The fungal priority pathogen and basidiomycete, Cryptococcus neoformans (Cn), causes lung and brain infection in predominantly immuno-compromised individuals and there is an urgent need for new treatment options. The pyrazolopyrimidine-based cyclin dependent kinase (CDK)7 inhibitor, BS-181, has anticancer properties, but its antifungal activity has not been investigated. We show that cryptococcal CDK7 more closely resembles the human enzyme than that of ascomycetes, and that BS-181 inhibits its activity. BS-181 inhibited growth of both Cn and Cryptococcus gattii (Cg), but not ascomycete fungi and delayed progression through the G2/M phase of the cell cycle. Transcriptomic analysis revealed that BS-181 induces splicing defects leading to elevated intron retention within the transcriptome and also suppresses translational processes. BS-181 displayed additive or synergistic activity with licensed antifungals against laboratory and clinical Cn and Cg strains, most notably with amphotericin B where synergy (2-4-fold reduction in the amphotericin B MIC) was achieved using low-sub micromolar concentrations of BS-181. Compared with either drug alone, BS-181-AmB combination therapy provided greater protection against Cn infection in a wax moth model (p ≤ 0.032) and extended survival of Cn-infected mice. These findings demonstrate that CDK7 inhibitors, already of interest as anticancer agents, could be repurposed to prevent or treat opportunistic fungal infections in cancer patients when combined with licensed antifungals limited by either toxicity or resistance.

IntroductionGlycolytic phenotype positively supports cancer cell migration and metastasis in various cancers including Triple negative breast cancers (TNBCs). In-depth understanding of molecular pathways associated with increased aerobic glycolysis in TNBCs could provide key insights into the drivers of TNBC progression.Methodsβ-catenin and glycolytic proteins (PFKP, LDHA, MCT1) were assessed by Immunohistochemistry (IHC) in TNBC patients (n = 98), with prognostic value evaluated by Kaplan-Meier and Cox regression. In vitro, the β-catenin inhibitor ie, XAV939 was tested for suppressing β-catenin-driven aerobic glycolysis in TNBC models using MTT for proliferation, Western blotting for protein expression, and wound healing, droplet invasion, and colony formation assays for physiological changes.Resultsβ-catenin and glycolytic markers (PFKP, LDHA, MCT1) were overexpressed in >50% of TNBCs. Kaplan-Meier and Cox regression analyses showed that combined expression of β-catenin with glycolytic markers correlated with reduced survival. In vitro, XAV939 suppressed β-catenin-driven aerobic glycolysis in TNBC cells, downregulating β-catenin and glycolytic proteins, reducing glycolytic activity, and impairing aggressive phenotypes (proliferation, migration, invasion, clonogenicity).ConclusionOverall, our results highlight the crucial role of β-catenin in controlling aerobic glycolysis via regulation of key glycolytic proteins, thereby positively driving the progression and metastasis of TNBCs. Additionally, our data strongly establish that XAV939 effectively inhibits glycolytic phenotype, thereby suggesting its therapeutic potential in TNBC patients.

Clinical use of stimulator of interferon genes (STING) agonists has challenges due to poor responsiveness and variable efficacy. Therefore, identifying tumor types that are sensitive to these agents and clarifying the underlying mechanisms are essential.

Liver cancer treatment with cisplatin is often hindered by drug resistance. This study aimed to identify key genes associated with cisplatin resistance in liver cancer and develop targeted inhibitors. Using genome-wide CRISPR-Cas9 screening, ATOX1 was identified as a critical gene for cisplatin resistance. ATOX1 was highly expressed in liver cancer tissues and associated with poor prognosis. Knockdown of ATOX1 in liver cancer cells enhanced cisplatin sensitivity in vitro and in vivo. Molecular dynamics simulation and virtual screening identified compound 8 as a potent ATOX1 inhibitor with high affinity (Kd = 12.5 μM) and exhibited synergistic effects with cisplatin on liver cancer cell growth. Mechanistically, compound 8 inhibits the activity of ATOX1, leading to intracellular copper accumulation. The elevated copper levels subsequently promote increased DNA methylation at the NOTCH1 promoter, resulting in suppression of the NOTCH1/HES1 signaling pathway and enhancing the sensitivity of liver cancer cells to cisplatin. In conclusion, ATOX1 is crucial for cisplatin resistance in liver cancer and linked to poor prognosis. Targeting ATOX1 with compound 8 may be a novel therapeutic strategy for overcoming cisplatin resistance.

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.

Intracerebral hemorrhage (ICH) is a cerebrovascular event associated with a high fatality rate, leading to a considerable health and economic burden. Tanshinone IIA (Tan IIA), a promising compound used to treat coronary artery disease, has recently been shown to exert significant neuroprotective effects. Therefore, whether Tan IIA can alleviate NETosis induced by LPS after ICH remains unclear. For this purpose, we explored the effects of Tan IIA on collagenase-induced ICH with peripheral inflammation and its potential mechanisms using an after-ICH infection animal model (male C57BL/6J mice) treated with Tan IIA for 5 days, starting at 2 months of age. Further analysis demonstrated that Tan IIA-treated ICH mice with peripheral inflammation exhibited improved motor and sensory dysfunction compared with untreated groups. Administration of Tan IIA in ICH mice with peripheral inflammation alleviated neuropathological alterations of the corpus striatum, including NETosis inhibition, glial inactivation, and inflammasome activity attenuation, and significantly decreased levels of PAD4 and H3 Cit in the corpus striatum of ICH mice with peripheral inflammation. In vitro investigations showed that Tan IIA suppressed neuroinflammation in LPS-stimulated glial cells by inhibiting the NLRP3/caspase-1 signaling pathway. Further molecular docking predicted that Tan IIA directly interacted with the NLRP3 protein. Collectively, these findings strongly indicate that Tan IIA is an effective compound for mitigating hemiplegia symptoms, NETosis, and neuroinflammation in the collagenase-induced ICH model with peripheral inflammation, primarily through the dual actions of inhibiting NET formation and suppressing the NLRP3/caspase-1 pathway.

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.

Jasmonate (JA) accumulation and signaling play key roles in regulating plant growth and flower development. The optimal timing of flowering is critical for the quality and yield of Brassica juncea. B-box (BBX) factors play roles in plant floral transition and stress response processes. However, whether BBX6-2 responds to JA signaling to regulate flowering time in B. juncea remains unclear. Here, we characterized the biological function of BjuBBX6-2 in response to JA in regulating the flowering time of B. juncea. Subcellular localization and transcriptional activation activity assays showed that BjuBBX6-2 localizes in the nucleus and exhibits transcriptional activation activity. Spraying BjuBBX6-2-overexpressing or BjuBBX6-2-silenced B. juncea plants with 50 μmol/L methyl JA significantly accelerated or delayed their flowering time, respectively. We demonstrated that BjuBBX6-2 interacts with the flowering factor Nuclear Factor Y, Subunit C4 (BjuNF-YC4), which interacts with Nuclear Factor Y, Subunit B2/3 (BjuNF-YB2/3). The BjuBBX6-2-BjuNF-YC4-BjuNF-YB2/3 multiple-protein complex bound to the promoter of the downstream flowering integrator gene SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (BjuSOC1) and promoted its expression. JASMONATE ZIM-DOMAIN 1 (BjuJAZ1), a key factor in the JA signaling pathway, interacted with BjuBBX6-2 to inhibit the activation of BjuSOC1 by BjuBBX6-2. In summary, BjuBBX6-2 cooperates with BjuNF-YC4 and BjuJAZ1 in response to JA signaling to participate in the flowering regulation of B. juncea. These findings highlight a previously uncharacterized mechanism of JA signaling-mediated flowering time regulation via interactions between BjuBBX6-2 and the integrator gene BjuSOC1, providing prospects for breeding enhanced B. juncea cultivars.

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.