ETV or 0.5 mg ETV monotherapy for 96 weeks. Results: In the mid-term, in Part A (n=122), greater least-squares mean (LSM) changes from baseline were observed in the GLS4/RTV plus ETV cohort than in ETV monotherapy cohort in HBV DNA (-6.28 vs -5.72 log10 IU/ml, p=0.0005), HBsAg (-0.87 vs -0.65 log10 IU/ml, p=0.0653), HBV pgRNA (-3.83 vs -1.91 log10 copies/ml, p<0.0001); The proportions of both HBV DNA and pgRNA negative patients were 17.3% (13/75, GLS4/RTV plus ETV) and 0% (0/30, ETV monotherapy). In Part B (n=123), greater mean LSM reductions in HBsAg (-0.17 vs -0.06 log10 IU/ml, p=0.0013), HBV pgRNA (-1.61 vs -0.28 log10 copies/ml, p<0.0001) were also observed in the GLS4/RTV+ETV cohort. the proportions of both HBV DNA and pgRNA-negative patients were 71.6% (48/67, GLS4/RTV plus ETV) and 18.9% (7/37, ETV monotherapy), respectively. No patients achieved HBsAg loss at week 48. GLS4/RTV + ETV were well tolerated, the most common adverse events were elevated alanine aminotransferase levels and hypertriglyceridemia, which were reversed by temporary GLS4/RTV discontinuation. Conclusions: The primary analysis at week 48 showed that the antiviral efficacy of GLS4/RTV with ETV was clearly superior to that of ETV monotherapy. GLS4/RTV with ETV was well tolerated; further studies evaluating its safety and efficacy are ongoing. (clinical trial identifier: NCT04147208). (c) 2025 The Author(s). Published by Elsevier Ltd on behalf of The British Infection Association. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Background and Aims:Chronic hepatitis B virus (HBV)-infected patients may exhibit liver fibrosis and other pathological changes despite normal alanine aminotransferase (ALT). This study aimed to assess the efficacy and safety of tenofovir amibufenamide (TMF) in chronic HBV-infected patients with normal ALT levels. Methods:The ongoing PROMOTE study (NCT05797714) is the first prospective, multicenter, randomized, open-label, blank-controlled clinical trial involving chronic HBV-infected patients with normal ALT levels. Participants were randomized in a 1:1 ratio to receive either TMF (TMF group) or no treatment (blank control group). The primary efficacy endpoint was the proportion of participants achieving HBV DNA levels <20 IU/mL at 48 weeks. Results:A total of 197 participants were enrolled, with 95 in the TMF group and 102 in the blank control group. At 48 weeks, a significantly greater proportion of participants in the TMF group achieved HBV DNA levels <20 IU/mL compared with the control group (74.2% vs. 9.0%, P < 0.001). The TMF group demonstrated more pronounced reductions in HBV DNA (-2.63 vs. -0.22 log10 IU/mL, P < 0.001), HBsAg (-0.07 vs. -0.04 log10 IU/mL, P = 0.02), and ALT levels (-14.09% vs. 0%, P = 0.003) compared with the blank control. In the TMF group, the proportion of participants with high-normal ALT levels (20-40 IU/L) was reduced. No significant differences were observed between the groups in creatinine, glomerular filtration rate, bone turnover biomarkers, lipid profiles, or phosphorus levels. Conclusions:TMF treatment demonstrates significant efficacy in chronic HBV-infected patients with normal ALT levels and shows a favorable safety profile regarding bone, renal, and lipid parameters. The PROMOTE study is ongoing, and further results at 96 and 144 weeks are expected to provide additional insights.