To observe the cytotoxic effects of the Blaps rynchopetera Fairmaire (BRF) ethyl acetate extract on the human ovarian carcinoma cell line SKOV3.
SKOV3 cells were exposed to 0 μg/mL (control), 5 μg/mL, 10 μg/mL, 25 μg/mL, 50 μg/mL, or 100 μg/mL BRF ethyl acetate extract for 2 h, 6 h, 12 h, 24 h, 36 h, or 48 h. SKOV3 cell viability was evaluated using a cell counting kit-8 (CCK-8) assay. β-catenin, cyclin D1, and p21 protein levels were detected via Western blotting. Expression of vascular endothelial growth factor (VEGF) and CD34 protein was detected via immunohistochemistry.
BRF ethyl acetate extract inhibited the viability of SKOV3 cells in a dose- and time-dependent manner. The IC50 of the BRF ethyl acetate extract for SKOV3 cells was 13.07 μg/mL. The cell viability was lowest after exposure to the BRF ethyl acetate extract for 48 h. β-catenin, cyclin D1, and p21 protein levels decreased in a time-dependent manner and were lowest after exposure to the 10 μg/mL BRF ethyl acetate extract. VEGF and CD34 protein staining intensity and the number of positive cells were the lowest after exposure to the 10 μg/mL BRF ethyl acetate extract.
BRF ethyl acetate extract may exert antitumor effects on ovarian cancer by suppressing tumor cell proliferation and angiogenesis.
Oxidative stress and inflammation are widely recognized as key mechanisms in the pathogenesis of Drug-Induced Liver Injury (DILI). Meanwhile, preclinical studies have demonstrated that tocotrienols (T3s), members of the vitamin E family, possess significant antioxidant and anti-inflammatory properties, suggesting a potential hepatoprotective role in various liver disorders. Clinical trials investigating Non-Alcoholic Fatty Liver Disease (NAFLD), a condition that shares important pathophysiological features with DILI, have also reported favorable outcomes associated with T3 supplementation. Notably, the overlap between the established mechanisms of T3s and the underlying pathophysiology of DILI provides a strong rationale for exploring the therapeutic potential of T3s in this context. Emerging evidence from studies on NAFLD further supports this approach, considering the common mechanistic pathways involved. Accordingly, this review aims to comprehensively evaluate current preclinical and clinical evidence on T3s in relation to DILI, elucidate the proposed mechanisms of action of this class of vitamin E analog, and identify key gaps in the literature that warrant further investigation.
The administration of neocuproine (NC), a copper(I)-binding agent, can induce phasic contractions in the neonatal rat bladder, while NC treatment can also promote tonic contractions in the adult rat bladder. These data may indicate that copper is important in regulating developmental contractile responses. Thus, investigating how responses to copper(I)-binding agents change during development could offer valuable insights into the mechanisms underlying overactive bladder. Therefore, this study aimed to induce developmentally different contraction responses in isolated rat whole-bladder tissues using NC.
Neonatal (1–2 weeks old) and adult (250–300 g) Wistar rats were used in the experiments. Isolated rat bladders were placed in organ baths containing 2 μM atropine and 2 μM guanethidine Krebs solution. The resulting tone changes in the preparation were recorded using a pressure transducer (cmH2O). Differences between groups were evaluated using one-way analysis of variance (ANOVA). The Student's t-test was used to assess the paired groups (GraphPad Prism); p-values smaller than 0.05 were considered statistically significant.
NC administration caused a significant increase in basal spontaneous contraction responses in neonatal rat whole-bladder tissue, whereas in adult whole bladder, NC treatment promoted a significant tonic contraction over basal tonus. In isolated whole-bladder tissue from neonatal rats, the increases in the amplitude and area under the curve (AUC) of spontaneous contraction responses induced by 50 μM NC were significantly reduced by the addition of 1 μM nifedipine, 50 μM adenosine triphosphate (ATP), 100–200 μM suramin, or 30 μM NS1619, a large-conductance Ca2+-activated K+ [BK] channel opener, to the medium. The number of spontaneous contractions decreased in the presence of NC; however, this finding was reversed in the presence of the aforementioned drugs. Moreover, the tonic contractions observed in the presence of NC in the adult bladder preparations were significantly reduced following the addition of 1 μM nifedipine, 50 μM ATP, 100 μM suramin, or 30 μM NS1619. However, the addition of 100 μM Nw-nitro-L-arginine (LA) to the medium had no significant effect on the amplitude, AUC, and frequency of NC-induced tonic contractions or spontaneous contraction responses.
These results suggest that copper may play an important role in the regulation of phasic/tonic contractions in the bladder during postnatal development.
Topotecan (TPT) is a novel class of anti-tumor drugs known for its broad-spectrum anti-cancer activity and low toxicity. This study aimed to investigate the potential mechanisms through which TPT mediates the phosphatase and tensin homolog/phosphatidylinositol 3-kinase/glycogen synthase kinase-3β (PTEN/PI3K/GSH-3β) signaling pathway to affect survival and tumor growth in Non-Small Cell Lung Cancer (NSCLC) xenograft nude mice.
A NSCLC nude mouse model was fabricated by subcutaneously injecting H1993 human NSCLC cells into the right axillary fossa. The mice were treated with cisplatin (DDP) and 0.5, 1.0 and 2.0 mg/kg of TPT. Tumor volume changes were monitored and assessments were performed on organ indices, immune function, tumor cell apoptosis, survival rates (SRs) and protein levels of components involved in the PTEN/PI3K/GSK-3β pathway in tumor tissues. One-way analysis of variance (ANOVA) or Chi-square tests were conducted using SPSS 23.0 to compare intergroup differences.
The SRs of nude mice treated with DDP and TPT markedly increased, with high-dose TPT treatment showing a drastically superior SR to DDP (p < 0.05). As the dose of TPT increased, the tumor volumes in the mice decreased markedly, and the indices of the thymus and spleen notably increased. Among T lymphocyte subsets, the proportion of CD4+ cells and the CD4+/CD8+ ratio increased, while the proportion of CD8+ cells decreased. Serum levels of interleukin-4, tumor necrosis factor (TNF)-α and interferon-γ increased. The apoptosis rate of tumor cells increased, and the relative expression level of PTEN in tumor tissue increased, whereas the levels of p-PI3K, p-PI3K/PI3K ratio, p-GSK-3β, and p-GSK-3β/GSK-3 ratio decreased (p < 0.05).
TPT dose-dependently inhibited NSCLC growth by modulating T-cell subsets, enhancing immune function, and exerting antitumor effects through the PTEN/PI3K/GSK-3β pathway. The high-dose group (2.0 mg/kg) demonstrated superior efficacy compared to the cisplatin and low-dose groups, validating the importance of concentration gradient design in determining the optimal therapeutic window.
Generic dydrogesterone products are widely indicated in various conditions such as infertility, menstrual disorders, and the prevention of miscarriage. Moreover, the therapeutic equivalence of generic dydrogesterone products has never been explored, despite reports of interindividual variability in the bioavailability of dydrogesterone. Therefore, this study aimed to compare two generic formulations of dydrogesterone (A and B) by evaluating the respective quality, pharmacokinetics, and endometrial tissue concentrations of these formulations in rats.
Differential scanning calorimetry (DSC), X-ray diffraction (XRD), and the drug content were compared in the quality analysis. Meanwhile, dissolution and rat plasma kinetic profiles were compared to determine the in vitro and in vivo equivalence. Endometrial drug levels were assessed by liquid chromatography–tandem mass spectrometry (LC-MS/MS) and in vivo imaging.
The DSC and XRD results revealed that a higher percentage of the dydrogesterone in formulation A was amorphous, compared to formulation B, where the dydrogesterone was more crystalline. The difference in drug content between the formulations was not significant (p > 0.05), and the dissolution profile and plasma bioavailability of formulations A and B were similar (p > 0.05). LC-MS/MS analysis showed that dydrogesterone levels in the endometrial tissue of rats treated with formulation A were markedly higher compared to those treated with formulation B. The in vivo imaging results corroborated with the LC-MS/MS analysis, with nearly 43.0% higher dydrogesterone accumulation observed in the uterus of animals treated with formulation A compared to formulation B.
These data exhibited that therapeutic inequivalence may exist between the generic formulations of dydrogesterone; hence, the generic formulation should be carefully selected.
Osteoradionecrosis of the jaw (ORNJ) is a common complication following radiotherapy for head and neck cancer. Thus, this study aimed to explore the effects of active components in Lycium barbarum on ORNJ through network pharmacology and to conduct experimental verification to identify potential therapeutic targets.
The main active ingredients in Lycium barbarum (Gouqi), a traditional Chinese herbal medicine, was used in this study. After identifying ferroptosis-related genes associated with ORNJ, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses and constructed an interaction network. A molecular interaction force analysis was then performed on eight binding conformations to identify the optimal conformation with the lowest binding free energy. We established a model of ORNJ in Sprague-Dawley (SD) rats and administered oral Lycium barbarum glycopeptide (LbGP) to the experimental group. Moreover, reverse transcription quantitative polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay (ELISA), and immunohistochemical staining techniques were then employed to detect the mRNA and protein levels of various relevant cytokines.
Based on network pharmacology predictions, this study identified three potential active components in Lycium barbarum, namely β-sitosterol, glycitein, and quercetin. The possible effects of these components in the treatment of ORNJ were analyzed. Seven hub genes related to ferroptosis and ORNJ were identified, and LbGP was selected for in vivo verification. The expression levels of TP53, EGFR, IL-6, and TNF were significantly altered in the LbGP group compared to the untreated group, as revealed by the qPCR, ELISA, and immunohistochemistry assays.
The use of Lycium barbarum extract may exert therapeutic effects on mandible injury following ORNJ by regulating the expression of TP53, EGFR, IL-6, and TNF.
Aerococcus urinae is an emerging pathogen associated with serious infections, particularly in immunocompromised individuals; however, no approved vaccines or targeted therapies currently exist. Thus, this study presents the rational design of a novel multi-epitope chimeric vaccine using integrative core-subtractive proteomics and advanced immunoinformatics.
Two conserved antigenic targets (cell division protein, filamenting temperature-sensitive protein Z (FtsZ), and single-stranded DNA-binding protein) were identified for epitope prediction through Geptop 2.0 essentiality screening, basic local alignment search tool for proteins (BLASTp) non-homology analysis against the human proteome, subcellular localization filtering using the subcellular localization predictive system (CELLO), and antigenicity scoring with VaxiJen (threshold ≥0.5). The final construct incorporated the carefully selected B cell, cytotoxic T lymphocyte (CTL), and helper T lymphocyte (HTL) epitopes to maximize immunogenicity and population coverage.
Comprehensive analyses demonstrated the stability, antigenicity, non-allergenicity, non-toxicity, and cytokine-inducing potential of the vaccine. Molecular docking and dynamics simulations confirmed strong binding affinity and structural stability with toll-like receptor 4 (TLR4), underscoring the ability of the vaccine to trigger innate immune activation. In silico cloning and codon optimization predicted efficient expression in Escherichia coli K12. Immune simulations further supported robust humoral and cellular responses.
These findings highlight a promising vaccine candidate warranting experimental validation as a potential strategy to protect against A. urinae infections.
Flavopiridol (Flavo), a synthetic flavonoid derived from Dysoxylum binectariferum, broadly inhibits cyclin-dependent kinases (CDKs) that regulate transcription in proliferative cells. In the peripheral nervous system, Schwann cells exhibit transcriptional changes during peripheral neurodegenerative processes (PNPs), involving c-Jun and Krox20 as key regulators. Additionally, hydrogen sulfide (H2S) promotes neuroprotection by activating the Wnt/β-catenin pathway, which induces cyclin D1 via β-catenin nuclear translocation.
To assess the therapeutic potential of Flavo in the PNPs, we conducted experiments targeting Schwann cell responses both in vivo and ex vivo. Moreover, we examined changes in transcriptional regulation, focusing on c-Jun and Krox20, and analyzed the effect of Flavo on the H2S/β-catenin/CDK signaling pathway using Western blot and morphological evaluation of demyelination, axonal degeneration, and Schwann cell proliferation.
Flavo modulated Schwann cell transcription by shifting the c-Jun/Krox20 balance and significantly suppressed abnormal Schwann cell proliferation. The in vivo analysis revealed that Flavo dose-dependently reduced demyelination and axonal degeneration. Mechanistically, Flavo inhibited the H2S/β-catenin/CDK axis, reducing β-catenin nuclear translocation.
Our findings suggest that Flavo provides multifaceted protection in peripheral nerves by correcting transcriptional dysregulation and attenuating key pathological features of Schwann cell dysfunction. Administering Flavo, in relation to the H2S/β-catenin/CDK pathway, may represent a promising therapeutic strategy for peripheral neurodegenerative diseases through multitarget transcriptional modulation.