Background:Poria cocos is a medicinal and edible fungus widely used in traditional Chinese medicine and functional food development. However, the global research structure and thematic evolution of this field remain insufficiently characterized. This study aimed to map publication trends, collaboration networks, knowledge bases, and emerging research themes in Poria cocos research. Methods: Publications related to Poria cocos were retrieved from the Web of Science Core Collection from 1 January 1990 to 1 April 2026. Articles and reviews were included. Bibliometric and visualization analyses were performed using Microsoft Excel and an open-source Python 3.11.8 workflow (NetworkX 3.6.1 and Matplotlib 3.10.8). Co-occurrence analysis was used to count the frequency of co-occurrence of certain elements (e.g., countries, regions, institutions, etc.); cluster analysis was used to classify keywords; and burst analysis was used to identify research trends and hotspots. Results: A total of 1404 records were included. Publication output increased markedly after 2014 and peaked in 2025, whereas 2026 data were incomplete. China contributed the largest share of publications and occupied the central position in international collaboration networks. The Journal of Ethnopharmacology and International Journal of Biological Macromolecules were major publication venues. Keyword and co-citation analyses indicated a thematic shift from phytochemical characterization and resource studies toward polysaccharides, pharmacological mechanisms, network pharmacology, and gut microbiota. Conclusions: This bibliometric analysis provides an overview of the evolving research landscape of Poria cocos. Future studies should strengthen data-driven mechanistic validation, quality standardization, translational pharmacology, and clinical evidence generation.
Background: Nigella sativa L. is a medicinal plant widely recognized for its diverse pharmacological properties. This study aimed to evaluate the antioxidant, diuretic, and cytotoxic effects of an aqueous seed extract and its effects on selected biochemical parameters in rats. The pharmacokinetic potential of identified bioactive compounds was also investigated in silico. Methods: The aqueous extract was prepared by maceration. Antioxidant activity was assessed using DPPH, reducing power, and total antioxidant capacity (TAC) assays. We evaluated the diuretic effect in vivo in Wistar rats by measuring urinary concentrations of Na+, K+, Cl−, and creatinine, using furosemide as a reference drug. Cytotoxicity was assessed in splenocytes and thymocytes. We analyzed seven identified compounds in silico for physicochemical properties, Lipinski compliance, intestinal absorption, cytochrome P450 inhibition, and blood–brain barrier permeability. Results: The extract showed significant antioxidant activity, with a DPPH IC50 of 0.254 ± 0.002 mg/mL and a TAC of 125.01 ± 4.220 mg AAE/g. It significantly increased urinary electrolyte and urinary concentration, indicating a diuretic effect lower than that of furosemide. Plasma analyses showed decreased electrolyte concentrations and increased creatinine levels. Cell viability exceeded 90%, indicating no significant cytotoxicity. Most compounds complied with Lipinski’s rule of five and showed favorable predicted intestinal absorption, with limited cytochrome P450 inhibition. Several compounds were predicted to cross the blood–brain barrier. Conclusions: The aqueous extract of N. sativa demonstrated antioxidant and diuretic activities without significant cytotoxicity. These findings support its pharmacological potential and warrant further investigation of its mechanisms of action and therapeutic relevance.
Background: Caffeine is a widely consumed psychostimulant that may affect anxiety-like, aggression-like, and social behaviors, potentially in association with oxidative imbalance. Objectives: This study evaluated the behavioral and biochemical effects of acute caffeine exposure in adult zebrafish and investigated the potential protective effects of N-acetylcysteine (NAC). Methods: Sixty adult wild-type AB zebrafish (Danio rerio), approximately 9–10 months old, were assigned to six groups (n = 10/group): control, caffeine (25 or 60 mg/L), NAC (1 mg/L), and NAC combined with either caffeine concentration. Behavioral effects were assessed using the Social Preference Test, Mirror-Biting Test, and Novel Tank Test. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were measured as oxidative stress-related markers. Results: Caffeine increased locomotor activity and induced anxiety-like and aggression-like behavioral alterations, including increased bottom-dwelling, thigmotaxis, and mirror-directed behavior, while reducing social preference. Caffeine significantly increased SOD activity at 25 mg/L (p = 0.010) and 60 mg/L (p = 0.008), and GPx activity at 25 mg/L (p = 0.008) and 60 mg/L (p = 0.006), compared with controls. NAC attenuated caffeine-induced hyperlocomotion and several anxiety-like and aggression-like alterations and modulated antioxidant enzyme responses, but did not fully restore social preference. Conclusions: Acute caffeine exposure produces behavioral alterations and changes in antioxidant enzyme activity in adult zebrafish. NAC partially attenuates these effects, particularly hyperlocomotion, anxiety-like behavior, and aggression-like responses, while showing more limited effects on caffeine-induced social alterations.
Background/Objectives: Advanced gastroesophageal adenocarcinoma (GEA) carries a poor prognosis, with median overall survival of 13–20 months despite standard chemotherapy. Since trastuzumab’s approval, biomarker-driven precision therapies have expanded rapidly. This review comprehensively summarises current and emerging targeted agents across the key molecular targets driving advanced GEA. Methods: A comprehensive literature review was conducted using PubMed, Google Scholar and Cochrane Library in accordance with PRISMA guidelines, searching English-language human studies published between February and July 2026, supplemented by updates during editing to reflect current standards. Results: HER2-directed therapy has progressed from trastuzumab through dual blockade, immunotherapy combinations, next-generation ADCs (notably trastuzumab deruxtecan) and bispecific antibodies such as zanidatamab, which has now overtaken trastuzumab in the first-line setting. CLDN18.2-targeted zolbetuximab has demonstrated survival benefit in biomarker-selected patients, with newer ADCs, BiTEs and the first approved solid-tumour CAR-T therapy (satricabtagene autoleucel) extending this target further. VEGFR2 inhibition with ramucirumab remains a cornerstone in later lines, while novel VEGF/PD-1(L1) bispecifics are under investigation. FGFR2b-targeted bemarituzumab showed early promise that weakened on phase 3 confirmation, and MET/EGFR-directed agents, including savolitinib and amivantamab, require stringent biomarker selection to demonstrate benefit amid tumour heterogeneity. Conclusions: Novel targeted agents, particularly to HER2 and CLDN18.2, have demonstrated survival benefit despite ongoing challenges. Other lines are more investigational.
Background: Colistin is increasingly being used as a last-line treatment option for multidrug-resistant Gram-negative infections; however, its clinical use remains limited due to its nephrotoxic side effects. This study investigated the potential nephroprotective effects of ethyl pyruvate (EP) and rutin hydrate (RH), alone and in combination, against colistin-induced renal damage. Methods: Forty-nine adult male Wistar albino rats were allocated into seven groups, and all agents were administered intraperitoneally once daily for seven days. On the eighth day, one kidney of each animal was perfused using a Langendorff system to measure perfusion pressure, while samples from the contralateral kidney and blood were used for biochemical, molecular, and histopathological analyses. Results: In the colistin group, urea, creatinine, total oxidant status, and perfusion pressure were significantly higher than in the control, EP, and RH groups (p < 0.05). Oxidative stress index, tissue MDA, KIM-1, caspase-3, and PUMA levels were also markedly increased. All of these parameters decreased in the treatment groups, with the lowest values for most parameters in the group receiving both EP and RH, which also showed the best-preserved renal architecture. Conclusions: These findings suggest that EP and RH may be promising adjuvant agents against colistin-induced nephrotoxicity. Combined EP and RH co-treatment showed numerically greater protection across most assessed parameters, although an overall advantage over single-agent co-treatment was not statistically established.
The aryl hydrocarbon receptor (AHR) and the vitamin D receptor (VDR) were long regarded as independent transcription factors governing distinct physiology—xenobiotic sensing and calcium–vitamin D homeostasis, respectively. AHR, a basic helix–loop–helix/PAS protein, heterodimerizes with ARNT and binds xenobiotic response elements (XREs) to drive cytochrome P450 genes such as CYP1A1; VDR, a nuclear receptor activated by 1,25-dihydroxyvitamin D3, heterodimerizes with RXR and binds vitamin D response elements (VDREs). Although their genes reside on separate chromosomes (AHR, Chr 7; VDR, Chr 12), an integrated view recognizes the two pathways as extensively cross-regulatory. This review synthesizes the molecular, immunological, and tissue-level evidence for VDR–AHR interplay. At the molecular level, the receptors cooperate at composite promoter architectures—most notably an everted-repeat VDRE positioned adjacent to an XRE in the CYP1A1 promoter—while AHR ligands reciprocally enhance CYP24A1-mediated catabolism of active vitamin D. Tryptophan metabolism provides a bidirectional hub: kynurenine and the UVB photoproduct FICZ serve as endogenous AHR ligands whose balance, modulated by VDR, shapes signaling output. The tumor suppressor p53 functions as a shared upstream regulator coupling genotoxic stress to both receptors, with convergence on the CDKN1A (p21) checkpoint. Functionally, AHR and VDR converge on the regulatory T cell (Treg)/Th17 axis to influence immune tolerance: sustained AHR activation by TCDD favors Foxp3+ Treg differentiation, transient FICZ-driven activation promotes Th17 responses, and VDR reinforces the tolerogenic arm while independently repressing IL-17. The receptors further cooperate in maintaining intestinal epithelial barrier integrity and NF-κB restraint, with parallel impairment in inflammatory bowel disease, and are co-activated in skin by solar UVB, which simultaneously generates vitamin D3 and the AHR ligand FICZ within keratinocytes. In cancer, VDR acts as a tumor suppressor, AHR exhibits context-dependent pro- and anti-tumor roles, and a three-way AHR–VDR–p53 interaction—inverted by mutant p53—forms a critical regulatory node. Throughout, the direction and magnitude of cross-talk prove highly dependent on cell type, ligand identity and kinetics, and species—distinctions often underappreciated in the literature. Clarifying these context-specific determinants is essential for translating AHR–VDR cross-regulation into rational therapies in autoimmunity, mucosal inflammation, dermatology, and oncology.
Background: Primary dysmenorrhea (PD) is a prevalent gynecological condition that significantly compromises the quality of life in adolescents and women of reproductive age. Within traditional Chinese medicine (TCM), Cold Coagulation and Blood Stasis Primary Dysmenorrhea (CCBS-PD) represents the most frequently observed syndrome pattern of PD. Wenjing Decoction (WJD), a classical TCM formulation, has been extensively employed for the treatment of CCBS-PD. However, given the multi-component and complex nature of WJD, the potential mechanisms underpinning its therapeutic effect have yet to be elucidated. Methods: A rat model of CCBS-PD was induced through ice-water bath stimulation in conjunction with estradiol benzoate and oxytocin. The pharmacological effects of WJD were evaluated by writhing response, hemorheological parameters, uterine index, histopathological examination, and biochemical assays. Serum-exposed constituents of WJD were characterized using UPLC-Orbitrap Exploris 120 MS. To study the mechanisms of WJD, methods from network pharmacology, molecular docking, and off-target metabolomics were used. Western blot analysis examined representative proteins in the signaling pathways predicted by network pharmacology. Network pharmacology and metabolomics were integrated to construct a pathway–metabolite–target–compound network, and representative targets were analyzed by RT-qPCR. Results: WJD treatment reduced writhing responses, improved hemorheological abnormalities, and alleviated uterine pathological changes in CCBS-PD rats. Serum pharmacochemistry identified 62 WJD-derived constituents. Metabolomics analysis indicated that WJD was associated with alterations in nitrogen metabolism, valine/leucine/isoleucine biosynthesis and arginine biosynthesis. Network pharmacology and molecular docking suggested several candidate compounds and targets, including Robinetin, Levistolide A, Pratol, 7,4′-dihydroxyflavone, EGFR, AKT1, ESR1, MMP9, MAPK3, and TNF. RT-qPCR showed that selected genes, including AKT1, EGFR, MAPK3, TNF, ESR1, MMP9 and CASP3, were changed in the model group and partially restored following WJD improvement. Western blot analysis demonstrated that WJD treatment decreased the phosphorylation levels of AKT, ERK1/2, and NF-κB, and down-regulated the protein expression of COX-2. Conclusions: WJD showed beneficial effects in a CCBS-PD rat model. Synthesized assessments indicate a potential link to the actions of serum-exposed constituents, alterations in amino acid metabolism, and modulation of inflammation-related signaling pathways. This research offers initial indications suggesting the multi-component and multi-target pharmacological actions of WJD, although the underlying mechanisms remain to be validated through targeted metabolomics and functional studies.
Background/Objectives: Oxidative stress, chronic inflammation, disrupted apoptosis, and altered autophagy are biological processes implicated in colorectal tumor development. Morin is a plant-derived flavonoid with antioxidant and anti-inflammatory properties, but its limited solubility and bioavailability may limit its biological activity. This study evaluated the effects of free morin and morin-loaded liposomes (MOR-Lips) on 1,2-dimethylhydrazine (DMH)-associated colonic biochemical, molecular, and histopathological alterations in rats. Methods: Rats were randomly assigned to six groups—vehicle control, MOR, MOR-Lips, DMH, DMH + MOR, and DMH + MOR-Lips—and treated for 10 weeks. Serum and colonic tissues were evaluated for cancer-associated biomarkers (CEA, CA19-9, CA125, HMG-CoA reductase), oxidative stress and antioxidant indices, nitrosative and oxidative DNA-damage markers (MDA, NO, 8-OHdG), inflammatory mediators (TLR4/NF-κB/COX-2, cytokines, MPO), proliferative indices (Ki-67, PCNA), apoptotic and autophagy-related regulators (Bax, caspase-3, p53, cytochrome c, BCL-2, p-AKT, LC3-II, Beclin-1, p62), and histopathological and immunohistochemical changes. Results: DMH exposure was associated with increased CEA, CA19-9, CA125, HMG-CoA reductase, MDA, NO, 8-OHdG, TLR4/NF-κB/COX-2, cytokines, MPO, Ki-67, and PCNA. DMH also reduced NRF2/HO-1 signaling and antioxidant defenses, shifted apoptosis-related markers toward a pro-survival profile, altered autophagy-related markers, and produced marked colonic histopathological abnormalities. Both free MOR and MOR-Lips attenuated several of these DMH-associated alterations, with MOR-Lips generally producing greater effects than free MOR. MOR-LIP treatment was associated with restoration of antioxidant marker profiles, reduced levels of inflammatory and proliferative markers, a shift toward a pro-apoptotic marker profile, partial normalization of autophagy-related markers, and improved colonic histopathological appearance. Conclusions: In DMH-exposed rats, MOR-Lips were associated with more favorable redox, inflammatory, proliferative, apoptosis-related, autophagy-related, and histopathological profiles than free MOR. These findings support further investigation of MOR-Lips as a formulation strategy for improving the biological activity of morin. Because quantitative preneoplastic and neoplastic endpoints were not measured, the results do not establish inhibition of colorectal carcinogenesis or chemopreventive efficacy.
Background: Triple-negative breast cancer (TNBC) remains a major cause of cancer mortality due to its aggressive behavior, metabolic adaptability, and high therapeutic resistance. Extracellular vesicles (EVs) within the tumor microenvironment contribute to tumor progression and metastasis by transferring pro-tumorigenic cargo. While conventional Tumor Treating Fields use high-frequency alternating fields to disrupt mitosis, low-energy triple-frequency bioelectromagnetic approaches remain poorly characterized. Methods: We evaluated a device–drug strategy combining triple-frequency low-intensity electromagnetic stimulation (EMS2: 396 Hz, 285 Hz, 528 Hz) with the pleiotropic drug Fingolimod (FTY720). Treatments were tested in MDA-MB-231 and ARM-G breast cancer cells, with Paclitaxel as a positive control. Cell proliferation was assessed by MTS assay, and extracellular vesicles were isolated following individual and combination treatments. Quantitative LC-MS/MS proteomics was used to characterize treatment-induced changes in EVs cargo. Results: EMS2 reduced proliferation in both cell lines and produced morphological changes consistent with altered cell-cycle progression. EMS2 alone triggered adaptive metabolic responses, whereas combination with Fingolimod suppressed these compensatory signatures. EVs proteomics revealed combination-specific alterations associated with mitochondrial stress, ER stress, NF-κB suppression, and autophagy-associated pathways. The combination also reduced levels of metastasis- and stroma-associated proteins, including Mitogen-Activated Protein Kinase 12 (MAPK12) and collagen-associated ECM components (Collagen Type I Alpha 1 Chain (COL1A1), Collagen Type VI Alpha 1 Chain (COL6A1), Collagen Type VI Alpha 3 Chain (COL6A3), and Matrilin 3 (MATN3)) in EVs. Bliss independence analysis identified a subset of metastasis-associated proteins suppressed in EVs beyond the level predicted by an additive model, an exploratory finding that will require further validation with dose–response and functional assays. Conclusions: Combined triple-frequency EMS2 and Fingolimod treatment altered the extracellular vesicle proteome, inducing signatures consistent with mitochondrial and endoplasmic reticulum stress, metabolic disruption, and reduced levels of metastasis-associated and stromal/ECM remodeling proteins, along with reduced proliferation. These findings suggest a coordinated anti-cancer effect of this tunable device–drug strategy, warranting further functional and in vivo validation to confirm therapeutic potential.
Background/Objectives: Sjögren’s Syndrome (SS) is an autoimmune disorder with impaired exocrine gland function. Its pathogenesis remains elusive, but aberrant innate/adaptive immunity and dysregulated interferon signaling are core features. Current SS research lacks exploration of innate immune mechanisms and therapeutic targets. Phloretin, a natural dihydrochalcone derivative, inhibits excessive innate immune activation, yet its role and mechanism in SS remain unreported. Methods: This study aimed to explore the therapeutic efficacy of phloretin against Sjögren’s syndrome (SS) and elucidate the regulatory mechanism underlying its effects on dendritic cells (DCs) and the Stimulator of Interferon Genes (STING)-driven innate immune cascade. Clinical specimens were utilized to analyze DC infiltration and STING expression in the labial glands of SS patients. In vivo experiments were conducted on NOD/Ltj SS mice with phloretin treatment, using hydroxychloroquine as a positive control. For in vitro studies, DCs were stimulated with poly I:C to verify the functional effects of phloretin and its regulatory role in the STING/TANK-binding kinase 1 (TBK1)/Interferon Regulatory Factor 3 (IRF3) signaling. Salivary secretion, glandular inflammatory responses, and systemic immune overactivation were evaluated in SS mice. In poly I:C-stimulated DCs, cell activation, migration, and co-stimulatory molecule expression were measured. The regulatory effects of phloretin on the STING signaling cascade and STING ubiquitin-mediated degradation were examined by Western blotting. Phloretin, initially identified as a candidate compound via virtual screening, was subjected to thermal shift assay (CETSA), drug affinity responsive target stability (DARTS) and molecular dynamics simulation (MD) for direct binding verification with STING. The STING overexpression rescue experiment further corroborates that phloretin modulates innate immune responses in a STING-dependent manner. Results: Clinical results revealed obvious infiltration of CD11c+STING+ DCs and CD11c+p-TBK1+ DCs in labial gland lesions of SS patients. In NOD/Ltj SS mice, phloretin treatment recovered salivary secretion, relieved glandular inflammatory injury, and restrained excessive activation of innate and adaptive immune responses, showing comparable therapeutic effects compared with hydroxychloroquine. In vitro experiments confirmed that phloretin could alleviate poly I:C-induced abnormal activation and migration of DCs, as well as reduce the expression of co-stimulatory molecules. Mechanistically, phloretin was found to interfere with the STING signaling cascade, alter the ubiquitination level of STING protein, and further inhibit the abnormal activation of DCs, which may contribute to its protective effects against Sjögren’s syndrome. Notably, results from MD, CETSA and DARTS collectively validate the interaction between phloretin and STING protein, providing solid molecular-level evidence for its regulatory effect on STING signaling transduction. Conclusions: DCs and STING-driven innate immunity critically contribute to SS progression. Phloretin effectively ameliorates SS-related immune disorders by restraining DC overactivation and STING signaling, supporting it as a promising candidate agent for SS treatment.
Background/Objectives: AC02 is a novel 39-amino-acid adrenocorticotropic hormone (ACTH) analogue designed for the treatment of infantile spasms. To support its clinical study, in which porcine ACTH1–39 served as the positive-control drug, reliable methods for the determination of AC02 and porcine ACTH1–39 in human plasma were required. Reported analytical methods for ACTH analogues are mainly immunoassays, which are easily affected by cross-reaction and the hook effect, necessitating a more selective analytical approach. Methods: Two LC-MS/MS methods were developed for the determination of ACTH analogues AC02 and porcine ACTH1–39 in human plasma. Human ACTH1–39 was included as a selectivity marker to confirm that endogenous ACTH does not interfere with the quantification of AC02. Based on the distinct concentration requirements and matrix challenges, two sample-preparation procedures were established: micro-solid-phase extraction coupled with protein precipitation for porcine ACTH1–39 (LLOQ 0.100 ng/mL), and acid-mediated protein precipitation for AC02 (LLOQ 0.500 ng/mL). The [M+6H]6+ ions were selected as precursor ions, and the corresponding 5+ fragment ions, formed by loss of the C-terminal phenylalanine, were used for MRM detection. Results: Despite a mass difference of only 0.98 Da between AC02 and human ACTH1–39, which are indistinguishable by mass spectrometry, baseline chromatographic separation was achieved. Both methods were fully validated in accordance with current bioanalytical guidelines. Conclusions: The validated methods were successfully applied to the phase I clinical study of AC02 and porcine ACTH1–39, enabling reliable quantification of the drug candidate and its active comparator, porcine ACTH1–39.
Background: Building on earlier work with methoxy-, dimethoxy-, bromo-, and nitro-substituted salicylaldehyde benzoylhydrazones, we synthesized and evaluated new 4-chloro- and 5-chloro derivatives bearing an additional chlorine substituent on the hydrazide-derived phenyl ring to further explore the impact of halogenation on cytotoxic activity. Methods: Chloro-hydrazones were obtained through a one-step condensation of 4- or 5-chlorosalicylaldehyde with benzhydrazide or 2-, 3-, and 4-chlorobenzhydrazides. Their physicochemical, pharmacokinetic, ADME, lead-likeness, and drug-likeness profiles were evaluated in silico using SwissADME, ACD/Labs v9.10, and MDL QSAR v2.2.0.0.446. The synthesized compounds were structurally characterized by IR, 1H NMR, 13C NMR, and HR ESI–MS, and their cytotoxic activity was subsequently assessed by the MTT assay in selected cancer cell lines. Molecular docking was performed against ABL1 tyrosine kinase (ABL1 TK), a potential molecular target relevant to two of the investigated leukemia cell lines, using GOLD v.5.2.2 (CCDC Ltd., Cambridge, UK). Results: The chloro-substituted salicylaldehyde benzoylhydrazones exhibited pronounced, cell-type-dependent cytotoxicity, with leukemia cells being substantially more sensitive than breast carcinoma cells. Most derivatives showed low- to sub-micromolar IC50 values against SKW-3 human T-cell prolymphocytic leukemia, K-562 human chronic myeloid (myelogenous) leukemia, and BV-173 human BCR::ABL1-positive leukemia cells, with K3 and K4 exhibiting the highest activity in SKW-3 cells (IC50 = 0.5 ± 0.1 µM). K5 showed particularly strong activity against K-562 and BV-173 cells (IC50 = 0.7 ± 0.1 and 0.9 ± 0.1 µM, respectively), compared with 26.9 ± 2.4 and 21.5 ± 3.3 µM for imatinib. HL-60 human acute promyelocytic leukemia cells showed intermediate sensitivity (IC50 = 1.3–13.9 µM), whereas the activity against MCF-7 estrogen receptor-positive and MDA-MB-231 triple-negative breast carcinoma cells was more variable (1.5–32.1 and 4.1–62.5 µM, respectively). The derivatives showed high selectivity toward malignant cells relative to non-malignant CCL-1 non-malignant mouse fibroblasts, with lower-bound SI values frequently exceeding 50 and reaching >200 in SKW-3 cells. Conclusions: The chloro-substituted salicylaldehyde benzoylhydrazones exhibited pronounced and selective cytotoxicity, particularly toward leukemia cell lines, identifying this scaffold as a promising starting point for further anticancer drug-discovery studies.
Background/Objectives: Magnetic iron oxide nanoparticles are investigated in cancer research; however, the direct biological effects of unloaded magnetic colloidal suspensions in renal cancer remain insufficiently characterized. This study compared two double-oleic-acid-coated formulations prepared from precursors: magnetite-based MCS 1 and maghemite-based MCS 2. Methods: The suspensions were characterized by vibrating-sample magnetometry, dynamic light scattering, bright-field scanning transmission electron microscopy, and energy-dispersive X-ray spectroscopy. Their effects on A704 renal adenocarcinoma cells were evaluated after 24 h of exposure to 1–5 µg/mL using MTT, neutral red uptake, JC-1, Hoechst 33342/MitoTracker Red CMXRos, acridine orange/propidium iodide staining, and a 7-day clonogenic assay. Results: MCS 2 showed higher volumetric saturation magnetization (1.541 vs. 1.059 Gs), a smaller Z-average hydrodynamic diameter (79.65 vs. 103.9 nm), and a more uniform volume-weighted distribution. Both formulations significantly reduced metabolic activity and neutral red uptake and induced mitochondrial depolarization, cell-death-associated morphological changes, and impaired clonogenic capacity. MCS 1 generally produced a moderate response with an apparent plateau, whereas MCS 2 showed a pronounced concentration-related effect at 4–5 µg/mL. At 5 µg/mL, MCS 2 reduced metabolic activity to approximately 49%, neutral red uptake to 50.04%, the JC-1 aggregate-to-monomer ratio to 46.02%, and colony formation to 31.44% of the control. Conclusions: The suspensions exhibited distinct physicochemical and biological profiles. MCS 2 produced greater effects at the highest concentrations, potentially related to its smaller hydrodynamic size and more uniform particle-size distribution. Further studies should establish tumor selectivity, cellular uptake, and the underlying molecular mechanisms.
Background: Pancreatic adenocarcinoma (PDAC) remains one of the most lethal malignancies, with limited survival gains despite advances in systemic therapy. However, rapid expansion of biomarker-directed therapies, immunotherapy, and novel treatment modalities has created an increasingly complex clinical trial landscape. We aimed to characterize contemporary PDAC clinical development and identify emerging therapeutic trends. Methods: We performed a narrative review with a scoping approach of active PDAC clinical trials registered on ClinicalTrials.gov. Eligible studies were initiated between January 1, 2021, and February 16, 2026, and included recruiting, active, or not-yet-recruiting phase I–III trials. Data extracted included trial phase, disease setting, therapeutic strategy, endpoints, enrollment, sponsorship, and late-phase development. Results: A total of 355 interventional trials were included. Most trials were phase I, phase I/II, or phase II trials, with fewer than 10% being evaluated in phase II/III or phase III development. Advanced or metastatic disease was the predominant setting. Chemotherapy remained the most frequently incorporated treatment modality, while molecularly targeted therapies were evaluated in 167 trials. KRAS/RAS-directed approaches represented the largest targeted subgroup, although only daraxonrasib and setidegrasib reached phase III evaluation. Immunotherapy was evaluated in 139 trials, although only a limited number progressed to late-phase development, reflecting the immune-resistant biology of pancreatic adenocarcinoma. Additional areas of active investigation included Claudin 18.2-targeted therapies, MTAP-associated approaches, homologous recombination deficiency-directed strategies, CD73 inhibition, radiotherapy, local interventions, surgery-focused optimization, imaging-guided approaches, and supportive-care interventions. Industry organizations were listed as the lead sponsor for approximately half of all studies, while non-commercial organizations supported most remaining trials. Conclusions: The contemporary PDAC clinical trial landscape is characterized by broad therapeutic diversification but limited late-phase maturity. Chemotherapy remains the dominant treatment backbone, whereas KRAS/RAS-directed therapies have emerged as the most advanced precision oncology strategy. Future progress will likely depend on successful integration of biomarker-selected therapies with established multidisciplinary treatment approaches, including optimized systemic therapy, local-control strategies, and supportive care.
Metabolic dysfunction-associated steatotic liver disease (MASLD) develops when hepatic lipid acquisition and synthesis exceed the capacity for oxidation and very-low-density lipoprotein export. Flavonoids act on several components of this network, yet their therapeutic development is constrained by poor aqueous solubility, extensive intestinal and first-pass metabolism, variable activity of circulating metabolites, and limited information on hepatic exposure. Experimental studies link representative flavonoids to AMPK–SREBP-1c and PPARα signaling, mitochondrial quality control, Nrf2-dependent redox defense, inflammatory pathways, and the gut–liver axis. By contrast, the available randomized trials of quercetin, hesperidin, anthocyanins, green-tea catechins, EGCG, and soy isoflavones show at most modest changes in liver fat or biochemical markers and do not demonstrate metabolic dysfunction-associated steatohepatitis (MASH) resolution or fibrosis regression. Liposomal, lipid-based, polymeric, and nanocrystal formulations have improved dissolution, systemic exposure, or liver distribution in preclinical models, but comparative pharmacokinetics, chronic safety, manufacturability, and clinical efficacy remain poorly defined. The evidence therefore supports viewing flavonoids as formulation-dependent investigational candidates rather than established MASLD therapies. Progress will depend on chemically standardized products, exposure–response studies, clinically relevant models, and adequately powered trials using validated imaging or histological endpoints.
Background/Objectives: Cutaneous fungal infections remain a major therapeutic challenge due to poor skin penetration and emerging antifungal resistance of current treatments. Drug repurposing offers a promising strategy to expand antifungal therapy, with ketoprofen (KPN) recently demonstrating antifungal activity. Methods: In this study, KPN-cationic aspasomes (Ca-ASPMs) were developed using a Quality by Design (QbD) approach to enhance topical delivery and maximize the therapeutic potential of repurposed KPN. A D-optimal experimental design was implemented to investigate the influence of ascorbyl palmitate amount (X1), ethanol concentration (X2), and cationic SAA type (X3) on the critical quality attributes of KPN-Ca-ASPMs, namely entrapment efficiency, particle size, and zeta potential. Results: Numerical optimization identified an optimum formulation comprising 10 mg ascorbyl palmitate, 5% ethanol, and didodecyldimethylammonium bromide (DDAB) as cationic SAA, with an overall desirability of 0.815. The optimal KPN-Ca-ASPM (F9) exhibited nanosized vesicles (213.24 ± 2.20 nm) and 91.30 ± 10.01% entrapment efficiency. TEM confirmed the spherical morphology of the vesicles, while DSC demonstrated successful incorporation of KPN within the aspasomal matrix. F9 exhibited enhanced in vitro drug release (68% after 6 h) and excellent storage stability. Confocal laser scanning microscopy demonstrated enhanced skin penetration of F9. In a murine cutaneous candidiasis model, F9 significantly enhanced the antifungal efficacy of KPN, producing a 2.989-log reduction in fungal burden compared with the untreated group and significantly outperforming the free drug (p = 0.0005). Histopathological examination confirmed restoration of normal skin architecture. Conclusions: Collectively, the QbD-guided development of KPN-Ca-ASPM provided a reproducible nanocarrier that significantly enhanced the topical antifungal efficacy of repurposed KPN, highlighting its potential for the treatment of cutaneous candidiasis.
Background: Alterations in glutamatergic homeostasis have been implicated in Alzheimer’s disease, but the relationship between total hippocampal tissue glutamate and Alzheimer-like molecular alterations remains unclear. This study evaluated the early neuroprotective potential of gentisic acid (GA) and erucic acid (EA) in an intracerebroventricular streptozotocin (icv-STZ)-induced rat model of sporadic Alzheimer-like pathology. Methods: Sixty-four female Sprague–Dawley rats were randomly assigned to eight groups: control, sham, STZ, STZ + GA (100 or 200 mg/kg/day), STZ + EA (25 or 50 mg/kg/day), and STZ + memantine (10 mg/kg/day). Treatments began immediately after STZ and continued orally for 21 days. Behavioral performance was assessed by Morris water maze and passive avoidance tests. Total hippocampal tissue glutamate was quantified by LC–MS/MS. Neuronal degeneration was assessed histopathologically, whereas tau, amyloid-β, and AChE immunoreactivity and tau-, APP-, and AChE-related in situ hybridization signals were evaluated. Results: Compared with controls, STZ-treated rats exhibited poorer behavioral performance, higher total hippocampal tissue glutamate, greater neuronal degeneration, and increased molecular signals. GA and EA administration was associated with attenuation of these alterations, with generally stronger effects at higher doses. EA at 50 mg/kg/day showed the lowest mean total hippocampal tissue glutamate among treatment groups. GA at 200 mg/kg/day and EA at 50 mg/kg/day showed the most consistent preservation across behavioral, histopathological, and molecular endpoints. Conclusions: Early GA and EA administration was associated with neuroprotective effects in the icv-STZ model. Because treatment began immediately after STZ administration, the findings support early or preventive neuroprotection rather than reversal of established Alzheimer-like pathology. Total tissue glutamate measurements neither distinguish extracellular glutamate nor directly demonstrate glutamate-mediated excitotoxicity.
Background/Objectives: In the search for potent non-sugar α-glucosidase inhibitors with improved safety profiles, a novel flavonoid glycoside was isolated for the first time from the stem bark of Albizia saponaria (Fabaceae). The objective of this study was to elucidate its chemical structure and evaluate its therapeutic potential as an anti-hyperglycemic agent compared to known related flavonoids and a standard clinical drug. Methods: Comprehensive structural elucidation was performed using high-resolution mass spectrometry and multidimensional 1D/2D NMR (1H, 13C, HSQC-DEPT, COSY, and CIGAR). To assess its inhibitory efficacy and pharmacokinetic profiles, an in silico comparative study was conducted against a database of related flavonoids (Quercitrin, Hyperoside, and Isoquercitrin) and the clinical drug Acarbose. This involved molecular docking simulations against human intestinal maltase-glucoamylase (PDB ID: 3TOP) alongside integrated ADMET modeling and toxicological screening. Results: The compound was successfully identified as 4′,7-dihydroxyflavan-3′-O-β-D-glucoside (1). Molecular docking revealed that Compound 1 exhibited a superior predicted binding affinity of −9.5 kcal/mol, outperforming Quercitrin (−9.3 kcal/mol), Hyperoside (−8.3 kcal/mol), Isoquercitrin (−7.9 kcal/mol), and Acarbose (−7.2 kcal/mol). This strong thermodynamic stability is driven by a robust conventional hydrogen-bonding network with key active site residues (Arg1377, Gln1372, and Gly1365), successfully overriding a localized electrostatic strain at Asp1279. Furthermore, ADMET modeling demonstrated a highly desirable local pharmacokinetic framework; its low Caco-2 permeability (−6.432) and low human intestinal absorption (HIA = 0.120) favor targeted luminal retention in the gastrointestinal tract, mirroring Acarbose while minimizing systemic exposure. Crucially, toxicological screening unveiled a significant safety advantage for Compound 1, marked by negligible CYP3A4 interaction (0.004) and a remarkably low risk of Drug-Induced Liver Injury (DILI = 0.213) compared to the high-risk hepatotoxic profile of Acarbose (DILI = 0.882) and the reference flavonoids (DILI > 0.69). Conclusions: These predictive findings establish Compound 1 as a highly promising, low-toxicity natural scaffold for anti-hyperglycemic drug development. Its superior binding affinity and minimized hepatotoxicity risk warrant subsequent in vitro and in vivo functional validation.
Purpose: To evaluate the incidence, predictors, and clinical impact of treatment-emergent epiretinal membrane (ERM) in diabetic macular edema (DME) treated with dexamethasone implant, aflibercept, ranibizumab, bevacizumab, or faricimab. Methods: In this multicenter, retrospective, nonrandomized study, 501 eyes with center-involving DME were assessed and 423 eyes without ERM on pretreatment OCT and with 24-month follow-up were included. Eyes were classified by the intravitreal agent selected by the treating physician in routine care and maintained throughout follow-up; eyes requiring a drug switch or retinal laser/vitrectomy during follow-up were excluded. The last eligible pre-index SD-OCT confirmed the absence of ERM; BCVA, central subfield thickness (CST), and vitreomacular-interface (VMI) morphology were assessed at baseline and 6, 12, 18, and 24 months. Crude comparisons, multivariable logistic regression, and Kaplan–Meier/log-rank analyses were performed. Results: ERM developed in 36/423 eyes (8.5%). Incidence was 16.0% with dexamethasone implant, 7.5% with aflibercept, 7.7% with ranibizumab, 7.4% with bevacizumab, and 4.3% with faricimab. Dexamethasone implant showed higher ERM odds than pooled non-dexamethasone therapy after adjustment (OR 2.24, 95% CI 1.02–4.92; p = 0.044). Baseline VMI abnormality, severe NPDR/PDR, prior PRP, and higher baseline CST were also associated with ERM. Eyes developing ERM had smaller BCVA gains (+2.8 vs. +7.1 ETDRS letters; p < 0.001) and CST reductions (88.6 vs. 130.8 µm; p = 0.004). Conclusions: Treatment-emergent ERM was associated with baseline ocular characteristics and treatment category and with poorer 24-month outcomes. These observational drug-group signals require prospective confirmation.
Chemically induced proximity (CIP) has revolutionized small-molecule pharmacology by moving beyond traditional occupancy-based inhibition and activation. Within this paradigm, monovalent nondegrading molecular glues (ndMGs) represent a structurally compact and mechanistically distinct class of linkerless compounds. These agents stabilize or induce selective biomolecular interactions, altering the functional state or cellular localization of macromolecular complexes without causing component degradation. Once considered rare phenomena unique to specific natural products, ndMGs are now being actively engineered across diverse therapeutic landscapes, with multiple candidates advancing into clinical trials and achieving regulatory validation. We systematically highlight their evolving capabilities to modulate oncogenic networks, intervene with immune activity, rectify metabolic signaling, control neurodegenerative trafficking and stress networks, and disrupt critical pathogen assemblies, while broadening the druggable landscape into non-canonical mechanisms. In each section, we analyze the therapeutic value of the targets, the molecular mechanisms of action, the latest progress, and the unique challenges faced by these strategies. In summary, the ndMG modality represents a highly versatile strategy to unlock the undruggable proteome.
