This study assesses the neuroprotective effects and molecular mechanisms of ciprofol against cerebral ischemia-reperfusion injury (CIRI) in rats. From July 2023 and July 2024, fifty male SD rats were randomly divided into five groups: control, model, ciprofol (Ci), erastin (Era), and compound C (CC). The model was induced by MCAO/R. Control group received identical surgical interventions without filament insertion. Rats neurological deficits were quantified using modified Garcia JH scores. Histopathological changes were evaluated through Nissl staining and TTC‒determined infarct volume. Mitochondrial ultrastructure was observed by transmission electron microscopy. Biochemical analyses quantified malondialdehyde (MDA), iron content and inflammatory cytokines (IL-1β, IL-6, TNF-α). Western blotting measured AMPK phosphorylation and ferroptosis proteins (GPX4, ACSL4). Relative to the model group rats, the Ci rats exhibited elevated modified Garcia JH scores (P < 0.05) accompanied by attenuated neuronal/mitochondrial damage and diminished infarct areas. Biochemical analyses revealed significant reductions in MDA, inflammatory cytokines, and ACSL4 protein expression in Ci rats, concurrent with enhanced GPX4 levels and elevated p-AMPK/T-AMPK ratios (P < 0.05). Notably, Era specimens demonstrated GPX4 downregulation with corresponding ACSL4 and iron accumulation compared to the Ci rats. Furthermore, the CC rats displayed decreased p-AMPK/T-AMPK activation relative to Ci rats (P < 0.05). Ciprofol ameliorates CIRI in rats by inhibiting ferroptosis and inflammatory factor through upregulating AMPK.

Although many DNA binding natural products exert their effects through non-specific mechanisms, a therapeutic opportunity exists for a subset of these compounds that alter the expression or activity of specific driver oncogenes in specific cell contexts. In this study, we integrate CUT&Tag with Global Run-On Sequencing (CUT, Tag, and GRO) to show that the minor groove binding compound, mithramycin (MMA), inhibits the Ewing sarcoma oncogenic driver, the EWS::FLI1 transcription factor. MMA causes either an increase or decrease in EWS::FLI1 binding to chromatin at downstream target response elements to poison nascent transcription. The reversal of EWS::FLI1 activity is limited by non-specific effects of the drug on RNAPII processivity but can be optimized by continuous administration at low concentration to cause more precise reversal of the oncogenic transcriptome and striking Ewing sarcoma xenograft regressions. The activity in vivo is further improved with a less-toxic second-generation analog, AIT-102.

BackgroundHypopharyngeal cancer is increasingly emerging as a disease that threatens global health, with poor prognosis and survival rates. However, clinical strategies and effective therapies remain limited.MethodsThe inhibitory effect of liensinine on tumor cells was detected through cell cycle, colony formation, and apoptosis assays. Changes in the expression levels of relevant proteins were detected and enrichment analysis of signaling pathways was performed through in vitro and RNA sequencing experiments. The transcription levels of relevant genes were further verified using reverse transcription polymerase chain reaction.ResultsWe previously discovered that the natural compound, liensinine, is effective in treating hypopharyngeal cancer. In this study, we found through in vitro and RNA sequencing experiments that liensinine can activate the Ras homolog family member B protein, thereby inhibiting the mitogen-activated protein kinase signaling pathway. Additionally, liensinine activates the nuclear factor kappa B signaling pathway and releases downstream inflammatory factors, effectively exerting its antitumor effects.ConclusionLiensinine induces cell death and inhibits hypopharyngeal cancer cell growth through multiple pathways, indicating that it is a potential chemotherapeutic agent for the treatment of hypopharyngeal cancer.

High myopia (HM) poses a growing public health challenge due to its increasing prevalence and the associated risks of blinding complications and psychological comorbidities. While traditionally considered an isolated ocular condition, emerging evidence implicates systemic mechanisms, notably through the gut-eye axis and immune factors, play important part in the pathogenesis of HM. Histone demethylase Kdm2a, the key H3K36me2 modification eraser, is critically involved in various inflammatory diseases, yet its specific role in the gut-eye axis and HM remains elusive. To address this, the HM model was successfully established. HM mice exhibited significant scleral thinning, reduced collagen protein, and prominent anxiety-like behaviors. Crucially, they were suffering from gut microbial dysbiosis and intestinal barrier impairment. Intriguingly, upregulated Kdm2a and correspondingly decreased H3K36me2 levels were observed in the intestinal epithelial cells (IECs) of HM mice. Treatment with Daminozide (DA), the selective inhibitor of Kdm2a, effectively suppressed myopia progression and ameliorated psychological comorbidities. Mechanistically, DA restored gut microbiota homeostasis, colonic morphology, and barrier integrity. The transcriptomic profiling further revealed the protective effects of Kdm2a inhibition on modulating key pathways involved in intestinal inflammation and tissue remodeling. Collectively, this work elucidates a novel gut-eye pathway in HM pathogenesis and identifies Kdm2a in IECs as a promising therapeutic target for HM and its associated psychological comorbidities.

Isolavonoides represent the second largest subgroup of flavonoids and have an influence on critical molecular pathways and restore cellular homeostasis, through the reprogramming of epigenetic regulatory mechanisms. This feature indicates a crucial therapeutic potential that could be better explored to attend cancer treatment. Isoflavonoids, acting as epigenetic modulators, could contribute to the development of new therapeutic approaches in cancer, especially in onco-hematological diseases. Pterocarpans are a subgroup of isoflavonoids that have been extensively studied for their biological properties. The molecule (+)-2,3,9-trimethoxypterocarpan demonstrates high gastrointestinal (GI) absorption and the ability to cross the blood-brain barrier (BBB) in silico without violating Lipinski's rule, making it a desirable candidate in leukemia treatment. The synthesis of this molecule dates back more than a decade. In silico models, such as SwissADME, corroborate the notion of good intestinal absorption and the ability to cross the BBB. Also, it is suggested that P-glycoprotein is a substrate, which is related to its potential for active efflux from both the BBB and GI. This review highlights the biological mechanisms of this class of natural products from a translational perspective, emphasizing their chemical properties and epigenetic biological activities, which offer new therapeutic perspectives, particularly in oncology.

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.

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.

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.

Inter-organellar communication is critical for cellular metabolism. One of the most abundant inter-organellar interactions occurs at the endoplasmic reticulum and mitochondria contact sites (ERMCSs). However, an understanding of the mechanisms governing ERMCS regulation and their roles in cellular metabolism is limited by a lack of tools that permit temporal induction and reversal. Through screening approaches, we identified fedratinib, an FDA-approved drug that dramatically increases ERMCS abundance by inhibiting the epigenetic modifier BRD4. Fedratinib rapidly and reversibly modulates mitochondrial and ER morphology, induces a distinct ER-mitochondria envelopment structure, and alters metabolic homeostasis. Moreover, ERMCS modulation depends on mitochondrial electron transport chain complex III function. Comparison of fedratinib activity to other reported inducers of ERMCSs revealed common mechanisms of induction and function, providing clarity to a growing body of experimental observations. In total, our results uncovered a novel epigenetic signaling pathway and an endogenous metabolic regulator that connects ERMCSs and cellular metabolism.

Arabidopsis halleri ssp. gemmifera is a promising Cd phytoremediation agent, however, the metal uptake and accumulation mechanism remain poorly understood. This study focused on the 2-hours early responses associated with Cd uptake and temporary Cd retention in roots. To distinguish Cd-specific responses from shared divalent metal responses, transcriptomic analyses were performed on roots exposed to Cd compared to excess Zn. Cd exposure induced a clearly larger number of differentially expressed genes than higher concentration of Zn exposure, indicating a distinct early response to Cd. Genes encoding transporters such as PCR2, DTX1, PDR8, PDR12, CAX4, MHX1, and ABCC2 were highly upregulated during the early exposure phase. Cd retention in roots may be mediated by these transporters, which could contribute to Cd efflux into the apoplast or vacuolar sequestration. Further, genes involved in intracellular Cd chelation, including those encoding glutathione, HIPPs, and HMPs protein, were upregulated rather than genes encoding phytochelatins. Additionally, upregulation of genes involved in cell wall biosynthesis and remodeling was observed, suggesting a structural modification occurs during early Cd exposure, contributing to reinforcement and temporary Cd storage before translocation. This hypothesis is supported by increased lignification in root tissues and the accumulation of Cd in the apoplastic region, indicating that cell wall serves as sequestration site in A. halleri.

This study explored the molecular mechanisms by which T7 peptide-modified liposomal irisin (T7@Lipo@Irisin) alleviates perioperative neurocognitive disorders (PND) via regulation of the AMPK/PGC-1α metabolic pathway. T7@Lipo@Irisin nanoparticles were prepared by thin-film hydration and ultrasonic dispersion and showed favorable physicochemical performance, with an encapsulation efficiency of approximately 85%. Serum analysis of healthy donors (n = 10) and PND patients (n = 6) showed higher IL-6 and TNF-α and lower brain-derived neurotrophic factor (BDNF) in PND. In vitro, T7@Lipo@Irisin restored mitochondrial membrane potential, reduced reactive oxygen species (ROS) accumulation, enhanced Neuro-2a hippocampal neuron viability, and activated the AMPK/PGC-1α axis under oxidative stress. In a PND mouse model, it improved Garcia neurological scores, preserved neuronal morphology, and decreased apoptosis. Multi-omic integration of scATAC-seq/scRNA-seq and TMT-based proteomics demonstrated enhanced neuro-glial crosstalk, epigenetic activation of metabolic/antioxidant genes (e.g., Sirt1, Nfe2l2), and upregulated pathways (mitochondrial function, NAD-dependent metabolism, synaptic homeostasis). Proteomics confirmed upregulation of SIRT1, NDUFS2, and BDNF, forming a network linked to energy metabolism and neural repair. Collectively, T7@Lipo@Irisin mitigates PND by activating AMPK/PGC-1α to enhance mitochondrial function and stabilize the neuro-microenvironment.

Cucumber sex differentiation is a complex process regulated by multiple factors, including calcium (Ca2⁺) signaling. Although Ca2⁺ has been implicated in flower sex determination, its precise regulatory mechanisms remain unclear. In this study, the androecious cucumber line Banna 31-10 was treated with CaCl₂ at the shoot apices at seven-day intervals for three applications (Ca-1W, Ca-2W, Ca-3W). CaCl₂ treatment significantly increased the number of female flowers, with the effect becoming more pronounced with prolonged application. Integrated transcriptomic and metabolomic analyses revealed extensive molecular and metabolic reprogramming in response to CaCl₂. Transcription factor (TF) analysis identified 347 TFs from 56 families, with ERF, bHLH, MYB, C2H2, and MADS-box families playing key roles. Integrated transcriptome-metabolome correlation highlighted the involvement of flavonoids and plant hormone pathways, including ethylene, abscisic acid, gibberellin, cytokinin, auxin, jasmonic acid, salicylic acid, and brassinosteroid. Notably, CsACS2 and multiple calmodulin-like (CML) genes were strongly associated with hormone biosynthesis and signaling, suggesting a central role of Ca2⁺-CML-mediated regulation in female flower differentiation. Weighted gene co-expression network analysis (WGCNA) identified hub genes in co-expression modules linked to CaCl₂ response, further supporting the regulatory network underlying sex differentiation. This study provides new insights into the role of the Ca2⁺ signaling system in cucumber sex differentiation.

Aflatoxin B1 (AFB1), a known mycotoxin and environmental hazard, has been linked to breast cancer, yet the exact biological pathways remain poorly characterized. We performed a comprehensive multi-omics assessment to investigate how AFB1 may influence breast tumor biology. This encompassed transcriptomic analysis, co-expression network modeling (WGCNA), immune landscape profiling, transcription factor regulatory mapping, and spatial plus single-cell transcriptomics. Predictive biomarkers were determined through a machine learning pipeline. Twenty-two genes were identified at the intersection of AFB1-predicted targets and disease-associated expression modules. A refined panel of seven biomarkers (EGFR, MIF, MET, PPARG, MME, NQO2, NR3C2) was established through model optimization. A composite classifier using glmBoost and StepGLM achieved high discriminative accuracy (area under the curve = 0.996). SHAP interpretability indicated PPARG may act protectively, while MIF showed risk-promoting characteristics. Expression heterogeneity was observed across cell populations and spatial regions. Our integrated analytical framework offers new insights into the oncogenic potential of AFB1 in breast cancer. The identified gene set may serve as both mechanistic mediators and diagnostic markers, underscoring the value of multi-omics and machine learning approaches in environmental carcinogenesis research.

Carya illinoinensis was a crop with high value and was widely cultivated in China. Particularly under the increasingly severe soil salinization background, the promotion of salt-tolerant C. illinoinensis varieties provided substantial economic benefits to local regions. However, the molecular mechanisms underlying C. illinoinensis's salt response remained unclear. Two-year-old C. illinoinensis saplings exhibited significant damage after treatment with 600 mM NaCl solution. Transcriptome data at nine salt treatment time points were obtained from two-year-old C. illinoinensis saplings using RNA-seq, and 7840 differentially expressed genes (DEGs) were identified to construct a time-ordered gene co-expression network (TO-GCN). DEGs in the TO-GCN were classified into 10 levels corresponding to salt treatment time points. Genes in Level 1 (L1) and Level 2 (L2) were enriched in Gene Ontology (GO) terms associated with photosynthesis and transport channel proteins. Cluster analysis identified three aquaporin-coding genes among highly expressed genes. Based on promoter cis-acting elements, A/T-rich sequence features, and antagonistic gene expression patterns in transcriptomic data, it was speculated that CiPIP2;8 might be transcriptionally repressed by the upstream transcription factor CiPLATZ23. This hypothesis was validated through a Dual-luciferase reporter (DLR) assay. The presence of ABA-responsive elements (ABREs) in ProCiPLATZ23 suggested its involvement in the abscisic acid (ABA) pathway. Transient expression of ProCiPLATZ23::GUS in Nicotiana benthamiana exhibited enhanced histochemical staining following ABA treatment. Within 24 h after spraying walnuts with ABA, the transcription level of CiPLATZ23 rapidly increased. Furthermore, 35S::CiPLATZ23 overexpression lines in Arabidopsis thaliana displayed reduced ABA sensitivity compared to Wild-type (WT) A. thaliana. Reverse transcription quantitative PCR (RT-qPCR), yeast one-hybrid, and DLR assays confirmed that CiPLATZ23 participated in regulating ABA sensitivity by binding to the promoter of AtPIP2;8, the homolog of CiPIP2;8 in A. thaliana. Characterization of canonical ABA pathway families confirmed that C. illinoinensis ABA responses were implicated under salt stress. This study employed TO-GCN analysis of RNA-seq data to elucidate salt stress response mechanisms in C. illinoinensis and identified a key module 'CiPLATZ23-PIP2; 8' that may be associated with ABA.

Cadmium (Cd) is a highly toxic heavy metal that substantially impacts crop productivity. Although γ-aminobutyric acid (GABA) has been shown to enhance plant resistance to abiotic stresses, the mechanism by which it mitigates Cd toxicity in maize (Zea mays L.) remains unclear. Using integrated physiological, transcriptomic, and metabolomic analyses, we found that exogenous GABA enhances maize seedling tolerance to Cd stress by simultaneously strengthening antioxidant defenses, regulating metal transport, and reinforcing cell walls. GABA significantly upregulates the activities of antioxidant enzymes, mitigating oxidative damage under Cd stress. Transcriptomic analysis confirmed induced expression of key antioxidant genes during the GABA-Cd interaction. Notably, Nramp and HMA transporter proteins were implicated in GABA-mediated suppression of Cd uptake and distribution. Transcriptional-metabolic co-analysis revealed that phenylpropanoid biosynthesis and the phenylalanine metabolism pathway, which are involved in cell wall biosynthesis, increase the GABA-mediated Cd tolerance. Exogenous GABA increased the activities of enzymes related to lignin synthesis. For example, the activity of phenylalanine lyase (PAL) increased by 41.48 % in roots at 4 days post-treatment, thus enhancing lignin deposition in the cell walls of maize roots. This structural reinforcement restricted Cd translocation from roots to shoots, thus reducing Cd accumulation in stem and leaf tissues. Conversely, pharmacological inhibition of endogenous GABA synthesis via 3-mercaptopropionic acid (3-MPA) decreased root lignin content and increased Cd shoot translocation, validating GABA's role in cell wall lignification. These findings clarify GABA's multifaceted role in mitigating Cd stress through coordinated physiological and molecular responses, highlighting its potential as a sustainable strategy to safeguard crop productivity under heavy metal contamination.