Quercetin, a well-known antioxidant, plays a crucial role in the response of plants to biotic and abiotic stresses. Triticum urartu is the donor of the A subgenome of common wheat. This study aimed to reveal the mechanism by which quercetin treatment alleviates salt stress in T. urartu. The quercetin treatment resulted in the reduction of Na+ content and enhancement of K+ content in the stressed leaves, while it elevated the Na+ content in the stressed roots. This indicated that under salt stress, quercetin treatment can promote transporting K+ to the leaves and retaining Na+ in the roots. Transcriptome analysis showed that in the roots, quercetin treatment increased the expression level of several genes encoding the rate-limiting enzyme for ethylene biosynthesis, core ethylene signaling proteins, and negative regulators of the core ABA signaling pathway (abscisic acid 8'-hydroxylase and protein phosphatase 2C) under salt stress, revealing that quercetin treatment may induce ethylene signal and suppress ABA signal. We observed that quercetin treatment increased the expression level of several respiration genes in roots under salt stress, including 36 glycolysis genes, 18 mitochondria-related genes, and 4 ATP synthase genes. This displayed that quercetin treatment may enhance the respiration of T. urartu. The enhancement of respiration will provide more energy and carbon sources for salt stress response. Overall, quercetin treatment alleviates the negative effect of salt stress in T. urartu plants via multiple mechanisms, offering potential for improving plant salt tolerance.

During citrus fruit development, exogenous gibberellin (GA) and 6-benzylaminopurine (6-BA, a synthetic cytokinin (CTK)) are both known to promote citrus peel thickness; however, the differences in their regulatory mechanisms on cell wall metabolism in citrus peels remain unclear. In this study, we found that GA treatment significantly increased cell wall polysaccharides in citrus peels, such as pectin and cellulose, whereas 6-BA treatment led to a notable accumulation of lignin. RNA-sequencing data revealed that several fruit ripening-related cell wall degradation genes, such as PME3, PL18, and EXPA1/8, exhibited decreased expression levels in both GA and 6-BA treatments. Additionally, a set of cell wall polysaccharide synthesis genes was upregulated in response to GA treatment but was largely downregulated in 6-BA-treated peels. Conversely, a group of lignin biosynthesis genes was upregulated in 6-BA-treated peels. GA treatment inhibited DELLA proteins (encoded by RGA and GAI) in the GA signaling pathway, whereas 6-BA treatment increased the expression of B-ARRs (ARR1 and ARR2) in the CTK signaling pathway. Furthermore, GA treatment elevated endogenous CTK levels, while 6-BA treatment also enhanced endogenous GA content, suggesting a reciprocal interaction between these two hormonal pathways.

Since the endoplasmic reticulum (ER) stress response is found in almost all tissues, its regulation and downstream factors have been vigorously explored.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma (RCC), with a rising incidence worldwide. However, the mechanisms by which ccRCC evades immune surveillance remain incompletely understood. Our findings indicate that fumarate hydratase (FH) expression is significantly downregulated in ccRCC, resulting in fumarate accumulation, which is correlated with a poor prognosis in ccRCC patients. RNA sequencing analysis suggests that dimethyl fumarate (DMF), an FDA-approved fumarate analogue, may impact tumor immunity. Our further investigation reveals that both DMF and the FH inhibitor (FHIN1) can promote immune evasion in ccRCC by upregulating PD-L1. Pre-treatment of tumor cells with DMF notably inhibits the cytotoxic effect of T cells. Mechanistically, fumarate induces PD-L1 expression through succination of HIF-1α at C800, facilitating its interaction with importin α3, p300, and PKM2, which promotes HIF-1α nuclear localization and transcriptional activity. Moreover, combining DMF with PD-L1 blockade therapy significantly enhances the efficacy of immunotherapy and prolongs the survival of tumor-bearing mice. Taken together, our study elucidates a mechanism by which FH downregulation promotes immune evasion through the fumarate-HIF-1α/p300/PKM2-PD-L1 axis, providing a novel target, drug, and strategy to improve immunotherapy for ccRCC.

Equine endometrial fibrosis is a leading cause of subfertility in aging mares. Fibrosis is a reparative response involving excess deposition of extracellular matrix (ECM) and increasing tissue stiffness. Augmented rigidity itself can drive fibrosis, by stimulating transition from fibroblasts to myofibroblasts. Myofibroblasts release latent transforming growth factor beta 1 (TGF-β1) from the ECM, thereby activating this profibrotic cytokine. Tissue culture polystyrene (TCP) is commonly used for in vitro experiments. The endometrium, however, is considerably softer than TCP. This study critically evaluated the use of hydrogels versus TCP. Differences in transcript abundance between equine endometrial fibroblasts cultured on TCP and hydrogels of decreasing stiffness (25 kPa to 2 kPa) and their transcriptional response to TGF-β1, were examined. Cells cultured on substrates of varying stiffness exhibited visual variations concerning adherence, morphology, and cell density, besides differences in basal gene expression and transcriptional response to TGF-β1. On stiffer substrates, the smooth muscle genes TAGLN and ACTA2, alongside the transcripts encoding the signaling proteins PDGFB, CCN2, and SERPINE1 were expressed at higher levels. This pattern was also evident for integrin receptor subunit ITGAV, while ITGB5 was expressed at lower levels on stiffer substrates. While ITGB3 demonstrated a response to TGF-β1 exposure independent of stiffness, an increase in transcript abundance of PDGFB, ITGAV, and ITGB5 towards TGF-β1 was only observed on softer hydrogels. The results highlight the importance of stiffness in cellular regulatory patterns, particularly relevant to fibrosis research. We recommend critically reconsidering the use of TCP when designing experiments in vitro.

This study investigates the mechanisms underlying acquired resistance to FGFR tyrosine kinase inhibitor (FGFR-TKI) in gastric cancer (GC), focusing on the interplay between ferroptosis and lipid metabolism of tumor cells.

The phytohormone methyl jasmonate (MeJA) enhances plant cold stress tolerance, but the underlying mechanisms remain elusive. Here, we discovered that MeJA induces a transient Ca2+ influx and elevated cytoplasmic free Ca2+ ([Ca2+]cyt) levels during the watermelon (Citrullus lanatus) response to cold stress. Conversely, silencing jasmonic acid carboxyl methyltransferase (ClJMT), encoding an enzyme that methylates JA to MeJA, led to contrasting effects compared with MeJA application. Upon cold exposure, MeJA rapidly and continuously upregulated 2 Ca2+-permeable channel genes, namely cyclic nucleotide-gated ion channel (ClCNGC) 2 and ClCNGC20. Silencing ClCNGC2 or ClCNGC20 attenuated MeJA-induced Ca2+ influx, [Ca2+]cyt accumulation, C-REPEAT BINDING FACTOR (CBF) pathway activation, and watermelon cold tolerance. Accordingly, ClCNGC2 or ClCNGC20 overexpression increased Ca2+ influx, [Ca2+]cyt levels, and expression of the CBF regulon and improved freezing tolerance in transgenic Arabidopsis thaliana plants. Multiple assays showed that ClCNGC2 and ClCNGC20 do not directly interact. Interestingly, silencing ClCNGC2 or ClCNGC20 abolished MeJA-induced upregulation of ClCNGC20 or ClCNGC2, respectively, in watermelon response to cold, demonstrating their reciprocal activation at the transcriptional level. Collectively, these findings suggest a mutual dependence between ClCNGC2 and ClCNGC20 in mediating MeJA-induced Ca2+ influx followed by [Ca2+]cyt elevation, subsequently activating the CBF pathway and enhancing cold tolerance in plants. This study provides insights into the molecular mechanisms underlying MeJA-mediated plant cold tolerance, holding potential for the breeding or engineering of cold-resistant cucurbit varieties.

Melatonin integrates multiple biological pathways to enhance rice drought tolerance with cooperation of WRKY, bHLH, ERF, MYB and NAC transcription factors, and crosstalk of salicylic acid, ethylene and auxin. Drought is one of the primary environmental threats, and detrimentally affects plant growth and development, impeding crop yield and food quality worldwide. Melatonin has recently emerged as a multifunctional biomolecule with promising aspects in plant stress tolerance. However, the way in which melatonin improves drought tolerance in rice has not been investigated systematically. Here, we demonstrate that melatonin minimized drought effects on rice, resulting in improved germination rate and growth performance. Application of exogenous 200 and 400 µM melatonin can significantly inhibit the accumulation of reactive oxygen species in rice, by enhancing the activity of ROS scavenging enzymes. The accumulation of osmolytes was also stimulated by melatonin to endure with a better preservation of leaf water status in drought-stressed rice. Genome-wide expression profiling by RNA sequencing reveals an increase of oxidoreductase activity, iron ion binding, hydrolase activity, cell wall biogenesis, root development, while a decrease of nitrogen compound metabolism and cellular biosynthesis. Furthermore, melatonin could modulate rice drought response through the cooperation of WRKY, bHLH, ERF, MYB and NAC transcription factors, and modulates salicylic acid, ethylene and auxin pathways. Our finding provides new insights into melatonin-mediated drought tolerance in rice, and facilitates the rational applications in stress management for agricultural, horticultural, and floricultural plants.

Salt stress adversely impacts crop growth and development, resulting in stunted growth and diminished grain yield. Therefore, this study explores the synergetic effects of seed priming with iron nanoparticles (FeNPs) integrated with a genome-wide association study (GWAS) on the phenotypic, biochemical, and agronomic traits of 138 barley accessions under control, salinity stress, and seed iron priming treatments. A normal phenotypic distribution was observed across all accessions under the tested conditions, with significant natural phenotypic variation in response to the treatments. Remarkably, seed priming with FeNPs showed a significant enhancement in superoxide dismutase (SOD) activity and selective modulation of catalase (CAT) and glutathione reductase (GR) activities, indicating a targeted oxidative stress response. Compared to control and salinity stress conditions, priming with FeNPs showed substantial increases in all agronomic traits, including spike length (SL), number of spikelets per spike (NSS), number of grains per spike (NGS), weight of grains per spike (WGS), and thousand kernel weight (TKW), suggesting its potential to mitigate the adverse effects of salinity and promote better crop performance. Based on GWAS analysis, sixteen highly significant marker associations/candidate genes were detected to be associated with antioxidant components. Using quantitative real-time PCR analysis (RT-qPCR), FeNPs seed priming effectively modulates the plant's transcriptional response to salinity stress by balancing rapid gene activation with sustained stress adaptation. This approach mitigates excessive defense responses while promoting long-term stability through controlled upregulation of key genes, such as PP2C, Phosphotransferase, Terpene Synthase Putative, and RWP-RK. The findings support the potential of FeNPs as a biotechnological tool to enhance crop resilience and optimize agronomic performance under adverse environmental conditions.

Grey hair, a common ageing-associated phenomenon in humans, is mainly attributed to the damage of melanocytes and the absence of melanin. Grey hair has long been treated with traditional medicine, and new research has shown that various plant-derived monomers can increase tyrosinase activity and melanogenesis, indicating that they may have therapeutic value in curing grey hair. In this study, we outlined the role of melanin and pigmentation during hair growth and collected various medicinal plant monomers with the potential value of grey hair reversal. Many active ingredients from medicinal plants, such as fraxinol, tribuloside, morin and naringenin, can upregulate melanogenesis and tyrosinase activity through different signalling pathways. Some of them can promote melanosome quantity, maturation and transportation as well. Monomers isolated from medicinal plants may act as stimulators of melanogenesis. Many plant-derived monomers perform as activators that upregulate melanin synthesis and tyrosinase activity through different signalling pathways. They are of great research value for the treatment of hair greying. Moreover, to further improve experimental effect, safety and reliability, a systematic and comprehensive evaluation system needs to be established in the future before studying their clinical efficacy.

Understanding the molecular mechanisms underlying cold and methyl jasmonate (MeJA) responses is vital for improving the cold tolerance of grapes. This study treated 'Pinot noir' plantlets with MeJA, screened key genes in the regulatory pathway using transcriptomics and proteomics analyses, and investigated their regulatory mechanisms under cold stress. The results showed that 50 μmol L-1 MeJA significantly inhibited the growth of grape roots length, increased the endogenous MeJA content and antioxidant enzyme activities, and reduced membrane damage under cold stress. In addition, 50 μmol L-1 MeJA and cold stress treatment greatly increased the number of differential genes and metabolites in the phenylalanine synthesis and hormone signal transduction pathways. The results indicated that VvPAL10, an important gene in the phenylalanine synthesis pathway, significantly improved transgenic Arabidopsis thaliana and grapevine callus tissue tolerance to low temperatures.

Flemingia macrophylla (Willd.) Merr. is commonly used as ethnic medicine in southwestern China, with genistein and genistin being the primary medicinal components. Magnesium has a significant impact on the growth, development, and accumulation of these active ingredients in F. macrophylla. To explore the effects of magnesium on the growth, development, and accumulation of active ingredients in F. macrophylla, tissue culture seedlings of F. macrophylla were treated with six different concentrations of magnesium, namely, T0 (0 g L-1), T0.5 (0.045 g L-1), T1 (0.09 g L-1), T2 (0.18 g L-1), T3 (0.27 g L-1), and T4 (0.36 g L-1), and then subjected to multi-omic analyses. Analysis of agronomic traits revealed that magnesium treatment significantly promoted rooting, whereas magnesium deficiency restricted root growth. Simultaneously, analysis of seedlings using high-performance liquid chromatography showed that 0.27 and 0.36 g L-1 magnesium significantly enhanced genistein synthesis and inhibited glycosylation (genistin). Furthermore, transcriptomic, proteomic, and metabolomic analyses indicated that differentially expressed genes and proteins under varying magnesium concentrations were primarily involved in phenylpropanoid, isoflavone, and flavonoid biosyntheses. Specifically, eight gene-protein pairs related to the synthesis of genistein and genistin were identified. Overall, our study identified essential genes/proteins involved in the synthesis of these two compounds and provided new ways to regulate genistein synthesis in medicinal plants.

Polycystic ovary syndrome (PCOS) is a common endocrine-metabolic disease in women of reproductive age. One of its core pathologies is ovarian granulosa cell (GC) dysfunction, and ferroptosis, as a novel cell death mode dependent on iron ions and lipid peroxidation, may be involved in the PCOS process, but the exact mechanism is unknown. Plumbagin (PLB) shows potential in PCOS treatment due to its antioxidant properties. The present study aimed to elucidate the molecular mechanisms by which PLB ameliorates mitochondrial dysfunction and ferroptosis in PCOS GCs through the YTH N6-methyladenosine RNA binding protein 1/L-type amino acid transporter 1 (YTHDF1/SLC7A5) axis.

Psoriasis is a common chronic inflammatory skin disease characterized by epidermal keratinocyte hyperproliferation and persistent immune activation. Histone deacetylase 3 (HDAC3), a member of the class I HDAC family, plays critical roles in regulating immunity and inflammation. However, its precise expression profile and functional contribution to psoriasis pathogenesis remain poorly defined.

Berbamine (BBM) has been reported to play an important role in the anti-inflammatory and anti-neoplastic activities. However, whether BBM mediates the anti-tumor efficacy in renal cell carcinoma (RCC) cells and the potential molecular mechanisms remain unclear.

Triple-negative breast cancer (TNBC) is associated with a poor prognosis due to insufficient molecular subtyping precision and limited actionable targets. Although metabolic reprogramming underlies TNBC chemotherapy resistance, establishing metabolic subtyping systems and investigating drug sensitivity across distinct metabolic subgroups could provide novel therapeutic avenues for breast cancer management. GSVA (Gene Set Variation Analysis) analysis of metabolic pathways reveals significant differences in TNBC (Triple-Negative Breast Cancer) patients. TNBC patients are classified into four metabolic subtypes through consensus clustering, based on their GSVA values of metabolic pathways. These subtypes are: MS_1, characterised by increased lipogenic activity; MS_2, characterised by increased carbohydrate and nucleotide metabolism; MS_3, a metabolism-active subtype with activation of all types of metabolism; and MS_4, characterised by suppressed metabolic activity across all types of metabolism. We next propose a novel method called MODIN (Multiomics-Driven Drug-Cell Interaction Network), which embeds multi-omics gene information (mRNA expression, copy number variation and DNA methylation) and drug SMILES data into a latent space, and then employs a multi-head attention-based interaction module to accurately predict the LN_IC50 values of 621 drugs in TNBC. Based on MODIN, noteworthy disparities in drug sensitivity emerge between the patient cohorts categorised as MS_2 and MS_3. MS_3 patients show a significantly higher sensitivity to chemotherapy regimens, especially for doxorubicin and docetaxel, while the MS_2 cohort displays marked resistance to these drugs. Our study reveals the metabolic heterogeneity of TNBC, and TNBC patients with increased carbohydrate and nucleotide metabolism exhibit the poorest prognoses and greater resistance to doxorubicin and docetaxel.

Antimicrobial peptides (AMPs) possess vaccine adjuvant activity; however, their specific targets and molecular mechanisms remain incompletely understood, which hinders their clinical application. This study aimed to elucidate the key targets and pathways through which the antimicrobial peptide BSN-37 modulates immune responses in macrophages, providing evidence for its potential clinical translation. In this investigation, Balb/c mice were administered BSN-37 for 12 h, after which total RNA was extracted from peritoneal macrophages to assess the mRNA expression levels of cytokines and key molecules on the cell surface, followed by transcriptomic sequencing. The results demonstrated that BSN-37 significantly upregulated the mRNA expression of these molecules and cytokines. A total of 228 differentially expressed long non-coding RNAs (lncRNAs) (121 upregulated, 107 downregulated) and 149 differentially expressed mRNAs (104 upregulated, 45 downregulated) were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed significant enrichment of differentially expressed mRNAs in immune response pathways, PI3K-Akt signaling, and NOD-like receptor signaling. Differentially expressed lncRNA target genes were associated with T cell receptor signaling, PD-1 checkpoint regulation, and other immune regulatory pathways. Protein-protein interaction network analysis identified core genes such as CCchemokine receptor 1 (CCR1) and Toll Like Receptor 8 (TLR8). Molecular docking studies confirmed that BSN-37 exhibited strong binding affinity to TLR8 and CCR1, with binding energies less than - 5 kcal/mol. RT-qPCR validation confirmed the reliability of the sequencing data. These findings indicate that BSN-37 activates multiple immune response pathways in macrophages by targeting immune-related genes such as TLR8 and CCR1, offering theoretical support for the development of novel immune adjuvants.

Hepatocellular carcinoma (HCC) presents a significant global health challenge, marked by high mortality and recurrence. This study investigated the synergistic potential of cisplatin and palbociclib (C + P) against HCC cell lines. RT-qPCR revealed that C + P significantly downregulated HCC-related genes, including Hsp90, β-catenin, and components of the PI3K/AKT/mTOR pathway, compared to cisplatin alone and controls. Western blotting confirmed a reduction in phosphorylated AKT (P-AKT) with palbociclib and C + P, while PTEN, a tumor suppressor, was upregulated in the C + P group. Annexin V-FITC assays demonstrated a substantial increase in apoptosis in palbociclib and C + P treated cells. Cell cycle analysis indicated S and G0-G1 phase arrest with C + P, suggesting a combined cytotoxic effect. Scratch wound assays showed that both palbociclib and C + P significantly inhibited cell migration compared to cisplatin and controls. These findings suggest a promising synergistic effect of C + P in overcoming cisplatin resistance in HCC. However, further research is needed to fully elucidate the complex interactions between these drugs.

Extracellular traps (NETs) released by neutrophils during inflammation play a role in clearing infection but also contribute to disease pathology. NETs consist of a DNA backbone containing histones, anti-microbial granule proteins, such as myeloperoxidase (MPO), and other proteins. MPO remains enzymatically active and generates hypochlorous acid (HOCl) to kill pathogens. However, HOCl also readily reacts with proteins, but whether histones and other NET proteins are modified by this oxidant is unknown. This is significant as post-translational modification of histones alters their intracellular and extracellular reactivity. In this study, we used a proteomic approach to characterise the protein composition of NETs and identify HOCl-induced oxidative modifications on histones and other proteins. NETs were collected from primary neutrophils and the PLB-985 cell line and stimulated with phorbol myristate acetate (PMA) or nigericin, a bacterial peptide derived from Streptomyces hygroscopicus. There was evidence for Lys nitrile and aminoadipic semialdehyde formation, Tyr and Trp chlorination, and Met oxidation on histones and other proteins, including quinone oxidoreductase. Chlorination of Tyr-88 on histone H4 was particularly abundant and occurred to a greater extent in NETs from neutrophils exposed to PMA compared to nigericin, consistent with nigericin triggering NET release via a non-oxidative pathway. Chlorination of histone H4 Tyr-88 was also observed in the nuclear and cytoplasmic cell extracts of stimulated cells and could be decreased on treatment of the neutrophils with the MPO inhibitor AZD5904. These findings provide the first evidence that HOCl modifies proteins within NETs, particularly histone H4, which may be relevant in disease.

Despite Myc oncoproteins being major causal factors in human cancer, they remain "undruggable." The MYCN oncogene is one of the most powerful prognostic markers for the childhood cancer neuroblastoma and represents an important target for developing novel therapeutics. Here, we report the finding and characterization of M606, a selective small molecule inhibitor of MYCN, which was identified by screening a diverse chemical library. M606 reduced MYCN protein levels in neuroblastoma cell lines and upregulated hypoxia-inducible factor 1 alpha (HIF1A). Using siRNA-mediated knockdown of MYCN, c-Myc, or HIF1A in HepG2 and BE(2)-C cells followed by M606 treatment, we demonstrated that Myc downregulation and HIF1A upregulation were two independent effects of M606 treatment. M606 selectively targeted neuroblastoma cell lines expressing higher levels of MYCN protein and delayed neuroblastoma development in the TH-MYCN transgenic mouse model. Metabolomic analysis showed that M606 modulated glucose metabolism, consistent with a hypoxic response and iron deprivation. Biochemical characterization of M606 not only confirmed its iron-chelating properties but also revealed its ability to downregulate MYCN promoter activity, which could be rescued by the addition of iron. Luciferase assays identified the minimal MYCN promoter region required for the M606 response, which contained overlapping E2F transcription factor binding sites. Further evaluation defined a key role for E2F3 in the M606-mediated response. The finding of a potent cell-permeable iron chelator that can chelate iron to directly downregulate MYCN transcription via an E2F3-mediated response represents a potentially valuable therapeutic approach in the treatment of cancers overexpressing Myc oncoproteins.