Vibrio parahaemolyticus is a foodborne pathogen that can cause severe gastroenteritis. After entering the human intestine through contaminated seafood (1.00% NaCl) V. parahaemolyticus will encounter a physiologically related dual pressure environment: low salinity and elevated bile salts (0.03%-0.30%). Although bile salts can affect V. parahaemolyticus under optimal salinity conditions (3.00% NaCl), little is known about their effects on paralysis under low salt conditions (0.90% NaCl) in the intestinal stress environment. This research uniquely simulated this intestinal niche using 0.90% NaCl-0.10% bile salts, revealing its effects on growth kinetics, motility, biofilm formation, and transcriptome responses. The main findings include: significant inhibition of growth (prolonged the lag time (LT)), decreased the maximum specific growth rate (µmax)), swimming ability, and biofilm formation; But it enhances the ability to swarming; And unique transcriptome reprogramming. In addition, transcriptome sequencing revealed that swarming related genes, biofilm related genes, and T3SS virulence genes were significantly down regulated, while iron metabolism and swimming related genes were significantly up-regulated. It is crucial that KEGG enrichment indicates that the ribosomal pathway may be the central regulatory hub for observed biofilm and motility inhibition. This research provides the first comprehensive analysis of the effects of bile salts on intestinal related low salinity, providing important insights into the intestinal adaptation and pathogenic mechanisms of V. parahaemolyticus.

The clinical application of donafenib in advanced hepatocellular carcinoma (HCC) is restricted by its limited therapeutic efficacy and a variety of treatment-associated adverse events. These factors collectively underscore the need for more effective and well-tolerated therapeutic strategies.

Our previous studies have demonstrated that Xinfeng Capsule (XFC) exerts therapeutic effects on hyperinflammation-associated hypercoagulability and self-perception of patients (SPP) with osteoarthritis (OA). However, the underlying molecular mechanisms remain unclear.

Palladium-catalysed reactions have emerged as indispensable tools in medicinal chemistry, enabling the precise construction of C-C and C-N bonds across a wide spectrum of drug-like molecular frameworks. This manuscript comprehensively examines advances reported over the past five years in palladium-catalysed methodologies applied to epigenetic drug discovery. The mechanistic diversity and synthetic adaptability of palladium catalysts for accessing scaffolds addressing the epigenetic targets have been highlighted. The robust drug design strategies and activity profile of the generated small molecule epigenetic inhibitors through palladium-assisted synthetic protocol are also presented in this compilation. Particular emphasis is placed on understanding the influence of ligand structure, base selection, and solvent optimisation in modulating catalyst reactivity. Collectively, this review offers a practical and forward-looking framework for the design and synthesis of next-generation epigenetic anticancer therapeutics (selective/non-selective/hybrid-inhibitors and degraders/PROTACS).

Glioblastoma (GB) is a highly aggressive brain tumour with a poor prognosis and limited responsiveness to standard chemotherapy, particularly temozolomide (TMZ), due to intrinsic resistance mechanisms. This study investigates the potential of Aesculus hippocastanum, known as horse chestnut extract (HCE), to enhance the therapeutic efficacy of TMZ in GB cells through modulation of the Wnt/β-catenin signalling pathway. Combined treatment of HCE (500 μg/mL) and TMZ (100 μM) significantly reduced cell viability and inhibited wound healing and colony formation compared to either agent alone at 48 h. Notably, the expression of β-catenin and Wnt-1 was significantly reduced in the combination group, followed by a significant downregulation of Nestin and β3-tubulin, markers of glioma stem-like cells and aggressiveness, respectively. Furthermore, apoptotic activity was significantly increased following the combined treatment. In a 3D U87-spheroid model, the combination therapy resulted in a substantial reduction in spheroid area, suggesting impaired tumour growth. Propidium iodide (PI) staining revealed increased membrane permeability in cells treated with the combination, which was accompanied by an increase in p53 expression, supporting the induction of apoptosis. Collectively, these findings demonstrate that HCE increases the cytotoxic effects of TMZ by inhibiting Wnt/β-catenin signalling, reducing tumour stemness, and promoting apoptotic pathways in GB cells.

Stomatal development has emerged as a valuable model for studying developmental processes. Examining gene function along the stomatal lineage often requires gene perturbation in a controlled and cell-stage-specific manner, but this remains tedious without a dedicated genetic tool. Here, we describe Stomatal XVE, a modular, two-component XVE-based inducible system that enables user-controlled gene overexpression and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based knockout at defined stomatal cell stages in Arabidopsis thaliana. The system consists of a collection of estrogen-responsive XVE driver lines under cell-stage-specific promoters and effector vectors responsive to activated XVE. This design simplifies cloning and allows users to scale their investigation. We validated the cell-stage specificity and inducibility of the XVE driver lines and characterized key induction parameters. To test the system functionally, we employed it to study MAPKKK YODA and a pathogen effector AvrPtoB. While YODA overexpression reproduced known early- and late-stage phenotypes, stage-specific knockouts argued against its late-stage role in guard cell (GC) differentiation. Furthermore, AvrPtoB expression during later stages triggered striking disruptions in GC morphology and viability, revealing cell-type-specific effects of the pathogen protein. Overall, our Stomatal XVE system enables precise functional analysis of genes across defined stages of stomatal development and is particularly well suited for investigating genes with pleiotropic effects.

The plant hormone abscisic acid (ABA) is historically recognized as a tuberization stimulator, with exogenous application significantly enhancing potato tuber formation. However, the physiological significance of endogenous ABA signaling in tuberization and its underlying molecular mechanisms remain poorly understood. Here, by using ABA-insensitive hypermorphic StHAB1G276D-overexpression mutant and StHAB1 knockdown mutant, we demonstrate that ABA signaling is essential for normal tuber formation in potato. Blocking of ABA signaling by StHAB1G276D-overexpression reduces underground stolon sensitivity to leaf-derived tuber-inducing signals. Notably, StHAB1 directly interacts with the tuber-forming signal StSP6A and reduces its phosphorylation level. Transcriptomic and quantitative phosphoproteomic analyses proved that StHAB1G276D modulates gene expression and phosphorylation of key players in the tuberization pathway and gibberellic acid signaling. Collectively, our findings uncover a critical role of endogenous ABA signaling in potato tuber formation and identify a mechanism linking the conserved FLOWERING LOCUS T and ABA pathways, offering molecular insights for accelerating potato tuberization by modulating hormone signaling.

Aluminum (Al), as a ubiquitous environmental pollutant, has been implicated in the pathogenesis and progression of various neurodegenerative diseases due to its neurotoxic properties. Recent studies indicate that aluminum exposure may induce aberrant epigenetic modifications, thereby disrupting gene expression patterns and cellular functions, ultimately leading to neuronal damage. This review focuses on examining the cross-hierarchical regulatory mechanisms of aluminum exposure on DNA methylation, histone modifications, noncoding RNAs, and RNA modifications. We point out an issue in current research-the isolated analysis of individual epigenetic modifications often neglects their network-based cascade effects. Future investigations should focus on integrating multi-omics approaches with dynamic modeling to elucidate the hierarchical transmission mechanisms underlying epigenetic crosstalk.

Aqueous humour (AH) contains relatively high concentrations of vitamin C (ascorbate). AH drains out of the anterior chamber through the trabecular meshwork (TM) and, therefore, TM cells in vivo are routinely bathed in this antioxidant. Yet, most TM cells are cultured in vitro in media without ascorbate. In this study, we investigated molecules expressed by TM cells cultured with and without ascorbate.

Transglutaminases (TGs) are calcium-dependent enzymes that cross-link proteins, contributing to apoptosis, extracellular matrix (ECM) stabilization, and inflammation. While TG2 has been extensively studied in hepatic injury, the role of TG7 in oxaliplatin-induced liver responses remains unclear. Oxaliplatin, a third-generation platinum chemotherapeutic, effectively treats solid tumors but can induce hepatic stress through oxidative and pro-inflammatory signaling. Adult rats received intraperitoneal oxaliplatin (10 mg/kg weekly) for 6 weeks. qRT-PCR, immunohistochemistry (IHC), immunofluorescence (IF), and a TG activity assay assessed hepatic TG7 expression, localization, and activity. Oxidative stress indicators (serum malondialdehyde [MDA] and reduced glutathione [GSH]) and pro-inflammatory cytokine transcription (CASP3, interleukin-6 (IL-6), tumor necrosis factor α (TNF-α)) were evaluated. Oxaliplatin exposure markedly increased TG7 mRNA and protein levels, elevated TG enzymatic activity, raised MDA (+49.4%), depleted GSH (-18.6%), and upregulated CASP3, IL-6, and TNF-α. DNA fragmentation and microscopic observations from IHC- and IF-processed sections were consistent with apoptosis-associated DNA degradation and subtle stress-related structural variations. Immunostaining revealed altered TG7 distribution within hepatocytes and sinusoidal regions. In this oxaliplatin-exposed rat liver model, TG7 upregulation and increased TG activity were associated with oxidative stress, inflammatory cytokine induction, and apoptotic signaling. These findings identify TG7 as a stress-associated marker during oxaliplatin exposure and support further studies to clarify its mechanistic role and evaluate its potential as utility as a biomarker under chemotherapy-associated hepatic stress conditions.

Non-small cell lung cancer (NSCLC) continues to be the primary contributor to deaths associated with cancer. Current treatments are often limited by drug resistance and toxicity, highlighting the need for novel therapeutic approaches. Building on previous findings demonstrating that Gamabufotalin (CS-6) is effective against hepatocellular carcinoma, this study explores its mechanism of action in NSCLC. The findings indicate that CS-6 suppresses the proliferation and migratory capacity of NSCLC cells in a concentration-dependent manner, while significantly inducing apoptosis. The 48-hour half-maximal inhibitory concentration (IC50) ranged from 30 to 80 nM. In xenograft models, CS-6 effectively suppressed tumor growth (P < 0.05) without causing significant systemic toxicity at effective doses (25 mg/kg and 50 mg/kg). Mechanistically, coiled-coil-helix-coiled-coil-helix domain-containing protein 2 (CHCHD2) was identified as the direct molecular target of CS-6 through Limited Proteolysis-Mass Spectrometry (LiP-MS), validated by cell thermal shift assay (CETSA), MicroScale Thermophoresis (MST), and Surface Plasmon Resonance (SPR). CHCHD2, also known as mitochondrial nuclear retrograde regulator 1 (MNRR1), is a bi-organelle regulator located primarily in the mitochondrial intermembrane space, where it controls respiratory chain stability and cristae structure, thereby regulating cell survival and apoptosis[1-3]. CHCHD2 is essential for NSCLC cell survival, as both its knockdown and overexpression reduced the efficacy of CS-6. Furthermore, transcriptomic analysis revealed that targeting CHCHD2 with CS-6 activates interferon signaling and significantly upregulates the tumor suppressor X-linked inhibitor of apoptosis (XIAP)-associated factor 1 (XAF1). In conclusion, these findings establish the mitochondrial CHCHD2-XAF1 axis as a key mediator of CS-6 activity, thereby highlighting CS-6 as a promising candidate for targeted therapy in NSCLC.

Paracetamol and di(2-ethylhexyl) phthalate (DEHP) are endocrine-disrupting chemicals (EDCs) known to impair prenatal gonadal development and inhibit testosterone synthesis in experimental models. We hypothesized that these substances may interact with the endocannabinoid system (ECS), potentially contributing to testicular and ovarian developmental abnormalities. Pregnant Wistar rats were exposed to paracetamol (50 or 250 mg·kg⁻¹·day⁻¹) or DEHP (750 mg·kg⁻¹·day⁻¹) from gestational days 15-18. We assessed fetal testicular testosterone production, early postnatal ovarian follicle counts, and the expression of genes involved in steroidogenesis, testis descent, ovarian development, and ECS signaling, synthesis, and degradation. In a parallel in vitro approach, fetal rat testes from unexposed animals were incubated for three hours with paracetamol or its metabolite AM404, with or without the CB1 antagonist rimonabant. A separate group was exposed to MEHP, DEHP's main metabolite. In vivo, DEHP reduced anogenital distance and testicular testosterone in males, increased Cnr2 expression in both gonads, and upregulated Napepld in fetal testes. MEHP increased testosterone secretion in vitro. In ovaries, high-dose paracetamol reduced the number of healthy primordial and transitional follicles and increased atresia in primary and secondary follicles. DEHP exposure also elevated atresia in early-stage follicles. These findings suggest greater ovarian sensitivity to paracetamol compared to the testis.Unlike paracetamol, DEHP altered the expression of key ECS genes, suggesting a possible interplay between phthalates and the ECS. This raises the possibility that ECS components may be involved in the mechanisms of phthalate toxicity and could represent potential biomarkers, warranting further investigation.

To compare a non-ester methacrylate-alternative dental resin (ER) with conventional ester-based methacrylate resin (MAR) in terms of (i) susceptibility to degradation by human neutrophils (hN), (ii) influence on hN pro-inflammatory phenotypes, and (iii) modulation of Streptococcus mutans (SM) virulence-gene expression.

Endometrial cancer (EC) is a common gynecological malignancy characterized by abnormal glucose metabolism. Paeonol (Pae), a natural phenolic compound derived from traditional Chinese medicine, exhibits broad-spectrum antitumor activity. However, its role in modulating glycolysis and the underlying molecular mechanisms in EC remain poorly understood. The effects on EC cell viability (CCK-8), proliferation (EdU), apoptosis (flow cytometry), invasion (transwell), migration (wound healing), and tube formation rate were assessed. Glycolytic parameters were measured using corresponding commercial kits. Protein and mRNA expression levels were determined by Western blotting and RT-qPCR. The interaction between tripartite motif protein 26 (TRIM26) and lactate dehydrogenase A (LDHA) was investigated through co-immunoprecipitation (Co-IP), cycloheximide (CHX) chase, and ubiquitination assays. A xenograft model was established to examine the in vivo efficacy of Pae. Pae inhibited proliferation, metastasis, tube formation, and glycolysis of EC cells, and induced apoptosis. Pae suppressed EC malignant behaviors by downregulating LDHA expression. TRIM26 promoted ubiquitination-mediated degradation of LDHA. Overexpression of LDHA reversed the tumor-suppressive effects of TRIM26 overexpression in EC cells. TRIM26 knockdown attenuated the antitumor effects of Pae. In vivo experiments demonstrated that Pae inhibited tumor growth and regulated TRIM26/LDHA expression. Pae was found to promote TRIM26 expression, which in turn enhanced TRIM26-mediated ubiquitination and degradation of LDHA, thereby contributing to glycolysis inhibition and suppression of EC progression. These results suggested that Pae might exert its effects by modulating the TRIM26/LDHA axis and supported its potential therapeutic value in EC.

Microcystin-LR (MC-LR) could be largely released in water environment during the cyanobacterial blooms, endangering the health of plants, animals, and even humans. Numerous evidences had demonstrated a strong correlation between the toxicities of MC-LR and the oxidative stress induced by MC-LR. Hydrogen peroxide (H2O2) is one of the primary constituents of reactive oxygen species (ROS) and tends to be overproduced under oxidative stress. Therefore, detecting the changes in H₂O₂ levels in organisms exposed to MC-LR can serve as an indicator of MC-LR-induced oxidative damage. However, the studies of directly detecting H₂O₂ levels in organisms exposed to MC-LR are lacking. In this work, we developed a novel near-infrared probe, DSP-B, to detect H2O2 under MC-LR-induced oxidative stress in organisms. DSP-B exhibited high sensitivity and specificity to H2O2, and the detection ability of DSP-B to endogenous and exogenous H2O2 has also been validated. Then DSP-B was applied to detect the H2O2 level in cells and zebrafishes treated with MC-LR to elucidate the effect of oxidative stress caused by MC-LR. Moreover, DSP-B was utilized for tissue visualization imaging in MC-LR-poisoned loaches model, enabling the upregulation of H2O2 to be successfully observed. This study offers a novel strategy for analyzing the MC-LR-induced oxidative stress and demonstrates the potential of using probe for MC-LR toxicity research. This probe is expected to provide assistances in evaluating the risks and hazards of MC-LR exposure to organisms in the environment.

Docetaxel is the first-line chemotherapy for metastatic prostate cancer (PC), but clinically meaningful mechanisms of resistance remain to be established. Here we show, in an in vivo model of docetaxel resistant PC patient-derived xenografts, increased expression of genes that drive development of multiciliated cells including FOXJ1 and its effectors, many of which regulate microtubules (MTs). Mechanistically, FOXJ1 overexpression confers docetaxel resistance in vitro and in vivo, which is associated with decreased docetaxel-mediated MT bundling. Overexpression of a MT-associated FOXJ1-regulated gene (TPPP3) has similar effects. Conversely, FOXJ1 knockdown impairs basal MT function, enhances taxane binding to MTs, and increases docetaxel sensitivity. These results establish mechanistic causality between the FOXJ1 signaling axis, MT biology, and taxane resistance. Clinically, FOXJ1 gene amplification is increased in taxane-treated PC patients. Moreover, in the CHAARTED clinical trial of docetaxel combined with androgen deprivation for metastatic PC, higher baseline FOXJ1 is predictive of decreased survival in PC patients treated with docetaxel, further supporting clinical relevance. Together, these findings identify a previously unrecognized clinically impactful mechanism of taxane resistance whose exploitation could stratify patients who will not benefit from taxane treatment.

Fungal metabolites represent a valuable but underexplored source of anticancer agents, in part due to poorly defined mechanisms of action. Kojic acid (KA) is a fungal secondary metabolite with reported anti-melanoma activity, but its mechanism of action remains unclear. Here, we show that KA inhibits melanoma progression by disrupting MYC-driven transcriptional programs. KA treatment reduced proliferation and induced apoptosis in melanoma cells in vitro, and suppressed tumor growth in xenograft models. Transcriptomic profiling revealed a dose-dependent repression of MYC target genes, with CCNA2 and KPNA2 identified as key effectors. Both genes were validated as direct MYC targets and were associated with poor prognosis in the melanoma cohort (TCGA-SKCM). KA did not alter MYC expression but impaired its promoter binding and transcriptional activation of CCNA2 and KPNA2. Single-cell analysis further localized this axis to a proliferative mitotic subpopulation, promoting melanoma progression. These findings uncover a previously unrecognized mechanism by which KA inhibits melanoma growth and suggest that targeting the MYC-CCNA2/KPNA2 pathway may provide a therapeutic strategy for melanoma.

Advanced castration-resistant prostate cancer (CRPC) is responsive to PARP inhibitors, but only in settings of defects in homologous recombination (HR). The oncogenic M1C protein drives CRPC progression; however, it is not known if M1C plays a role in the response to PARP inhibition. The present work demonstrates that M1C is induced by olaparib treatment of HR-competent CRPC cells. As a result, M1C drives (i) ATM expression, (ii) phosphorylation of KAP1(S824) and (iii) activation of STING, which have been linked to derepression of the LINE-1 (L1) retrotransposon. In this way, M1C is necessary for induction of (i) L1-5'UTR, L1-ORF1 and L1-ORF2 transcripts and (ii) the encoded ORF1p RNA binding protein. Activation of retrotransposons induces genomic instability and drug resistance. By extension, we show that M1C also activates HERV-K102/108 gag, pol and env genes and expression of the HERV-K ENV protein. Our work further demonstrates that M1C integrates L1 and HERV-K activation with induction of APOBEC3 (A3) genes that evolved to restrain genomic instability induced by these retrotransposons. Of translational relevance, these findings demonstrate that M1C (i) is essential for inducing L1, HERV-K and A3 expression and resistance of CRPC cells to olaparib, and (ii) is a target for advancing the treatment of HR-competent CRPC with PARP inhibitors.

Elevated homocysteine (Hcy) levels are well established as an independent risk factor for atherosclerosis and its associated cardiovascular diseases. Macrophage pyroptosis- mediated inflammation plays a crucial role in the progression of atherosclerosis. Notably, glycoprotein non-metastatic melanoma protein B (GPNMB) expression is increased in macrophages within atherosclerotic plaques; however, whether GPNMB participates in Hcy-induced macrophage pyroptosis remains elusive. In the present study, we found that GPNMB expression was upregulated in Hcy- treated THP-1- derived macrophages. Consistently, serum GPNMB levels were significantly higher in patients with hyperhomocysteinemia (HHcy) compared with healthy controls. Functional experiments showed that silencing GPNMB reduced Hcy-triggered pyroptosis in THP-1-derived macrophages, whereas GPNMB overexpression exerted the opposite effect. Mechanistically, GPNMB upregulated the NOX2/NF-κB signaling pathway in THP-1-derived macrophages. Importantly, the pro-pyroptotic effect of GPNMB overexpression in Hcy-treated THP-1-derived macrophages was counteracted by either inhibition of NADPH oxidase 2 (NOX2) using the specific inhibitor gp91ds-tat or blockade of NF-κB activation with the inhibitor BAY11-7082. Moreover, serum GPNMB levels were correlated with serum Hcy levels and lipid profiles in both healthy individuals and HHcy patients. Collectively, these findings demonstrate that GPNMB facilitates Hcy-induced macrophage pyroptosis associated with the upregulation of the NOX2/NF-κB signaling pathway, highlighting the potential relevance of GPNMB as a candidate target for the clinical management of HHcy-related atherosclerotic cardiovascular disease.

Enzalutamide resistance remains a critical obstacle in the treatment of castration-resistant prostate cancer (CRPC), with potassium calcium-activated channel subfamily N member 4 (KCNN4) emerging as a key mediator of therapeutic failure. Here, we demonstrate that KCNN4 is significantly upregulated in enzalutamide-resistant PCa cells and clinical tissues, correlating with poor prognosis. Mechanistically, p300, a histone acetyltransferase, directly binds to KCNN4 and mediates its acetylation at lysine 16, which competitively inhibits ubiquitination-mediated degradation, thereby stabilizing KCNN4 protein. Notably, p300 inhibition disrupts KCNN4 acetylation, restores its proteasomal degradation, and resensitizes resistant cells to enzalutamide both in vitro and in vivo. Moreover, KCNN4 knockdown suppresses tumor growth and synergizes with enzalutamide in xenograft models, underscoring the therapeutic potential of targeting the p300-KCNN4 axis. Collectively, our findings reveal a previously unrecognized epigenetic regulatory mechanism coupling p300-mediated acetylation to potassium channel stability, providing a promising therapeutic strategy to overcome chemoresistance in advanced prostate cancer.