Dengue virus (DENV) infection remains a major global health threat. Its incidence is increasing, with frequent outbreaks affecting millions of people each year. Although vaccines are available, their limited effectiveness and the absence of targeted antiviral therapies highlight the critical need for alternative treatment approaches. In this study, we investigated the antiviral activity of bidebiline A and heteropsine, two dimeric aporphine alkaloids isolated from Trivalvaria costata, against DENV-infected Huh7 liver cells. Our findings reveal that heteropsine inhibits DENV production and infection in Huh7 cells more effectively than bidebiline A. Heteropsine also exerts antiviral activity in endothelial and lung cell lines. Mechanistic studies, including time-of-addition assays and molecular docking, elucidate that heteropsine targets early steps during cellular infection, possibly by binding to domain III of the dengue virus envelope protein (EDIII). Viral binding and internalization assays confirmed that heteropsine disrupts viral entry. Furthermore, heteropsine suppresses DENV-induced immunopathogenesis by downregulating the expression of cytokine/chemokine genes (TNF-α, IL-6, RANTES, and IP-10) and attenuating nuclear translocation of the p65 subunit of the nuclear factor-kappaB (NF-κB) transcription factor. These findings highlight the importance of heteropsine as a promising antiviral candidate with the potential for further development to address the urgent need for effective dengue therapeutics.

The prevalence of nonalcoholic fatty liver disease (NAFLD) is rising at an alarming rate, making it a major global public health problem. The main pathophysiology of NAFLD is elevated de novo lipogenesis (DNL) in hepatocytes which leads to lipid accumulation. Because of their function in controlling DNL, sirtuin 1 (SIRT1) and AMP-activated protein kinase (AMPK) have been considered viable therapy targets for reduce lipid accumulation.

In recent years, metabolic reprogramming has emerged as a significant breakthrough in elucidating the onset and progression of gastrointestinal (GI) malignancies. As central regulatory hubs for lipid metabolism, sterol regulatory element binding proteins (SREBPs) integrate dietary metabolic signals and carcinogenic stimuli through subtype-specific mechanisms, thereby promoting malignant tumour phenotypes. In this review, we first present the molecular background, structural characteristics, and posttranscriptional regulatory networks associated with SREBPs. We subsequently describe a systematic analysis of the distinct activation patterns of SREBPs in liver, gastric, colorectal, and other gastrointestinal cancers. Furthermore, we explore targeted intervention strategies for different SREBP subtypes, including small molecule inhibitors (such as fatostatin, which inhibits SREBP cleavage), natural compounds (such as berberine, which modulates the AMPK/mTOR pathway), and statin-mediated inhibition of the mevalonic acid pathway. These strategies may enhance tumour cell sensitivity to chemotherapeutic agents (such as 5-FU, gezil, and tabine) and improve the response to synergistic chemoradiotherapy by reversing adaptive metabolic resistance driven by the tumour microenvironment. Through this review, we hope to provide new insights into precise interventions targeting various subtypes of the SREBP molecule.

Chronic non-healing foot ulcers are a major complication in diabetic patients, contributing to significant morbidity and mortality. Microorganisms in these wounds form biofilms, conferring greater virulence and enhanced protection from antibiotics. Hence, we examined naringenin, and other natural compounds like chlorofuranone, 4-nitropyridine N-oxide, and quercetin as a positive control against the major pathogenic organism that forms biofilm in foot ulcers. Here, we focused on Pseudomonas aeruginosa, which produces high levels of biofilm in diabetic foot ulcers. Naringenin (47.10 µg/ml for PA21; 124.7 µg/ml for PA333) and other natural compounds were tested for their ability to inhibit biofilm formation and virulence in vitro, and their effect on biofilm-associated gene expression was studied. The biofilm inhibitory mechanism of naringenin was elucidated using in silico analysis and in vitro reporter gene assay. In vitro biofilm assays, confocal and scanning electron microscopy showed that natural compounds effectively inhibited biofilm, without causing cell death. Treatment with these compounds significantly altered the expression of genes associated with quorum sensing in P. aeruginosa, such as lasR, pslA, algA, gacS, and pelA. Naringenin decreased the production of major virulence factors in P. aeruginosa. Molecular docking showed that naringenin exhibited the strongest binding affinity to LasR, and the same was validated by reporter gene assay using plasmid pSB1142 indicating its role as a competitive inhibitor in the las quorum sensing system in P. aeruginosa. The findings of this study could be extrapolated to in vivo diabetic wound infection models to help optimize the use of naringenin in effective biofilm control for better wound management in diabetic patients.

The incidence of prostate cancer (PCa) is high among elderly men. Cepharanthine hydrochloride (CH) is recognized for its important role in the prevention and treatment of various diseases. However, its effects and mechanisms of action in the context of PCa remain unclear. Our study aims to examine the therapeutic role and mechanisms of action of CH in PCa. Targets of CH and PCa-related genes were identified using different databases, and the biological processes through which CH might exert its therapeutic effects were predicted via protein-protein interaction (PPI) network and enrichment analyses. Subsequently, the PCa cell lines PC-3 and DU145 were used to assess the concentration- and time-dependent effects of CH on cell viability, proliferation, and migration. Transcriptomic sequencing and differential expression analysis were used to identify the key target protein of CH and the key signaling pathways involved in its therapeutic effects against PCa. Molecular docking was used to analyze the binding between CH and its target protein. Additionally, quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, gene knockout, pharmacological intervention, and tumor formation experiments were performed to validate the therapeutic effects and mechanisms of action of CH against PCa in vitro and in vivo. Network pharmacology showed that CH, a Chinese herbal medication, might prevent PCa by regulating protein phosphorylation-related biological processes. In vitro experiments showed that CH inhibited the proliferation and migration of PCa cells in a concentration-dependent manner. In addition, integration of transcriptomic sequencing, differential expression analysis, and GO enrichment analysis suggested that the ERK protein played a crucial role in the anti-tumor activity of CH. Molecular docking and molecular dynamics simulations revealed strong binding affinities between CH and ERK1/2. Further experimental verification, involving qRT-PCR, western blotting, gene knockout, pharmacological intervention, and tumor formation experiments, demonstrated that CH upregulated dual-specificity phosphatase (DUSP) 1 and suppressed the phosphorylation of ERK, thereby inhibiting the development and progression of PCa in vivo and in vitro. In conclusion, the findings of this study suggest that CH suppresses the ERK signaling pathway by enhancing the expression of DUSP1, thereby exerting anti-tumor effects against PCa in vitro and in vivo. Therefore, CH may serve as a novel therapeutic agent for PCa, showing remarkable potential for clinical application.

Compound 5, a cinnamanilide derivative, upregulates brain derivated neurotrophic factor (BDNF) expression but with low soluablity. In this study, PX5-9, a N-oxide derivative of 5, demonstrated significant protective effects in the HT22 glutamate-induced toxicity model and showed no significant toxicity at 30 μM. Western blot analysis confirmed that PX5-9 increased BDNF levels similar to 5. Solubility tests revealed a significant improvement in PX5-9 (37.10 ± 0.33 μg/mL) compared to 5 (< 15 ng/mL). Pharmacokinetic studies of PX5-9 revealed favorable properties, fast absorption and also can be transformated into parent compound 5, suggesting it is a potential candidate for these diseases involving with BDNF. The N-oxide modification might be a good prodrug design to enhance solubility while preserving biological activity.

Anti-angiogenic therapies (AATs) exhibit limited efficacy, as most patients with cancer inevitably develop resistance to them. In this study, data generated using a nasopharyngeal carcinoma orthotopic mouse model, combined with clinical data, reveal compensatory vasculogenic mimicry (VM) formation during AAT treatment and the association of VM with poor prognosis in nasopharyngeal carcinoma. Additionally, data-independent acquisition mass spectrometry-based proteomics shows that upregulation of a disintegrin And metalloprotease 10 (ADAM10) contributes to VM. Mechanistically, epigenetic and high-resolution chromatin interaction landscape analyses demonstrate that although ADAM10 does not interact with either the proximal or distal enhancers, DEAD-box helicase 5 (DDX5), a transcription factor of ADAM10, is regulated by long-range looping enhancer-promoter interactions. Further analyses identify transcription factors binding to critical constituents of the DDX5 super-enhancer. Ingenol mebutate, which docks excellently with DDX5, reverses ADAM10-mediated gene expression changes, thereby effectively suppressing compensatory VM formation and metastasis and improving prognosis. Collectively, these findings provide insights into the clinical application of AATs.

Pancreatic cancer is a highly aggressive gastrointestinal tumor with limited treatment options, such as surgery and chemotherapy. Thus, further research into its pathogenesis and new treatments is necessary.

Helium, previously described as "biologically inert", displays great medicinal potential in a number of respiratory ailments via influencing redox signaling. However, whether or how this gas functions in plant biology is still elusive. Here, hydroponic and pot experiments showed that mimicking the responses achieved by nitric oxide (NO)-releasing compound, exogenous helium supply with helium-enriched solution or its fumigation significantly prevented NaCl-induced growth inhibition of alfalfa seedlings. Lower level of Na+/K+ ratio in both root and shoot parts caused by lower efflux of net K+ and higher efflux of net Na+, as well as higher abundances of ion transport genes (SOS1, SOS2, SOS3, NHX1, HKT1, and AKT1) were simultaneously observed. Consistently, reactive oxygen species (ROS) accumulation and oxidative injury were abolished by helium. These were further supported by stimulating antioxidant machinery and reprogramming gene expression related to antioxidant defence and related transcriptional factors (MYB4, WRKY33, ERF8, and ERF11) against salinity toxicity. Further experiments showed that helium supplementation strengthened endogenous NO production by up-regulating the expression of nitrate reductase (NR2) and its enzymatic activity, and NO-based S-nitrosylation was further influenced. Above responses were remarkably impaired by the removal of endogenous NO when its scavenger was added. Overall, these results revealed that helium control of salinity tolerance is partially mediated by a NO signaling cascade governing redox and ion homeostasis reestablishment, two important defense strategies against salinity stress.

Studies have demonstrated that adult male fish exposed to estrogenic endocrine-disrupting compounds (EDCs) exhibit reduced sperm motility and fertility. However, the effects of prepubertal estrogenic EDC exposure on testicular development and subsequent reproductive capacity remain poorly understood. In this study, prepubertal male (XY) tilapia at 30 days after hatching (dah) were exposed to 17β-estradiol (E2) for 60 days, covering the prepubertal period, to investigate its impact on testicular development and fertility at sexual maturity. Morphological and histological analyses revealed that prolonged E2 exposure reduced number of spermatogenic cells, increased testicular cell apoptosis, and significantly decreased the gonadosomatic index in E2-treated XY (E2-XY) fish. Fertility analysis of sexually mature E2-XY fish at 180 dah showed a significant reduction in sperm fertilization capacity compared to Control-XY fish. Sperm quality analysis indicated that E2-XY fish had an increased proportion of immotile sperm and a decreased proportion of progressive sperm. Additionally, the straight-line velocity, curvilinear velocity, and beat frequency of sperm flagella were significantly reduced in E2-XY fish sperm. Scanning electron microscopy (SEM) and immunofluorescence analysis revealed a significant reduction in flagellum length in E2-XY sperm compared to Control-XY sperm. Transcriptome sequencing of 180 dah E2-XY testes demonstrated a significant downregulation of genes associated with motor activity, microtubule binding, microtubule motor activity, and dynein complex formation. Specifically, the expression of kinesin and dynein axonemal heavy chain gene family members was significantly reduced. qPCR and Western blot (WB) assays confirmed that the expression of kinesin family members (kifc1, kif2c, kif11, kif16b, kif17, kif20a, kif20ba, kif20bb, kif22) and dynein heavy chain domain-containing protein family members (dnah2, dnah5, dnah7, dnhd1), which are involved in flagellar movement and assembly, was significantly decreased in E2-XY testes compared to Control-XY testes. These findings demonstrate that prepubertal E2 exposure downregulates genes critical for sperm flagella formation and motility in tilapia, resulting in shortened flagella, reduced motility, and irreversible fertility impairment, in contrast to the recoverable fertility observed in adult fish following exposure to estrogenic EDCs. This study provides novel insights into the mechanisms underlying reduced fertility in male fish exposed to estrogenic EDCs.

Breast cancer, the most prevalent malignant tumour in women, is characterised by high metastatic potential and frequent recurrence, both of which significantly impact patient prognosis following metastasis. To address this challenge, identifying novel therapeutic target combinations is critical for improving metastatic breast cancer treatment. This study investigates the mechanism by which asparaginyl endopeptidase (AEP) regulates breast cancer metastasis. Bioinformatics analysis revealed a potential interaction between AEP and CD74, which was subsequently confirmed through co-immunoprecipitation (co-IP) experiments. Further investigations demonstrated that AEP activates ERK pathway phosphorylation via CD74 regulation, thereby enhancing epithelial-mesenchymal transition (EMT) progression and promoting breast cancer cell migration. Compared to controls, dual inhibition of AEP and CD74 effectively reduced EMT markers and the migratory capacity of cancer cells in vitro. Subsequent in vivo experiments showed that this combinatorial strategy significantly suppressed breast cancer lung metastasis in mice without observable toxicity. These findings elucidate the molecular mechanism through which AEP promotes metastasis via CD74 regulation, while validating the therapeutic efficacy and safety of dual AEP/CD74 targeting. This study provides a novel conceptual framework and potential therapeutic targets for metastatic breast cancer intervention.

Sphingosine-1-phosphate receptor 3 (S1PR3) has been implicated in promoting tumor progression in various cancers. However, the role and molecular mechanisms of S1PR3 in oral squamous cell carcinoma (OSCC) remain poorly understood. The aims of this study were to investigate the function of S1PR3 in OSCC progression and its potential as a therapeutic target.

Alfalfa (Medicago sativa L.) has the benefits of high yield and nutritional value as a sustainable forage. However, the water deficit significantly limits its growth and yield performance. Nitric oxide (NO) is a signal molecule that can enhance plant tolerance. The majority of previous studies focus on the role of exogenous NO in plant tolerance. However, the underlying mechanism of endogenous NO in alfalfa drought tolerance remains largely unexplored.

Phytohormones play pivotal roles in regulating floral development and secondary metabolite synthesis in saffron (Crocus sativus L.).

The resilience of plants against environmental challenges, particularly salinity and dehydration, is crucial for global food security. This study delves into the intricate interaction between NaCl-induced salinity and Azolla aqueous extract (AAE). In a pot trial, wheat kernels were primed with deionized water or 0.1% AAE for 21 h. Seedlings underwent various treatments; tap water, 250 mM NaCl, AAE priming and spray, and combined AAE with NaCl treatments. Seedlings were analyzed for ionic balance, secondary metabolism, antioxidant efficacy, and molecular response to experimental treatments.

The expression of ubiquitin-like molecule interferon-stimulated gene 15 kDa (ISG15) and its post-translational modification (ISGylation) are significantly activated by interferons or pathogen infections, highlighting their roles in innate immune responses. Over 1100 proteins have been identified as ISGylated. ISG15 is removed from substrates by interferon-induced ubiquitin-specific peptidase 18 (USP18) or severe acute respiratory syndrome coronavirus 2-derived papain-like protease. High ISGylation levels may help prevent the spread of coronavirus disease 2019 (COVID-19). Polyamines (spermidine and spermine) exhibit anti-inflammatory, antioxidant, and mitochondrial functions. However, the relationship between nutrients and ISGylation remains unclear. This study assessed the effects of spermine and spermidine on ISGylation. MCF10A and A549 cells were treated with interferon-alpha, spermine, or spermidine, and the expression levels of various proteins and ISGylation were measured. Spermine and spermidine dose-dependently reduced ISGylation. Additionally, spermidine directly interacted with ISG15 and USP18, enhancing their interaction and potentially reducing ISGylation. Therefore, spermidine may prevent ISGylation-related immune responses.

Curcumin, a plant derived natural product isolated from Curcuma longa. The aim of this study is to investigate the anti-proliferative effects and the underlying mechanisms of curcumin in Cutaneous T cell lymphoma (CTCL), a type of non-Hodgkin lymphoma that primarily affects the skin. The study found that curcumin induced apoptosis in CTCL cells by activating mitochondrial signaling pathways and caspases leading to growth inhibition. Furthermore, Curcumin treatment downregulated the expression of S-phase kinase protein (SKP2) with concomitant upregulation of GADD45A, CDKN1A and CDKN1B. Curcumin also suppresses the expression of anti-apoptotic molecules including XIAP and cIAPs. Curcumin treatment of CTCL cells generates reactive oxygen species (ROS) and depletion of glutathione. Pretreatment of CTCL with N-acetyl cysteine prevented curcumin-mediated generation of ROS and prevention caspase activity. Co-treatment of CTCL with subtoxic doses of curcumin and bortezomib potentiated the anticancer action. Co-treatment of CTCL with subtoxic doses of curcumin and bortezomib potentiated the anticancer action. Molecular docking studies revealed a strong binding affinity of curcumin to the active site of SKP2, primarily involving key residues crucial for its activity. Altogether, our results suggest that targeting SKP2 and GADD45A signaling by curcumin could be an attractive strategy for the treatment of CTCL.

Epithelial-mesenchymal transition (EMT) describes a process by which epithelial cells acquire mesenchymal properties associated with increased migration, invasion, and resistance to therapy. In pancreatic ductal adenocarcinoma (PDAC), targeting the molecular and intercellular communication pathways that drive EMT represents a promising therapeutic strategy. Here, we investigate the effects of combined treatment with gemcitabine (G), paricalcitol (P), and hydroxychloroquine (GPH) in KPC-Luc orthotopic mouse models of PDAC, using single-cell RNA sequencing (scRNA-seq), high-dimensional weighted gene co-expression network analysis (hdWGCNA), and cell-cell communication analysis. GPH treatment reduces EMT, which is associated with the downregulation of the essential gene fibronectin (Fn1). Collagen and Fn1 pathways co-expression decreases in GPH-treated KPC-Luc tumors. Cancer-associated fibroblasts (CAFs) appear dominant in collagen signaling, whereas macrophages mediate Fn1 signaling. GPH treatment reduces the expression interaction strength between ligands and receptors (collagen-integrin and Fn1-Cd44 or Fn1-Sdc4) compared to sham, PH, and G. Altogether, this study presents a comprehensive single-cell resolution map of the molecular and cellular mechanisms by which GPH treatment impairs EMT in PDAC, identifying potential therapeutic targets within the fibronectin and collagen signaling axes.

Pathogenesis of Staphylococcus aureus is largely associated with its biofilm formation, that protects the cells from host immune system and antimicrobial threats. Considering the concern over the emergence of antimicrobial resistant S. aureus strains, this study was aimed to explore an effective alternative therapeutant. Trigonelline, an alkaloid, was evaluated for its antibiofilm and antivirulence activities against S. aureus. Trigonelline efficiently inhibited and eradicated biofilm, and abled to decrease the production of protease and hemolysin, the major virulence factors of S. aureus. Inhibition of biofilm formation and eradication of mature biofilm on the catheter surface suggested its potentiality in clinical application. The observed reduction in biofilm formation and virulence factor production following trigonelline treatment may be attributed to its ability to alter the expression of key regulatory genes such as agrA, sarA, saeR, arlR, icaR, and sigB, which control quorum sensing network and biofilm development. Additionally, molecular docking analysis revealed a substantial binding affinity of trigonelline to these regulatory proteins, further supporting its possible inhibitory mechanism. Thus, trigonelline might be a promising alternative chemical lead to manage biofilm-associated bacterial infections caused by S. aureus.

Numerous studies have highlighted the anti-cancer effects of nonsteroidal anti-inflammatory drugs (NSAIDs), although the underlying mechanisms remain unclear. This study focuses on elucidating the impact of the NSAID ibuprofen (IBU) on cancer cells exposed to hypoxia, as the hypoxic microenvironment significantly influences tumor progression, metastatic potential, and therapy resistance. Given that carbonic anhydrase IX (CA IX) is a key hypoxia-associated protein and a promising therapeutic target due to its tumor-specific expression, we primarily examined the impact of IBU on CA IX and the transcription factors regulating CA IX expression. We found that IBU downregulates expression and protein level of CA IX in hypoxic colon carcinoma and head and neck cancer cells, resulting in a reduction of membranous CA IX. To elucidate the mechanism of this phenomenon, we analyzed the key CA IX-regulating transcription factor HIF-1 and found decreased levels of the HIF-1α subunit in IBU-treated cells, leading to its impaired binding to the CA9 promotor. Analysis of another transcription factor involved in CA IX expression, NFκB, showed suppressed NFκB pathway under IBU treatment. Moreover, we demonstrated IBU-mediated induction in apoptosis in cancer cells, as well as a decrease in their ability to migrate. Our study is the first to demonstrate that ibuprofen downregulates carbonic anhydrase IX expression in hypoxic colon and head and neck tumor cells by decreasing HIF-1α levels. Additionally, ibuprofen impairs key transcription factors NFκB and STAT3, leading to reduced adaptation to hypoxic stress, decreased tumor cell viability, and migration. This indicates its potential as a therapeutic agent in combination therapy for colon carcinoma or head and neck cancer.