Natural products and their derivatives have long served as a prolific reservoir for drug discovery. All-trans retinoic acid (ATRA) regulates lipid metabolism and shows anti-atherosclerotic potential, but its application is limited by poor drug-like properties and retinoid-related toxicity. Here, an allene-based Chem-Stamp technology has empowered the efficient construction of a diverse non-natural ATRA library by fine-tuning the number of isoprene units, the side chain, and the terminal group, leading to the identification of G301-WSS-2003 as a novel ATRA-based LDLR up-regulator. In vivo, G301-WSS-2003, with favorable exposure and bioavailability (e.g., t1/2 > 5 h, F > 30%), reduced serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) by 27%, 36%, and 26%, respectively, and significantly attenuated atherosclerotic lesion formation with no liver function abnormalities. These results highlight that Chem-Stamp is a powerful technology platform in the discovery, which results in identifying G301-WSS-2003 for further development as a promising candidate for the treatment of atherosclerosis.

Brassica juncea is a promising candidate for the phytoremediation of heavy metal - contaminated soils. MicroRNAs (miRNAs) act as key regulators in plant heavy metal stress responses and detoxification. However, the physiological adaptation mechanisms and the specific roles of miRNAs in B. juncea under cadmium (Cd) stress remain to be fully elucidated. This study aimed to investigate the physiological changes and expression profiles of five candidate miRNAs in leaves and roots under varying Cd concentrations and exposure durations.

SlERF26 acts as an essential downstream node in melatonin signaling, forming a novel regulatory module that integrates ethylene and carotenoid metabolism to drive tomato fruit ripening. Melatonin is an emerging regulator of plant development and stress responses, yet the specific transcriptional mechanisms by which it modulates climacteric fruit ripening remain largely unclear. In this study, we investigated the physiological effects of exogenous melatonin on tomato (Solanum lycopersicum cv. 'Micro Tom') fruit ripening and elucidated the underlying molecular regulatory network. Our results showed that exogenous application of melatonin, particularly at 100 μmol·L⁻1, significantly accelerated ripening progression, characterized by earlier color transition, elevated soluble sugar levels, and enhanced carotenoid accumulation (especially lycopene). This phenotypic change was accompanied by an earlier and stronger increase in ethylene production and respiration rate at the ripening transition stage. Integrated transcriptomic profiling and weighted gene co-expression network analysis (WGCNA) identified SlERF26, a member of the AP2/ERF superfamily, as a key melatonin-responsive transcription factor that is strongly and positively correlated with ethylene and pigment accumulation traits. Functional characterization using virus-induced gene silencing (VIGS) demonstrated that suppression of SlERF26 significantly delayed ripening, impaired chlorophyll degradation, and attenuated the expression of key genes involved in carotenoid (SlPSY1, SlPDS) and ethylene (SlACS2, SlACO1) biosynthesis. In addition, SlERF26 silencing caused enzyme-specific and stage-dependent changes in ripening-related enzymes, including reduced PDS and ACO levels at the 40 DAP transition stage, while PSY levels were increased, possibly reflecting a compensatory response. Furthermore, melatonin supplementation failed to fully rescue the ripening defects in SlERF26-silenced fruits, suggesting that SlERF26 contributes to melatonin-associated regulation of ethylene biosynthesis and carotenoid accumulation. Collectively, these findings support a working model in which SlERF26 contributes to melatonin-associated regulation of ethylene and carotenoid metabolism during tomato fruit ripening.

Numerous studies have demonstrated an association between diesel particulate matter (DPM) exposure and cardiotoxicity; recent evidence further suggests that cardiomyocytes may directly internalize DPM. In the present study, we investigated the cardiotoxic effects of SRM 1650b, a representative heavy-duty diesel emission particulate standard, which is considered a major contributor to the ongoing air pollution crisis. Isolated male Wistar rat hearts were perfused with different concentrations of SRM 1650b following stabilization, followed by 30 min of ischemia and 60 min of reperfusion. Results demonstrated deteriorated cardiac hemodynamics and elevated tissue injury compared with normal controls across different concentrations of SRM 1650b (HC, highest concentration = 300 μg/mL; MC, medium concentration = 100 μg/mL; LC, low concentration = 10 μg/mL). Administration of SRM 1650b significantly increased oxidative stress in both cardiac tissue and mitochondria. It also induced a decline in mitochondrial bioenergetic enzyme activities and corresponding respiratory efficiency compared with normal controls. The expression of mitochondrial quality-control-associated genes, including Pgc-1α, Tfam, Polg, Fis1, Mfn1, and Pink1, was significantly reduced, along with a decline in mitochondrial DNA copy number. Collectively, these alterations resulted in heightened myocardial sensitivity to ischemia-reperfusion injury. Furthermore, expression of the PI3K/Akt signaling pathway was reduced following SRM 1650b administration and decreased further after ischemia-reperfusion challenge. These findings suggest that SRM 1650b-mediated cardiotoxicity is associated with impaired mitochondrial functional integrity and suppressed PI3K/Akt signaling, thereby reducing the cardiac capacity to withstand ischemia-reperfusion injury.

Esophageal cancer is a type of malignant tumor with high incidence and mortality rates worldwide, and esophageal squamous cell carcinoma (ESCC) is the predominant pathological subtype in China. Despite continuous advancements in treatment methods, the prognosis of ESCC patients remains poor and there is an urgent need to develop new therapeutic strategies. Recent epigenetic frontiers have identified the dysregulation of lysine acetylation, a dynamic and reversible post-translational modification, as a pivotal driver of ESCC pathogenesis. This process is orchestrated by a dynamic interplay between "writers" (lysine acetyltransferases, KATs), "erasers" (lysine deacetylases, KDACs), and "readers". In this review, we systematically delineate the molecular landscape of lysine acetylation in ESCC and emphasize how these epigenetic modulators reshape the transcriptional program of tumor cells. Specifically, we highlight the oncogenic or tumor-suppressive roles of key acetylating enzymes and their non-histone substrates, which bridge the gap between epigenetic alterations and malignant phenotype. And we summarize the latest research progress and clinical application potential of KAT inhibitors (KATi), KDAC inhibitors (KDACi), and inhibitors of lysine acetylation readers in various tumor. Additionally, we discuss the challenges of drug resistance in epigenetic therapy and opportunities in targeting acetylation modification. By providing a comprehensive synthesis of the acetylation-ESCC axis, this review aims to offer theoretical guidance for the development of novel epigenetic biomarkers and targeted interventions to improve the prognosis of ESCC patients.

Carbon tetrachloride-induced liver injury is a well-established model of toxic hepatitis, characterized by marked oxidative and inflammatory damage. It remains a relevant experimental system because broadly effective therapeutic options are still limited. Juglone, a natural naphthoquinone derived from Reynoutria japonica, has demonstrated antioxidant and anti-inflammatory potential. However, mouse evidence combining standard liver injury measures with stress- and cell-death gene profiling remains limited. Building on juglone studies in injury and fibrosis, we tested juglone in a CCl4 model. Male C57BL/6 mice were assigned to normal, vehicle, CCl₄, silymarin (200 mg/kg), and juglone (8 mg/kg) groups, and CCl₄ (in corn oil) was administered intraperitoneally. Endpoints included body weight, serum liver enzymes and lipid profile, H&E histology, and RT-qPCR panels for inflammatory, pyroptosis-associated, ER stress and autophagy-linked, apoptotic, oxidative-stress, and profibrotic transcripts. CCl₄ exposure was associated with weight loss, enzyme elevation, dyslipidemia, and architectural disruption, whereas juglone treatment was associated with improved biochemical indices and histology alongside coordinated downregulation of inflammatory/pyroptosis-related transcripts (including Il-1β, Il-6, Nlrp3, and Gsdmd) and moderation of ER stress/autophagy and apoptosis-linked transcripts, with downregulation of profibrotic transcripts (Acta2, Mmp2, and Tgfβ1). In this mouse CCl₄ model, juglone treatment was associated with reduced serum and tissue injury signatures together with changes in stress and inflammation-linked transcriptional programs. Because the molecular findings are primarily based on RT-qPCR, these results are best interpreted as gene-level associations compatible with modulation of oxidative stress, inflammasome- and pyroptosis-related mRNA expression, ER stress responses, apoptosis-linked transcripts, and remodeling-associated transcripts, rather than as confirmed pathway inhibition or established antifibrotic activity. These data support juglone as a hepatoprotective candidate and identify protein-level and functional validation as important next steps.

Shuxin oral liquid (SX) is a traditional Chinese medicine formulation widely used for coronary heart disease (CHD) characterized by "Qi deficiency and blood stasis". Because myocardial fibrosis (MF) contributes to adverse cardiac remodeling in ischemic heart disease, investigating the anti-fibrotic potential of SX may provide pharmacological evidence related to its traditional use.

Sorafenib resistance remains a major challenge in the treatment of hepatocellular carcinoma (HCC). Through an in vivo CRISPR/Cas9 screen, we identified protein SUMOylation as a key pathway enriched in sorafenib-resistant HCC tumors. SUMO1 expression was significantly upregulated in resistant tumors and cell lines, and its modulation directly influenced sorafenib sensitivity both in vitro and in vivo. Proteomic analysis revealed that SUMO1 overexpression enhanced glycolysis, and metabolic assays confirmed increased extracellular acidification rate (ECAR) and decreased oxygen consumption rate (OCR) in SUMO1-high cells. We further identified PKM2 as a key SUMOylation target, mediated by the E3 ligase TRIM28. The SUMOylation of PKM2 increased its enzymatic activity, promoted aerobic glycolysis, and conferred sorafenib resistance. The inhibition of PKM2 with Compound 3k reversed glycolytic flux and restored sorafenib sensitivity. Clinically, SUMOylated PKM2 was highly expressed in HCC tumors and was associated with the expression of markers of glycolysis and sorafenib resistance in HCC. Our study revealed a novel SUMO1-PKM2 axis that drives glycolysis and sorafenib resistance in HCC, suggesting a potential therapeutic target for overcoming drug resistance.

Allyl isothiocyanate (AITC), a bioactive compound found in various cruciferous vegetables including wasabi, exhibits potent antimicrobial properties against foodborne pathogens. This study demonstrates that AITC significantly inhibits the growth of Vibrio parahaemolyticus in both culture media and seafood matrices (minced shrimp). For mid-log phase planktonic and biofilm-embedded V. parahaemolyticus, AITC exerts potent bactericidal effects that intensify at higher concentrations (≥50 μg/ml) and longer exposure times. Crucially, AITC disrupts pre-formed biofilms on abiotic (e.g., polystyrene) and biotic (shrimp shell) surfaces, degrading extracellular matrices and dispersing embedded cells, as visualized by scanning electron microscopy (SEM). Sublethal AITC exposure (25 μg/ml) triggers extensive transcriptional reprogramming in V. parahaemolyticus RIMD2210633, downregulating 1299 genes, including key virulence determinants (e.g., T3SS1, T3SS2, T6SS2, and TDH), flagellar assembly components, exopolysaccharide (EPS) biosynthesis machinery, and type IV pili. Paradoxically, despite inhibiting swimming motility and biofilm formation, AITC upregulates cyclic di-GMP (c-di-GMP) synthesis, suggesting a stress-adaptive response. Collectively, these findings reveal that AITC combats V. parahaemolyticus through direct bactericidal activity, biofilm dismantling, and global downregulation of virulence and colonization-associated genes, with the observed c-di-GMP increase representing a stress-adaptive response whose functional significance requires further investigation. This work supports the potential of AITC as a natural strategy to enhance seafood safety and provides a mechanistic framework for its multi-target action.

α-fluoro-β-alanine (FBAL), a catabolite of the chemotherapeutic agent 5-fluorouracil (5-FU), has attracted significant attention due to its cardiotoxicity and neurotoxicity. However, its association with 5-FU resistance, a major obstacle in cancer treatment, has rarely been reported. In this study, FBAL was identified as a β-arrestin1-biased ligand of sphingosine 1-phosphate receptor 2 (S1PR2), which upregulates dihydropyrimidine dehydrogenase (DPD) expression and drives 5-FU resistance in colorectal cancer. Mechanistically, following exposure to FBAL, S1PR2, a G protein-coupled receptor (GPCR), is phosphorylated by recruiting G protein-coupled receptor kinase 6 (GRK6). Phosphorylated S1PR2 coupled with β-arrestin1, but not G proteins, to activate the MEK/ERK/AP-1 pathway and promote DPYD transcription. Ser343 was identified as the key phosphorylation site of S1PR2 using IP-MS analysis. This residue within the C-terminal domain mediates receptor interaction with β-arrestin-1 to activate the β-arrestin-1-dependent ERK pathway, as was confirmed in HCT116S1PR2KO-ΔC cells and HCT116S1PR2KO-S343A cells. In vivo, mice bearing orthotopic xenografts of HCT116S1PR2KO-S343A cells exhibited significantly enhanced higher sensitivity to 5-FU treatment compared to those HCT116S1PR2KO-WT cells following FBAL. Taken together, this study showed that exposure to FBAL induces S1PR2 phosphorylation at Ser343 within the C-terminal mediated by GRK6, activating the β-arrestin1-dependent ERK pathway to upregulate DPD expression. This study not only delineates a phosphorylation-dependent signaling pathway in chemoresistance but also establishes S1PR2 as a promising therapeutic target for overcoming 5-FU resistance in cancer.

Quercetin is recognized for diverse pharmacological activities. However, the mechanism underlying its broad-antiviral effects has not been elucidated. Herein, we identified quercetin as a potent inhibitor of both double-stranded DNA virus Bombyx mori nucleopolyhedrovirus (BmNPV) and single-stranded RNA virus porcine reproductive and respiratory syndrome virus (PRRSV). Surface plasmon resonance (SPR) revealed that quercetin targets host ubiquitin-activating enzyme 1 (Uba1) homologs. Uba1 knockdown reduced viral proliferation and enhanced the antiviral effect of quercetin, whereas Uba1 overexpression functioned oppositely. Quercetin bound Uba1 homologs with high affinity. Notably, mutation of two binding residues, Q977 and G978, significantly disrupted the binding between BmUba1 and quercetin, and abolished quercetin's antiviral activity. Quercetin obstructed the transfer of ubiquitin from Uba1 to the E2 enzyme Ubc6, impairing the ubiquitination process. Similarly, quercetin inhibited PRRSV proliferation via targeting Uba1 in mammals. These findings elucidate the molecular mechanism underlying the pharmacological effects of quercetin, providing a theoretical basis for the development of novel antiviral agents against both DNA and RNA viruses.

Non-small cell lung cancer (NSCLC) is among the most lethal cancers globally, partly due to dysregulated STAT3 signaling, which is related to tumor growth. Combination strategies involving STAT3 inhibitors with chemotherapeutic agents may improve therapeutic efficacy. Therefore, the current study assessed the effects of Stattic (a STAT3 inhibitor) in combination with vinblastine (a microtubule-destabilizing agent) against A549 lung cancer cells.

Lung cancer presents complex etiopathology involving a mix of genetic predispositions and environmental factors. The best treatment modality is surgical resection. However, it becomes ineffective in the advanced metastatic stage. Thus, cisplatin-based chemotherapy, though restricted by an intrinsic and/or acquired chemo-resistant phenotype, remains the first-line therapy for advanced non-small-cell cancer (NSCLC).

Fungal metabolites with diverse structures serve as valuable natural resources for green biopreparations and immune elicitors in sustainable agricultural biocontrol. As a small-molecule metabolite derived from the edible fungus Pholiota adiposa, 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (MCA) has been reported to enhance stress tolerance and plant growth, yet its potential and mechanism against fruit tree fungal diseases remain unclear. This study investigated MCA-mediated resistance to Glomerella leaf spot (GLS) in apples. MCA pretreatment significantly alleviated lesion formation in apple leaves and fruits, upregulated immunity associated genes and increased defense-related enzyme activities. Transcriptomic analysis revealed that MCA activated the phenylpropanoid pathway, elevated lignin biosynthetic gene expression, and promoted lignin accumulation and cell wall reinforcement, while also boosting apple seedling growth. As the first application of MCA as a novel immune elicitor in fruit trees, these findings confirm its dual functions of disease resistance and growth promotion, expanding MCA's application scope and providing theoretical support for green sustainable apple production.

In this study, Salmonella Typhimurium (ST) vaccine diluent effects on caecal tonsils gene regulation in layer chicks were investigated. A live attenuated ST vaccine was reconstituted in Marek's and/or water diluents, chicks vaccinated and caecal tonsils were assessed for global gene expression at days 7 and 14 post-vaccination. Differentially expressed genes (DEGs) involved in the activation of the intestinal immune network for IgA production were upregulated earlier (e.g. day 7 post-vaccination) in the water diluent group, while in the Marek's diluent group, this pathway was enriched by the upregulated DEGs at day 14 post-vaccination. The commonly upregulated biological pathways between the two diluents were cytokine-cytokine receptor interaction, toll-like receptor signaling pathway and cytosolic DNA-sensing pathway. At day 7 post-vaccination, 28.5% and 40.2% of the significantly upregulated genes were unique to Marek's and water diluents, respectively, whereas 31.3% were common between the Marek's and water diluent groups, showing the role of the diluent in priming the vaccine for host immune system modulation. At day 14 post-vaccination, 52.7% and 15.4% of the upregulated DEGs were unique to Marek's and water diluents, respectively. Overall, the data showed that the live attenuated ST vaccine reconstituted in Marek's diluent affected the regulation of immune system-related genes in the caecal tonsils of chicks, with a difference in pattern when water was used as a diluent.

UFMylation, a recently identified ubiquitin-like modification, is essential for cellular stress homeostasis, particularly endoplasmic reticulum (ER) stress regulation. However, its biological and therapeutic exploration has been hindered by the absence of potent small-molecule inhibitors. Here, we report the first discovery of two compounds targeting the UFMylation E3 ligase complex core protein DDRGK1: Osimertinib, originally designed as an EGFR T790M selective inhibitor, acting through a previously unrecognized covalent mechanism, and CP-24, a novel non-covalent inhibitor. Both compounds disrupt the DDRGK1-UFL1 interaction, globally suppress UFMylation, inhibit ER-phagy, and induce ER stress. In glioblastoma (GBM), pharmacological UFMylation inhibition markedly reduces tumor cell viability and sensitizes cells to Temozolomide and radiotherapy. Both compounds also exert strong immunomodulatory activity, promoting macrophage polarization toward an anti-tumor M1 state. In vivo, Osimertinib, benefiting from superior pharmacokinetics, significantly suppresses tumor growth in immunodeficient models and achieves tumor-free outcomes in 65% of immunocompetent mice. These tumor-free mice develop durable anti-GBM immune memory, rapidly clearing tumors upon rechallenge, an outcome unattainable by previous GBM treatments. Mechanistically, Osimertinib enhances anti-tumor immunity by promoting macrophage M1 polarization, T cell expansion, and reducing PD-1 protein levels. Collectively, our study introduces Osimertinib and CP-24 as valuable chemical probes for dissecting UFMylation biology and highlights Osimertinib's potential for off-label use in EGFR-wildtype GBM. More broadly, we establish UFMylation inhibition as a dual-targeting therapeutic strategy that disrupts tumor survival pathways and reprograms the immune microenvironment, offering a promising avenue for durable GBM control.

Thyroid hormone (TH) is an important regulator of human brain development, and maternal TH imbalance is linked to adverse outcomes such as reduced IQ and decreased motor function in children. As part of the VHP4Safety initiative, using the human neural progenitor test (hNPT), we investigated the molecular effects of elevated levels and absence of T3. Neural progenitor cells were differentiated for 10 days under no (0 nM), normal (4.1 nM; the reference), or high (410 nM) triiodothyronine (T3) conditions. RNA-Seq was used to assess gene expression, and a molecular Adverse Outcome Pathway (AOP) network was applied to interpret transcriptomic changes compared to the reference alongside Gene Ontology (GO) enrichment analysis. Data show that normal T3 levels induce key neurodevelopmental processes which are not active without T3 exposure, based on significantly altered gene expression in pathways related to neurogenesis, synaptogenesis, differentiation and metabolism, including pathways of brain-derived neurotrophic factor (BDNF) signalling, proliferation, oligodendrocyte specification, and glycolysis. Key Events (KEs) such as reduced BDNF and impaired proliferation were, compared to normal levels of T3, significantly affected in the molecular AOP network in absence of T3, while high T3 levels showed minimal differences in transcriptional effects. Our findings demonstrate that absence of T3 inhibits brain development since no T3 condition affects key neurodevelopmental processes in vitro. This study illustrates the utility of integrating transcriptomics with molecular AOPs to provide a structured, mechanistic framework for evaluating the regulation of neuronal cell proliferation and differentiation in brain development. The framework may be useful for chemical risk assessment.

MYB is a key transcriptional regulator of hematopoietic cell proliferation, survival, and lineage commitment. Aberrant MYB activity has been linked to hematologic malignancies and clinically relevant features, including poor prognosis and drug resistance. However, the mechanisms by which MYB contributes to drug resistance in leukemia remain unclear.

The photorespiratory metabolism safeguards photosynthesis against abiotic and biotic stress. Nitric oxide (NO) and reactive oxygen species (ROS) levels rise in plants during abiotic stress. Low concentrations of NO or ROS are beneficial as signalling molecules, but they can be toxic to plant cells at high concentrations. ROS are known to modulate photorespiration; however, it is unclear whether NO affects photorespiratory enzymes and photochemical components simultaneously. We therefore used sodium nitroprusside (SNP) under dark, moderate light (ML), or high light (HL) conditions to simultaneously investigate its impact on photorespiratory enzymes and photochemical components. The NO levels were increased upon SNP exposure in Pisum sativum leaves, particularly under HL conditions. The NO release in leaves was confirmed when the NO scavenger cPTIO (2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt) was present, since it decreased the majority of elevated NO. The nitrosative/oxidative stress in Pisum sativum leaves was confirmed by the increase in nitrosothiols and tyrosine-nitrated proteins, as well as reduced aconitase activity after SNP exposure at HL. The protein levels, mRNA levels, and the enzyme activities of the following four photorespiratory enzymes: glycolate oxidase (GO), hydroxypyruvate reductase (HPR), glycerate kinase (GK), and phosphoglycolate phosphatase (PGLP) were markedly increased under elevated NO conditions. Catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD) also showed increased activity, elevated protein and transcript levels upon exposure to SNP. Parallel studies on chlorophyll a fluorescence confirmed that NO restricted electron transport at both PSII and PSI, inhibited photosynthesis and respiration, and damaged photosynthetic pigments. We concluded from this study that NO at high concentrations upregulated photorespiratory enzymes while inhibiting photochemical components such as photosystem II and I (PSII/PSI) simultaneously.

Antimony (Sb), a toxic metalloid with significant phytotoxicity and potential carcinogenicity, poses a threat to agricultural safety due to its extensive use. Soybean (Glycine max L.), a globally important source of edible oil and protein, has been investigated for heavy metal resistance; yet its specific responses to Sb stress remain poorly understood. In this study, integrated phenotypic, physiological, transcriptomic, and metabolomic analyses were applied to elucidate the molecular defense network underlying soybean response to Sb(III) stress. Soybean exhibited dose-dependent phytotoxic effects, including leaf wilting, root damage, and reduced biomass, while tolerating Sb stress up to 90 mg/kg, alongside a pronounced capacity for Sb accumulation and translocation. Multi-omics analysis revealed a coordinated molecular defense network comprising three key pathways, flavonoid biosynthesis, alanine, aspartate, glutamate metabolism, and isoquinoline alkaloid biosynthesis, that collectively contribute to Sb tolerance. Key components of this network, including genes (e.g., ABC transporters, PALs, CHIs, and PERs) and metabolites (e.g., isoliquiritigenin, chlorogenic acid, L-aspartate, L-glutamate), were identified as critical mediators. Functional validation via yeast heterologous expression demonstrated that GmCHI4, GmPER52, and GmPAL1 enhanced Sb tolerance, further supporting their roles within this defense network. These findings delineate a multi-layered molecular defense network and provide precise genetic and metabolic targets for breeding Sb-tolerant soybean cultivars.