The poor redissolubility of freeze-dried Spirulina protein extract (SPE) powder severely limits its application in functional foods. Based on mild and efficient SPE extraction (93% yield while preserving macromolecular subunit integrity), this study elucidated the molecular mechanism of freeze-drying-induced redissolubility loss by analyzing particle-size distribution, molecular-weight/subunit distribution, secondary structure, and intermolecular forces. After freeze-drying, SPE showed approximately 65% protein solubility, a polydisperse particle-size distribution, and a predominantly ordered secondary structure (α-helix + β-sheet > 60%). Hydrophobic interactions (25.2 mg L−1) and hydrogen bonds (48.5 mg L−1) mainly maintained the compact aggregates. Optimizing aqueous dissolution conditions alone was insufficient; even at pH 12.0, solubility reached only 87%. Intermolecular-force-targeted analysis confirmed hydrophobic interactions as the key factor limiting SPE redissolubility. Unlike high-concentration SDS (0.5 mol L−1), which caused non-specific strong disruption, SDS (0.005 mol L−1), DTT (0.005 mol L−1), and urea (2 mol L−1) disrupted aggregates through a cascade synergistic mechanism: urea broke down the hydrogen-bond network and relaxed the rigid framework, whereas DTT reduced disulfide bonds and opened molecular crosslinks. These changes exposed buried hydrophobic regions, enabling low-concentration SDS to precisely disrupt dominant hydrophobic interactions. Consequently, protein conformation shifted toward random coils (63.4%), uniformly sized soluble complexes formed, and protein dissolution rate increased to 96%. These findings provide a mechanistic basis for improving highly hydrophobic freeze-dried protein powders.

The textural properties of food colloidal systems are highly dependent on the gelling characteristics of animal-derived proteins (such as egg protein, EP) and polysaccharides (such as starch, pectin, and gums). The development of plant-based protein alternatives faces challenges such as insufficient gel strength and poor structural stability. This study focused on purified Chlorella protein extract (PCPE). To address the insufficient gel performance of PCPE, a synergistic optimization strategy of "modification combined with calcium ion regulation" was proposed. Results revealed that alkali treatment strengthened ionic interactions while weakening hydrogen bonds and hydrophobic interactions, promoting protein molecular rearrangement and resulting in a narrower and more concentrated particle-size distribution. This increased the initial storage modulus to 136 Pa, comparable to that of EP. TGase treatment catalyzed protein-protein and protein-polysaccharide covalent cross-linking, forming a dense network structure and significantly increasing gel hardness to approximately 1.5 times that of EP. Furthermore, the addition of calcium ions synergistically enhanced gel strength via salt bridge formation, further elevating the initial storage modulus to 297 Pa. However, a high calcium concentration (25 mM) led to an approximately 20% reduction in water-holding capacity for TGase-treated PCPE gels. This study elucidates the multidimensional regulatory mechanisms underlying enhanced plant-protein gelation and provides a strategy for developing sustainable plant-based gelled foods.

Low-moisture foods (LMF) such as sesame seeds are frequently contaminated by Salmonella, posing food safety risks. To overcome the increased thermal resistance of Salmonella in LMF and the quality issues with traditional thermal treatments, this study investigated the efficacy of sequential gaseous chlorine dioxide (ClO2) and 405 nm blue light against Salmonella on sesame seeds, along with effects on product quality and residues. The results showed that Salmonella remained stable on sesame seeds after short-term adaptation. Individual treatment yielded limited inactivation, with blue light achieving 1.9 log reductions after 20 min and high-concentration ClO2 (similar to 69.06 mg/L) achieving 3.7 log reductions after 60 min. In contrast, the combination of ClO2 followed by blue light generated a strong synergistic bactericidal effect, achieving more than 6.1 log reductions of Salmonella within 60 min, significantly higher than the reverse sequence (about 4.1 log reductions). The blue light exposure after ClO2 treatment not only increased the bacterial inactivation efficacy but also helped reduce ClO2 residues. The Weibull model best described the non-linear inactivation kinetics. The combined treatment reduced color bleaching, antioxidant loss, and lipid oxidation compared with ClO2 alone. Residual ClO2 was significantly reduced to below 1.3 mg/kg, meeting safety standards. SEM revealed severe bacterial membrane rupture and structural damage under the combined treatment. The sequential ClO2-blue light strategy presents a promising, safe, and quality-preserving non-thermal decontamination method for LMF.

To explore the effects of various pretreatment methods on the extraction, separation, and processing properties of Chlorella protein, freezing, freeze-drying, and spray-drying were applied. Proteins were subsequently extracted by high-pressure homogenization and separated using chitosanase combined with ethanol precipitation, followed by determination of extraction yield, purity, amino acid composition, and molecular weight distribution. The protein isolates underwent different storage conditions-refrigeration, freezing, hot-air drying, and freeze-drying-after which their functional properties were evaluated. Results indicated that spray-drying significantly improved the protein extraction rate to 67.3% (ethanol concentration was 80%) and increased purity to 48.8% (ethanol concentration was 50%), outperforming both freezing and freeze-drying while better preserving RubisCo and essential amino acids, with total amino acids reaching 74.8% and essential amino acids accounting for 40.8%. Refrigerated storage resulted in superior functional performance, yielding an emulsifying activity index (101.4 m2/g) 3.4 times higher than that of whey protein, along with significantly improved solubility (52.9%) and foaming capacity (137.5%). These findings demonstrate that spray-drying enhances extraction efficiency through effective cell wall disruption, while refrigeration maintains structural stability, reduces hydrophobic group exposure, and optimizes functional properties, providing a theoretical basis for efficient extraction and industrial application of Chlorella protein.

Chlorella pyrenoidosa is a protein-rich microalga (> 40% dry weight), and its protein extraction is challenging due to its multilayered cell wall. In this study, the protein extraction efficiency and structural disruption of high-pressure disruption (HPD), alkaline, ultrasonication, and enzymatic methods were compared. At 40 kpsi, HPD achieved 88% protein yield-outperforming alkaline (70%), ultrasonication (64%), and enzymatic (34%) methods, without evident low-molecular-weight fragmentation (based on SDS-PAGE), and maintaining a biological value of 70.1. Instantaneous pressurization fragmented cells into 0.1-10 mu m particles, releasing both intracellular proteins and cell wall components. Monosaccharide analysis indicated the concurrent release of structural sugars, with HPD extracts containing 191 g & centerdot;kg(-1) total monosaccharides (4 & times; aqueous extracts), including cell wall-specific muramic acid (14.7 g & centerdot;kg(-1)) and N-acetylglucosamine (18.4 g & centerdot;kg(-1)), alongside intracellular glucose and galactose. These findings are consistent with extensive physical disruption of the multilayered cell wall and enhanced intracellular protein release. In contrast, alkaline treatment degraded the outer sheath (N-acetylglucosamine: 17.4 g & centerdot;kg(-1)) but resulted in protein aggregation, while ultrasonication created surface pores and released protein content. Enzymatic methods (e.g., Viscozyme (R) L) selectively hydrolyzed surface polysaccharides without penetrating deeper layers. Scanning electron microscopy and particle size analysis confirmed extensive structural disintegration after HPD treatment. Overall, these results demonstrate the effectiveness of HPD under the tested conditions for promoting protein extraction from C. pyrenoidosa. The potential integration of mechanical disruption and enzymatic hydrolysis warrants further investigation to evaluate possible complementary effects and process efficiency.

Electrospinning enables edible films with high specific surface area and tunable nanostructure, yet many food-related systems still rely on synthetic carriers (e.g., polyvinyl alcohol or polyethylene oxide) or animal proteins (e.g., whey protein isolate (WPI)) and remain mechanically fragile. This study used high-purity Spirulina protein (SPP90) and pullulan to fabricate fully food-grade electrospun films and showed that SPP90 outperformed WPI in interfacial activity, rheological behavior, and spinnability, enabling compact, uniform membranes with superior texture. SPP90 exhibited an emulsifying activity index approximately twice that of WPI, foam stability of 90%, viscosity of 77,846 mPa & sdot;s and a storage modulus (G ') of 213 Pa in heat-induced gels, reflecting strong intermolecular network formation. These characteristics are associated with improved solution stability and electrospinning performance. Enrichment of hydrophobic amino acids and the formation of lamellar structures may enhance protein-polysaccharide entanglement and a dense elastic network that increased fiber strength. Compared with WPI, SPP90-based membranes displayed significantly higher hardness (23.4 N) and chewiness (16.3 N), together with finer fiber morphology (mean diameter 194.9 nm), suggesting a distinct structure-function synergy. In contrast, residual pigments and acidic polysaccharides in lower-purity Spirulina protein (SPP70) appeared to interfere with chain alignment and entanglement, producing heterogeneous fibers with defects and much lower hardness (6.4 N) and chewiness (2.0 N). These findings identify Spirulina protein as a functionally promising, clean-label alternative to animal-derived proteins and synthetic carriers, enabling structurally robust edible films with potential relevance for sustainable edible packaging applications.

Mitochondrial fission is mediated by dynamin-related protein 1 (gene name DNM1L) and fusion by mitofusins (MFN1 and MFN2) and optic atrophy 1. The role of mitochondrial dynamics in liver disease and cancer remains poorly understood. We analyzed single, double, and triple liver-specific KO mice lacking mitochondrial fission and fusion proteins using systematic analyses of mitochondrial morphology, untargeted metabolomics, RNA-seq, hydrodynamic tail vein injection of oncogenes, and human hepatocellular carcinoma samples. Liver-specific Dnm1l-KO (L-Dnm1l-KO) mice showed increased alanine aminotransferase levels and hepatic fibrosis, with spontaneous liver tumors developing by 12 to 18 months of age. L-Mfn1- and L-Mfn2-KO mice showed no significant liver damage or tumor development, although a small percentage of L-Mfn1, Mfn2 double KO mice developed tumors. Dnm1l, Mfn1, and Mfn2 triple KO (TKO) mice experienced significantly reduced liver injury and fibrosis, along with decreased spontaneous and oncogene-induced tumorigenesis. L-Dnm1l-KO mice showed increased activation of the cGAS/STING/interferon pathway and pyrimidine metabolism, which were significantly normalized in TKO mice. Deletion of hepatic cGas reduced both basal and oncogene-induced liver injury and tumor development in L-Dnm1l-KO mice. These findings indicate that mitochondrial dynamics are crucial for maintaining hepatic pyrimidine metabolism and regulating the cGAS/STING-mediated immune response to prevent liver tumorigenesis.

Background Despite surgical and intravesical chemotherapy interventions, non-muscle invasive bladder cancer (NMIBC) poses a high risk of recurrence, which significantly impacts patient survival. Traditional clinical characteristics alone are inadequate for accurately assessing the risk of NMIBC recurrence, necessitating the development of novel predictive tools.Methods We analyzed microarray data of NMIBC samples obtained from the ArrayExpress and GEO databases. LASSO regression was utilized to develop the predictive signature. We combined gene signature and clinicopathological factors to construct a clinical nomogram for estimating NMIBC recurrence in a local cohort. Finally. the biological functions and potential mechanisms of SDCBP in bladder cancer were investigated experimentally in vitro and in vivo.Results An 8-gene signature was developed, and its efficiency for predicting NMIBC recurrence was evaluated using Kaplan-Meier and time-dependent ROC curves in both training and validation datasets. Immunohistochemical testing revealed elevated levels of ACTN4 and SDCBP in recurrent NMIBC tissues. We integrated the two proteins with clinical factors to develop a nomogram model, which showed superior accuracy compared to individual parameters. Gene Set Variation Analysis and Gene Set Enrichment Analysis unveiled SDCBP exerted cancer-promoting biological processes, such as angiogenesis, EMT, metastasis and proliferation. Experimental procedures demonstrated that silencing SDCBP attenuated cell growth, glucose metabolism and extracellular acidification rate, accompanied by decreased expression of p-AKT, p-ERK1/2, LDHA and Vimentin.Conclusions The established 8-gene signature holds promise as a tool for predicting NMIBC recurrence, while targeting SDCBP may represent a potential strategy for delaying disease relapse.

Collecting tea residue mesophyll cells (TRMCs) pioneers a novel process enabling industrial-scale, cost-effective leaf protein recovery with high yields, surpassing conventional extraction barriers. However, their food applications are constrained by large particle size and poor fluidity, leading to low interfacial adsorption efficiency in emulsions. In this study, high-pressure homogenization (HPH) technology, outperforming airflow ultrafine grinding and ultrasonic crushing, was used to enhance the performance of TRMCs. The effects of different intensities of HPH treatment (100-500 bar for 1-5 min) on the basic composition, microstructure, physicochemical properties, and functional characteristics of TRMCs were investigated, and the underlying mechanism of their emulsification improvement was analyzed. The results showed that HPH treatment only slightly affected basic components (such as protein and total sugar) and the color of TRMCs but significantly reduced the particle size (D(3,2) as low as 5.1 mu m). The treatment also eliminated the aggregation and improved the dispersion stability by enhancing electrostatic repulsion. In addition, HPH treatment balanced surface hydrophilicity-hydrophobicity (contact angle increased from 46.6 degrees to 69.2 degrees), exposing hydrophobic domains and more protein and polysaccharide structures, increased the emulsification activity index by 1.3 times, and decreased creaming index by 1.7 times. Unlike traditional Pickering emulsion stabilizers requiring chemical modification or multi-step assembly, HPH-treated TRMCs achieved superior emulsification through synergistic physical fragmentation and chemical group exposure, maintaining natural biocompatibility.

Tea residue, a tea processing by-product, yields alkaline-extracted proteins with limited emulsifying capacity due to strong interactions with polysaccharides such as pectin and cellulose. A polysaccharide-targeted enzymatic strategy was developed by systematically comparing carbohydrases, proteases, and transglutaminase (TG). Selected carbohydrases (Viscozyme (R) L, pectinase, cellulase) effectively disrupted protein-polysaccharide complexes, enhancing the emulsifying activity index (EAI) by 2.4-fold compared to untreated controls (P < 0.05), and significantly outperforming proteases and TG. Mechanistic analysis showed that polysaccharide degradation reduced ionic and hydrogen bonding (>90 %), exposed hydrophobic groups (220 % increase in free sulfhydryl), and promoted beta-sheet formation (up to 56 %), facilitating dense interfacial film formation. Dialysis experiments revealed that removing small peptides and pectin fragments (<3 kDa) increased alpha-helix content (to 27 %) but impaired emulsification, while retention of oligosaccharides improved stability via viscosity and steric hindrance. This enzymatic approach offers a sustainable and structure-guided pathway to convert food processing waste into clean-label, plant-based emulsifiers, contributing to the functional valorization of agro-industrial by-products.

BACKGROUND:Little is known regarding the joint effect of multiple essential metals (EMs) on metabolic syndrome (MetS). This study aimed to investigate individual and overall correlations of EMs with MetS and its components among Chinese community-dwelling older adults. METHODS:Six urine EMs, including molybdenum (Mo), vanadium (V), selenium (Se), calcium (Ca), cobalt (Co), and magnesium (Mg), were measured using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in 2222 older adults. Logistic regression and restricted cubic spline (RCS) models were utilized to assess single EM effects. Quantile-based g-computation (Qgcomp) and Bayesian kernel machine regression (BKMR) models were applied to estimate the overall effects of multiple EMs. RESULTS:After controlling for potential confounders, Mo was significantly inversely connected to MetS (OR = 0.852, 95 % CI: 0.750 ∼ 0.967). RCS displayed a negative linear association of Mo with MetS (Pfor overall = 0.039, Pfor nonlinearity = 0.942). Both BKMR and Qgcomp models showed that EM mixture was inversely related to MetS, with Mo having the highest weight. Furthermore, Mo was inversely linked with high blood pressure, and V and Se were inversely linked with low high-density lipoprotein cholesterol (HDL-C), whereas Mg was positively linked with low HDL-C. The EM mixture was negatively correlated with high blood pressure and low HDL-C. CONCLUSIONS:Urine Mo is in a linearly inverse correlation with MetS, individually and as a mixture. The EM mixture is connected to decreased MetS risk, mainly driven by Mo, which decreases the risk of high blood pressure.

Although Spirulina protein can be effectively extracted by ultrasonic treatment, the subsequent ultrafiltration for protein purification is always inefficient. Several carbohydrases and/or ethanol were tested to improve the efficiency of ultrafiltration, and the key components that influence the purification were analyzed. The results showed that Viscozyme® L treatment enhanced the Spirulina protein extraction yield from 89 % to 99 %. Treated by Viscozyme® L, protein-polysaccharide complexes with a molecular weight of approximately 175 kDa had reduced leading to a reduction of particle size from 4800nm to 460nm. In comparison, individual carbohydrase had little effect on the crude extracts. In the subsequent ultrafiltration (100kDa MWCO), the Viscozyme® L treated crude extract had a purity of 74 % in the retentate only 10 % higher than that without enzyme. When alcohol-soluble components, including 97 % of chlorophyll, were removed by ethanol, the complex of the protein-polysaccharide complex (156 kDa) in Viscozyme® L treated crude extract was depolymerized to two fractions (protein of 132 kDa and polysaccharide of 99 kDa). This depolymerization facilitated the follow-up ultrafiltration, yielding a Spirulina protein fraction with a purity exceeding 96 %. The high-purified spirulina protein contained more than 45 % essential amino acids, which can be applied to functional foods and nutritional supplements.

This investigation examined focal adhesion kinase (FAK)'s role in trophoblast cellular processes during early-onset preeclampsia (EOPE). We analyzed FAK and its phosphorylated form (pY397FAK) expression patterns in both normal (n = 15) and EOPE (n = 15) placental tissues, including first trimester samples, using immunohistochemistry and Western blot techniques. Next, Y15 was used to inhibit FAK activity. CCK-8 detection, Western blotting, wound healing assay, Transwell assays and flow cytometry were employed to systematically evaluate FAK's impact on trophoblast cell line HTR8/SVneo. Through transcriptomic and bioinformatics analyses, we identified Rap1 as a potential downstream mediator of FAK signaling in trophoblasts. In a mouse model of preeclampsia, we found decreased expression of both FAK and Rap1 in placental tissues, supporting our in vitro findings. These results suggest that FAK may contribute to preeclampsia development by regulating trophoblast invasion and proliferation through the Rap1 signaling pathway. Our study provides insights into the molecular mechanisms underlying EOPE and identifies FAK as a potential therapeutic target for preeclampsia treatment.

Green tea residue (GTR) contains a high protein content. However, the protein in GTR can't be effectively extracted using traditional methods. Thus, a novel method using ethylenediamine tetraacetic acid (EDTA), ammonium oxalate, or Celluclast (R) 1.5 L were used to disperse leaf tissues and to collect mesophyll cells to enrich the protein. Compared with EDTA or ammonium oxalate treatment, Celluclast (R) 1.5 L treatment achieved the highest amounts of mesophyll cells, about 2.7 x 106 g- 1 of GTR. The number of collected mesophyll cells was positively and linearly correlated with the extraction rate of glucose and xylose, indicating that cellulose and hemicellulose were key components influencing cell collection. Celluclast (R) 1.5 L treatment enriched the protein content by 1.65 times in collected mesophyll cells to 50% protein content with a protein recovery of 88%, providing a novel scheme to obtain high-quality leaf protein for the food industry.

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Despite lignocellulose hindering the extraction of intracellular components, tea residue can serve as an excellent substrate for fungal fermentation owing to their lignocellulose-degrading abilities. Thus, the fermentation efficiencies of Lentinus edodes, Lentinus sajor-caju (Fr.), Flammulina filiformis, Hericium erinaceus, Pleurotus pulmonarius, and Monascus kaoliang B6 were evaluated using tea residue as a medium. P. pulmonarius and L. sajor-caju (Fr.) exhibited the fastest growth rates, with colony radii of 33.1 and 28.5 mm, respectively. M. kaoliang B6 demonstrated substantial degradation abilities for cellulose, hemicellulose, and lignin, with decolorization radii of 12.2, 0.9, and 8.5 mm, respectively. After a 9-days liquid fermentation, M. kaoliang B6 achieved the highest conversion efficiency at 27.8%, attributed to its high cellulase (191 U center dot mL- 1) and lignin peroxidase (36.9 U center dot L- 1) activities. P. pulmonarius and L. sajor-caju (Fr.) showed lower conversion rates of 8.6% and 3.8%, despite having high hemicellulase activities (67.1 and 70.9 U center dot mL- 1). Fermentation by M. kaoliang B6 resulted in a reduction of protein and total sugar content in the tea residue by 174 and 192 mg g- 1, by which the mycelium's protein and total sugar content increased by 73 and 188 mg g- 1. Co-fermentation of these three strains had little effect on the improvement of conversion efficiency, which might owe to the antagonistic interactions among the strains. Generally, utilizing tea residue for edible fungi fermentation is a sustainable process for bio-waste treatment, enabling efficient nutrient conversion under mild conditions without adding chemicals.

Although ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) has been obtained from green tea residue mesophyll cells (TRMCs), its intact release has not yet been achieved. To release RuBisCO, this study employed a combination or sequential treatments using urea, β-mercaptoethanol, sodium dodecyl sulfate (SDS), and enzymes. Factors that hindered RuBisCO release from TRMCs were investigated through SDS-PAGE analysis, protein release quantification, and electron microscopy techniques. Alkali treatment of TRMCs at 95 °C facilitated protein release, while also causing protein modification or degradation. Conversely, the combined treatment of β-mercaptoethanol with urea and/or SDS could effectively disrupt the disulfide bonds, hydrogen bonds, and/or hydrophobic interactions within the cells, leading to the release of 40% or more of the proteins. This treatment showed strong electrophoretic bands at 55 and 15 kDa, indicating that RuBisCO was completely released. No protein was released during the treatment with SDS and pepsin/papain/alkaline protease, indicating that RuBisCO was hindered by the presence of cellulose and hemicellulose. Sequential treatment with SDS and Viscozyme L dissolved TRMC lignocellulose without releasing RuBisCO, suggesting the low solubility of RuBisCO. Overall, the presence of lignocellulose in the cell wall and the low solubility of RuBisCO were identified as key factors hindering its release from the TRMCs.

Alzheimer’s disease (AD) is the most common form of progressive dementia and there is no truly efficacious treatment. Accumulating evidence indicates that impaired autophagic function for removal of damaged mitochondria and protein aggregates such as amyloid and tau protein aggregates may contribute to the pathogenesis of AD. Epidemiologic studies have implicated alcohol abuse in promoting AD, yet the underlying mechanisms are poorly understood. In this review, we discuss mechanisms of selective autophagy for mitochondria and protein aggregates and how these mechanisms are impaired by aging and alcohol consumption. We also discuss potential genetic and pharmacological approaches for targeting autophagy/mitophagy, as well as lysosomal and mitochondrial biogenesis, for the potential prevention and treatment of AD.

Epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) treatment prolongs the survival of lung cancer patients harbouring activating EGFR mutations. However, resistance to EGFR-TKIs is inevitable after long-term treatment. Molecular mechanistic research is of great importance in combatting resistance. A comprehensive investigation of the molecular mechanisms underlying resistance has important implications for overcoming resistance. An accumulating body of evidence shows that lncRNAs can contribute to tumorigenesis and treatment resistance. By bioinformatics analysis, we found that LINC00969 expression was elevated in lung cancer cells with acquired gefitinib resistance. LINC00969 regulated resistance to gefitinib in vitro and in vivo. Mechanistically, gain of H3K4me1 and H3K27Ac led to the activation of LINC00969 expression. LINC00969 interacts with EZH2 and METTL3, transcriptionally regulates the level of H3K27me3 in the NLRP3 promoter region, and posttranscriptionally modifies the m6A level of NLRP3 in an m6A-YTHDF2-dependent manner, thus epigenetically repressing NLRP3 expression to suppress the activation of the NLRP3/caspase-1/GSDMD-related classical pyroptosis signalling pathways, thereby endowing an antipyroptotic phenotype and promoting TKI resistance in lung cancer. Our findings provide a new mechanism for lncRNA-mediated TKI resistance from the new perspective of pyroptosis via simultaneous regulation of histone methylation and RNA methylation. The pivotal role of LINC00969 gives it the potential to be a novel biomarker and therapeutic target for overcoming EGFR-TKI resistance in lung cancer.

Background and Aims: The aim of the study was to investigate the role and mechanisms of tuberous sclerosis complex 1 (TSC1) and mechanistic target of rapamycin complex 1 (mTORC1) in alcohol-associated liver disease. Approach and Results: Liver-specific Tsc1 knockout (L-Tsc1 KO) mice and their matched wild-type mice were subjected to Gao-binge alcohol. Human alcoholic hepatitis (AH) samples were also used for immunohistochemistry staining, western blot, and quantitative real-time PCR (q-PCR) analysis. Human AH and Gao-binge alcohol-fed mice had decreased hepatic TSC1 and increased mTORC1 activation. Gao-binge alcohol markedly increased liver/body weight ratio and serum alanine aminotransferase levels in L-Tsc1 KO mice compared with Gao-binge alcohol-fed wild-type mice. Results from immunohistochemistry staining, western blot, and q-PCR analysis revealed that human AH and Gao-binge alcohol-fed L-Tsc1 KO mouse livers had significantly increased hepatic progenitor cells, macrophages, and neutrophils but decreased HNF4α-positive cells. Gao-binge alcohol-fed L-Tsc1 KO mice also developed severe inflammation and liver fibrosis. Deleting Tsc1 in cholangiocytes but not in hepatocytes promoted cholangiocyte proliferation and aggravated alcohol-induced ductular reactions, fibrosis, inflammation, and liver injury. Pharmacological inhibition of mTORC1 partially reversed hepatomegaly, ductular reaction, fibrosis, inflammatory cell infiltration, and liver injury in alcohol-fed L-Tsc1 KO mice. Conclusions: Our findings indicate that persistent activation of mTORC1 due to the loss of cholangiocyte TSC1 promotes liver cell repopulation, ductular reaction, inflammation, fibrosis, and liver injury in Gao-binge alcohol-fed L-Tsc1 KO mice, which phenocopy the pathogenesis of human AH.