Aging is characterized by interconnected disturbances in oxidative stress, chronic inflammation, mitochondrial dysfunction, and nutrient-sensing pathways that collectively contribute to progressive metabolic and vascular decline. Among the central molecular mechanisms implicated in these processes are dysregulation of the AMPK-mTOR-SIRT1 axis, persistent NF-κB activation, impaired Nrf2-mediated antioxidant defense, and mitochondrial redox imbalance. Pharmacological strategies capable of modulating multiple components of this network have therefore gained increasing interest in aging-related research. Empagliflozin, a sodium-glucose cotransporter-2 (SGLT2) inhibitor, has demonstrated pleiotropic metabolic effects extending beyond glycemic control, including modulation of AMPK signaling, attenuation of oxidative stress, and suppression of inflammatory activation. Nebivolol, a third-generation β1-selective adrenergic blocker with nitric oxide-mediated vasodilatory properties, has been associated with endothelial protection, vascular redox regulation, and anti-inflammatory activity. Although both agents influence pathways relevant to aging biology, current evidence is derived predominantly from experimental models of cardiovascular or metabolic disease rather than from physiological aging studies. Moreover, direct evidence evaluating their combined effects in aging remains unavailable. This review critically examines the mechanistic roles of empagliflozin and nebivolol within the context of aging-associated metabolic and vascular dysfunction. Particular emphasis is placed on their reported interactions with AMPK-mTOR-SIRT1 signaling, Nrf2-dependent antioxidant responses, NF-κB-mediated inflammation, mitochondrial function, and endothelial homeostasis. In addition, the review discusses current limitations in the evidence base, including the predominance of reductionist experimental approaches, limited translational validation, and the absence of direct combinational investigations. Comparative consideration is also given to established aging-relevant candidates, including metformin, rapamycin, resveratrol, GLP-1 receptor agonists, NAD+ modulators, and senolytic strategies, in order to contextualize the potential relevance and limitations of metabolic-vascular pathway modulation in aging research. Rather than proposing definitive evidence of aging-relevant efficacy, the available literature supports a biologically plausible framework in which modulation of metabolic and vascular pathways may influence interconnected mechanisms underlying aging-related decline. Further integrative experimental and translational studies are required to determine whether coordinated targeting of these pathways may provide meaningful therapeutic benefit in aging and age-associated disorders.
Understanding the neuropharmacological effects and abuse liability (AL) of electronic nicotine delivery systems (ENDS) is essential for evaluating their potential role in tobacco harm reduction (THR). Nicotine delivered via ENDS engages central nervous system (CNS) pathways involved in reinforcement, craving, and dependence. This research characterized nicotine pharmacokinetics (PK) and AL of Vuse Alto ENDS across multiple nicotine concentrations and flavor and contextualized these findings relative to combustible cigarettes and nicotine replacement therapy (NRT) gum. Two randomized, open-label, crossover clinical studies were conducted under controlled human-use conditions. Study 1 evaluated eight Vuse Alto ENDS flavors containing 1.5% nicotine, with sequential plasma sampling over 240 min following product use. Study 2 assessed four Vuse Alto ENDS (1.5%-5% nicotine) and incorporated validated subjective effect assessments and PK sampling, alongside participants' usual brand cigarettes and NRT gum. Adult smokers and dual users were enrolled under strict eligibility criteria to ensure participant safety and data integrity. All Vuse Alto variants demonstrated rapid systemic nicotine uptake, with PK profiles consistent across flavors, indicating flavor-independent nicotine delivery. Observed Cmax ranged from 4.27 to 6.04 ng/mL, AUC0-15 from 46.09 to 64.63 ng × min/mL, and AUC0-240 from 420 to 509 ng × min/mL. Higher nicotine concentrations produced proportionally greater Cmax and AUC values, though all remained lower than those from cigarettes. Tmax for ENDS were consistently 6 min, aligning with the rapid onset typical of inhaled nicotine, while NRT gum exhibited delayed absorption (45 min). Subjective measures including product liking, positive effects, and intent to use again were lower for ENDS compared with cigarettes and generally comparable to NRT gum. Urge-to-smoke decreased within 15 min for all ENDS variants, similar to NRT gum but less rapidly than cigarettes. Findings demonstrate that Vuse Alto ENDS deliver nicotine more slowly and at lower levels than cigarettes, reflecting a lower AL while maintaining sufficient CNS-relevant nicotine exposure to potentially support transitions away from smoking. Flavor did not materially influence PK or AL outcomes. Collectively, the results indicate an intermediate neuropharmacological profile consistent with reduced dependence potential and support the potential utility of Vuse Alto ENDS within THR strategies.The clinical studies were registered at ClinicalTrials.gov; NCT05239884 and NCT05210699.
Chronic pain is a global healthcare issue. Mechanistic insight into pain mechanisms in the brain is needed. Increasing evidence has demonstrated a critical role for neuroimmune signaling factors in the pathogenesis of chronic pain. The amygdala, a bilateral limbic structure, is involved in the emotional-affective dimensions of pain and pain-modulation. There is good evidence for pain-related hemispheric lateralization in the central nucleus of the amygdala (CeA) in different pain models, but pain-related lateralization of neuroimmune signaling in the CeA remains to be determined. This study addressed the question if the well-documented right-hemispheric lateralization in pain conditions also occurs with the exogenous activation of neuroimmune signaling, which would suggest intrinsic differences between right and left CeA as the basis for pain-related hemispheric lateralization. To do so, either lipopolysaccharide (LPS), a toll-like receptor 4 (TLR4) agonist, or polyinosinic:polycytidylic acid (PolyI:C), a TLR3 agonist, was injected stereotaxically into the CeA. Mechanosensitivity, emotional-affective responses, and anxiety-like behaviors were evaluated in male and female rats 3 and 7 days post drug administration. Left and right CeA were collected at the end of the experiments for Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) analysis from female rats. Our results suggest that both right and left CeA are capable of generating sensory and emotional pain-like behaviors when activated exogenously by immunostimulants at the early stage (day 3) of neuroimmune activation in both sexes. At the later stage (day 7), sex-, treatment- and hemisphere-specific differences emerged with a TLR3-mediated right hemispheric CeA lateralization in females but not in males. qRT-PCR analysis confirmed local neuroimmune activation in both right and left CeA, with treatment-dependent gene expression changes occurring mainly ipsilateral to the injection site. Therefore, exogenous neuroimmune activation is possible in both the left and right CeA and generates pain-like behaviors initially, whereas females, but not males, develop lateralization subsequently, perhaps suggesting resilience developing in the left but not right CeA in terms of output and coupling to pain modulatory systems. Mechanistic insights into the hemispheric lateralization of neuroimmune signaling-related pain modulation in the amygdala may aid the development of therapeutic strategies for chronic pain relief.
Whether pulmonary arterial hypertension (PAH)-targeted therapies, particularly endothelin receptor antagonists (ERAs), are associated with disproportionate sepsis reporting in real-world pharmacovigilance data remains insufficiently explored. The monocyte transcriptional states that characterize sepsis-related immune dysregulation and may provide biological context for such reporting signals are also incompletely defined. We constructed an integrated, hypothesis-generating analytical framework incorporating: (i) FDA Adverse Event Reporting System (FAERS) disproportionality analysis coupled with XGBoost-based modeling for pharmacovigilance signal detection; (ii) single-cell RNA sequencing (scRNA-seq) analysis of peripheral blood mononuclear cells with intercellular communication inference; (iii) weighted gene co-expression network analysis (WGCNA) and cytoHubba-based topological prioritization, combined with an ensemble machine learning framework for diagnostic signature construction; and (iv) molecular docking and 100-nanosecond all-atom molecular dynamics (MD) simulation. FAERS analysis identified Maitentan and ambrisentan as PAH-targeted therapies with positive reporting signals for the MedDRA Preferred Term "Sepsis," with adjusted reporting associations persisting after adjustment for available demographic variables. Sex-stratified analysis showed marked heterogeneity in reporting signals, although these findings may be influenced by the sex distribution of PAH populations and other unmeasured confounders. ScRNA-seq resolved seven monocyte subpopulations, among which the interferon-responsive Mono_IFN subset-marked by IFIT1, ISG15, and IFITM3 expression-occupied a signaling hub position within the IFN-γ communication network and expanded in sepsis-associated states. Systematic comparison of 112 integrated machine learning algorithm combinations based on cytoHubba-prioritized genes yielded a 29-gene diagnostic model with cross-cohort discrimination for sepsis. Transcriptional co-expression analysis nominated IFITM3, a marker of the interferon-responsive monocyte state, as a candidate node connecting the diagnostic signature with interferon-related immune dysregulation. Molecular docking and 100-nanosecond MD simulation suggested a structurally stable riociguat-IFITM3 interaction in silico. This finding remains exploratory and requires biochemical and functional validation. This integrated pharmacovigilance and transcriptomic study identifies sepsis-reporting signals associated with selected endothelin receptor antagonists and characterizes an IFITM3-associated interferon-responsive monocyte state in sepsis datasets. These findings should be interpreted as reporting associations and transcriptomic hypotheses rather than evidence of causal drug-induced sepsis. The predicted riociguat-IFITM3 interaction provides a computational hypothesis for future experimental validation.
Although most cases of estrogen receptor-positive (ER+) breast cancer initially respond to endocrine therapy, many patients ultimately develop resistance. A major contributor to endocrine resistance in metastatic disease is the acquisition of constitutively active somatic mutations in the estrogen receptor ligand-binding domain (LBD). We previously identified midasin (MDN1), a ribosome biogenesis protein, as significantly overexpressed in letrozole-resistant MCF-7 cells. Because these cells are ERlow/- and represent only a subset of endocrine-resistant tumors, we hypothesized that ESR1 mutations cooperate with MDN1 dysregulation to confer a survival advantage. To address this, the cBioPortal database was queried to assess correlations between breast cancer subtypes and MDN1 expression levels. MCF-7 cell lines harboring ER point mutations were evaluated by RNA sequencing and immunoblot analysis to measure ER and MDN1 expression. To identify pharmacologic inhibitors of midasin, computational docking analyses were performed using a panel of ribozinoindole (Rbin) analogs, followed by biological evaluation studies. Initial analyses demonstrated that MDN1 expression is elevated in human breast cancer tumors, including luminal, HER2+, and triple-negative breast cancer. RNA sequencing of parental MCF-7 cells and ESR1-mutant derivatives, MCF-7 Y537S and MCF-7 D538G, revealed comparable MDN1 transcript levels across all cell lines. In contrast, immunoblot analysis showed mutation-dependent differences in MDN1 protein expression, with the highest levels observed in MCF-7 D538G cells, followed by MCF-7 Y537S cells and then parental MCF-7 cells. Computational docking analyses of Rbin analogs led to the selection of Rbin-1 and Rbin-2 for biological evaluation. Viability assays revealed minimal activity for Rbin-1, whereas Rbin-2 reduced proliferation by 30%-55% across all three cell lines, with the most pronounced effects observed in MCF-7 D538G cells at 24 and 48 h. Consistent with these findings, Rbin-2 treatment decreased MDN1 protein expression by approximately 50% in all cell lines, while ER levels remained largely unchanged. Collectively, these results establish the feasibility of pharmacologically targeting midasin in mammalian cell lines and support a functional link between MDN1 expression and ESR1 mutation-driven endocrine resistance. This work provides a foundation for future mechanistic studies of midasin as a potential therapeutic vulnerability in ER-mutant breast cancer.
Mature brain-derived neurotrophic factor has long been known to as an epigenetic regulator. It is excessively secreted after status epilepticus, leading to a variety of permanent structural and functional changes in the brain. While its precursor forms proBDNF regulates nerve development, neural transmission and cognitive function, it is not fully known whether the expression of proBDNF is changed in vivo or how it influences neuronal signaling and function ultimately leading to spatial cognitive impairments. Here, we investigated changes in proBDNF levels in the hippocampus of the pentylenetetrazole (pentylenetetrazole)-induced epileptic rat model. Functional blocking of proBDNF singling and its related pathway in the hippocampal regions was conducted to explore the potential mechanisms. Meanwhile, local field potentials were recorded in the hippocampal CA3-CA1 pathway and the directionality of neural information flow (NIF) between two regions was evaluated. We found that seizures were induced by PTZ-treated rats, which exhibited excessive proBDNF expression in the hippocampus only after undergoing behavioral training. Intra-hippocampal infusions of anti-proBDNF antibody into the CA1 but not the CA3 region could mitigate the PTZ-induced memory consolidation deficits and we confirmed the involvement of p75NTR rather than TrkB signaling. The excessive proBDNF could act on both presynaptic and postsynaptic sites through p75NTR signaling to exaggerate neural activity of putative fast-spiking interneurons. This was evidenced by increased spontaneous excitatory postsynaptic current frequency and amplitude, and further corroborated by action potential-independent miniature EPSC (mEPSC) recordings, which revealed concurrent increases in both mEPSC frequency and amplitude specifically in the epilepsy group. Importantly, this over-expression of proBDNF disrupted phase synchronization and directional coupling strength at the CA3 to CA1 synapses. However, blocking proBDNF or inactivation of the p75NTR signaling could effectively enhance the phase-locked value and neural information flow at the gamma and high-frequency oscillations, and significantly alleviate the PTZ-induced impairments in memory processing. Our findings are consistent with the hypothesis and provide the first direct evidence that the over-activation of proBDNF signaling represents a potential mechanism involved in neural dysfunction and NIF disruption leading to memory impairments in kindled animals.
Vancomycin, primarily excreted through the urine, is used for complicated urinary tract infections (cUTIs) caused by Gram-positive bacteria. Although serum therapeutic drug monitoring (TDM) is usually performed in vancomycin therapy, its benefits and risk factors in patients with cUTIs remain unclear. Adults with Gram-positive bacterial cUTIs receiving serum vancomycin TDM were enrolled from three prospective, multicenter trials. Minimal inhibitory concentration (MIC) was measured by agar dilution for pathogens collected from all patients. Clinical characteristics and pharmacokinetic/pharmacodynamic (PK/PD) indices were analyzed between the vancomycin treatment success and failure groups. A total of 74 adult patients with cUTIs were enrolled. Median initial daily dose of vancomycin was 1.0 g (interquartile range [IQR], 1.0-2.0 g), given in divided doses every 12 h or as a once-daily regimen. Most concomitant antibiotics targeted Gram-negative bacteria or fungi, with very limited anti-Gram-positive co-therapy. The median serum trough concentration (Cmin) of vancomycin was 9.22 mg/L (IQR, 4.36-14.33 mg/L) and 24-h area under the concentration-time curve to MIC (AUC24/MIC) was 455 (IQR, 268-627). Despite low attainment of the AUC24/MIC 400-600 target (27/74, 36.5%), the treatment success rate was 86.5% (64/74) and the nephrotoxicity rate was 4.1% (3/74). Urinary pathogens isolated included Enterococcus spp (55/74), Streptococcus spp (10/74), and Staphylococcus aureus (9/74), including eight methicillin-resistant S. aureus [MRSA]). Both solid tumor and S. aureus infection showed exploratory associations with vancomycin treatment failure. Patients with solid tumor had a lower probability of attaining the target AUC24/MIC, likely due to the elevated MIC of the predominant Enterococcus strains in this population. In contrast, patients with S. aureus-induced cUTIs achieved higher AUC24/MIC levels. All isolates exhibited low MICs (≤1 mg/L), no heteroresistance was detected, and the predominant molecular type was clone complex 5 (CC5). Vancomycin was effective in Enterococcus-dominant cUTIs and had a modest response in a few MRSA cases, despite low PK/PD target attainment. Serum Cmin and AUC24/MIC showed limited association with clinical outcomes, whereas solid tumor and S. aureus infection showed exploratory associations with treatment failure. However, TDM remains valuable for safety monitoring and individualized dosing. These findings should be validated in larger cohorts.
Diabetic retinopathy remains a major cause of vision loss, and therapeutic strategies beyond anti-vascular endothelial growth factor treatment are still needed. This study aimed to identify genetically supported druggable targets for diabetic retinopathy and to evaluate finerenone as a candidate therapeutic intervention in experimental retinopathy. We performed druggable Mendelian randomization by integrating druggable-gene resources, blood cis-expression quantitative trait locus data, and a large genome-wide association dataset for diabetic retinopathy. Significant genes were further evaluated using colocalization analysis, functional enrichment, protein-protein interaction network analysis, drug prediction, and molecular docking. Finerenone was subsequently assessed in db/db mice and in the oxygen-induced retinopathy model. Thirty candidate druggable genes were associated with diabetic retinopathy after false discovery rate correction. Among them, CDH2 was the only candidate showing significant colocalization with diabetic retinopathy risk, with a posterior probability for a shared causal variant of 0.85. Protein-protein interaction analysis showed relatively high connectivity of CDH2 within the candidate network, and molecular docking suggested a favorable predicted interaction between finerenone and N-cadherin. In vivo, finerenone reduced avascular and neovascular areas in oxygen-induced retinopathy retinas and decreased retinal expression of TNF-α, IL-1β, and N-cadherin in db/db mice. These findings prioritize CDH2/N-cadherin as a genetically supported candidate target in diabetic retinopathy and support finerenone as a potential therapeutic candidate. The retinal protective effects of finerenone may be associated with suppression of inflammatory responses and downregulation of N-cadherin, although the direct mechanistic link requires further validation.
Osteoarthritis (OA) is a degenerative musculoskeletal disease characterized by cartilage degradation and inflammation. Sophoridine (SR), a quinolizidine alkaloid from traditional Chinese herbs, has demonstrated anti-inflammatory properties, but its role in OA is unknown. The effects of SR on chondrocyte viability were assessed using the Cell Counting Kit-8 (CCK-8) assay. Interleukin-1β (IL-1β)-stimulated murine primary chondrocytes were used as an in vitro OA model. Extracellular matrix (ECM) synthesis was evaluated by toluidine blue staining. Key molecules involved in ECM metabolism, inflammation, and pyroptosis, as well as nuclear factor kappa B (NF-κB) signaling activity, were analyzed using quantitative reverse transcription PCR (RT-qPCR), Western blotting, and immunofluorescence. Chondrocyte morphological changes were examined by scanning electron microscopy. To validate causal mechanisms, a functional rescue experiment was performed in vitro using a specific NF-κB activator. For in vivo studies, an anterior cruciate ligament transection (ACLT)-induced murine OA model was established to evaluate the therapeutic efficacy of SR. OA progression was assessed by functional tests, micro-computed tomography (micro-CT), and histopathological analyses. Immunohistochemistry and immunofluorescence were further performed to evaluate molecular and signaling alterations in cartilage tissues. In IL-1β-stimulated chondrocytes, SR (20 and 40 μg/mL) significantly inhibited ECM degradation and suppressed the expression of pro-inflammatory cytokines. In vivo studies utilizing a well-established murine OA model induced by ACLT demonstrated that intraperitoneal SR administration (7.5 and 15 mg/kg/day for 8 weeks) markedly ameliorated cartilage destruction and inflammation. Notably, SR prominently reduced chondrocyte pyroptosis and suppressed NF-κB pathway activation. Mechanistically, co-treatment with an NF-κB activator significantly reversed the SR-mediated protection of ECM components and abolished the downregulation of key pyroptosis-related proteins. Our study indicates that SR effectively attenuates OA progression, which was associated with the suppression of chondrocyte pyroptosis and inflammation via NF-κB signaling pathway inhibition, suggesting SR as a promising therapeutic agent for OA treatment.
Prescription errors remain a significant challenge to medication safety in hospital settings. Forced Interception (FI) systems, which automatically flag and block potentially problematic prescriptions, serve as critical safeguards against adverse drug events. However, the specific characteristics and underlying causes of intercepted prescriptions, particularly in Chinese hospital contexts, require further investigation to inform targeted quality improvement strategies. This study aimed to analyze the characteristics and interception reasons of FI prescriptions in a hospital setting, with the goal of identifying patterns that could guide system upgrades, clinical training, and policy interventions. This study conducted a retrospective analysis of FI prescriptions intercepted by the hospital's electronic prescribing system. Prescriptions were analyzed for interception reasons, drug categories, specific medications, and prescribing department patterns. Data were collected and categorized to identify the most frequent issues and drug types involved in forced interceptions. A total of FI prescriptions were analyzed. The most common interception reasons were "Treatment duration exceeded" (54.89%) and "Exceeding Dosage" (28.11%), together accounting for the majority of interceptions. Traditional Chinese medicine and central nervous system drugs were the most frequently intercepted drug categories. The top three intercepted medications were Duloxetine Hydrochloride Enteric Capsules (3.63%), Atorvastatin Calcium Tablets (3.11%), and Tandospirone Citrate Capsules (2.81%). Departmental analysis revealed distinct prescribing patterns: the cardiology department showed high interceptions for hyperlipidemia-related drugs, while the mental health department had numerous interceptions for long-term antidepressant and anxiolytic prescriptions. The study suggests that targeted interventions, including upgrading electronic prescribing systems, department-specific training, and forming special review panels, are necessary to reduce prescription errors and improve medication safety. Future work should focus on multicenter studies and, as a longer-term direction, explore the potential of artificial intelligence for dynamic risk prediction to enhance the continuous optimization of medical quality.
Migraine in children and adolescents not only impacts academic pursuits and family life but also have secondary psychological effects. Determining the role of acute medication for migraine treatment in this population can reduce the burden associated with migraine. This network meta-analysis aimed to identify the relative efficacy and safety of acute migraine drug in children and adolescents migraine populations. The Cochrane Register of Controlled Trials and MEDLINE via PubMed and Embase databases were searched from inception to August 2025, only published studies in English. Double-blind randomized clinical trials evaluating the currently available acute treatments for childhood and adolescent migraines were included. The primary efficacy endpoint was pain freedom at 2 hours. Secondary efficacy endpoints included the proportion of participants with pain relief at 2 hours, pain freedom from two to 24 h, and the proportion using rescue drugs after 2 hours and up to 24 h. Adverse events (AEs) were also evaluated. The analysis included 30 studies (involving 8,914 participants and 13 pharmacological interventions). All treatments included demonstrated higher odds ratios (ORs) compared with the placebo for pain freedom at 2 hours. Dihydroergotamine was associated with the highest ORs, but its confidence interval included null values. Sumatriptan/naproxen sodium, ibuprofen, zolmitriptan nasal spray, sumatriptan nasal spray, and rizatriptan showed statistical significance. Sumatriptan/naproxen sodium yielded the highest odds (OR: 2.91, 95% CI: 1.87-4.53), followed by ibuprofen (OR: 2.88, 95% CI: 1.47-5.64), and rizatriptan showed the lowest (OR: 1.51, 95% CI: 1.23-1.86). Only sumatriptan/naproxen sodium was associated with a significantly higher OR compared with placebo for pain freedom from two to 24 h (OR: 2.31, 95% CI: 1.31-4.07). Ibuprofen exhibited the highest effect size for pain relief at 2 hours (OR: 3.21, 95% CI: 1.10-9.34). None of the included drugs was found to reduce the use of rescue drugs from two to 24 h. Zolmitriptan was associated with the highest risk of AEs among all treatments. Acetaminophen appears to have the lowest risk of adverse events, comparable to that of a placebo. Ibuprofen can effectively relieve symptoms, characterized by a favorable benefit-risk profile. Sumatriptan and zolmitriptan nasal sprays also exhibited robust efficacy, specifically among populations with prominent nausea and vomiting. Sumatriptan/naproxen sodium merits consideration, especially in patients exhibiting an inadequate response to monotherapy. Dihydroergotamine demonstrated potential benefits in refractory and chronic migraine; however, high-quality studies are warranted to validate these findings.
Pancreatic ductal adenocarcinoma (PDAC) is the fifth most common malignancy globally, with tumor uncontrolled angiogenesis being major causes of therapeutic failure and patient death. Isoalantolactone (IATL), a natural compound, exhibits antioxidant, anti-inflammatory, anti-proliferative, and anti-tumor properties. However, its role in inhibiting tumor angiogenesis in pancreatic cancer and the underlying mechanisms remain unclear. This study aims to investigate the anti-angiogenic effects of IATL in PDAC and elucidate the associated molecular pathways. In vitro experiments were performed using PDAC cell lines (Panc02, PANC-1, and SW 1990) and HUVECs. In vivo studies were conducted using an orthotopic pancreatic cancer model in C57BL/6 mice. Cell viability, wound-healing, tube formation, in vivo imaging system analysis, laser speckle contrast imaging, immunohistochemistry, immunofluorescence, RT-qPCR, ELISA, and Western blot assays were used to evaluate the effects of IATL on tumor growth, angiogenesis, and inflammatory responses. Molecular docking and cellular thermal shift assay were performed to assess the interaction between IATL and NLRP3. IATL significantly inhibited the proliferation and migration of Panc02, PANC-1, and SW1990 cells. In the orthotopic pancreatic cancer model, IATL dose-dependently suppressed tumor growth, as evidenced by reduced IVIS fluorescence signals and tumor volume. IATL also markedly inhibited angiogenesis, as shown by reduced HUVECs migration and tube formation, decreased tumor blood perfusion detected by laser speckle imaging, and downregulated CD34 and VEGFA expression both in vivo and in vitro. Network pharmacology, molecular docking, and cellular thermal shift assay identified NLRP3 as a direct target of IATL. Mechanistically, IATL suppressed NLRP3 inflammasome activation, reduced ASC speck formation, inhibited the NLRP3/IL-1β signaling axis, and decreased the expression of inflammatory cytokines, including IL-6, TNF-α, IL-1β, and IL-18. NLRP3 knockdown mimicked the effects of IATL, whereas NLRP3 overexpression partially reversed its anti-tumor, anti-angiogenic, and anti-inflammatory effects, further supporting the target specificity of IATL. IATL functions as a novel NLRP3 pathway inhibitor, suppressing angiogenesis through anti-inflammatory mechanisms, thereby effectively inhibiting PDAC progression. These findings suggest that IATL holds potential as a therapeutic agent for pancreatic cancer.
Despite intensive multimodal therapy, high-risk neuroblastoma remains associated with poor clinical outcomes because of treatment resistance, recurrence, and progressive disease, underscoring the need for new therapeutic strategies. We investigated whether proteasome-associated deubiquitinating enzymes are therapeutically relevant in neuroblastoma by assessing ubiquitin C-terminal hydrolase L5 (UCHL5) and ubiquitin-specific protease 14 (USP14) expression in neuroblastoma tissues and evaluating the antitumor activity of VLX1570 in preclinical models. Immunohistochemical (IHC) analysis showed stronger UCHL5 and USP14 immunoreactivity in neuroblastoma tissues than in normal peripheral nerve tissue. In human neuroblastoma cell lines IMR-32, SK-N-SH, and SH-SY5Y, VLX1570 reduced cell viability in a dose- and time-dependent manner, induced apoptosis, and triggered G2/M arrest. These effects were accompanied by induction of CCAAT/enhancer-binding protein homologous protein (CHOP) and suppression of proliferating cell nuclear antigen (PCNA)-associated proliferative signaling, as reflected by reduced expression of PCNA, phospho-histone H3, and Bcl-2 together with increased p21 and p53 expression. PCNA knockdown experiments further supported inhibition of PCNA-associated proliferative signaling as a functionally relevant component of VLX1570-induced cytotoxicity. In addition, VLX1570 enhanced cisplatin-induced apoptosis in vitro and potentiated cisplatin antitumor activity in neuroblastoma xenograft models. Together, these findings support proteasome-associated deubiquitinating enzyme inhibition as a pharmacologic strategy in neuroblastoma and provide a rationale for further evaluation of VLX1570, particularly in combination with cisplatin.
Gegen Qinlian Decoction (GQD), a classic traditional Chinese medicine formula recorded in Shang Han Lun, has been used clinically for approximately two thousand years. In recent years, its potential roles in ulcerative colitis (UC) and colorectal cancer (CRC) have attracted increasing attention because of the multi-component, multi-target, and multi-pathway regulatory characteristics of traditional Chinese medicine (TCM). UC is a chronic relapsing inflammatory disease and an important risk factor for CRC. Although emerging immunomodulators, biologics, and targeted therapies have achieved certain efficacy in the treatment of UC and CRC, limited response, drug resistance, adverse effects, and disease recurrence remain major challenges for some patients. Persistent intestinal inflammation can disrupt the epithelial barrier, alter gut microbiota, induce immune imbalance, enhance oxidative stress, and activate pro-tumorigenic signaling, thereby contributing to the transition from UC to CRC. Within this inflammation-to-cancer framework, preclinical evidence suggests that GQD may exert comprehensive regulatory effects on multiple pathological processes. It may suppress inflammatory signaling pathways such as TLR4/NF-κB, IL-6/JAK2/STAT3, and the NLRP3 inflammasome, regulate Th17/Treg balance and macrophage polarization, restore epithelial barrier integrity, reshape gut microbiota homeostasis, and modulate oxidative stress, abnormal cell death, and metabolic dysregulation. These effects may act synergistically to limit the transition from an inflammatory microenvironment to a pro-tumorigenic microenvironment. In established CRC, GQD appears more suitable as a potential adjunctive therapy rather than a primary cytotoxic anticancer agent, with possible roles in reducing inflammation-associated tumor-promoting signals, remodeling the tumor immune microenvironment, enhancing responses to chemotherapy or immunotherapy, and mitigating treatment-related intestinal toxicity. In summary, GQD may have potential important value in the treatment of UC and CRC, and this review provides a reference for future research and clinical application.
Metabolic dysfunction-associated fatty liver disease (MAFLD) is recognized as the hepatic manifestation of metabolic syndrome. Hepatic steatosis resulting from impaired energy metabolism constitutes the core of its pathogenesis, and this disease has become a global public health concern. AMP-activated protein kinase (AMPK) is widely expressed in high-energy-consuming organs and functions as a vital energy sensor that maintains systemic energy homeostasis. Current preclinical in vitro and in vivo evidence demonstrates that AMPK activation modulates multiple MAFLD-related pathological processes, including enhancing cellular autophagy, regulating lipid metabolism, reducing inflammatory and oxidative damage, improving IR, and mitigating mitochondrial dysfunction. Plant metabolites have attracted increasing research attention for targeting the AMPK pathway in MAFLD basic research, due to their multi-target regulatory characteristics and low adverse reaction profiles. Accordingly, based on the intrinsic connection between the AMPK signaling pathway and MAFLD, this review summarizes the research progress regarding the pharmacological mechanisms of plant metabolites acting on the AMPK pathway in MAFLD intervention. Notably, most studies included in this review are preclinical experiments conducted in vitro and in vivo, with scarce supporting clinical data. Further efforts are still needed to advance the translation of these findings and confirm their clinical potential. This review systematically summarizes recent basic research progress and identifies unresolved issues, so as to offer a theoretical basis for subsequent mechanistic studies and translational exploration of traditional Chinese medicine against MAFLD.
Imperata cylindrica is a perennial botanical drug with both ecological impacts and diverse utilization values. As a traditional Chinese medicine, its rhizome is widely applied in traditional medical systems of various countries, which can cool blood to arrest bleeding and clear away heat to induce diuresis. Modern research has isolated and characterized 141 chemical metabolites from Imperata cylindrica, mainly including flavonoids and triterpenoids. These metabolites have been associated with multiple bioactivities, including anti-inflammatory, antioxidant, and immunomodulatory effects, and many proprietary Chinese medicines containing its rhizome have been applied in clinical practice. Imperata cylindrica also shows good application prospects in food processing, agriculture and animal husbandry, and ecological restoration. However, current research is limited by insufficient characterization of active metabolites, inadequate studies on isolated metabolites, and a predominance of in vitro and preclinical evidence. In the future, efforts should focus on the in-depth development and pharmacological verification of active metabolites, improving the cell-animal experimental system, promoting industrial application in multiple fields, and counteracting its invasion status through resource utilization to achieve a win-win situation of ecology and economy.
Aristolochic Acid I (AAI) is a potent nephrotoxin and Group 1 carcinogen. Despite stringent regulatory restrictions, AAI-containing herbal remedies are still sporadically used to treat respiratory symptoms, presenting a previously underappreciated exposure risk for patients with lung adenocarcinoma (LUAD). However, the specific tumor-promoting effects of AAI on preexisting LUAD remain to be fully elucidated. In this study, we integrated network toxicology, TCGA transcriptomic analysis, and molecular docking to identify core oncogenic networks potentially affected by AAI. The associated pro-tumor phenotypes and transcriptional regulatory abnormalities were subsequently investigated through a series of in vitro and in vivo experiments. Network analysis and TCGA data mining identified a cluster of seven hub genes-including ERBB2, SERPINE1, CCNA2, and CHEK1-that are crucial to LUAD progression and significantly associated with poor clinical prognosis. Molecular docking simulations suggested potential binding affinities between AAI and these target proteins. Functional assays demonstrated that acute AAI exposure significantly accelerated the proliferation, migration, and invasion of LUAD cell lines (PC9 and NCI-H1299) in vitro, while promoting macroscopic xenograft tumor growth in vivo. The RT-qPCR and WB results showed that AAI exposure was accompanied by an increase in the expression of core genes. Our findings suggest that, beyond its established chronic mutagenic toxicity, AAI may act as a potent tumor promoter in LUAD. By potentially influencing key oncogenic networks, AAI accelerates the malignant progression of LUAD, underscoring the severe clinical hazards of AAI exposure in patients with preexisting lung malignancies.
To investigate the potential impact of plasticizers on infertility and elucidate the underlying mechanisms, we employed an integrated approach combining network toxicology, molecular docking, and in vitro experimental validation. Initially, we identified potential targets by intersecting plasticizer-related targets with infertility-associated targets, yielding 134 candidate targets. These targets were subsequently analyzed using the STRING database and Cytoscape software to construct protein-protein interaction (PPI) networks, from which 20 hub genes were identified. Functional enrichment analysis conducted using the R software revealed that these targets participate in multiple biological pathways. Molecular docking simulations performed with AutoDock Vina and visualized using PyMol demonstrated strong binding affinities between key hub targets and three representative plasticizers: diethyl phthalate (DEP), dimethyl phthalate (DMP), and dioctyl phthalate (DOP). For experimental validation, we examined the effects of plasticizers on the human ovarian granulosa cell lines. CCK-8 assays revealed significant inhibition of cell proliferation following exposure to plasticizer. Flow cytometry and western blot analyses further showed that plasticizer treatment upregulated the anti-apoptotic protein BCL-2 while downregulating pro-apoptotic BAX, as well as key reproductive markers, including CYP19A1, ER-α, and FSHR. Collectively, these findings demonstrate that plasticizers may contribute to infertility through multiple molecular pathways, as predicted by network toxicology and confirmed by in vitro experiments. This integrated approach provides compelling evidence for the reproductive toxicity of plasticizers and offers insights into their potential mechanisms of action.
Social isolation (SI), attributable to modern lifestyles and fast-growing technology, is a leading cause of depression. Selenium nanoparticles (Se-NPs) are neuroactive agents owing to their antioxidant and anti-inflammatory activities. Additionally, physical and mental activities (Ph&M) exert neuroprotective effects by optimizing the release of both neurotransmitters and growth factors. However, their neuroprotective effects against SI-induced depression are still poorly investigated. We aim to explore the neuroprotective effect of Se-NPs, Ph&M, and their combination to guard against the harmful effects of SI-induced depression in a rat model. Fifty Sprague Dawley rats were randomly allocated into five groups: control, SI, Ph&M, orally administered Se-NPs (0.1 mg/kg), and a combination group. Neuroprotective activity was quantitatively estimated pharmacologically, biochemically, histologically, and behaviorally. SI caused behavioral and biochemical alteration in the rat model, decreasing neurotransmitter levels, increasing the transcription of inflammatory response genes (TLR4 and NF-κB), and consequently increasing the production of the cytokines TNF-α and IL-1β. It also activated the proinflammatory NLRP3/caspase-1 pathway. The ER stress parameters PERK, CHOP, and GRP78 were significantly elevated. Impairment of autophagy and increased neurodegeneration were detected via the decline of AMPK/SIRT-1/Beclin-1 PI3K/AKT gene expression and m-TOR overexpression. Decreased expression of TrkB and CREB mRNA and consequent decline in brain-derived neurotropic factor were recorded. SI caused a drastic drop in Wnt3a and β-catenin levels and increased GSK3β activity affecting neuroplasticity and cognitive functions. Administration of Se-NPs and/or application of Ph&M, especially their combination, provided significant protection against prior SI effects. Se-NPs and Ph&M, especially their combination, showed promising protective effects against neuroinflammation, oxidative stress, apoptosis and subsequent alterations of test animal behaviors precipitated by SI.
Hemophilia B is a rare X-linked congenital bleeding disorder characterized by a deficiency in coagulation factor IX (FIX). Standard management relies on exogenous factor replacement; however, the development of neutralizing alloantibodies (inhibitors) against infused clotting factor can significantly reduce therapeutic efficacy and complicate long-term management. Consequently, patients turn to bypassing agents or non-factor therapies to help achieve adequate hemostatic control. Concizumab is a novel subcutaneous non-factor therapy for hemophilia A and B that targets the tissue factor pathway inhibitor (TFPI). While it is approved for patients >12 years old, data remains sparse in younger children. Prophylactic treatment with concizumab reduces several limitations associated with inhibitor development and variability in treatment response with conventional therapies, especially in pediatric populations. We report a toddler with hemophilia B who developed a low-titer inhibitor following treatment with recombinant coagulation factor IX agents, resulting in recurrent bleeding complications and consistent subtherapeutic hemostatic control on standard bypassing agents. The patient was transitioned to concizumab for long-term prophylaxis, with dose adjustments based on clinical response and concizumab drug levels. Following the initiation of concizumab, the patient demonstrated a marked reduction in bleeding episodes. We highlight an individualized early drug-level-guided dose escalation approach in a very young child, demonstrating that pharmacokinetic monitoring can be utilized proactively to optimize therapeutic response in this age group, leading to effective drug levels and excellent clinical response. This case supports the role of concizumab in potentially reducing treatment burden and improving hemostatic outcomes in young patients with severe hemophilia B and inhibitors.