Frankenia hirsuta (FH) is an underexplored halophytic plant used for managing inflammatory and metabolic disorders. However, its phytochemical composition and pharmacological mechanisms remain largely uncharacterized. Thus, this study aimed to comprehensively elucidate the phytochemical profile, molecular mechanisms, and pharmacological efficacy of FH using an integrated approach combining liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolite characterization, in vitro bioassays, in-vivo validation, and computational analyses. Metabolite profiling was performed using LC-MS/MS, followed by molecular docking and network pharmacology analyses to predict potential targets involved in neuropathic, inflammatory, and metabolic pathways. In vitro cytotoxicity and anticoagulant activities of the aerial and root ethanolic extracts were assessed using MTT and activated partial thromboplastin time (aPTT) assays, respectively. The in vivo antidiabetic and neuroprotective effects were evaluated in streptozotocin-induced diabetic mice, using behavioral assays (hot plate and von Frey filaments), biochemical markers of oxidative stress and inflammation, and histopathological and immunohistochemical (IHC) examinations. LC-MS/MS analysis revealed a diverse bioactive constituents, including phenolic acids, flavonoid glycosides, and feruloyl derivatives. Network pharmacology and docking studies highlighted strong binding affinities toward key enzymes such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), cyclooxygenase-2, and AKT1, indicating multimodal anti-inflammatory and neuroprotective potential. In vitro, FH extracts exhibited significant cytoprotective effects and prolonged aPTT in a dose-dependent manner, confirming their safety and mild anticoagulant properties. In vivo, FH root extract (75 mg/kg) and its isolated feruloyl glycoside (FG) (5-10 mg/kg) significantly restored nociceptive thresholds, improved glycemic control, normalized oxidative and inflammatory biomarkers, and preserved pancreatic and sciatic nerve histoarchitecture. IHC analyses further demonstrated downregulation of TNF-α and IL-6 and upregulation of IL-10 in treated groups, confirming their anti-inflammatory and regenerative effects. This study provides the first comprehensive evidence that FH exerts potent neuroprotective and antidiabetic effects mediated through antioxidant, anti-inflammatory, and cytokine-modulating pathways. The integrated LC-MS/MS, computational, and experimental validation framework highlights FH and its FG as promising candidates for developing safe, plant-derived therapeutics targeting diabetic neuropathy and related metabolic dysfunctions.
Lycopodium clavatum L. has a long history of use in various traditional medicine systems. This review aims to systematically examine its ethnobotanical applications, phytochemical constituents, and pharmacological activities, with a particular focus on the innovative drug delivery potential of its sporopollenin microcapsules. A systematic literature search was conducted to gather and synthesize relevant information on the ethnobotany, chemistry, and pharmacology of L. clavatum. The key bioactive components include lycopodane-type alkaloids, triterpenoids, flavonoids, and phenolics. These compounds contribute to validated pharmacological effects such as anti-parasitic immunomodulation, induction of apoptosis in cancer cells, anticholinesterase activity, and hepatoprotection. The spores are a rich source of sporopollenin, which serves as a biocompatible platform for fabricating microcapsules with controlled release and enhanced bioavailability for therapeutic agents. Some clinical reports suggest efficacy for conditions like urolithiasis and hyperuricemia. The phytochemical profile of L. clavatum supports its traditional uses and highlights its potential in phytotherapy. Future research should focus on developing standardized extracts, conducting rigorous clinical trials, exploring synergistic formulations, and further exploiting sporopollenin-based nanocarriers to bridge ethnopharmacology with modern medicine.
Comprehensive pain management in children requires a specialized skillset, with a limited number of clinicians possessing the level of expertise required to successfully navigate the complexities of holistic care. The emergence of pediatric anesthesia fellowship programs in sub-Saharan Africa presents an opportunity to embed a pediatric pain curriculum for trainees, improving the availability of specialist skill and knowledge in the field. Existing pain curricula fall short in addressing the sociocultural aspects of pediatric pain identified through research as being unique to the African context, and do not include elements of leadership and advocacy training required to navigate the complexities of resource-constrained healthcare settings. A Delphi survey including literature review, iterative rounds of surveys and expert consensus was used to establish a pediatric pain curriculum for pediatric anesthesia fellows undertaking advanced training in sub-Saharan Africa. The 22-member expert panel included anesthetists, nurses, surgeons, pharmacists, pediatricians, a physiotherapist and a patient-caregiver dyad with a lived experience of pain. After completing three rounds of surveys, a steering committee of five members was assembled to resolve outstanding items to achieve the final curriculum. The process yielded a curriculum containing 20 knowledge items and 23 skills items. Attitudes are a key component of the curriculum and were grouped into six themes. A further aspect of the process was the identification of foundational knowledge with which trainees should enter a fellowship training program. This was termed the foundational curriculum. Using a Delphi method, consensus has been achieved on a pediatric pain curriculum for pediatric anesthesia fellows in sub-Saharan Africa with potential to meet the identified need for transformative pain care in this patient population.
This study aimed to develop polylactic-co-glycolic acid (PLGA) microspheres encapsulating dezocine and evaluate their efficacy in managing postoperative pain and mitigating cognitive impairment. Dezocine-loaded PLGA microspheres (DEZ@PLGA MS) were fabricated using a single-emulsion solvent evaporation technique and were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and in vitro release studies. A rat model of open reduction and internal fixation for right hindlimb tibial plateau fracture was established in healthy male rats. Following modeling, rats were randomly divided into four groups (n = 6 per group) and received intramuscular injections of equal volumes of normal saline, blank PLGA microsphere suspension, dezocine injection (5 mg/kg), or DEZ@PLGA MS suspension (5 mg/kg). Pain thresholds of the left hind paw and cognitive behavioral tests were assessed at specified time points post-administration. After testing, rats were euthanized. Muscle tissue at the injection site was collected for hematoxylin and eosin (H&E) staining to assess inflammatory responses, and hippocampal tissue was harvested for Western blot analysis. The SEM results revealed spherical microspheres with a smooth surface and uniform particle size. FTIR and DSC analyses confirmed stable encapsulation of dezocine (DEZ) in the PLGA matrix, with potential intermolecular interactions and amorphous drug dispersion. In vitro release studies demonstrated a sustained release profile of DEZ from the microspheres. Animal experimental results indicated that the DEZ@PLGA MS group exhibited a significantly prolonged duration of effective analgesia compared to the dezocine injection group. Additionally, DEZ@PLGA MS improved cognitive function, as evidenced by cognitive behavioral tests and Western blot results. H&E staining confirmed no significant inflammatory responses in muscle tissue across all groups, highlighting good biocompatibility of the formulation. Our study demonstrated that DEZ@PLGA MS effectively manages acute postoperative pain over an extended duration and improves postoperative cognitive function.
Atherosclerosis (AS) is a chronic inflammatory vascular disorder driven by endothelial dysfunction, oxidative stress, lipid dysregulation, mitochondrial injury, and maladaptive immune activation. Metformin, the first-line therapy for type 2 diabetes, has vasculoprotective effects beyond glycemic control. This review summarizes current evidence on metformin as a multi-target modulator of AS through regulation of nuclear erythroid 2-related factor 2 (Nrf2)/Krüppel-like factor 2 (KLF2) signaling, AMP-activated protein kinase (AMPK)/sirtuin-1 pathways, cyclic GMP-AMP synthase and stimulator of interferon genes (cGAS-STING) signaling, mitochondrial homeostasis, lipid metabolism, and inflammation. Relevant studies were identified through PubMed, Web of Science, Google Scholar, and China National Knowledge Infrastructure databases, focusing on the effects of metformin on atherosclerosis and related pathways involving oxidative stress, mitochondrial function, ferroptosis, inflammation, and innate immunity. Metformin enhances antioxidant defenses by activating Nrf2 through Kelch-like ECH-associated protein 1 degradation and increasing cytoprotective mediators such as heme oxygenase-1 and NAD(P)H quinone oxidoreductase 1. It also modulates cGAS-STING signaling and promotes AMPK-mediated KLF2 activation, thereby improving endothelial quiescence, increasing endothelial nitric oxide synthase-derived nitric oxide, and suppressing inflammatory signaling and adhesion molecules. Furthermore, metformin protects against ferroptosis by stabilizing mitochondrial function, reducing lipid peroxidation, and limiting iron-dependent oxidative injury. It also exerts systemic anti-atherogenic effects by modulating gut microbiota, lowering trimethylamine-N-oxide, and increasing short-chain fatty acids and glucagon-like peptide-1. Metformin attenuates plaque formation, inflammation, expansion of the necrotic core, and instability.
BACKGROUND: The morbidity and mortality related to wound infections and microbial resistance are real public health concern in low-income settings where data are lacking, empirical antibiotic use is common and microbiological diagnostics are limited. This study aimed to describe the prevalence and antimicrobial resistance (AMR) patterns resulting from wound infections. METHODS: We conducted a one-year multicenter cross-sectional study across hospitals in Goma, Democratic Republic of the Congo. Consecutive patients with clinically infected wounds were enrolled. Wound swabs were processed using standard culture and susceptibility testing. Firth’s bias-reduced logistic regression was used to assess factors associated with surgical site infection (SSI) and AMR, with analyses stratified by Gram staining. RESULTS: Most patients were below 40 years old, with a median age of 27 years (IQR: 22–39), predominantly males (64.5%). Gunshot injuries (33.2%), road traffic accidents (24.5%) and cesarean section  (14.8%) were the leading cause of wounds. SSIs accounted for 21.25% of all clinically diagnosed infected wounds; they were significantly associated with female sex (OR = 3.65, 95% CI: 1.18–11.92, p = 0.03) and abdominal surgery (OR = 272.92, 95% CI: 85.66–1208.58, p < 0.001). In female patients, a high rate of SSIs was observed following cesarean section  (86.8%). Overall microbial swab cultures were negative in 21.0%. Among 18 isolated species, Gram-negative bacteria were predominant with Proteus mirabilis (24.1%), Pseudomonas aeruginosa (13.6%), and Escherichia coli (12.7%), as the leading pathogens. In Gram-stratified Firth models, empirical antibiotic therapy was not independently associated with increased in vitro AMR. Amikacin showed a protective association among Gram-negative isolates, while estimates for Gram-positive cocci were imprecise due to small sample size. Pseudomonas aeruginosa exhibited reduced susceptibility to multiple drug classes. CONCLUSION: Wound infections in Goma are largely linked to trauma and cesarean deliveries, with Gram-negative microorganisms dominating. The microbial ecology comprises ESKAPE pathogens and species known for their emerging resistance patterns such as Proteus mirabilis, Pseudomonas aeruginosa and Escherichia coli. Empirical antibiotic therapy was not independently associated with AMR after adjustment. These findings underscore the urgent need for enhanced diagnostic strategies, strengthened infection prevention measures, and effective antimicrobial stewardship in this resource-limited and conflict-affected setting.
Geraniol (GI), an acyclic monoterpene alcohol, exhibits diverse anti-cancer activities. However, its potential effects against gastric cancer remain poorly understood. The present study aimed to explore whether GI promotes apoptosis in gastric cancer cells, and decipher the possible mechanism underlying this effect. The cell viability and cell cycle distribution of SGC-7901 cells and MKN45 cells were evaluated using MTT assay, MB assay, and flow cytometry. The expression of Bax, Bcl-2, and glycogen synthase kinase-3 (GSK-3β) proteins, the mitochondrial membrane potential (MMP), and cell apoptosis in SGC-7901 cells were determined using Western blotting, JC-1 staining, immunofluorescence analysis, and Annexin V/propidium iodide double staining. In addition, cell apoptosis and the expression of proliferating cell nuclear antigen, Cleaved-caspase-3, Bax, Bcl-2, and GSK-3β proteins in the xenografts were determined using terminal deoxynucleotidyl transferase biotin-dUTP nick-end labeling, immunohistochemistry, and Western blotting. GI significantly inhibited the viability of gastric cancer cells and arrested the cell cycle at the G0/G1 phase in a dose-dependent manner. GI also promoted apoptosis and decreased the MMP in gastric cancer cells. Moreover, GI significantly upregulated the Bax/Bcl-2 ratio and downregulated the expression of pGSK-3β and β-catenin both in vitro and in vivo, while increasing the expression of Cleaved-caspase-3 in SGC-7901 cell xenografts. Furthermore, GI reversed the anti-apoptotic effect of the GSK-3β inhibitor-LiCl, confirming its pro-apoptotic role. GI suppresses gastric cancer progression both in vitro and in vivo, by inducing apoptosis through inhibition of the Wnt/β-catenin pathway.
Cerebral ischemia-reperfusion injury (CIRI) presents a major therapeutic challenge in stroke management, where oxidative stress and neuroinflammation synergistically exacerbate neuronal damage. This study examined whether exogenous hydrogen sulphide (H₂S) protects against CIRI by simultaneously modulating the nuclear erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) antioxidant response and inhibiting nuclear factor kappa-B (NF-κB) and Nod-like receptor (NLR) family, pyrin domain-containing 3 (NLRP3) inflammasome engagement in mice. Animal models were subjected to middle cerebral artery occlusion and reperfusion (MCAO/R) in mice, and neurological deficits were subsequently assessed using a rating scale and 2,3,5-triphenyltetrazolium chloride (TTC) staining to evaluate MCAO/R damage. The expression level of reactive oxygen species (ROS) was detected using an assay kit. The levels of Nrf2/HO-1 pathway and NLRP3 inflammasome were examined by Western blot and immunohistochemical staining. The experimental data showed that exogenous administration of H₂S was effective in improving behavioral disorders and reducing infarct volume. Treatment with exogenous H₂S regulated the expression of the Nrf2/HO-1 signalling pathway, while H₂S effectively inhibited the expression of NF-κB and the formation of the NLRP3 inflammasome, as evidenced by reduced levels of phosphorylated NF-κB subunit p65 (p-p65), NLRP3, apoptosis-associated speck-like protein (ASC), and activated Caspase-1, and led to decreased maturation of the proinflammatory cytokines interleukin (IL)-1β and IL-18. This study demonstrated that exogenous administration of H₂S exerted neuroprotective effects by regulating the Nrf2/HO-1 antioxidant pathway and inhibiting NLRP3 inflammasome activation.
Palbociclib (PBB) is an oral cyclin-dependent kinase 4/6 (CDK4/6) inhibitor approved for the treatment of HR+/HER2- breast cancer. However, poor adherence and limited tolerability of oral administration often compromise its therapeutic effectiveness, especially in palliative care for metastatic conditions. Dose reductions are frequently required to manage toxicity, but lower doses can still provide effective tumour control with reduced neutropenia risk, thereby improving quality of life and progression-free survival. Developing a long-acting injectable (LAI) formulation of PBB offers significant advantages for sustained therapy in advanced-stage cancer management. This study focuses on dose selection, release profile optimisation, and the design of sterile PBB-loaded PLGA microsphere suspensions for intramuscular (IM) administration using physiologically based pharmacokinetic (PBPK) modelling and simulations. The PBPK model, developed and validated with data from oral and intravenous routes, enabled the prediction of IM pharmacokinetics. Clinical target product profiles were defined based on IC50 and minimum steady-state concentration (Css, min). The dose optimisation study revealed that the rational selection of dose for both strengths, with an optimised sustained-release profile (Target 2, showing minimal initial burst and controlled release reaching ~55% by day 10.5 and ~90% by day 21), achieved the desired clinical quality target product profiles. The developed model will further support polymer selection, specification setting, and drug-to-polymer ratio. Incorporating PBB's physicochemical properties and host response helps guide rational formulation design.
To elucidate the therapeutic mechanism of Qufengzhitong Pills (QFZTP) in the treatment of rheumatoid arthritis (RA). Blood-absorbed active components of QFZTP and its potential therapeutic targets for RA were screened via liquid chromatography-tandem mass spectrometry (LC-MS/MS) and network pharmacology. Core targets were analyzed using protein-protein interaction networks, with key pathways identified via Gene Ontology/Kyoto Encyclopedia of Genes and Genomes enrichment. Molecular docking was performed for major components and core targets. An IL-1β-induced MH7A cell-based RA model was established; CCK-8, ELISA (IL-6/TNF-α), wound-healing/Transwell, flow cytometry, EdU, and western blot (PI3K/AKT/NF-κB/MAPK pathways) were used to assess cell viability, cytokines, migration, apoptosis, proliferation, and protein expression. LC-MS/MS identified 92 active components in rat serum, and network pharmacology identified 369 potential QFZTP targets for RA, enriched mainly in TNF signaling, PI3K/AKT, MAPK, and apoptosis pathways. Core components (crocetin, epigallocatechin, bergapten) showed stable binding (<-5.0 kcal/mol) to AKT1, EGFR, and PI3K, occupying similar active pockets as native inhibitors. In vitro, QFZTP-containing serum significantly reduced IL-1β-induced MH7A cell viability; inhibited IL-6/TNF-α secretion (comparable to native QFZTP solution); attenuated migration/proliferation; induced apoptosis; downregulated EGFR, p-PI3K, p-AKT, p-p38, p-NF-κB, and Bcl-2; and up-regulated Bax. QFZTP synergistically regulates the PI3K/AKT, NF-κB, and MAPK pathways to inhibit synovial cell proliferation and migration, induce apoptosis, and attenuate inflammation. This study provides a robust theoretical and experimental basis for its clinical application in RA.
To match the disintegration time of conventional oral films with 3D printed fast-dissolving oral films (FDFs), micro-composites have been used in the formulation. However, in certain cases the 3D-printer failed to produce desired films. The viscoelastic properties were evaluated for polyvinyl alcohol and polyvinylpyrrolidone filaments containing chitosan micro-ribbons and cellulose microfibres as micro-composites with the hypothesis that intermittent nozzle blockage was the mechanism responsible for the observed printing failures. Domination of loss modulus over storage modulus was observed for successful printing. Micro-composites improved the viscoelastic properties of filaments including filaments that failed to print. The novelty of this research was that poor viscoelastic properties could not be accounted for the failure of FDF 3D printing for formulations with high micro-composite contents. Filaments of these formulations exhibited rough surfaces with visible aggregates. These observations suggested intermittent nozzle blockage by aggregated micro-composites could have been the cause of 3D printing failure. This hypothesis was supported by successful printing when printer nozzle diameter increased. The domination of loss modulus over storage modulus was essential for filaments to achieve successful FDF 3D printing. However, micro-composites at high concentrations in the formulation may induced nozzle blockage leading to printing failures.
Hepatotoxicity associated with Isoniazid (INH) and Rifampicin (RIF) is one of the major impediments in antituberculosis therapy. We aimed to investigate the protective effects of the cornel iridoid glycoside (CIG) on liver injury caused by INH and RIF in vivo and in vitro. Liver injury model was induced by INH and RIF (150 mg/kg). Then the mice were orally administered with CIG (50, 100, and 200 mg/kg) and silymarin (100 mg/kg) for 2 weeks. HepG2 cells were treated with 0.1 mg/ml INH and 0.2 mg/ml RIF to establish injury model. Blood samples and cell lysis were used for biochemical analysis and western blot assay. Liver tissues were harvested for histological examination. Treatment of CIG significantly restored liver function, reduced the levels of inflammatory factors, enhanced the antioxidant activity, and alleviated lipid metabolism disorder compared with the model group in vivo. CIG restored cell viability and activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in vitro. Furthermore, CIG inhibited the protein expression of JAK/STAT pathway. CIG is effective in preventing INH and RIF-induced hepatotoxicity in vivo and in vitro. These findings provide scientific basis for the clinical application of cornel in treating liver injury.
This study investigates Panax notoginseng saponins (PNS) as a potential treatment for gp120-induced HIV-associated neurocognitive disorder (HAND), in light of the limited efficacy of current therapies. A rat model of gp120-induced HAND was established. Cognitive function was evaluated using the Morris water maze and novel object recognition tests. Hippocampal metabolomics and network pharmacology were integrated to identify critical pathways and targets. The expression of core targets was validated by enzyme-linked immunosorbent assay (ELISA). PNS administration significantly mitigated gp120-induced cognitive deficits, demonstrated by shortened escape latency and an increased number of platform crossings in the Morris water maze. Network pharmacology analysis pinpointed the NOD-like receptor signaling pathway as the principal target, a finding corroborated by metabolomics data. ELISA confirmed that PNS decreased hippocampal levels of the pro-inflammatory cytokines IL-1β, IL-6, and CCL2 while upregulating the anti-apoptotic protein Bcl2. PNS confers neuroprotection against gp120-induced cognitive impairment by modulating the NOD-like receptor signaling pathway, which suppresses neuroinflammation and inhibits apoptosis.
The aim of this study was to evaluate the antiallodynic and antihyperalgesic effects of rutin alone and in combination with pregabalin in a chronic constriction injury-induced neuropathic pain model. The potential antiallodynic and antihyperalgesic effects of combining rutin at doses of 25, 50, and 100 mg/kg with 3 mg/kg of pregabalin were evaluated in a rat neuropathic pain model induced by chronic constriction injury. Assessments were conducted over a 180-minute period using an electronic von Frey device and the plantar test. Spontaneous locomotor activity was evaluated using a Plexiglas activity cage. All combination groups exhibited statistically significant antiallodynic and antihyperalgesic effects at specific time intervals compared to the groups where rutin or 3 mg/kg pregabalin were administered alone. Furthermore, the efficacy of the combinations was found to be comparable to that of the group treated with 30 mg/kg pregabalin. The 25 mg/kg rutin combination was most effective and independent of locomotor activity. Given rutin's low side-effect profile and the dose-dependent risks of pregabalin, their combination at low-dose pregabalin may represent a safe and effective alternative for neuropathic pain treatment.
Staphylococcus epidermidis is a public health concern due to its antimicrobial resistance, causing infections that can worsen in immunosuppressed patients. Geopropolis (GEO) is produced by Meliponines and contains anacardic acid (AA). This study evaluated the antibacterial action of GEO and AA, and their modulatory effects on macrophages, both with and without dexamethasone (DEX) treatment. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of GEO, AA, ceftaroline (CPT), and gentamicin (GEN) were evaluated. The effect of the treatments was evaluated on the bacteria's ability to produce biofilm and on THP-1 cells differentiated into macrophages with or without DEX, evaluating cell viability, TLR-2 expression, eicosanoids (PGE2 and LTB4), cytokine (TNF-α, IL-1β, IL-6, and IL-10), and the bactericidal activity against S. epidermidis. GEO exhibited no antibacterial activity. AA had a more efficient MIC and MBC than the antimicrobials and inhibited biofilm formation. AA did not affect TLR-2 expression. It increased the bactericidal capacity of macrophages suppressed or not by DEX, and inhibited IL-10 production. Our findings suggest AA practical applications, as antibacterial coatings, wound care formulations, and infection control. AA has antibacterial and immunomodulatory properties and could be an effective adjuvant in treating S. epidermidis infection.
This study investigated α-phellandrene (PE)'s therapeutic effects on hypoxia-induced pulmonary hypertension (HPH), with emphasis on pulmonary vasoconstriction and vascular remodeling. A rat model of HPH was successfully established and then the hemodynamic indexes were assessed. HE and Masson's staining were performed to observe tissue morphological changes and fibrosis. Immunohistochemistry and immunofluorescence were used to quantify Collagen I/III, and alpha-smooth muscle actin (α-SMA). malondialdehyde, glutathione, superoxide dismutase, and glutathione peroxidase assays for antioxidant status. Western blotting to analyse the protein expression levels in lung tissue of HPH rats. Our results indicate that PE significantly mitigated HPH, as evidenced by reductions in right ventricular (RV) systolic pressure, RV hypertrophy (evaluated via the RV/BW ratio and Fulton index), and key structural changes. PE effectively diminished pulmonary vascular remodeling, demonstrated by decreased vascular wall thickness and area, along with downregulation of Collagen I/III and α-SMA expression. Mechanistically, the protective effects of PE were associated with modulation of the AKT/eNOS/sGC/PKG pathway, a critical regulator of vascular tone and remodeling, as well as a reduction in oxidative stress and apoptosis. These findings highlight that PE alleviates HPH through a multifaceted approach targeting vasoconstriction and vascular remodeling, underscoring its potential as a novel therapeutic agent.
The present study explored whether allyl isothiocyanate from the traditional Chinese medicine mustard seed, ameliorated the arthritis by modulating immune cell responses, particularly by inhibiting mast cell activation and the Pim1 pathway. Micro-CT, hematoxylin and eosin staining, and flow cytometry were performed to evaluate pathological changes in collagen-induced arthritis mice. To investigate the mechanism underlying allyl isothiocyanate-mediated mast cells activation, degranulation, intracelluar calcium measurement, RNA sequencing, real-time polymerase chain reaction and western blotting were carried out. Allyl isothiocyanate reduced the number of mast cells in the ankle joints of collagen-induced arthritis mice, and it inhibited the degranulation of mouse bone marrow-derived mast cells and RBL-2H3. RNA sequencing revealed that allyl isothiocyanate significantly inhibited Pim1 expression. Additionally, it inhibited the expression of Pim1, p-STAT5, p-AKT and c-myc in RBL-2H3 and RAW264.7 cells and the ankle joints of collagen-induced arthritis mice. Adoptive transfer of mouse bone marrow-derived mast cells early in collagen-induced arthritis exacerbated joint inflammation, which was similarly inhibited by allyl isothiocyanate. These findings support mast cell as a major cellular target of AITC in CIA and suggest that Pim1-related signaling contributes to AITC-mediated suppression of mast cell activation.
This study aimed to elucidate potential mechanisms associated with the cardioprotective efficacy of methanolic Delonix regia extract against the pathophysiological diabetic cardiac remodeling. Phytochemical analysis of the D. regia extract was conducted utilizing liquid chromatography-tandem mass spectrometry. Diabetes was induced using a high fructose/salt/fat diet, with a concurrent low dose streptozotocin. The extract was administered orally during the final six weeks. The analysis identified fifty-nine phytoconstituents within the floral extract. Functionally, the extract was associated with improvements in diabetes-induced alterations in cardiac performance and normalization of electrocardiographic indicators of ventricular activity. Structurally, the extract was associated with attenuation of cardiac remodeling, including reduced cardiomyocyte hypertrophy, improvement of diabetes-induced structural distortions, and decreased fibrotic deposition. Additionally, the extract was associated with modulation of apoptotic markers and increased levels of endothelial nitric oxide synthase (eNOS), tetrahydrobiopterin, cyclic guanosine monophosphate, and glutathione. Molecular docking suggested that these effects may be attributable to the combined actions of phenolic and flavonoid constituents. Methanolic extract of D. regia may have therapeutic potential in the attenuating left ventricular remodeling associated with diabetic cardiomyopathy. These effects appear to be associated with modulation of eNOS/NO/cGMP and NF-κB/iNOS/TNF-α trajectories, along with anti-apoptotic and antifibrotic actions.
The concept of pharmacokinetic volume of distribution (Vd) was first described > 90 years ago. Vd can provide a quantitative estimate of drug distribution to a hypothetical 'peripheral' space outside the blood or plasma (the measurement site), assuming concentration equality between blood and the peripheral space. After single intravenous doses, calculation of Vd by the area method [Vd(area), also termed Vz or Vß] is the most stable and valid approach, since Vd(area) reflects drug distribution at all times after distribution equilibrium is attained, and depends only on observed properties of drug behaviour. In contrast, Vd by the steady-state method [Vd(ss)] is applicable only at a single time point, underestimates the actual extent of drug distribution, and is dependent on 'invisible' hypothetical rate constants. The notion that 'Vd(area) depends on elimination' is incorrect. Rather, Vd(area) and clearance-variables independent of each other-separately determine elimination half-life (T½), which is a dependent variable. T½ is an important property of drugs used in clinical practice, and can be altered-via changes in Vd(area) without changes in clearance-by factors such as obesity, age, and gender. Vd (area) is the most appropriate metric reflecting in vivo drug distribution.
The application of Quality-by-Design approaches in the development of analytical methods has changed the way drugs are manufactured, switching from trial-and-error and one-variable-at-a-time methods to a structured, risk-based scientific framework. The Analytical Target Profile is at the heart of Quality-by-Design. It clearly lays out the criteria for how well the method will perform. The identification of Critical Quality Attributes and Critical Method Parameters through systematic risk assessment tools like the Ishikawa diagram and Failure Mode and Effect Analysis supports this. Statistical methods, particularly Design of Experiments, have been crucial for identifying and optimizing key variables. This leads to the development of a Method Operable Design Region (MODR). The MODR sets the limits for the analytical method's reliable results, which means that changes can be made after approval without having to go through the whole process again. Box-Behnken and Central Composite are two common designs that are used to determine how various factors interact in order to ensure that methods perform effectively. Quality-by-Design-based control strategies combine lifecycle management and real-time monitoring to make sure that quality continues to improve more effectively. Literature screening and data organization were performed using Microsoft Excel (Microsoft Corporation, Redmond, WA, USA). Reference management and duplicate removal were carried out using EndNote (Clarivate Analytics). Database searches were conducted across PubMed, Web of Science, Elsevier, and Google Scholar using predefined keywords.