Iron-based contrast agents (IBCAs) have potential as alternatives to Gd-based contrast agents (GBCAs), intending to address the long-term safety concerns associated with gadolinium. We investigated [Fe-EOB-tCDTA] as a potential alternative to [Gd-EOB-DTPA]2- for liver magnetic resonance imaging (MRI). [Fe-EOB-tCDTA] was synthesized by reacting the monoanhydride of tCDTA with 4-ethoxybenzylamine followed by iron chelation. Its kinetic stability was spectrophotometrically evaluated using a zinc stress test. The T1 relaxivity was measured in water and serum at 1.41 T, 37 °C and 3 T, 23 °C. Cellular cytotoxicity against liver-derived BRL-3A cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. The uptake of [Fe-EOB-tCDTA] by liver cells was investigated using LA-ICP-MS, in competition with [Gd-EOB-DTPA]2-. T1 contrast effects in BALB/c mice were evaluated by DCE-MRI. [Fe-EOB-tCDTA] exhibited higher kinetic stability than [Fe-(tCDTA)]- and demonstrated a r1 of 1.94 and 2.45 mM-1s-1 at 1.4 and 3 T in serum. No significant differences in the short-term cytotoxicity were observed between [Gd-EOB-DTPA]2- and [Fe-EOB-tCDTA]. [Fe-EOB-tCDTA] inhibited [Gd-EOB-DTPA]2- uptake in BRL-3A liver cells. [Fe-EOB-tCDTA] (0.2 mmol/kg) demonstrated a comparable blood peak RE% compared to [Gd-DO3A-butrol] (0.1 mmol/kg). However, RE of [Gd-EOB-DTPA]2- in liver at a clinical dose was significantly higher than that of [Fe-EOB-tCDTA] at both injection doses. [Fe-EOB-tCDTA] provides comparable blood enhancement to [Gd-DO3A-butrol] and exhibits hepatobiliary excretion like [Gd-EOB-DTPA]2- but without a comparable liver contrast. [Fe-EOB-tCDTA] may serve as an alternative to nonspecific GBCAs, particularly for patients with renal insufficiency and a contraindication to GBCAs. This study reports on the synthesis and in vitro and in vivo characterization of the iron complex [Fe-EOB-tCDTA]. This complex demonstrates strong blood contrast and liver excretion, though it lacks strong liver contrast. This complex could serve as an extracellular contrast agent, particularly for patients with reduced kidney clearance. [Fe-EOB-tCDTA] was synthesized as a novel iron-based MRI T1 contrast agent with liver excretion. [Fe-EOB-tCDTA] generated a strong blood enhancement effect and exhibited hepatobiliary excretion. Negative charge is crucial to the hepatic uptake and long-lasting liver enhancement.
Leptospirosis, caused by pathogenic Leptospira species, is a globally significant zoonotic disease with high morbidity and mortality. However, the organs or cells mainly involved in capturing circulating leptospires and the related mechanisms remain poorly understood. In this study, we firstly proved that the liver was the primary organ that captured leptospires during the very early intravascular phase of infection in mice. Then, we used unbiased flow sorting of leptospires-positive cells and intravital microscopy of mice infected with leptospires, and found that liver macrophages were the main leptospires-capturing immune cells. The depletion of liver macrophages abolished the ability of liver to capture leptospires and prompted leptospiral spread in other organs. The C-type lectin receptor signaling pathway and Clec4d were identified as the differential pathways and gene through RNA-seq analysis, respectively. The ectopic expression of Clec4d in HEK-293T cells or treatment with a Clec4d inhibitor, mannan proved that Clec4d functioned as a capture receptor of leptospires. Mechanistically, the transcription factor CCAAT/enhancer-binding protein beta (C/EBPβ) was activated and directly bound to the promoter of Clec4d to promote the expression of Clec4d in liver macrophages, thereby enhancing leptospiral capture. Mice treated with C/EBPβ inhibitor showed a significant inhibition of liver macrophages in capturing leptospires and increased leptospiral load in other organs. Our findings identify a novel mechanism by which the liver macrophages restrict leptospiral dissemination through C/EBPβ-Clec4d axis, and suggest a therapeutic strategy to prevent leptospiral dissemination through enhancing liver macrophages functions.
Inhibiting the activation of hepatic stellate cells (HSCs) represents a key therapeutic strategy for alleviating liver fibrosis induced by schistosomiasis. Diverse cell populations secrete pro-inflammatory cytokines and chemokines, which induce HSC activation and thereby promote hepatic fibrosis progression. Tristetraprolin (TTP) exerts a pivotal role in the post-transcriptional regulation of pro-inflammatory cytokines by either accelerating mRNA degradation or suppressing translation, a regulatory mechanism closely associated with the pathogenesis of various hepatic disorders. However, the pathological roles of TTP in Schistosoma japonicum-induced liver fibrosis remain largely elusive. Here, we report that TTP is upregulated in the liver during S. japonicum-induced liver fibrosis, and its overexpression markedly ameliorates this fibrotic pathology in vivo. We further identify that TTP negatively regulates TGF-β1 mRNA stability by promoting N6-methyladenosine (m6A) RNA methylation, thereby inhibiting HSC activation. Mechanistically, TTP enhances transcription of the WT1-associated protein (WTAP) gene through its interaction with SMAD2/3. Furthermore, treatment with an m6A RNA methylation inhibitor confirms that TTP-mediated protection against S. japonicum-induced liver fibrosis, an effect associated with increased m⁶A RNA methylation in vivo. Thus, our findings uncover a novel and critical role of TTP in exerting its anti-fibrotic function via the WTAP/m6A epitranscriptomic machinery in the pathogenesis of S. japonicum-induced liver fibrosis. This finding provides a rationale for the therapeutic targeting of TTP-mediated m6A RNA methylation in S. japonicum-induced liver fibrosis.
Preoperative chemotherapy increases resectability in patients with resectable colorectal liver metastasis who undergo curative hepatectomy; however, there is no consensus regarding patient selection. The aim of this study was to identify high-risk patients with synchronous colorectal liver metastasis and compare the outcomes of preoperative chemotherapy with those of upfront surgery. This multi-institutional, retrospective study enrolled patients who underwent their first liver resection for synchronous colorectal liver metastasis from January 2010 to July 2019. We retrospectively studied the clinicopathological features and prognoses of synchronous colorectal liver metastasis subjected to curative liver resection. Surgical outcomes were compared between the preoperative chemotherapy and upfront surgery groups. We analyzed 215 patients categorized into the preoperative chemotherapy (n=111) and upfront surgery (n=104) groups. The distribution of primary colorectal cancer locations significantly differed, and the preoperative chemotherapy group had more and larger tumors than did the upfront surgery group. There were no significant between-group differences in recurrence-free survival and overall survival. Primary lymph node metastasis ≥N2, tumor burden score >5, positive carbohydrate antigen 19-9 on treatment, and the Kirsten rat sarcoma viral oncogene homolog mutation were independent risk factors for recurrence-free survival. The recurrence-free survival rate was significantly higher in the preoperative chemotheapy group than in the upfront surgery group for patients with ≥2 risk factors. Our study suggests new criteria for identifying high-risk patients with synchronous colorectal liver metastasis and shows that preoperative chemotherapy may be an effective option for high-risk patients with ≥2 risk factors.
Coronavirus (CoV) disease 2019 (COVID-19) deteriorates existing hepatopathies, such as liver cirrhosis and metabolic-associated syndrome liver disease (MASLD), which in turn increases the risk of serious complications. The pathophysiology of COVID-19 includes inflammation, proinflammatory cytokine storms, oxidative stress and fibrosis. Notably, pentoxifylline (PTX) blocks nuclear factor-κB activity, thus inhibiting the secretion of proinflammatory cytokines, and is an antioxidant and antifibrotic agent. The present study reported on the use of PTX in 20 patients with liver cirrhosis (13 men, 7 women) of different etiologies and in 25 patients with MASLD (16 women, 9 men) infected with severe acute respiratory syndrome-CoV-2; age range of all patients, 29-83 years. Firstly, the detection of CoV-2 was confirmed by PCR. All patients received PTX (400 mg twice daily; per os) for 8 weeks, alongside standard care provided for COVID-19 symptoms and for their liver condition. In all patients, the following inflammatory markers were assessed at the beginning and at the end of the study: C-reactive protein, D-dimer, ferritin, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase (LDH), platelet count and oxygen saturation. Statistical analysis was performed using the Wilcoxon signed-rank test. At the end of the study, no patient required admission to the intensive care unit and no patient fatalities were noted. Notable improvement was noted in seven of the eight inflammatory markers in both liver pathologies, and the only sole parameter that worsened was LDH. Notably, no serious adverse events were observed in these patients. In conclusion, PTX treatment was associated with favorable clinical outcomes and improved inflammatory marker levels in patients with liver cirrhosis and MASLD with COVID-19.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is a systemic condition associated with compromised bone integrity. Emerging evidence suggests that disturbances in hepatic vitamin D metabolism may contribute to these skeletal impairments. However, the hepatic mechanisms driving bone deterioration remain poorly defined. This study aimed to establish a human 3D in vitro model of MASLD and demonstrate that hepatic vitamin D dysregulation adversely affects bone homeostasis. Liver spheroids composed of HepaRG cells, LX-2 stellate cells, and HUVECs were stimulated with 600 µM free fatty acids (2:1 oleic: palmitic acid) to induce MASLD-like features, validated by BODIPY staining and gene expression. MASLD model induction led to downregulation of hepatic genes regulating lipid and vitamin D metabolism. ELISA confirmed significantly reduced 25-hydroxyvitamin D levels, aligning with downregulation of CYP2R1 and CYP27A1. Transcriptomic profiling of human MASLD liver biopsies validated these molecular changes. To evaluate MASLD's systemic impact on bone, THP-1-derived macrophages and SCP-1 mesenchymal stem cells were seeded onto bone scaffolds and co-cultured with spheroids. Bone scaffolds co-cultured with MASLD spheroids showed impaired mineralization and elevated expression of bone resorption markers. These findings mirror skeletal dysfunction observed in MASLD patients and suggest a mechanistic link between hepatic vitamin D dysregulation and bone pathology. This study introduces a pioneering 3D human liver-bone co-culture model that reveals MASLD-driven disruption of hepatic vitamin D metabolism as a direct contributor to bone deterioration. This 3D model develops a powerful translational platform for decoding systemic disease mechanisms and targeting the liver-bone axis therapeutically.
This study seeks to investigate the underlying mechanism of glycolytic key gene bisphosphoglycerate mutase (BPGM) in nonalcoholic fatty liver disease (NAFLD). qRT-PCR and immunohistochemistry were utilized to detect BPGM levels in clinical NAFLD samples. HepG2 cells and liver organoids were treated with free fatty acid. (FFA). The role of BPGM in NAFLD was explored at cellular, organoid, and animal levels. Metabolomics was performed to analyze differential metabolites and metabolic pathways. Furthermore, we examined the regulatory mechanisms of BPGM by HIF-1α in NAFLD. Results indicated that high expression of BPGM in NAFLD samples was correlated with NAFLD progression. Moreover, Severe group had higher BPGM expression than Mild group. FFA treatment induced time-dependent steatosis and BPGM upregulation in HepG2 cells and liver organoids, whereas BPGM knockdown attenuated lipid accumulation, cellular injury, and oxidative stress. At the animal level, knockdown of BPGM reversed high-fat diet (HFD) induced lipid accumulation and liver tissue injury. Metabolomics studies showed significant changes of metabolic pathways including glycolysis/gluconeogenesis and pyruvate metabolism. Verification experiment showed FFA increased pyruvic acid levels, and knockdown of BPGM decreased pyruvic acid levels. Pyruvic acid further reversed the changes in NAFLD progression caused by BPGM knockdown at the cellular and organoid levels. Finally, HIF-1α regulated the expression of BPGM in NAFLD. Together, our findings suggest that BPGM contributes to abnormal glucose metabolism and promotes hepatic steatosis, thereby driving NAFLD progression.
Oral delivery of bioactive proteins remains a challenging area, as it demands effective gastric protection and intestinal site-specific release. We design two intestinal site-specific microgels via Fe3 +-COO- coordination or STMP-conjugated cross-linking of eco-friendly and biocompatible Artemisia sphaerocephala Krasch. polysaccharides (ASKP) extracted from desert sand-fixing plant; these microgels release lactoferrin (Lf) precisely in the small intestine or colon on-demand. Both microgels protect Lf from gastric digestion, preserve the Lf receptor (LfR)-binding domain, and facilitate efficient LfR-mediated endocytosis of Lf by intestinal epithelial cells. Small intestine-targeted microgels regulate Lf metabolism, boost Lf bioavailability by ≈6-fold, and drive marked hepatic Lf accumulation. These effects ameliorate alcoholic liver injury via the Nrf2-mediated antioxidant pathway and CPT1A-amplified fatty acid β-oxidation after oral uptake. Colon-targeted microgels alleviate ulcerative colitis by suppressing the TLR4/MyD88/NF-κB inflammatory axis and restoring gut microbiota homeostasis. Notably, this is the first demonstration of site-specific intestinal delivery achieved by using different cross-linking chemistries on the same ASKP backbone, enabling distinct gut-regional specific therapeutic actions. This work establishes food-grade microgel platforms as promising intestinal site-specific delivery systems for spatiotemporally controlled, on-demand delivery of bioactive proteins.
Elevated bilirubin serves as a subtle yet early indicator of liver dysfunction, highlighting the need for a rapid, accessible, and reliable point-of-care solution. To the best of our knowledge, this study presents the first portable electrochemical platform for point-of-care bilirubin sensing for liver diagnostics. The portable device features onboard power management and a customized analog front end with precise signal acquisition, conditioning, and filtering to support Cyclic Voltammetry (CV), Differential Potential Voltammetry (DPV), and amperometric techniques for the electrochemical detection of any analyte. Herein, the optimised gold nanoparticles/SPCE enables effective bilirubin adsorption and subsequent oxidation at ∼0.35 V. An analyte-selective nafion-chitosan membrane suppressed interferences, achieving a 0.3 μM detection limit at pH 7.4. The on-device detection was evaluated by utilising the standard electrochemical setup, followed by high selectivity, repeatability, and reproducibility. Real-sample testing revealed ∼9.5% deviation from clinical values, confirming strong agreement with conventional methods. The on-device bilirubin detection through the amperometry technique delivers results both on-device and via a wireless user interface. In the future, the developed device can potentially detect other bioanalytes with minor modifications in terms of suitable chemistry.
Colorectal cancer (CRC) is a leading malignancy, and colorectal liver metastases (CRLM) are common. Accurate detection is critical for staging and treatment planning. Although contrast-enhanced MRI is the reference standard, its use is limited by cost, longer examination time, and contraindications such as renal impairment or prior severe allergic reactions. This study aimed to evaluate the diagnostic performance of an abbreviated non-contrast liver MRI protocol for detecting CRLM and to compare performance between experienced and in-training readers. In this retrospective study, 87 patients with CRC who underwent liver MRI between January 2014 and March 2024 were included. The abbreviated non-contrast protocol comprised T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), and diffusion-weighted imaging (DWI). Two independent readers evaluated the images using a 4-point confidence scale, which was dichotomized for analysis. Sensitivity, specificity, predictive values, and accuracy were calculated, and interobserver agreement was assessed using weighted kappa. Contrast-enhanced MRI served as the reference standard. The abbreviated non-contrast MRI demonstrated high diagnostic performance. Sensitivity and specificity were 94.1% and 88.7% for the abdominal radiologist, and 91.2% and 96.2% for the oncologic imaging fellow. The area under the receiver operating characteristic curve (AUROC) was 0.93 and 0.94, respectively, with no significant difference between readers (p = 0.494). Interobserver agreement was substantial (weighted κ = 0.74). Non-contrast MRI demonstrated reliable diagnostic performance with substantial interobserver agreement for CRLM detection, suggesting its potential clinical applicability. While contrast-enhanced MRI remains the standard, non-contrast MRI may be a feasible alternative in select cases. Further large-scale validation is necessary before broader clinical adoption.
metabolic dysfunction-associated steatotic liver disease (MASLD) is one of the leading causes of global liver morbidity and liver transplantation. In this context, dietary patterns emerge as a key therapeutic tool for modulating the cardiometabolic and hepatic profiles in this condition. to evaluate the effectiveness of therapeutic approaches based on dietary patterns on the cardiometabolic and hepatic factors in adult individuals diagnosed with MASLD. a comprehensive search was conducted in PubMed, EMBASE, and Web of Science from inception to October 1, 2025. Eligibility criteria were restricted to randomized controlled trials (RCTs). Furthermore, the Risk of Bias 2 (RoB-2) tool was applied for the assessment of bias. a total of 11 RCTs were selected, representing a total population of 737 adult participants. The effectiveness of Mediterranean Diet (MD) prescriptions was evaluated in 36.4 % (n = 4) of the studies, while 27.3 % (n = 3) assessed time-restricted eating (TRE) patterns. The evidence analyzed suggests that metabolic improvements were concentrated in dietary patterns such as the MD, with the DASH diet reporting marked cardiovascular effects. Our results suggest that these patterns not only act on hepatic fat likewise, mitigate concomitant cardiovascular risk factors, with moderate evidence particularly evident in relation to hepatic enzymes. dietary patterns characterized by a high intake of fiber, polyphenols, and polyunsaturated fatty acids, such as the MD and DASH, demonstrate high effectiveness in the treatment of MASLD. Future dietary interventions should compare approaches involving additional olive oil supplementation or specific combinations of polyunsaturated fatty acids.
Sepsis is characterized by an imbalance between excessive oxidative stress and endogenous antioxidant responses, leading to multi-organ dysfunction. The aim of this study was to investigate the protective effect of argan oil (AO) against lipopolysaccharide (LPS)-induced oxidative stress, with particular focus on peroxisomal-related antioxidant implications in liver, brain, kidney and heart. Phenolic and pigment contents (chlorophylls and carotenoids) of AO and olive oil (OO) were quantified, and antioxidant capacities determined in vitro using DPPH, ABTS, and FRAP assays. Mice were supplemented daily with AO or OO for 28 days prior to challenge with intraperitoneal injection of LPS (5 mg/kg) to induce acute oxidative stress. Although OO exhibited a higher phenolic content and more potent in vitro antioxidant activity across all assays, in vivo outcomes noted tissue-specific antioxidant efficacy. Antioxidant enzyme activities (superoxide dismutase, catalase, glutathione peroxidase) and glutathione levels as well as lipid peroxidation (malondialdehyde) were measured in the four target organs. LPS challenge initiated marked alterations in antioxidant defenses and increased oxidative damage in a tissue-dependent manner, with liver exhibiting the greatest response. AO supplementation efficiently mitigated LPS-induced redox imbalance, restoring antioxidant enzyme activities and limiting lipid peroxidation across organs, whereas OO displayed more variable and organ-dependent effects. These findings demonstrate that AO exerts significant in vivo antioxidant protection against LPS-induced oxidative stress despite lower in vitro antioxidant capacity than OO. Data show importance of tissue-specific and peroxisome-associated mechanisms in modulating oxidative stress responses and support AO as a promising nutritional strategy for attenuating sepsis-related organ dysfunction.
Aspergillus species are opportunistic fungi that can cause severe, potentially life - threatening infections, particularly in immunocompromised individuals. Invasive aspergillosis most commonly affects the lungs (70 % -90 % ) but may also involve other organs, including the central nervous system (10 % -25 % ), heart (5 % -10 % ), kidneys (5 % -10 % ), liver and spleen (5 % -8 % ), and gastrointestinal tract (2 % -5 % ). The incidence of aspergillosis is markedly increased among solid-organ transplant recipients due to prolonged immunosuppression. Thyroid involvement is extremely rare ( <1 % ) and is most often reported in autopsy studies. Infection of the thyroid gland may occur through hematogenous dissemination, direct extension from adjacent structures, or iatrogenic routes. Herein, we describe a liver transplant recipient who developed thyroid aspergillosis, emphasizing the importance of considering fungal infections in the differential diagnosis of thyroid lesions in immunosuppressed patients.
Prevention remains a key strategy to address the growing burden of metabolic dysfunction-associated steatotic liver disease (MASLD), highlighting the importance of exploring modifiable risk factors. Accumulating evidence suggests a close link between physical frailty and MASLD. However, how frailty interacts with metabolic syndrome to affect MASLD and the causality and direction of the association remain largely unknown. Leveraging data from 405,224 UK Biobank participants with a 13.65-year follow-up, we found that physical frailty was associated with an increased risk of clinically diagnosed MASLD and exacerbated the adverse effect of metabolic syndrome on MASLD incidence, implying that frail people may be more vulnerable to this disease because of metabolic syndrome. Mendelian randomization provided evidence for a potential causal effect of physical frailty on MASLD but not the reverse direction. Moreover, the metabolome-wide association analysis revealed widespread associations of plasma metabolites with both frailty and MASLD, suggesting a shared metabolomic foundation between them. Some metabolites, including fatty acids and triglyceride-rich lipoprotein biomarkers, partially explained the frailty-MASLD relationship, indicating a potential metabolomic mechanism. If confirmed in further studies, frailty screening may help identify high-risk individuals and inform early prevention for MASLD, especially for those with metabolic syndrome.
The current study aimed to evaluate the modulatory effects of chemically prepared and Rhus coriaria (sumac) green-synthesized zinc oxide nanoparticles (ZnO-NPs) on circulating neutrophils and the histological structure of liver in carbon tetrachloride (CCl4) treated rats. CCl4 rats showed biochemical changes and histopathological damage, together with increased neutrophil markers [myeloperoxidase (MPO) and citrullinated histone (H3cit)] as well as decreased neutrophils' phagocytic activity and miR-223 expression compared to control. ZnO-NPs treatment was significantly associated with biochemical and histopathological improvement, decreased neutrophil markers, and increased both phagocytic activity and miR-223 expression. In addition, green-synthesized ZnO-NPs were associated with greater histopathological improvement and a significant increase in miR-223 expression in comparison to chemically prepared ZnO-NPs. It can be concluded that ZnO-NPs may have immunomodulatory effects on circulating neutrophils that have a biological impact on disease progression. In addition, green-synthesized ZnO-NPs may be more efficient than chemically synthesized ones at providing a greater modulatory effect on circulating neutrophils and better hepatoprotective effects.
Prolyl hydroxylase domain enzyme 1 (PHD1) is a key regulator of hypoxic adaptation and metabolic homeostasis, playing an important role in tissue damage and repair. To enable precise pharmacological interrogation of PHD1 function, we developed the first PHD1 degrader using proteolysis-targeting chimera (PROTAC) technology. Our lead compound, SH-26, a cereblon (CRBN)-recruiting PROTAC, induced PHD1 degradation in a concentration-, time-, and ubiquitin-proteasome system (UPS)-dependent manner across multiple cell lines. In an acetaminophen (APAP)-induced acute liver injury (ALI) model, SH-26 demonstrated protective effects, attenuating hepatic inflammation and necrosis without detectable cytotoxicity. Mechanistically, SH-26-mediated PHD1 degradation attenuated APAP-triggered reactive oxygen species (ROS) accumulation, mitochondrial dysfunction, and NLRP3 inflammasome activation, leading to robust in vivo protection against ALI. Collectively, our work identifies SH-26 as the first effective PHD1 degrader and demonstrates its utility as a chemical tool to dissect the pathological role of PHD1 in ALI.
This review explores the metabolic pathways dysregulated in both atherosclerotic cardiovascular disease (ASCVD) and metabolic dysfunction-associated steatotic liver disease (MASLD), focusing on lipid, carbohydrate, amino acid, and energy metabolism and the specific alterations within major contributing cell types. In the setting of metabolic syndrome, lipid and carbohydrate overload impair hepatic metabolism, resulting in the accumulation of lipotoxic species and ensuing cellular damage, inflammation, oxidative stress, and cardiovascular consequences. Amino acid metabolism is emerging as a key regulator of cell fate and function in both MASLD and ASCVD. Mitochondrial dysfunction and cellular stress promote a pseudo-Warburg effect, shifting cells from efficient oxidative phosphorylation to anaerobic glycolysis and impairing homeostasis. Emerging therapies targeting hepatic metabolism to reduce cardiovascular risk and MASLD burden hold promise for future dual treatments. MASLD and ASCVD arise from common metabolic derangements that converge on shared cellular and molecular pathways. Defining these cross-tissue mechanisms may enable the development of integrated therapeutic approaches aimed at jointly mitigating hepatic and vascular injury, thus redefining treatment paradigms in cardiometabolic disease.
To investigate the clinical characteristics and prognosis of patients with biliary atresia (BA) who undergo reoperation after initial Kasai portoenterostomy, and to evaluate the feasibility and clinical value of the reoperation. We retrospectively analyzed the clinical data of 19 patients with BA who underwent reoperation at Beijing Tsinghua Changgung Hospital from August 2022 to September 2025. Data collected included age at initial Kasai, jaundice clearance status after the first surgery, preoperative liver function and imaging findings, and reoperation techniques. Follow-up was conducted via outpatient visits, telephone calls, or WeChat. The primary outcome was anicteric native liver survival (total bilirubin < 2.0 mg/dL). Clinical characteristics and prognosis were analyzed. The median age at the initial Kasai and reoperation was 59 and 320 days. Of the 19 patients, 12 achieved jaundice clearance after the first surgery. Preoperative imaging before reoperation revealed intrahepatic cystic anechoic areas in 12 patients. Surgical techniques included intrahepatic cyst-jejunostomy (n = 8), re-anastomosis after resection of residual fibrous mass (n = 8), and hepatic duct enlargement plasty with re-anastomosis (n = 3). At a median follow-up of 494 days, eight patients (42.1%) maintained anicteric native liver survival. Univariate analysis identified initial postoperative jaundice clearance as the only significant prognostic factor (P = 0.013). Kaplan-Meier survival analysis showed that the two-year anicteric native liver survival rate after reoperation was 70.7% in the clearance group (median not reached), whereas the non-clearance group had a median survival of only 1.342 years, with a significant between-group difference in survival (χ²=6.572, p = 0.010). Successful initial postoperative jaundice clearance is a critical prognostic indicator for reoperation in BA. Reoperation effectively prolongs anicteric native liver survival in patients with prior clearance but recurrent jaundice, whereas it offers limited benefit to those without initial clearance, who should be prioritized for liver transplantation assessment.
Citrullinemia type I (CTLN1) is a severe urea cycle disorder caused by pathogenic variants in the ASS1 (argininosuccinate synthetase 1) gene, for which liver transplantation remains the only curative option. Here, we used prime editing to correct the Ass1fold mouse model of CTLN1. Adeno-associated virus (AAV)-mediated delivery of the PE7 prime editor with an optimized prime editing guide RNA (pegRNA) achieved 71 and 54% correction of the pathogenic Ass1 mutation in hepatocytes of neonates and juveniles, respectively. Delivery of mRNA-encoded PE7 and synthetic pegRNA via lipid nanoparticles (LNPs) resulted in 24% correction after a single 3 mg kg-1 dose in neonates and 13% after three 4 mg kg-1 doses in juveniles. All treated groups showed full normalization of survival and of blood citrulline and ammonia concentrations, with restored urea cycle function and correction of natural behavior defects. Consistent with these findings, immunostaining demonstrated restoration of wild-type-like ASS1 protein localization in functionally relevant periportal and intermediate-zone hepatocytes. Editing was confined to the liver, with minimal indel formation and off-target activity and only transient elevations in liver enzymes. In a cellular reporter system, 6 of 15 recurrent human pathogenic ASS1 mutations studied, including the most common ASS1G390R variant, were corrected with similar or higher efficiencies than Ass1fold. These findings highlight prime editing as a precise and potentially curative treatment strategy for individuals with CTLN1 and other genetic liver diseases.
MASLD is linked to dyslipidemia, but it remains unclear whether this alteration depends more on disease activity or fibrosis. To determine whether dyslipidemia is primarily associated with activity and/or fibrosis through standard lipid profile and advanced lipidomics. A cross-sectional study was conducted including patients with suspected MASLD at a single center. Intraoperative liver biopsies were obtained. Only patients with MASLD were included in the analysis. Histological activity was defined as a NAFLD Activity Score (NAS) ≥5 and fibrosis was staged according to standard criteria. Clinical, biochemical, lipoprotein, and fatty acid profiles were assessed, including nuclear magnetic resonance (NMR) spectroscopy for lipoprotein subfractions and gas chromatography for fatty acids. Multivariable linear regression models were used to evaluate independent associations of activity and fibrosis with lipid parameters. A total of 49 patients were analyzed (mean age 48 years, 46% men, 94% with BMI > 35). Liver biopsies revealed a 30.6% prevalence of activity (NAS ≥ 5) and 36.7% of fibrosis. Higher lipid levels in patients with activity were related to apolipoprotein B, non-HDL cholesterol, and triglycerides in both univariate and multivariate models. Fibrosis was not associated with any lipid parameter. Lipidomics confirmed associations of activity with VLDL+IDL cholesterol (β = 0.474, 95% CI 0.204-0.745, p = 0.001), VLDL+IDL triglycerides (β = 0.454, 95% CI 0.198-0.710, p = 0.001), and VLDL particle concentration (β = 0.409, 95% CI 0.155-0.663, p = 0.002), and showed a trend toward an inverse association with the percentage of large VLDL particles (β=-0.298, 95% CI -0.602-0.006, p = 0.054). Besides, the omega-6/triglycerides ratio was inversely associated with activity (β=-0.345, 95% CI -0.630 to -0.060, p = 0.019). Hepatic inflammatory activity showed strong associations with a non-traditional atherogenic lipidomic profile. These findings suggest that activity, rather than liver fibrosis, may be a key factor in the MASLD atherosclerotic momentum.