Osteoporosis (OP) is a prevalent degenerative musculoskeletal disorder in middle-aged and elderly populations, characterized by reduced bone mineral density and an increased risk of fragility fractures. Its pathogenesis is closely linked to oxidative stress imbalance; however, effective long-term therapeutic options remain limited. Here, we report that diacerein, a clinically used drug for osteoarthritis, significantly ameliorates OVX (ovariectomy)-induced bone loss and abnormal bone metabolism. Mechanistically, we demonstrate that diacerein activates the transcription and activity of NRF2 (nuclear factor erythroid 2-related factor 2), a central regulator of the antioxidant response, leading to the restoration of cytoprotective protein expression and thereby exerting protective effects against osteoporosis. The beneficial effects of diacerein on bone protection were largely abolished in Nfe2l2 knockout mice, further underscoring the essential role of NRF2. Further investigation revealed that diacerein is metabolized in vivo into rhein, which exerts potent epigenetic activity. The upregulation of NRF2 appears to be primarily mediated through rhein's inhibition of DNA methyltransferases (DNMTs) 1 and 3a. In summary, our study suggests that diacerein, a drug with a well-established safety profile and suitability for long-term use, represents a promising therapeutic candidate for the treatment of osteoporosis.

Chronic stress is associated with inflammatory activation and oxidative stress responses leading to endothelial dysfunction, which promotes the development of atherosclerosis (AS). SGLT2 inhibitors, such as Dapagliflozin (DAPA), exhibit a protective effect against cardiovascular diseases. However, the effects and mechanisms of DAPA on chronic stress-induced AS are largely unknown. The aim of this study was to determine whether DAPA confers a protective effect against chronic stress-induced AS and to elucidate its further molecular mechanisms. The combined high-fat diet-fed and chronic unpredictable mild stress in ApoE-/- mice and lipopolysaccharides- and corticosterone-induced human umbilical vein endothelial cells (HUVECs) were employed to evaluate the antiatherosclerotic effect of DAPA under chronic stress in vivo and in vitro. Histological staining, western blot analysis, siRNA transfection, reactive oxygen species (ROS) staining, and apoptosis assessment were used to investigate the potential mechanisms of DAPA against AS under chronic stress. The results indicate that DAPA significantly improved plaque size and increased plaque stability in the aorta under chronic stress and reduced inflammation and oxidative stress and inhibited apoptosis in the aorta and HUVECs. Chronic stress upregulated regulated in development and DNA damage response 1 (REDD1) expression, which exacerbated cellular inflammation, oxidative stress, and apoptosis levels, leading to endothelial dysfunction. In contrast, DAPA downregulated REDD1 expression and activated the AKT/FoxO1 pathway. In addition, p53 was a transcriptional regulator of REDD1 under chronic stress. More importantly, p53 agonists prevented DAPA from downregulating REDD1 and inhibited AKT/FoxO1 activation, thereby exacerbating chronic stress-induced endothelial dysfunction. These results suggest that DAPA effectively attenuates chronic stress-induced endothelial dysfunction and AS by downregulating REDD1 to activate the AKT/FoxO1 pathway.

Pharmacological enhancement of endoplasmic reticulum (ER) proteostasis is an attractive strategy to mitigate pathology linked to etiologically diverse protein misfolding diseases. However, despite this promise, few compounds have been identified that enhance ER proteostasis through defined mechanisms of action. We previously identified the phenylhydrazone-based compound AA263 as a molecule that promotes adaptive ER proteostasis remodeling through mechanisms including preferential activation of the ATF6 signaling arm of the unfolded protein response (Plate et al., 2016). However, the protein target(s) of AA263 and the potential for further development of this class of ER proteostasis regulators had not been previously explored. Here, we employ chemical proteomics to demonstrate that AA263 covalently targets a subset of ER protein disulfide isomerases, revealing a potential molecular mechanism for the activation of ATF6 afforded by this compound. We then use medicinal chemistry to establish next-generation AA263 analogs showing improved potency and efficacy for ATF6 activation, as compared to the parent compound. Finally, we show that treatment with these AA263 analogs enhances secretory pathway proteostasis to correct the pathologic protein misfolding and trafficking of both a destabilized, disease-associated α1-antitrypsin (A1AT) variant and an epilepsy-associated GABAA receptor variant. These results establish AA263 analogs with enhanced potential for correcting imbalanced ER proteostasis associated with etiologically diverse protein misfolding disorders.

Colorectal cancer (CRC) ranks among the leading causes of cancer-related mortality worldwide. Hydrogen sulfide (H2S) has been found to possess a characteristic of anticancer, which may offer a potential novel treatment for CRC. Here, we discover the potential targets and mechanism of H2S intervention in CRC employing multiomics analysis and experimental validation. The key targets of H2S intervention in CRC were identified by integrating differentially expressed genes (DEGs) from tumor and normal tissues, the CRC-associated genes, and the targets of H2S. The STRING and Cytoscape tools were explored to obtain hub genes. Functional enrichment analysis, assessment of diagnostic and prognostic significance, single-cell datasets, and cell experiments were used to explore the impact of core targets on CRC and the potential mechanism through which H2S exerts regulatory effects on CRC. Our results identified 9250 genes closely linked to CRC from DEGs and CRC-associated genes, 505 targets for H2S, and 322 potential targets of H2S intervention in CRC. Subsequently, five hub genes were filtered, including MAPK1, MAPK3, JUN, ESR1, and AKT1. The 322 common targets were enriched in the cellular stress responses and IL-17 signaling pathway. Additionally, MAPK3 had good diagnostic and prognostic value for CRC. JUN was highly expressed in immune cells. Cell experiments showed that sodium hydrosulfide (NaHS), a donor of H2S, prominently inhibited cell proliferation, promoted cell apoptosis for CRC, and downregulated the expression of MAPK1, MAPK3, AKT1, and JUN. Taken together, this study elucidates the possible genes and therapeutic mechanisms underlying exogenous H2S intervention in CRC, thereby laying a foundation for the further development of H2S-based therapeutic strategies in CRC management.

We identified the characteristics of CBL and CIPK families of Magnolia biondii and the MbCBL4-overexpressed Arabidopsis plants conferred salt tolerance. Calcineurin B-like protein (CBLs) and CBL-interacting protein kinase (CIPKs) are important components of the Ca2⁺-mediated signal pathway. These proteins play a key role in plant growth, development, and response to environmental stress. Magnolia biondii is a woody plant valued for ornamental and medicinal uses and is frequently exposed to abiotic stresses during its growth cycle. Nevertheless, there are still gaps in the study of CBL and CIPK gene families in M. biondii. In this study, 6 CBL and 20 CIPK genes were identified from the M. biondii genome. Phylogenetic analysis divided these genes into 4 CBL and 7 CIPK subgroups, and cross-species comparisons across 34 plants indicated that monocotyledons generally harbor more CBLs/CIPKs than Magnoliaceae. Quantitative real-time PCR (qRT-PCR) analysis revealed that MbCBLs and MbCIPKs showed different transcription levels under drought, cold, and salt stress. Protein-protein interaction assays (Y2H and LCI) verified physical interaction of MbCBL1/MbCIPK18 and MbCBL4/MbCIPK18. Functionally, MbCBL4 overexpression in Arabidopsis conferred enhanced salt tolerance: primary root length and chlorophyll content increased by 2.74-fold and 2.71-fold relative to wild type; fresh weight increased by up to 60%, SOD and CAT activities rose by 47% and 28%, while H₂O₂ and O₂⁻ levels declined by 46% and 38%. These results indicate that MbCBL4 enhances salt tolerance by promoting growth, antioxidant capacity, and reactive oxygen species scavenging. These findings provide important insights into the functional roles of MbCBL and MbCIPK genes and the regulation of MbCBL4 under salt stress.

Particulate matter (PM) poses risks to environmental and human health, yet its toxicity mechanisms in aquatic organisms remain unclear. This study investigated the effects of 2 PM types, a standard reference material (S-PM10, NIST, USA) and particulates from the Mae Moh Power Plant, Thailand (MMPS), on zebrafish embryo development and gene expression. Embryos were exposed to various concentrations, and mortality, hatching rates, and morphological abnormalities were assessed. S-PM10, with irregular morphology and broad particle size, induced developmental defects and reduced hatching. MMPS, characterized by uniform, spherical particles, caused higher mortality. qRT-PCR revealed that S-PM10 significantly upregulated oxidative stress (sod1, gstp2) and apoptosis (bax, casp3a) genes. In contrast, MMPS downregulated oxidative stress markers but upregulated apoptosis-related genes. These results suggest particle morphology and size influence toxicity profiles, S-PM10 triggers developmental disruption, while MMPS induces acute lethality. This study underscores the importance of particle characteristics and molecular responses in evaluating PM toxicity.

Cisplatin resistance remains a major challenge in bladder cancer. Although the tumor suppressor ASPP2 is a critical co-factor for TP53-mediated apoptosis, its role in metabolic reprogramming and cisplatin response remains unclear. This study aimed to delineate the mechanism by which ASPP2 regulates cisplatin sensitivity through metabolic reprogramming. We first assessed the clinical significance of ASPP2 using patient tissues and public databases, finding that its downregulation in bladder cancer is associated with poor patient survival. Through gain- and loss-of-function studies in vitro and in vivo, we further demonstrated that ASPP2 inhibits the mevalonate (MVA) pathway independently of TP53 status, thereby sensitizing cells to cisplatin-induced DNA damage and apoptosis. This chemosensitizing effect was specifically reversed by the addition of MVA pathway metabolites. Moreover, WWP2 was identified as the E3 ubiquitin ligase responsible for ASPP2 degradation via K48-linked ubiquitination. Finally, WWP2 silencing was shown to stabilize ASPP2, suppress the MVA pathway, and synergize with cisplatin to impede tumor growth in mouse models. Overall, the WWP2-ASPP2-MVA pathway axis is identified as a novel driver of cisplatin resistance in bladder cancer. These results establish a mechanistic basis for targeting this axis to restore chemosensitivity, offering a promising therapeutic strategy for recalcitrant disease.

Low temperature combined with low light stress affects off-season pepper cultivation by impairing growth and reducing yield. Betaine acts as osmoregulatory, improving plant stress resistance, but how it mediates m6A methylation remains unknown. This study investigates effects of betaine on low temperature combined with low light conditions by treating pepper seedlings and comparing m6A methylation patterns in the transcriptome, alongside validating methylase gene functions through VIGS and overexpression techniques. Results indicated m6A modification sites affected by betaine are located within coding sequences and 3' untranslated regions under LL stress. Post-treatment with betaine upregulated m6A peaks significantly outnumbered downregulated ones. Transcriptome analysis revealed that betaine enhanced m6A methylation, leading to the degradation of genes like auxin-induced protein AUX28, acid phosphatase-1-like, calmodulin-7, U-box domain-containing protein-21-like, late embryogenesis abundant protein D-34-like, and Protein TAR1. However, it stabilized genes of bHLH87-like, pyrophosphate-energized vacuolar membrane proton pump-like, ABC transporter C family member 4-like, F-box domain-containing protein with hypo-m6A methylation, which are linked to peroxidase synthesis and O-methyltransferase metabolic pathways. Additionally, CaMTC enhanced the tolerence of pepper seedlings to low temperature combined with low light by increasing the m6A methylation level, outlining a regulatory network associated with m6A methylation and betaine. Overall, these findings provides a theoretical framework for the role of betaine in m6A modification under low temperature combined with low light stress, and offers a potential strategy for betaine to enhance plant resistance through the regulation of m6A methyltransferases.

Glutathione (GSH) serves as a redox-active molecule and the predominant non-protein sulfhydryl compound in plants and a critical regulator in alleviating abiotic stress. Our previous research has demonstrated that foliar application of exogenous GSH can enhance the salt tolerance of tomato seedlings. However, the underlying molecular mechanism remains unexplored. In this study, RNA-seq analysis revealed that exogenous GSH significantly influenced plant hormone signal transduction, MAPK signaling pathway, and starch and sucrose metabolism. In addition, the transcription factor SlMYB48 was identified. The expression of SlMYB48 was strongly induced under salt stress but suppressed when GSH was applied simultaneously. Transgenic overexpression (OE) and knockout mutant lines of SlMYB48 were generated and exposed to salt stress, demonstrating that SlMYB48 functioned as a negative regulator of salt tolerance in tomato seedlings. Foliar GSH application increased endogenous GSH content, enhanced the activity and expression of key enzymes in GSH metabolism and the antioxidant system, and reduced ROS accumulation and oxidative injury in OE lines subjected to salt stress. Furthermore, exogenous GSH significantly elevated the expression of starch and sucrose metabolism related genes and increased the corresponding sugar content in OE plants under salt stress. Importantly, GSH application suppressed SlMYB48 expression in the OE lines exposed to salinity. Collectively, these findings indicate that exogenous GSH enhances salt tolerance in tomato seedlings by repressing SlMYB48 expression, thereby modulating the antioxidant system and osmotic adjustment. This study establishes a theoretical framework for elucidating GSH regulated molecular breeding for salt resistance, highlights SlMYB48 breeding potential, and guides practical applications.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy that has a poor survival rate of ∼13 % with limited options for effective therapies. DDX3 is a member of the DEAD-box RNA helicase enzyme family. It acts as an adapter protein that interacts with several transcription factors, enhancing their binding ability to the promoters of genes involved in cancer progression. Previously, we demonstrated that PAF1, a component of the RNA polymerase II-associated factor 1 complex, interacts with DDX3 to promote PDAC stemness. Here, we investigated the therapeutic efficacy of RK-33, a small-molecule inhibitor targeting DDX3, in combination with gemcitabine (GEM) and 5-fluorouracil (5FU), which enhanced therapeutic efficacy in KRAS-driven PDAC.-DDX3 and PAF1 exhibit progressively increased expression in various stages and correlate well with poor survival of PDAC. Targeting DDX3/PAF1 significantly mitigated clonogenic, EMT, and stemness phenotypes in PDAC cells. It also reduced tumor growth, proliferation, and increased apoptosis in xenograft and PDAC organoid models. Finally, MXRA5, EDIL3, COL13A1, and SLC16A2 were identified as top downstream response genes upon RK-33 treatment and potential new targets to mitigate extracellular matrix remodeling, angiogenesis, cell migration, and cell cycle progression, thereby enhancing the therapeutic efficacy of GEM and 5FU Overall, our data indicate that RK-33 enhances the therapeutic efficacy of GEM and 5FU in mitigating PDAC aggressiveness. Consequently, these findings open new avenues for developing efficacious therapeutic adjuvants to treat advanced pancreatic cancer.

Biliverdin reductase A (BLVRA) is a key enzyme in bilirubin metabolism, where it reduces biliverdin to bilirubin. Bilirubin is a potent antioxidant that protects cells from oxidative stress. Therefore, reduced or deregulated BLVRA activity may contribute to increased oxidative DNA damage, which is one of the factors leading to the neoplastic transformation of cells.

G-quadruplex structures within the promoter regions of some oncogenes diminish transcriptional activity. The suppression and downregulation of Cyclooxygenase-2 (COX-2), which is encoded by the prostaglandin-endoperoxide synthase 2 (PTGS2) gene, could control the size of colorectal cancer tumors. This study aimed to investigate the impact of a mononuclear octahedral cobalt(III) Schiff base complex [CoL3] (L = 2-((allylimino)methyl)-6-methoxyphenol) on the G-quadruplex structures in the PTGS2 promoter region and to assess its downregulation effects in the human colorectal cancer cell line HT-29. At first, molecular docking was used to evaluate the binding of [CoL3] to the PTGS2 promoter region. Then, the potential and stabilization of G-quadruplex formation in the PTGS2 promoter motifs were investigated using circular dichroism (CD) spectroscopy, polyacrylamide gel electrophoresis, and polymerase chain reaction (PCR) stop assays. COX-2 expression was also assessed by real-time PCR and western blot. Molecular docking analysis indicates that [CoL3] has a potent interaction with the PTGS2 promoter region. Additionally, [CoL3] has an intense inhibitory effect on the PTGS2-PCR stop assay. The CD measurements confirmed the conformational property of G-quadruplex DNA structures induced by [CoL3]. Treatment of colorectal cancer cells with the [CoL3] resulted in a modest increase in PTGS2 expression by a mean factor of 1.69 (p < 0.05). Conversely, the expression of COX-2 proteins was downregulated significantly, with a mean protein intensity of 0.47 (p < 0.001). Therefore, [CoL3] induces G-quadruplex formation in the promoter region of PTGS2, which ultimately inhibits protein expression of COX-2. This means that [CoL3] can be an effective agent for colorectal cancer treatment.

Timosaponin AIII (Tim-AIII), a steroidal saponin derived from Anemarrhena asphodeloides, has emerged as a promising antitumor agent, yet its precise molecular targets and mechanisms in breast cancer remain poorly defined. Here, we identify fibroblast growth factor 2 (FGF2) as a direct binding target of Tim-AIII using a combination of network pharmacology, CETSA, and surface plasmon resonance assays. Mechanistically, Tim-AIII exhibits a dual therapeutic mode of action. First, it induces reactive oxygen species (ROS)-mediated endoplasmic reticulum (ER) stress, activating the eIF2α-ATF4-CHOP axis and initiating apoptosis. Second, it dampens the FGF2-FGFR1-PI3K/AKT signaling cascade, thereby inhibiting epithelial-mesenchymal transition (EMT) and suppressing cell migration and invasion. RNA sequencing and enrichment analyses confirm that Tim-AIII regulates critical oncogenic pathways, including ER stress, calcium signaling, and PI3K/AKT. In vivo evaluations demonstrate that Tim-AIII significantly reduces tumor growth without detectable systemic toxicity in breast cancer-bearing mice. This study not only elucidates the molecular basis of Tim-AIII's antitumor efficacy but also positions it as a potential targeted therapeutic for breast cancer, with dual action on ERS-induced apoptosis and EMT suppression.

Osteoporosis is a common skeletal metabolic disorder. Cardamonin (CAR) is a natural chalcone compound with multiple activities. However, the role and mechanism of CAR in osteoporosis progression remain largely unknown.

Although immunotherapy has reshaped the treatment landscape for gastrointestinal (GI) malignancies, durable clinical benefit is achieved in only a subset of patients, largely due to the emergence of immune resistance. Accumulating evidence highlights histone lactylation, an epigenetic modification driven by excessive lactate accumulation, as a pivotal mediator linking tumor metabolism to immune suppression. By reprogramming transcriptional networks within the tumor microenvironment, histone lactylation skews macrophages toward an immunosuppressive phenotype and compromises T-cell effector function, thereby facilitating tumor immune escape. In metabolically active GI tumors with pronounced lactate production, aberrant histone lactylation contributes to the establishment of a profoundly immunosuppressive niche that undermines responses to immune checkpoint inhibitors. Targeting lactylation-related pathways has therefore gained attention as a novel therapeutic avenue, with the potential to restore antitumor immune activity, enhance cytotoxic lymphocyte function, and sensitize tumors to immunotherapy. This article summarizes current understanding of the crosstalk between lactate metabolism, histone lactylation, and immune regulation, and highlights therapeutic approaches targeting this epigenetic axis to enhance immunotherapy efficacy in GI cancers.

Triple-negative breast cancer (TNBC) is distinguished by high invasiveness and a tendency for recurrence. Recent studies have suggested that E3 ubiquitin ligases play a crucial role in the initiation and progression of various tumors. However, there is still an absence of systematic understanding regarding the specific function and molecular mechanisms of its member gene RNF130 in TNBC.

Alcohol is a major cause of hepatocellular carcinoma (HCC), accounting for 30% of cases worldwide. Sorafenib, a tyrosine kinase inhibitor (TKI), was the standard first-line treatment for advanced HCC until 2021, but sorafenib resistance is common. We explored the impact of chronic alcohol exposure (CAE) on sorafenib response and sought to identify associated resistance mechanisms. Huh-7 HCC cells were chronically exposed to alcohol for 6 months. Sorafenib resistance was assessed by measuring cell viability (IC50) and by evaluating the protein expression of signaling pathways involved in resistance using immunoblotting. RNA sequencing was performed to identify mechanisms of resistance. Sorafenib response was assessed using the RECIST 1.1 criteria in HCC patients. A retrospective study of 86 HCC patients from the CHIEF cohort (alcohol-related vs. non-alcohol-related etiologies) evaluated overall survival (OS) and progression-free survival (PFS) using the log-rank test. CAE significantly decreased cell sensitivity to sorafenib (p = 0.006), indicating increased resistance. The ERK pathway was involved. RNA sequencing of our cells identified a total of 80 differentially expressed genes associated with drug resistance and aggressiveness. Clinically, alcohol-related HCC patients were less responsive to sorafenib (35% responders vs. 65%, p = 0.014) and had significantly different OS (p = 0.0234). Median OS was 10 months (95% CI = [6.1, 15.7]) for alcohol-related HCC and 12.1 months (95% CI = [7.7, 64.9]) for other etiologies. PFS was lower in the alcohol group (5.72 months (95% CI = [4.63, 12.8]) vs. 9.66 months (95% CI = [4.40, 39.9], p = 0.0298). Sorafenib resistance due to chronic alcohol consumption is consistent in both in vitro models and clinical settings. KEY MESSAGES: Chronic alcohol exposure reduces the effectiveness of sorafenib in hepatocellular carcinoma (HCC), as demonstrated in both in vitro and clinical settings. In vitro, alcohol-exposed HCC cells showed increased sorafenib resistance, associated with activation of the ERK signaling pathway and differential expression of 80 genes linked to drug resistance and tumor aggressiveness. Clinically, patients with alcohol-related HCC had poorer responses to sorafenib and shorter overall and progression-free survival compared to patients with non-alcohol-related HCC. These findings suggest that alcohol-related HCC may require alternative or personalized therapeutic strategies beyond standard TKI treatments.

Uveal melanoma (UM) is among the most prevalent intraocular malignant tumors worldwide. Celastrol exhibits broad-spectrum anticancer properties; however, its underlying therapeutic mechanism in UM is yet to be elucidated. In this study, a network pharmacology approach was employed to identify potential common targets of celastrol and UM. These targets were further analyzed in conjunction with transcriptomic data and machine learning algorithms, which led to the identification of CTNNB1 and STAT3 as key molecular targets. The functional roles of these targets were investigated through immune infiltration analysis and single-cell RNA sequencing (scRNA-seq), while the binding stability between celastrol and CTNNB1/STAT3 was assessed using molecular docking (MD) and molecular dynamics simulation (MDS). Subsequently, celastrol was administered to B16-F10 and C918 cell lines, demonstrating that it significantly suppresses cell proliferation and migration by downregulating CTNNB1 and STAT3 expression, while simultaneously inducing apoptosis and cell cycle arrest. Moreover, real-time quantitative PCR (qPCR) and western blot (WB) analyses corroborated the modulation of target expression levels. Therefore, celastrol exerts potent anti-tumor effects in UM by inhibiting the CTNNB1 and STAT3 signaling pathways, thereby suppressing tumor cell proliferation and metastasis, as well as promoting cell cycle arrest and apoptosis.

Bufei Formula (BFF), a traditional Chinese medicine, has been widely utilized in the clinical treatment of chronic obstructive pulmonary disease (COPD), though its underlying mechanisms remain unclear and warrant further investigation.

In melanoma, SOX9 and SOX10 are markers of the mesenchymal and melanocytic state, respectively. Using a panel of BRAFV600E positive YUMM lines, we find that, following chronic vemurafenib treatment, SOX10 is lost whereas SOX9 is induced. Overexpression or knock-down of either SOX9 or SOX10 had no impact on vemurafenib sensitivity. However, we find that SOX9 is necessary to program a vemurafenib-resistance memory state following a drug holiday in vitro. RNA-Seq studies show that the loss of Sox10 represents an intermediate state that is accompanied by the loss of Sox6 and the induction of Sox7, Sox9 and other phenotype switching markers. However, SOX7 expression is not sufficient to induce vemurafenib resistance. Upon acquired drug resistance, we observed differential chromatin accessibility in the Sox9 and Sox10 upstream regions, supporting their activation and repression, respectively. Overall, our data show that the loss of SOX10 and SOX9 induction are critical to program drug resistance. Furthermore, we show that the YUMM cell lines represent a good murine model to investigate transitions to an acquired drug resistant state.