The role of hepatic insulin resistance (HIR) in the development of fatty liver, diabetes and cardiovascular diseases is well known, yet the molecular basis of HIR remains unclear, limiting targeted therapeutic strategies. Here we show that insulin signalling-inactivated phosphorylated GSK-3β (p-GSK-3β) is revitalized via reactive oxygen species-mediated sulfenylation, leading to glycogenesis termination and gluconeogenesis initiation, two hallmarks of HIR. Mechanistically, sulfenylated or 'oxidatively activated' p-GSK-3β regains the enzymatic activity to phosphorylate liver glycogen synthase, thereby blocking glucose storage. This activated p-GSK-3β can further phosphorylate insulin-suppressed Forkhead box O1, thus liberating its transcriptional activity to promote the expression of gluconeogenic enzymes. Notably, this dual-pathway mechanism is conserved in clinically relevant human liver samples and organoids. These findings elucidate the molecular mechanism by which HIR is formed and provide potential strategies against HIR by targeting sulfenylated or 'oxidatively activated' p-GSK-3β.

Ischemic stroke (IS) is a common and refractory cerebrovascular disease that imposes a heavy burden on patients' families and society due to its high incidence, complex pathological mechanisms, and high mortality rate. Corilagin, the primary active component of the traditional Tibetan medicines Terminalia chebula and Phyllanthus emblica, is widely used in Tibetan medicine for treating cerebrovascular diseases. This study aims to comprehensively elucidate the mechanism of Corilagin in treating cerebral ischemia in IS through RNA transcriptome sequencing, validate its neurorestorative effects in cerebral ischemic rats, and explore the therapeutic potential of Corilagin for IS. METHODOLOGY:In vitro, a glucose-oxygen deprivation (OGD) model of neural stem cells (NSCs) was established. Cell viability was assessed using the CCK8 assay, followed by RNA extraction for transcriptome analysis.In vivo, the therapeutic efficacy of Corilagin was validated using a rat model of cerebral ischemia. RESULTS:In the rat MCAO model, Corilagin was found to improve neurological deficits, promote the survival of damaged neurons in the cortex and hippocampal CA1 region, and exert neuroprotective effects. Transcriptome sequencing revealed that Corilagin acts on NSCs by regulating mitochondrial function, energy metabolism, and oxidative stress levels.Western blotting further validated its modulation of the HIF-α/ALDOA pathway. In the in vitro OGD model, Corilagin effectively mitigated NSC injury. CONCLUSION:Corilagin exhibits therapeutic potential for ischemic stroke through neural stem cell-mediated neurorepair.

Epigenetic modifications, such as DNA methylation, histone methylation, and RNA methylation, dynamically regulate gene transcription and play critical roles in cellular differentiation, development, and disease, especially cancer. Inhibitors targeting the enzymes responsible for these modifications have emerged as promising cancer therapies. For instance, DNMT inhibitors (e.g., azacitidine, decitabine) can reactivate tumor suppressor genes via demethylation; HMT inhibitors like tazemetostat and EPZ-5676 modulate chromatin structure to exert anti-tumor effects; and RNA methyltransferase inhibitors such as STM2457 disrupt RNA metabolism to suppress tumor growth. Despite encouraging preclinical and clinical results, challenges including toxicity and drug resistance remain obstacles to broader clinical application. This review summarizes recent advances in epigenetic inhibitor development to support the design of safer and more effective targeted cancer therapies.

BACKGROUND AND OBJECTIVES:Neural stem cells (NSCs) are important for endogenous brain repair but are highly vulnerable to mitochondrial dysfunction during cerebral ischemia-reperfusion injury (CIRI). Preliminary bioinformatics analysis suggested that immune microenvironment remodeling and JAK/STAT signaling activation are involved in CIRI pathology. This study investigated whether protocatechuic acid (PCA) protects NSCs after ischemia-reperfusion injury and explored the underlying mechanisms focusing on the JAK2/STAT3 pathway and mitophagy. RESULTS:Bioinformatics analysis identified JAK/STAT signaling as a potential regulatory node in CIRI. Transcriptome sequencing further showed that PCA markedly altered the gene expression profile of NSCs, with differentially expressed genes enriched in the JAK/STAT pathway. In vitro, PCA suppressed OGD/R-induced JAK2/STAT3 activation, restored mitochondrial membrane potential, reduced DCFH-DA and MitoSOX Red oxidation-sensitive fluorescence signals, promoted TOMM20/LC3 colocalization, enhanced PINK1/Parkin-related mitophagy, and improved NSC survival. Co-treatment with the JAK2 inhibitor AG490 further supported the protective and pro-mitophagic effects of PCA, whereas co-treatment with the JAK/STAT pathway activator RO8191 weakened PCA-mediated mitochondrial protection and mitophagy restoration. In vivo, PCA improved neurological function, reduced infarct volume, attenuated hippocampal neuronal injury, preserved endogenous NSCs and NeuN-positive cells, and enhanced mitophagy-related signals in MCAO/R rats. Importantly, in vivo Western blot analysis showed that PCA suppressed MCAO/R-induced JAK2/STAT3 phosphorylation, whereas RO8191 co-treatment attenuated this inhibitory effect and weakened the protective effects of PCA, supporting the involvement of JAK2/STAT3 inhibition in PCA-mediated neuroprotection. CONCLUSION:PCA alleviates CIRI and protects NSCs partly by suppressing JAK2/STAT3 activation, restoring mitochondrial function, and promoting PINK1/Parkin-mediated mitophagy. These findings provide experimental evidence supporting mitochondrial quality control and NSC protection as potential therapeutic targets for ischemic stroke.

Ischemic stroke accounts for ∼ 87% of all strokes. Existing reperfusion therapies are limited by narrow time windows. Small extracellular vesicles (sEVs) can cross the blood-brain barrier and deliver microRNAs (miRNAs) that modulate post-stroke inflammation, apoptosis, angiogenesis, and neurogenesis. However, most evidence is from preclinical studies (MCAO models), and no approved clinical protocols exist. This review summarizes ischemic stroke pathogenesis and critically evaluates preclinical advances in sEV-miRNA therapy. We highlight key translational barriers: lack of standardized sEV isolation (MISEV2023), miRNA cargo heterogeneity, dosing uncertainties, limited biodistribution data, off-target risks, regulatory bottlenecks, and absence of stroke-specific trials. We also evaluate exosomal miRNAs from various cell sources (e.g., bone marrow mesenchymal stem cells). This review provides a critical synthesis of preclinical evidence to establish a foundation for future translational research in precision stroke therapy.

Idiosyncratic Drug-Induced Liver Injury (IDILI) challenges drug development and clinical use. Its unpredictable nature complicates mechanistic research. Polygonum multiflorum Thunb (PM), a traditional tonic herb, has been used for centuries in China and East Asia. Now, PM-induced IDILI is a focus of traditional Chinese medicine safety research. Nevertheless, the underlying mechanisms of PM-triggered IDILI require further investigation. Previous clinical studies identified elevated Tumor Necrosis Factor-alpha(TNF-α) levels in patients susceptible to PM-induced liver injury. This finding prompted us to employ integrated network pharmacology, targeted lipidomics, and transcriptomics to investigate the combined effect and mechanism of PM and the susceptibility factor TNF-α in C57 mice. Histochemical staining, biochemical assays, and inflammatory cytokine analysis revealed significant hepatic inflammatory infiltration in the TNF-α+PM co-exposure group versus controls. This group exhibited markedly elevated levels of liver injury markers (ALT, AST, DBIL) and inflammatory mediators (IL-6, TNF-α) (P < 0.05, P < 0.01). Subsequent administration of the TNF-α antagonist etanercept restored hepatic architecture and significantly reduced cytokine levels (P < 0.01), approaching baseline values. Notably, neither TNF-α nor PM monotherapy induced liver injury in mice. Network pharmacology analysis identified 33 bioactive PM components and 11 core targets. Targeted lipidomics revealed 32 differential lipid metabolites (DLMs), primarily glycerolipids (GL) and glycerophospholipids (GP). Transcriptomics demonstrated cooperative regulation of 199 differentially expressed genes(DEGs) by TNF-α and PM. Network enrichment analysis further uncovered lipid dysregulation mediated by Pparg, Cyp4a12a, Cyp4a12b, and Cyp4a31 genes, with RT-qPCR validation confirming these findings. In summary, TNF-α plays a central driving role in PM-induced liver injury. When co-administered with PM, it activates the PPAR signal ing pathway via downregulation of Cyp4a12a, Cyp4a12b, and Cyp4a31, thereby dysregulating lipid metabolism, promoting inflammatory mediator production, and ultimately causing liver injury.

This study aimed to explore the mechanism by which the ethanol extract of Cyanotis arachnoidea(ECA) ameliorates ischemic stroke(IS) in rats, based on metabolomics and transcriptomics. A rat model of middle cerebral artery occlusion(MCAO)-induced IS was established using the modified suture occlusion method. Ninety Sprague-Dawley(SD) rats were randomly divided into the sham operation(sham) group, the model(model) group, ECA low-, medium-and high-dose groups(ECA-L, ECA-M, ECA-H), and the positive drug nimodipine(NMDP) group. The therapeutic effects of ECA were evaluated by neurological deficit scores, 2,3,5-triphenyltetrazolium chloride(TTC) staining, serum biochemical assays, hematoxylin-eosin(HE) staining, and Nissl staining. Transcriptomic and metabolomic analyses were performed on brain tissues from the sham, model, and ECA-H groups. Core gene expression was verified using real-time fluorescence quantitative polymerase chain reaction(RT-qPCR). The results showed that ECA significantly reduced neurological deficit scores and cerebral infarct volume, ameliorated pathological damage in the cerebral cortex, and dose-dependently downregulated the inflammatory factor levels and oxidative stress markers in IS rats. Transcriptomic analysis revealed that ECA-H regulated the expression of 328 differentially expressed genes(DEGs) in the model group, with 129 genes identified as core regulatory targets. Gene Ontology(GO) enrichment analysis showed that these DEGs were mainly involved in IS-related processes, including cerebral cortex development and neuronal development. Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis showed that the DEGs were primarily enriched in pathways such as the calcium signaling pathway and the cyclic adenosine monophosphate(cAMP) signaling pathway. RT-qPCR verification demonstrated that ECA significantly upregulated the relative expression levels of 10 genes related to these two signaling pathways. Metabolomic analysis revealed that ECA-H significantly regulated the levels of 11 differential metabolites in rat brain tissues, mainly involving metabolic pathways such as folate-mediated one-carbon metabolism, lysine degradation, and cysteine and methionine metabolism. In summary, the ECA exerts neuroprotective effects in IS by synergistically activating the calcium and cAMP signaling pathways, targeting the expression of core genes, and regulating key metabolic pathways, thereby inhibiting neuroinflammation, balancing oxidative stress, and alleviating neuronal damage.

Glycolysis and protein lactylation both drive hepatocellular carcinoma (HCC) progression, yet their mechanistic interplay remains unclear. Through integrated single-cell and spatial transcriptomic analyses stratified by glycolytic activity, we identified alanyl-tRNA synthetase 1 (AARS1), a recently characterized protein lactyltransferase, as a key metabolic-immune regulator in HCC. Clinically, AARS1 is upregulated in tumors, correlates with elevated glycolytic flux measured by 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT), poor prognosis, and immunotherapy resistance. In murine models, hepatocyte-specific knockout of AARS1 suppressed tumor growth and reduced the abundance of regulatory T cells (Tregs). Mechanistically, AARS1 catalyzes the lactylation of activating transcription factor 6 (ATF6) at lysine 424, preventing its degradation and leading to transcriptional activation of TDO2. This process promotes L-kynurenine production and supports Treg differentiation and function. Furthermore, L-kynurenine-AHR signaling drives eNAMPT secretion from Tregs, which augments tumor cell glycolysis and lactate production, thereby reinforcing a feedback loop that sustains AARS1-catalyzed ATF6 lactylation. Pharmacological inhibition of AARS1 with β-alanine sensitized tumors to PD-1/PD-L1 blockade.

Through a comprehensive analysis combining network pharmacology prediction and transcriptomics, this study systematically explained the multi-target mechanism of Cyanotis arachnoidea(CA) Gel in improving melasma. A melasma model was induced in female SD rats by progesterone injection combined with ultraviolet B(UVB) irradiation for 40 consecutive days, while the blank control group was only fed routinely. After successful model establishment, the rats were randomly divided into five groups and administered different doses of CA ethanol extract gel(high, medium, and low doses) or arbutin Gel(positive control), which were applied once daily for 28 consecutive days. Subsequently, the levels of superoxide dismutase(SOD), malondialdehyde(MDA), and tyrosinase(TYR) in the skin, serum, and liver tissues were measured. Hematoxylin-eosin(HE) staining and Masson-Fontana staining were used to observe the pathological changes in the tissues. Network pharmacology combined with transcriptomics was employed to identify core targets and pathways, and the differential gene expression was validated by quantitative real-time PCR(qPCR). Pharmacodynamic experiments showed that CA Gel significantly increased SOD activity and decreased MDA and TYR levels in the skin, serum, and liver of model rats. It also improved epidermal thickening, inflammatory infiltration, collagen loss, and melanin deposition. Network pharmacology analysis showed that CA mainly regulated core targets such as signal transducer and activator of transcription 3(STAT3), epidermal growth factor receptor(EGFR), and interleukin-6(IL-6), and modulated the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT) and interleukin-17(IL-17) signaling pathways. Transcriptomic analysis showed that CA Gel significantly downregulated the gene expression of heat shock protein 90β family member 1(Hsp90b1), heat shock protein 90α family member 1(Hsp90aa1), and the key steroid synthesis enzyme cytochrome P450 family 17 subfamily A member 1(Cyp17a1), while upregulating thioredoxin 1(Txn1). qPCR results confirmed that CA Gel regulated oxidative stress and inflammatory response by inhibiting the IL-17 signaling pathway and steroid hormone synthesis. This study, for the first time, reveals the molecular mechanism of CA Gel in improving melasma through multi-target synergistic regulation of oxidative stress, inflammatory response, and hormone metabolism pathways, providing a scientific basis for the treatment of pigmentation diseases with traditional Chinese medicine.

Immunotherapy has demonstrated significant efficacy in colorectal cancer (CRC), but its therapeutic effects remain limited in microsatellite stable (MSS) patients, indicating the critical role of the tumor immune microenvironment (TIME) in regulating immune responses. Lipid rafts, dynamic membrane microdomains enriched in cholesterol and sphingolipids, have emerged as potential targets for TIME remodeling through their integration of immune signal transduction, enrichment of cell death receptors, and regulation of immune cell functionality. This review outlines the pivotal mediating roles of lipid rafts in cellular survival, death, and tumor progression. Specifically, MSS-type CRC exhibits lipid raft structural remodeling driven by dysregulated lipid metabolism, which fosters multiple immune escape mechanisms through exosome-mediated immunosuppressive signaling, promotion of tumor-associated macrophage (TAM) M2 polarization, enhanced infiltration of regulatory T cells (Tregs), and functional exhaustion of effector cells, such as CD8+ T cells and NK cells. Finally, we discuss targeted therapeutic strategies based on lipid raft characteristics and CRC molecular profiles, proposing an innovative multidimensional treatment framework combining immune checkpoint inhibitors with lipid raft-targeted interventions and chemoradiotherapy. This approach provides theoretical and strategic support for overcoming CRC immunotherapy resistance and advancing clinical translation.

Hepatic fibrosis (HF) is an important pathological state in the progression of chronic liver disease to end-stage liver disease and is usually triggered by alcohol, nonalcoholic fatty liver, chronic hepatitis viruses, autoimmune hepatitis (AIH), or cholestatic liver disease. Research on novel therapies has become a hot topic due to the reversibility of HF. Research into the molecular mechanisms of the pathology of HF and potential drug screening relies on reliable and rational biological models, mainly including animals and cells. Hence, a number of modeling approaches have been attempted based on human dietary, pathological, and physiological factors in the development of HF. In this review, classical and novel methods of modeling HF in the last 10 years were collected from electronic databases, including Web of Science, PubMed, ScienceDirect, ResearchGate, Baidu Scholar, and CNKI. Animal models of HF are usually induced by chemical toxicants, special diets, pathogenic microorganisms, surgical operations, and gene editing. The advantages and limitations of hepatic stellate cells (HSCs), organoids, and 3D coculture-based HF modeling methods established in vitro were also proposed and summarized. This information provides a scientific basis for the discovery of the pathological mechanism and treatment of HF.

Regulatory T cells (Tregs) contribute to the immune escape of hepatocellular carcinoma (HCC). However, the drivers of the accelerated Treg accumulation in HCC remain unclear. In this study, Treg infiltration-related genes were analysed, and proteasome activator subunit 3 (PSME3) was identified as a pivotal driver using bioinformatics analysis. Functional experiments were performed to investigate the correlation between PSME3 expression and programmed cell death-1 (PD-1) monoclonal antibody therapy resistance in HCC. Elevated levels of PSME3 lead to metabolic reprogramming towards glycolysis and upregulation of osteopontin (OPN) expression. This glycolysis-induced OPN secretion by HCC cells promotes the differentiation and enrichment of Tregs while inhibiting CD8+ T cells in vivo and in vitro. Mechanistically, PSME3 enhanced PTEN-FBXL7 binding and promoted PTEN degradation through FBXL7-mediated ubiquitination, thereby enhancing glycolysis. The combination of PSME3 inhibition and PD-1 blockade is a promising strategy for HCC treatment. In conclusion, our data showed the critical role played by PSME3 in triggering Treg infiltration and inducing anti-PD1 tolerance.

The role of tumor-resident microbiota in modulating tumor immunity remains unclear. Here, we discovered an abundance of intra-tumoral bacteria, such us E.coli, residing and resulting in Colorectal cancer liver metastasis (CRLM). E.coli enhanced lactate production, which mediated M2 macrophage polarization by suppressing nuclear factor-κB -gene binding (NF-κB) signaling through retinoic acid-inducible gene 1 (RIG-I) lactylation. Lactylation of RIG-I suppressed recruitment of NF-κB to the Nlrp3 promoter in macrophages, thereby reducing its transcription. This loss of Nlrp3 affected the immunosuppressive activities of regulatory T cells (Tregs) and the antitumor activities of and CD8+ T cells. Small-molecule compound screening identified a RIG-I lactylation inhibitor that suppressed M2 polarization and sensitized CRLM to 5-fluorouracil (5-FU). Our findings suggest that tumor-resident microbiota may be a potential target for preventing and treating CRLM.

The problems of uncontrolled stem cell differentiation and the limited number of cells available represent significant obstacles to the advancement of stem cell research in the medical field. It is therefore imperative to control the potential of stem cells and to develop effective methods to regulate stem cell fate. A number of natural products derived from TCM have been shown to provide valuable insights into the regulation of biological signals and epigenetic mechanisms in stem cells. Furthermore, the structural modification of these natural products by synthetic means promises to unlock a wealth of possibilities in the field of precision regenerative medicine. However, a systematic and comprehensive review of these natural products has yet to be conducted. This article presents an overview of the natural products that have been identified as regulating the fate of stem cells. It includes a discussion of the classification, mechanism of action, and current prospects for the application of these natural products as lead compounds. This provides valuable insights for the advancement of TCM modernization and internationalization, precision regenerative medicine involving natural products and stem cells, the discovery of green lead compounds, and the development of new drugs.

Background:Polygonum multiflorum (PM), known as a traditional Chinese herb renowned for its tonic properties, has been used medicinally for millennia. However, it has drawn attention significantly due to the potential to induce idiosyncratic drug-induced liver injury (IDILI) in recent years. Previous studies identified the TNF-α, the pro-inflammatory cytokine, as a key factor contributing to susceptibility to PM induced-IDILI (PM-IDILI). However, the effects by which TNF-α mediates PM-IDILI remain poorly understood. Methods:This study sought to elucidate the role of TNF-α in PM-IDILI using a TNF-α-sensitized C57BL/6J mouse model, integrating analyses of the gut microbiota and metabolomics We employed biochemical analysis, inflammatory markers, inflammatory liver histopathological, sequencing of 16S rRNA gene, as well as untargeted metabolomics based on LC-MS to systematically evaluate the extent of liver injury and characterize alterations in gut microbiota and liver metabolites following PM administration in TNF-α pre-treated mice. Results:The results demonstrated that PM treatment in TNF-α-sensitized mice significantly elevated levels of indicators as AST (3.6-fold compared to the control group, P < 0.05) and ALT (3.9-fold compared to the control group, P < 0.01), increased plasma levels of IL-6 and IL-1β (P < 0.05 or P < 0.01), induced infiltration of inflammatory cell substantially in the liver. TNF-α-mediated PM disrupted the intestinal microbiota structure, characterized by reduced abundance of Akkermansia and increased abundance of Lachnospiraceae_NK4A136_group, Bacteroides, Alloprevotella, and Blautia. Furthermore, hepatic metabolomics analysis revealed that significant perturbations in TNF-α + PM treated mice, particularly affecting glutathione metabolism, purine metabolism, and arachidonic acid metabolism pathways. Conclusion:These findings suggest that TNF-α sensitization predisposes mice to PM-IDILI, potentially by disrupting gut microbial homeostasis and altering host hepatic metabolism. This research provides critical theoretical and experimental evidence relevant to the safe and effective clinical application of PM.

Mechanisms of adaptation of regulatory T cells (Tregs) to harsh tumor metabolic microenvironments for suppression of anti-tumor immunity remain largely unclear. Here, using spatial metabolomics and transcriptomics, we show that human hepatocellular carcinoma harbored metabolically heterogeneous subregions characterized by high glutaminolysis and ammonia contents, where Tregs were frequently present but CD8+ and CD4+ effector T cells die. We found Tregs used the urea cycle to detoxify ammonia by upregulating argininosuccinate lyase (ASL); meanwhile, ammonia was also converted to spermine by the FOXP3 transcription factor regulated spermine synthase (SMS). A direct interaction between spermine and PPARγ was verified by X-ray crystallography, leading to comprehensively modulating the transcription of multiple mitochondrial complex proteins to enhance oxidative phosphorylation and immunosuppression of Tregs. Clinically, anti-PD-1-treated dying tumor cells used transdeamination to release ammonia, which reinforced Treg function, leading to immunotherapeutic resistance. Targeting ammonia production to suppress Tregs presents a potential strategy for anti-tumor immunotherapy.