Nitric oxide (NO) is a key signaling molecule that plays a vital role in maintaining homeostasis of physiological processes such as immune responses and neurotransmission. However, excessive NO production during inflammatory responses to infection can lead to cytotoxicity and tissue damage. The nasal epithelial barrier is a crucial first line of immunological defense against viral infections, and it is likely exposed to excessive NO levels during chronic inflammation. Therefore, clarifying the effects of NO on this barrier is thus critical. In this study, we investigated the biological effects of sustained NO exposure on RPMI2650 human nasal epithelial cells. Post-NO exposure transcriptomic analyses revealed significant upregulation of genes involved in the p53 signaling pathway. RT-qPCR analyses confirmed the temporal upregulation of p53 target genes associated with apoptosis and cell cycle regulation. These gene expression changes downregulated cell proliferation and induced cell death. Our findings suggest that excessive NO exposure induces nasal epithelial cell death via the p53 pathway, which over the long term can result in tissue damage and dysfunction under inflammatory conditions. These results provide new insights into how prolonged NO exposure affects the nasal epithelial cells and may contribute to the progression of chronic infectious diseases.

Melanoma, a malignant tumour originating from melanin-producing melanocytes, poses a significant threat to human health, including cutaneous melanoma and uveal melanoma (UM). Although surgical resection remains a primary treatment modality, radiotherapy has emerged as another therapeutic option, particularly for UM. Nevertheless, the adverse effects induced by radiotherapy are considerably pronounced. In this study, we identified WT-161, a selective histone deacetylase 6 (HDAC6) inhibitor, as a potent radiosensitizer for melanoma therapy through high-throughput drug screening. Mechanistically, inhibition of HDAC6 disrupted its interaction with DNA damage repair proteins (DDB2 and DEK) and suppressed gene expression in the DNA damage repair pathway, leading to the accumulation of irradiation-induced DNA damage and tumour regression. Our findings establish HDAC6 as a predictive biomarker for radiation response in melanoma and demonstrate that pharmacological inhibition of HDAC6 with WT-161 could expand the clinical utility of radiotherapy in patients with UM.

Fufang Baizhi Tincture (FFBZD) is a traditional topical Chinese medicine. This study demonstrates that FFBZD regulates melanocytes by modulating apoptosis via Caspase 7 (CASP7) and transcriptional regulation via E1A-associated protein p300 (EP300), thereby influencing vitiligo. These findings further links its traditional use with specific genetic targets and mechanism-based preventive and therapeutic potential.

Activation of hypoxia-inducible factors (HIFs) supports cancer cell survival, yet how HIFs govern cell death remains unclear, despite evidence that HIF-1 acts as a tumor suppressor in cell renal cell carcinoma (ccRCC). Here, we report a cell death-priming role for HIF-1/2 in ccRCC. Through cell viability screens with chemical libraries, we identify SGI1027 and its analog MS1129 as HIF-1/2-dependent cell death inducers that specifically kill VHL-deficient ccRCC cells in vitro and patient-derived xenografts in mice. Mechanistically, SGI1027 and MS1129 induce proteasomal degradation of DNMT1/DNMT3A/DNMT3B proteins, leading to the loss of promoter methylation and subsequent upregulation of TRAIL, DR4, and DR5 in ccRCC cells. HIF-1/2 induces procaspase-10 expression serving a commitment point to activate TRAIL-induced apoptosis in VHL-deficient ccRCC following SGI1027 or MS1129 treatment. Notably, recombinant TRAIL protein synergizes with SGI1027 or MS1129 to kill VHL-deficient ccRCC in mice. Collectively, our study unveils an apoptosis induction strategy that involves hijacking HIFs for ccRCC treatment.

The pathological mechanism of poor osseointegration of implants caused by aseptic loosening of implants is unclear. This study investigated the changes in implant osseointegration induced by titanium ions and explored the relationship between titanium implants, immune cells and osseointegration and the underlying mechanism of aseptic loosening of titanium implants. In vitro, mouse osteoblasts and mouse macrophages were co-cultured in a titanium ion environment that did not affect osteogenic activity. The results showed that the presence of macrophages in this titanium ion environment caused a decrease in osteogenic capacity. In addition, titanium ions lead to M1 polarization of macrophages and the production of inflammatory mediators, the inflammatory response can affect the ability of osteoblasts. Through transcriptome sequencing analysis of osteoblasts, it was found that the angiotensinogen (Agt) was up-regulated in osteoblasts added to the titanium ion group and knockdown or overexpression of Agt could change the effect of titanium ions on osteoblasts through macrophages. These results indicate that the application of gene Agt may provide new ideas for solving aseptic loosening of implants.

The escalating prevalence of multidrug-resistant Salmonella enterica serovar Typhimurium (S. Typhimurium) necessitates the urgent development of innovative therapeutic strategies. Targeting bacterial virulence factors has emerged as a promising alternative to conventional antibiotics. Among these factors, type I fimbriae serve as essential structural determinants for host colonization and pathogenesis. In this study, we investigated the anti-virulence potential of Tectorigenin (TE), a natural flavonoid isolated from the traditional medicinal herb Belamcandae Rhizoma. Our results demonstrate that TE significantly inhibits swarming motility, biofilm formation, and hemagglutination capacity of S. Typhimurium by directly influence key structural and regulatory genes within the fim operon, including fimA, fimD, fimF, fimH, fimI and the regulator STM0551. In vivo experiments further confirmed that TE treatment alleviates clinical symptoms in S. Typhimurium-infected mice, effectively reducing intestinal inflammation and preserving epithelial barrier integrity. Collectively, these findings highlight TE as a potent anti-virulence agent that targets type I fimbriae, offering a novel non-bactericidal approach for the management of S. Typhimurium infections.

Osteoarthritis (OA) is a prevalent degenerative joint disease with no curative treatment currently available. Recent evidence suggests that chondrocyte ferroptosis contributes to OA progression. Chondroitin sulfate (CS), widely used in OA management, exhibits anti-inflammatory and antioxidant properties, yet its role in modulating ferroptosis remains unclear. In this study, we investigated whether CS alleviates OA by inhibiting chondrocyte ferroptosis and explored the underlying mechanisms. Using an in vitro ferroptosis model induced by RSL3 in rat chondrocytes, we found that CS significantly restored cell viability and ameliorated ferroptosis-related changes, including reduction of intracellular and mitochondrial ROS, lipid peroxidation, and iron overload. CS also downregulated the expression of ferroptosis markers PTGS2 and ACSL4, while upregulating SLC7A11 and HSPA8 in a dose-dependent manner. Network pharmacology and transcriptomic analysis identified HSPA8 as a key overlapping gene among CS targets, OA-related differentially expressed genes, and ferroptosis-related genes. In a rat OA model induced by modified Hulth surgery, CS treatment attenuated cartilage degradation, as evidenced by improved OARSI scores, restored COL2A1 expression, and suppressed MMP13. Immunohistochemistry confirmed that CS upregulated SLC7A11 and HSPA8 while downregulating ACSL4. These findings demonstrate that CS mitigates OA progression by inhibiting chondrocyte ferroptosis, potentially through upregulation of HSPA8 and subsequent enhancement of SLC7A11 expression. Our study provides novel insights into the mechanism of CS in OA treatment and highlights ferroptosis as a promising therapeutic target.

Lignin deposition in stone cells is critical for pear fruit quality. However, calcium ions (Ca2+) exert critical regulatory effects on fruit growth and development. Nevertheless, the molecular mechanisms underlying Ca2+-mediated stone cell formation in pear remain poorly characterized. Our study revealed that exogenous application of CaCl2 decreased lignified stone cell formation in "Nanguoli" (Pyrus ussuriensis) fruits and significantly downregulated the expression of lignin biosynthesis-related genes laccase7 (PuLAC7) and peroxidase42 (PuPRX42). Transcriptome sequencing (RNA-seq) identified a transcription factor, PuMYB73, which was significantly inhibited by CaCl2 in pear fruit stone cell formation and lignin accumulation. Yeast one-hybrid (Y1H) and β-glucuronidase (GUS) activity analysis revealed that PuMYB73 directly binds and activates lignin biosynthesis genes PuPRX42 and PuLAC7 promoters, thereby decreasing PuPRX42 and PuLAC7 expression after CaCl2 treatment. Strikingly, PuMYB73 interacts with PuNAC21 to form a Ca2+-responsive module, lowering the transcription of PuPRX42 and PuLAC7 after Ca2+ treatment, which contributed to decreasing pear stone cell production. Collectively, exogenous CaCl2 treatment inhibits stone cell and lignin biosynthesis in pears mediated by the PuMYB73-PuNAC21 regulatory module. Our results revealed that the Ca2+-PuMYB73-PuNAC21-PuLAC7/PuPRX42 regulatory module inhibits lignin biosynthesis, providing important insights into reducing stone cell content in pear via molecular breeding.

The m6A modification plays key roles in RNA metabolism and function and is implicated in various human diseases. In this study, we reported a novel molecular glue strategy for transcript-specific m6A editing using synthetic bifunctional molecules containing an RNA-targeting moiety and a ligand that recruit an endogenous m6A erasing enzyme. Through cyclic γ-AApeptide library screening, we identified a novel peptidomimetic binder to the long noncoding RNA MALAT1 A2577 region, which has a high m6A level. We developed a bifunctional molecular glue by coupling the identified MALAT1-binding cyclic γ-AApeptide to fluorescein, a reported binder to the m6A eraser FTO. We demonstrated that this bifunctional molecular glue successfully recruited FTO to the target RNA site, achieved the m6A erasing, disrupted HNRNPC-MALAT1 binding, and destabilized MALAT1. We anticipate that this novel molecular glue strategy will offer a new direction in developing molecules to regulate RNA modifications.

Bone fracture healing is a multifaceted process that involves different stages and intercellular interactions. In this study, we aimed to investigate the effect of Taohong Siwu decoction (TSD) on bone fracture healing and the underlying mechanisms.

The addition of CDK4/6 inhibitors to endocrine therapy has significantly improved outcomes in HR+/HER2- breast cancer (BC). However, variable patient responses and acquired resistance remain a clinical challenge. We therefore defined the comprehensive molecular response to palbociclib, the most clinically used CDK4/6 inhibitor. Global analyses of gene expression, protein abundance, splicing, and chromatin accessibility revealed broad patterns and specific changes that result from CDK4/6-inhibition in BC cells. We uncovered unexpected feedback between CDK4/6 and estrogen-response signaling, which has clear clinical implications. We also revealed a widespread alternative splicing program that partially overlapped with genes whose expression is regulated, and which is expected to impact protein function. These molecular changes nominated combination therapies that interfere with the activation of CDKs or ERα. Accordingly, co-targeting CDK7, which regulates CDK2, CDK4/6, and ERα, additively impacted cell fitness. Collectively, these data reveal a complex, multitiered response to CDK4/6 inhibition, with implications for therapeutic efficacy.

Inhibiting telomerase could lead to telomere shortening, chromosomal instability, and eventually cell death in cancer cells. The use of steroids in cancer treatment is significant; besides, drug repurposing can be a quicker and less expensive method of drug discovery than de novo drug development. Accordingly, diverse 280 steroidal drugs-following a pharmacophore-based rationale-were docked against telomerase receptor and compared to the co-crystallized inhibitor (BIBR1532) as a reference standard. Additionally, a molecular dynamics simulation for 200 ns was performed to confirm the docking results. Moreover, the six steroidal drugs (Betamethasone disproportionate, Beclomethasone disproportionate, Clobetasone butyrate, Desonide, diflucortolone valerate, and Hydrocortisone butyrate) were selected for further in vitro investigation. The antitumor activities of the six steroidal drugs against H1299, HuH7, HCT116, A549, MDA-MB-231, MCF7, PC3, MG63, and A375 cancer cell lines, besides OEC and HSF normal cell lines, were evaluated and compared to doxorubicin (Dox) as a reference positive standard. Especially, Clobetasone butyrate represented the better cytotoxicity against seven cancer cell lines (H1299, HuH7, HCT116, MDA-MB-231, PC3, MG63, and A375) with IC50 values of 12.56, 13.95, 12.00, 17.51, 11.69, 20.64, and 20.21 µg/mL, respectively. The outstanding antitumor six steroidal drugs were further investigated for their telomerase downregulatory potentials. All analogs recorded outstanding downregulatory results against telomerase, especially Clobetasone butyrate, Desonide, Diflucortolone valerate, and Hydrocortisone butyrate, where the protein expression for telomerase was downregulated up to 0.66-, 0.76-, 0.56-, and 0.73-fold change for clobetasone butyrate, desonide, diflucortolone valerate, and hydrocortisone butyrate, respectively, compared to the control.

Despite mounting evidence that commensal microbes enhance host defenses, whether and how they directly suppress pathogen virulence remains elusive. Here, we investigate metabolites from the gut microbiota of infection‑resistant Tibetan chickens for their ability to reduce Salmonella virulence gene expression and elucidate the molecular mechanism by which these compounds inhibit the LuxS/AI‑2 quorum‑sensing system.

Sedum plumbizincicola is an efficient cadmium (Cd) hyperaccumulator with strong potential for phytoremediation. Although SpCTP3 has been identified as a key Cd-responsive gene, its functional contribution to Cd tolerance remains partially characterized. Here, SpCTP3-overexpressing (OE) plants displayed increased Cd accumulation, improved growth, and enhanced photochemical efficiency under Cd exposure, providing phenotypic evidence supporting a positive role of SpCTP3 in Cd response. Transcriptomic profiling further indicated that SpCTP3 overexpression modulates pathways related to redox homeostasis and stress signaling. Moreover, we identified a Cd-inducible transcription factor, SpMYB108, that binds to and activates the SpCTP3 promoter. Yeast one-hybrid and dual-luciferase assays confirmed this direct regulatory interaction. Collectively, these results provide integrated phenotypic, transcriptional, and regulatory insights into the role of SpCTP3 in Cd hyperaccumulation.

Transforming growth factor (TGF)-β1 is a pro-fibrotic cytokine that affects extracellular matrix (ECM) deposition and fibroblast activity. 17β-Estradiol (E2), the dominant ovarian steroid during the follicular phase (FLP) of the estrous cycle, can also influence ECM remodeling and fibrosis, through prostaglandin (PG) synthesis. PGs have opposing roles in fibrosis, with PGE₂ showing anti-fibrotic effects and PGF₂α promoting fibrosis. Equine endometrosis, whose main pathological feature is fibrosis, is marked by chronic inflammation and ECM accumulation, and may involve mediators like TGF-β1, PGs, and E2. This study aimed to assess how TGF-β1, E2, and their combination affect PG synthase and receptors transcription (qPCR) and PG concentrations (ELISA) in equine endometrial explants during the FLP, after 24 and 48 h. Prostaglandin-endoperoxide synthase 2 (PTGS-2) mRNA was reduced with TGF-β1 and combination treatments at 24 h. Estradiol and combined treatments downregulated microsomal prostaglandin E synthase1 (PGES) mRNA at 24 h, while prostaglandin F synthase (PGFS) mRNA reduced with TGF-β1 at 24 h and with E2 at 48 h. The PGE₂ concentration was lower in TGF-β1 +E2 group than in controls and TGF-β1 alone at 48 h. In contrast, PGF₂α concentration increased with E2 at 24 h and TGF-β1 and TGF-β1 +E2 treatments at 48 h. Prostaglandin E receptor (EP)2 and 4 mRNA upregulated with the combination treatment, while prostaglandin F receptor (FP) mRNA decreased in all treated groups. These findings suggest that TGF-β1 and E2 interact to regulate PG pathways, with potential to drive fibrotic changes in the equine endometrium, by shifting the balance between anti- and pro-fibrotic mediators like PGE₂ and PGF₂α.

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental pollutants increasingly linked to human disease. Perfluorohexanesulfonic acid (PFHxS), a widespread PFAS detected in human serum, has an uncharacterized role in gastric cancer (GC), one of the leading causes of cancer mortality worldwide. Here, we employed an integrated multi-omics approach combining network toxicology, large-scale transcriptomic analyses from TCGA and GEO cohorts, single-cell RNA sequencing, and molecular simulations, followed by in vitro and in vivo validation at environmentally relevant concentrations. We identified 667 common targets of PFHxS and GC and developed an 11-gene gradient boosting machine (GBM) prognostic signature that robustly stratified patient survival across independent cohorts and correlated with distinct tumor immune microenvironments. Mechanistically, PFHxS was predicted and experimentally confirmed to directly bind Kelch-like ECH-associated protein 1 (KEAP1), a key regulator of oxidative stress. Chronic low-dose PFHxS exposure downregulated KEAP1 protein expression, disrupted KEAP1/NRF2 antioxidant signaling, and promoted GC cell proliferation, migration, invasion, and tumor growth in vivo. Together, these findings provide the first molecular evidence that PFHxS acts as an environmental driver of GC progression by targeting KEAP1, while also delivering a clinically relevant prognostic tool. This work highlights a previously unrecognized environmental risk factor for gastric cancer and offers new perspectives for risk assessment, prevention, and therapeutic intervention.

Emerging environmental health issues posed by micro- and nanoplastics (M/NPs) have raised significant concerns. Accumulating evidence suggested that M/NPs can bioaccumulate in gonads and impair fertility in animals, yet the underlying cellular mechanisms and tissue-specific responses remain poorly understood. In this study, we employed in vivo and in vitro models to systematically investigate the impact of polystyrene micro- and nanoplastics (PS-M/NPs, 100 nm and 5 µm) on ovarian development and function in pubertal female mice. Following 35-day exposure, we observed size-dependent reproductive toxicity, with 100 nm PS-NPs causing reduced body weight gain and ovarian size, disrupted folliculogenesis, and altered hormone levels. Leveraging single-cell RNA-sequencing (scRNA-seq), we uncovered profound alterations in intracellular communication networks across seven ovarian cell types. Granulosa cells (GCs) were identified as the primary target of PS-NPs, exhibiting marked transcriptional changes, including dysregulation of FSCN1, a critical actin cytoskeleton regulator. In vitro experiments confirmed that only 100 nm PS-NPs were internalized by GCs, leading to cell cycle arrest, necroptosis, and hormonal dysfunction. Mechanistically, PS-NPs triggered F-actin cytoskeleton remodeling, increasing cell stiffness and histone modifications (H3K4me3, H3K27ac) associated with chromatin accessibility. Integrated ATAC-seq and RNA-seq analyses implicated STAT1 as a key transcriptional regulator driving PS-NP-induced epigenetic and transcriptional changes. Overall, our findings establish the first single-cell resolution atlas of PS-NP-mediated ovarian toxicity, revealing that NPs disrupt reproduction through cytoskeletal damage and epigenetic reprogramming. This work provides unprecedented insights into the molecular and epigenetic consequences of M/NPs in mammalian reproduction, emphasizing the potential health risks of environmental M/NP exposure.

5-Aminolevulinic acid (ALA) is a plant growth regulator that enhances salt tolerance, yet the underlying mechanisms remain unclear. In this study, we investigated the function and molecular mechanism of the Protein Phosphatase 2A (PP2A) catalytic subunit MdPP2AC in ALA-induced salt tolerance in detached leaves, calli, and rooted plantlets of 'Gala' apple (Malus × domestica). Results showed that pretreatment with ALA before NaCl stress significantly alleviated NaCl-induced damage, enhanced antioxidant enzyme activities, and improved chloride (Cl-) interception in the roots with less transport to the aboveground. Further analysis revealed that the PP2A activity and MdPP2AC expression were induced by NaCl and further by ALA. Functional studies showed that overexpressing (OE)-MdPP2AC enhanced salt tolerance, whereas RNA interference (RNAi)-MdPP2AC or application of cantharidin (CT, a specific inhibitor of PP2A activity) compromised salt tolerance, but exogenous ALA mitigated CT-aggravated salt injury. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), luciferase complementation imaging (LCI), sequence truncation and site-directed mutagenesis assays confirmed that MdPP2AC can interact with the chloride channel protein MdCLC-c2, and the C-terminal Glu684 residue of MdCLC-c2 is indispensable for the molecular interaction and Cl- transport activation. Function identification with Δgef1 mutant yeast showed that MdCLC-c2 transformation promoted intracellular Cl- accumulation as well as cell salt tolerance, and the effects were further promoted by MdPP2AC transformation and exogenous ALA. Collectively, we propose that MdPP2AC mediates ALA-induced salt tolerance in apple by interacting with MdCLC-c2, which promotes Cl- sequestration into root vacuoles with less transport to the aboveground, thus maintaining Cl- homeostasis at the cellular and plant levels. These findings reveal a novel mechanism whereby PP2AC modulates chloride channel function to maintain ion homeostasis within the ALA signaling pathway, providing critical insights into ALA-mediated apple salt tolerance.

Chronic low-grade inflammation underlies many microvascular complications of diabetes, including diabetic kidney disease (DKD). Lipoxins (LXs), an endogenously produced family of lipid mediators, resolve inflammation and protect against renal scarring as occurs in DKD. This study examined the mechanism by which LXs protect against DKD, focusing on the regulation of VCAM-1 and the recruitment of macrophages to the diabetic glomerulus. LXA4 and two fourth-generation mimetics were assessed in diabetic ApoE knockout mice, followed by in vitro studies in the main renal cell populations, including podocytes, proximal tubular, mesangial, and glomerular endothelial cells. LXs attenuated albuminuria, mesangial expansion, and collagen and fibronectin deposition as both a preventive and delayed intervention in experimental DKD. LXs also consistently attenuated the TNF-α-induced expression of VCAM-1 in all the human and mouse renal cell populations examined. Further analysis identified that the renoprotection was in part mediated by an epigenetic modification of the VCAM-1 gene through H3K4 monomethylation, which did not appear to be dependent on NF-κB activation in human glomerular endothelial cells. LXs protect against DKD by modulating glomerular endothelial cell inflammation and via a novel LX-mediated epigenetic mechanism regulating the VCAM-1 promoter in these cells.

The growing global demand for protein, combined with the urgent need for effective strategies to manage conditions such as obesity and diabetes, highlights legume proteins as valuable sources of derived bioactive peptides with health-promoting properties. In this study, we employed the Caenorhabditis elegans model to investigate the effects of supplementation with hydrolysates derived from fava bean and pea protein on healthspan. Supplementation with fava bean and pea protein hydrolysates reduced fat accumulation and age-related lipofuscin pigment in the worms, without impairing their development. The fava bean protein hydrolysate significantly decreased total reactive oxygen species levels and enhanced stress tolerance to juglone exposure, suggesting the modulatory activity of the mitochondrial oxidative stress response. In contrast, pea protein hydrolysate improved the heat stress resistance of C. elegans, and gene expression and mutant analyses revealed the involvement of the endoplasmic reticulum unfolded protein response (ER-UPR) pathway in mediating its health-promoting effects. Together, our data demonstrate that fava bean and pea protein hydrolysates support healthspan in C. elegans by modulating distinct cellular stress response pathways and pave the way for further investigation in more complex animal models.