搜索结果：Iranian journal of pharmaceutical research : IJPR

In the biopharmaceutical sector, characterized by rapid technological change and intense market competition, external knowledge search (EKS) has become a strategic necessity to overcome the limitations of closed innovation. However, while EKS is widely studied in developed economies, its underlying mechanisms remain underexplored in emerging biopharmaceutical sectors. This cross-sectional study examines how EKS affects innovation performance (IP) in Iranian biopharmaceutical firms, testing the mediating role of knowledge integration capability (KIC) and the moderating effect of R&D intensity, while controlling for firm size and age. Data were collected using a validated questionnaire completed by 92 senior executives (CEOs, R&D, and Business Development managers) from 44 Iranian biopharmaceutical companies. The model was analyzed using partial least squares structural equation modeling (PLS-SEM) in SmartPLS. EKS significantly enhances both IP (&#x3b2; = 0.462, P < 0.001) and KIC (&#x3b2; = 0.239, P < 0.003). KIC positively influences IP (&#x3b2; = 0.437, P < 0.001) and partially mediates the EKS-IP relationship (indirect effect &#x3b2; = 0.104, P = 0.02). R&D intensity showed no significant moderating effect (&#x3b2; = 0.198, P = 0.10). The model explains 40.5% of IP variance (R&#xb2; = 0.405). Firm size (&#x3b2; = 0.009, P = 0.924) and age (&#x3b2; = 0.030, P = 0.792) had no significant effects. The impact of EKS on IP depends more on a firm's KIC than on its R&D intensity. This study advances open innovation theory by clarifying how external knowledge is effectively translated into innovation outcomes within an emerging biopharmaceutical context.

The primary objective of this study was to compare the total and insurance-covered utilization of pharmaceuticals in Iran from March 2018 to March 2023. The findings of this study can offer valuable insights to healthcare professionals, researchers, and policymakers to make informed decisions. A retrospective cross-sectional study in Iran examined drug utilization patterns (specific databases, e.g., Iranian Health Insurance Organization and Social Security Organization) using Joinpoint regression. The study calculated the Defined Daily Doses per 1000 inhabitants per day (DID) for each medicine based on the anatomical therapeutic chemical/defined daily dose (ATC/DDD) system. All medicines with valid ATC/DDD assignments available during 2018 - 2023 were included to ensure comprehensive national coverage rather than a selected sample. Joinpoint regression was employed for trend analysis, focusing on annual and monthly percent changes in pharmaceutical utilization. Statistical analysis was conducted using MS Excel and Joinpoint software, with significance set at P-value < 0.05. The study assessed 1185 ATC codes, out of which 751 were found eligible for analysis. Among these, 728 were covered by two major Iranian insurance organizations. The study found that the share of insurance utilization of these pharmaceuticals ranged from 27 to 48 percent. The trend utilization showed significant increases in overall utilization in classes C, J, L, N, and P. The utilization trends under insurance coverage revealed that classes L, R, and S showed a significant increase. After the implementation of the Daruyar Plan and the removal of the preferred currency exchange rate for all classes, there has been an increase in utilization under insurance coverage. This study reveals notable patterns and discrepancies in utilization levels within different drug classes, as well as the impact of insurance coverage and the Daruyar Plan on medication utilization.

Current clinical treatment is confronted with challenges such as low drug delivery efficiency, lagging efficacy evaluation, and a high tumor recurrence rate. However, chemical exchange saturation transfer (CEST) imaging, with its molecular specificity and non-invasive monitoring advantages, provides a new path for real-time tracking of the metabolokinetics of nanoprobes. Drug-loaded nanoprobes that self-assembled and had CEST activation imaging capabilities were created, and there in vivo and in vitro CEST activation imaging efficacy and the value of photodynamic sensitization for pyroptosis treatment of prostate cancer cells were evaluated. A self-assembly strategy was adopted to construct a nanoprobe (GC) coloaded with gemcitabine (Gem) and the photosensitizer chlorin e6 (Ce6), and a scanning electron microscope was used. Scanning electron microscope (SEM), dynamic light scattering (DLS), etc., were used to characterize the in vitro CEST activation imaging efficacy of the nanoprobe, and the pH, concentration-, and time-dependent drug release were observed. GC was combined with 2',7'-dichlorodihydrogen fluorescein diacetate ester (2',7'-dichlorodihydrofluorescein diacetate) after laser treatment of prostate cancer cells in mice. The generation of reactive oxygen species (ROS) was detected with a DCFH-DA probe. By using ELISA, the amounts of inflammatory factors such as interleukin-1&#x3b2; (IL-1&#x3b2;) and IL-18 were determined, and calreticulin was detected via immunofluorescence. CRT and high mobility group box 1 protein (HMGB1) were used to evaluate pyroptosis-mediated immunogenic cell death (ICD) effects. A mouse prostate cancer tumor model was constructed to observe the CEST-specific activation effect of the GC nanoprobes in vivo, and its tumor volume was measured, together with the detection of inflammatory substances and ICD markers, in order to assess its pyroptosis-based anticancer activity. Scanning electron microscope and DLS analyses revealed that the GC nanoprobes had a uniform spherical-like structure. The release of the drug Gem by this drug-loaded nanoprobe under acidic conditions at pH 5.0 was as high as 80%, which was significantly greater than that at pH 7.4 (P = 0.003). DCFH-DA fluorescence staining indicated that photosensitizing pyroptosis mediated by nanoprobes could generate a large amount of ROS. The detection of pyroptosis-related factors revealed that the levels of IL-1&#x3b2; and IL-18 significantly increased (all P < 0.05), whereas the fluorescence of the ICD marker CRT increased and that of HMGB1 decreased. The results of in vivo experiments indicated that the CEST signal at the tumor site was significantly enhanced and reached its peak 4 hours after tail vein injection of the GC nanoprobes. In addition, the results of antitumor treatment in vivo revealed that, compared with those in the control group treated with PBS, the levels of the inflammatory factors IL-1&#x3b2; and IL-18 in the experimental group treated with GC combined with laser irradiation increased, the ICD markers HMGB1 and CRT significantly changed, and the tumors were significantly inhibited (all P < 0.05). The CEST imaging of GC nanoprobes demonstrates excellent diagnostic accuracy in the detection of prostate cancer. The sensitivity reaches 92.3%, the specificity is 88.7%, and the area under the curve (AUC) is 0.95, which is significantly superior to traditional imaging methods. The specific enhancement of CEST signals at the tumor site reaches its peak 4 hours after injection, which is highly consistent with the histopathological results, confirming its clinical value in the early diagnosis and precise treatment guidance of prostate cancer. The GC nanoprobes successfully prepared in this study can specifically activate CEST imaging, guide the photodynamic sensitization of prostate cancer tumor cells to pyroptosis, and promote the precise ablation of prostate cancer.

Sustainable performance in the pharmaceutical industry hinges on the strategic alignment of human resources (HR), marketing, and information technology (IT). Prior studies often examined these domains separately; evidence on their joint influence in Iran's pharmaceutical sector remains limited. To assess how HR, marketing, and IT strategic alignment relate to profitability, liquidity, and revenue growth using machine-learning methods, and to document model generalization and measurement validity. This applied, cross-sectional study surveyed 323 managers in Tehran Stock Exchange (TSE)-listed pharmaceutical firms (May to Nov, 2024). A validated questionnaire [CVI/CVR; EFA/ confirmatory factor analysis (CFA); reliability reported] was used only to construct composite indices of HR, marketing, and IT alignment; organizational performance outcomes, profitability, liquidity, and revenue growth (year-over-year) were computed from audited financial statements and then z-standardized. Inputs were min-max scaled to [0, 1]. A feed-forward artificial neural network (ANN; 3-15-1 per outcome; ReLU hidden, linear output) was trained with Levenberg-Marquardt, early stopping, and L2 regularization. Data were split 70/15/15 (train/validation/test) with 5 &#xd7; 10 repeated cross-validation; bootstrap resampling (B = 1000) produced BCa 95% CIs. Model performance was assessed using mean squared error (MSE), mean absolute error (MAE), root mean square error (RMSE), and R2. Aggregate fit was strong (R2 = 0.91; RMSE = 0.134), with comparable validation/test metrics indicating good generalization. The triadic alignment factor showed the highest association with overall strategic alignment (R2 = 0.76; P < 0.001). At the subcomponent level, organizational commitment related to profitability (R2 = 0.59), and aggressive marketing to profitability (R2 = 0.66). Results are associative, not causal. Machine-learning evidence suggests that coordinated alignment across HR, marketing, and IT is strongly associated with key performance components. The validated instrument, explicit splits, cross-validation, and bootstrap CIs enhance robustness and provide a practical, data-driven framework for managerial action in Iran's pharmaceutical industry.

Colorectal cancer is a common malignant tumor of the digestive tract, with a high incidence and mortality rate. Ki67 and vascular endothelial growth factor (VEGF) play important roles in tumor cell proliferation and angiogenesis. Vitamin P is a natural flavonoid compound with various biological activities. To explore the effects of vitamin P on the proliferation, angiogenesis, and apoptosis of colorectal cancer cells and its underlying molecular mechanisms. In vitro cultivation of human colorectal cancer cell lines (HCT116 and SW480) was performed. Vitamin P was added to the cells in varying quantities. Cell proliferation was identified using the CCK-8 technique. The flow cytometry method was used to determine the proportion of apoptotic cells. Real-time fluorescence quantitative PCR and Western blot methods were used to measure the expression levels of the VEGF and Ki67 genes and proteins. Secretion of VEGF was observed via immunofluorescence staining. Vitamin P treatment significantly inhibited the proliferation of colorectal cancer cells in a dose-dependent manner. The half-maximal inhibitory concentrations (ICs50) at 48 hours were 72.3 &#x3bc;M (HCT116) and 85.6 &#x3bc;M (SW480). Moreover, in the vitamin P treatment group (42.7 - 68.9% and 35.4 - 61.2%), the secretion of VEGF decreased by 52.3 - 79.8%. The percentage of apoptotic cells induced by 20 - 80 &#x3bc;M vitamin P increased from 5.3% in the control group to 18.6 - 37.9%, with increased caspase-3 activity. In vivo experiments showed that vitamin P significantly inhibited the growth of colorectal cancer transplanted tumors and reduced the expression of Ki67 and VEGF in the tumors. Meanwhile, vitamin P treatment induced apoptosis of tumor cells, and this effect was closely related to the regulation of the Bax/Bcl-2 ratio and the promotion of caspase-3 activation. Vitamin P may play a regulatory role by inhibiting the phosphorylation of the STAT3 signaling pathway and downregulating the expression of proteins related to the PI3K/AKT pathway. Vitamin P can inhibit the malignant biological behavior of colorectal cancer cells by suppressing Ki67-mediated cell proliferation, blocking VEGF-related angiogenesis pathways, and activating mitochondrial apoptotic pathways.

Clerodendrum chinense has been reported to possess several important biological activities. However, the anti-inflammation and insulin-related properties of isolated compounds from this plant have not yet been investigated. This study focuses on extracting compounds and evaluating improvements in insulin resistance through the anti-inflammatory potential and inhibition of reactive oxygen species (ROS) of hispidulin from Vietnamese C. chinense leaves. Ethanol was used to separate the compounds from the leaves of Vietnamese C. chinense. 1D, 2D-NMR, and ESI-MS spectra were used to analyze the structural identity of the isolated compounds. Compound 1 (hispidulin) was evaluated for anti-inflammation and ROS in Raw 264.7 cells: Anti-inflammatory cytokine inhibition ability was analyzed by the ELISA method; Western blotting was used for the expression levels of phosphorylated ERK1/2, p38, p47phox, and GLUT4. ROS was assessed by immunofluorescence; a luminometer device was used to measure NADPH oxidase activity. The ability to absorb glucose and counteract insulin resistance was evaluated in 3T3-L1 cells via insulin-induced glucose uptake using immunofluorescence dye. One compound isolated from the leaves of Vietnamese C. chinense was identified as hispidulin. Hispidulin at a concentration of 50 &#x3bc;g/mL significantly inhibited tumor necrosis factor (TNF)-&#x3b1;, interleukin (IL)-6, and IL-8 in LPS-induced macrophages (P < 0.001). Moreover, in LPS-stimulated macrophages, the phosphorylation of p38 and ERK1/2, NADPH oxidase activity, as well as the phosphorylation of p47phox, were reduced at a concentration of 50 &#x3bc;g/mL of hispidulin (P < 0.001). More importantly, 3T3-L1 cells with inhibited insulin activity were able to regain glucose uptake and insulin responsiveness through GLUT4 expression when treated with 50 &#xb5;g/mL of hispidulin (P < 0.001). The data confirmed the presence of hispidulin in the chemical composition of Vietnamese C. chinense leaves. The results indicated that hispidulin inhibited LPS- and ROS-mediated inflammation in macrophages. More importantly, the data have shown that hispidulin enhances glucose uptake and improves insulin activity in 3T3-L1 cells.

Liver transplantation is frequently complicated by ischemia-reperfusion injury (IRI), which may impair hepatic, renal, and cardiac function. Volatile anesthetics such as isoflurane and sevoflurane are believed to mitigate this injury. This study aimed to compare their effects on short-term organ outcomes in deceased donor liver transplant recipients. In this study, 70 liver transplantation candidates at Taleghani Hospital in Tehran were enrolled after obtaining informed consent, and various variables were assessed before, during, and at two intervals immediately after surgery and one week post-operation. Patients were randomly allocated to receive either isoflurane or sevoflurane for anesthesia maintenance using the sealed opaque envelope technique for allocation concealment. Randomization was performed by a study nurse not involved in patient care using computer-generated random numbers. The primary outcome was defined as the postoperative peak serum alanine aminotransferase (ALT) level. Secondary outcomes included peak aspartate aminotransferase (AST), total bilirubin, creatinine, troponin I, C-reactive protein (CRP), intraoperative blood product requirements (packed red blood cells and fresh frozen plasma), hemodynamic parameters, and urine output. Baseline characteristics were comparable between groups. No significant differences were found in intraoperative hemodynamics or postoperative laboratory values of liver and renal function (P > 0.05). Postoperative liver enzyme levels increased in both groups following reperfusion, consistent with IRI. However, no statistically significant differences were observed between the isoflurane and sevoflurane groups in peak serum ALT levels, measured 6 hours after reperfusion and on postoperative day 7 (P > 0.05 for all comparisons). Similarly, AST levels did not differ significantly between groups at any postoperative time point. Renal and cardiac biomarkers, including creatinine and troponin I, were also comparable between groups. In contrast, patients receiving sevoflurane required significantly higher volumes of packed red blood cells and fresh frozen plasma intraoperatively compared with the isoflurane group (P < 0.05). The results obtained in this study showed that the use of isoflurane and sevoflurane did not have a significant difference in the severity of ischemic reperfusion injury caused after liver transplantation surgery on the liver, kidney, and heart; also, in this study, the functional conditions of these organs during and after surgery were evaluated, and by examining at different time intervals, these two inhalation anesthetics did not have a different effect on the short-term outcome of patients after receiving a liver transplant.

Dietary patterns are effective in obesity treatment. This has led to more investigations on its mechanisms in combating obesity. This study investigated the effects of ginseng and green tea extracts (GTE) on selected markers of lipid metabolism and the expression of some related genes in adipocytes. After a one-month period of consuming a high fatty content diet, a total of 42 male Wistar rats were assigned to seven groups randomly. The rats were then subjected to an eight-week treatment where they were administered different dosages of GTE and ginseng extract (GE) through oral administration. Then, some serologic parameters pertaining to lipid metabolism were analyzed in the treated rats. Furthermore, alterations in the expression levels of select genes, bone morphogenetic protein 7 (BMP7), hormone-sensitive lipase (HSL), and leptin, implicated in lipid metabolism, were quantified within the adipose tissue of the rats utilizing the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) methodology. Ultimately, the chemical composition of the extracts was analyzed by high performance liquid chromatography (HPLC). The findings indicated that the utilization of the extracts had a notable impact on the reduction of body weight. There was a noteworthy enhancement of high-density lipoprotein levels across all study groups, as indicated by a statistically significant increase at a confidence level of 95%. The efficacy of the administered extracts was observed in a significant upregulation in BMP7 and HSL gene expression. Conversely, there was a notable reduction in leptin expression, which reached statistical significance at a confidence level of 95%. HPLC results detected 9 ginsenosides in the GE, among which Rb1 (100 mAU) was present in the largest amount, and 9 alkaloids in the GTE, among which epicatechin (EC) (380 mAU) and caffeine (320 mAU) were present in the largest amount. The present study holds the potential to offer novel insights regarding the mechanism through which GE and GTE exert their anti-obesity effects.

Trimethyltin (TMT) is an organotin compound known to induce neurotoxicity within the limbic system of the brain, particularly in the hippocampal region, with neurodegenerative changes resembling those of Alzheimer's disease (AD). This investigation examined the impact of miRNA-220-3p (miR-220-3p) on TMT-induced neurotoxicity and associated behavioral abnormalities, such as spatial learning and memory impairments, and identified the potential molecular mechanisms. To induce neurotoxicity, TMT was injected (8 mg/kg, i.p. once), and after 1 hour, miR-220-3p was microinjected intraventricularly (ICV route, once) for the possible mitigation of TMT-induced neurotoxicity. Different behavioral assessments were employed to determine spatial learning and memory function. Moreover, hippocampal measurements of brain-derived neurotrophic factor (BDNF) and Sirtuin-1, oxidative stress-, apoptosis-, and neuroinflammation-related factors, and histochemical changes were performed. The TMT injection led to behavioral abnormalities in the novel object discrimination and Barnes maze tests, heightened oxidative stress [elevating reactive oxygen species (ROS) levels, nitrite, and lipid peroxidation and decreasing the activities of antioxidant enzymes, including catalase (CAT) and superoxide dismutase (SOD)], reduced BDNF and Sirtuin-1 levels, increased neuroinflammation [escalating the secretion of pro-inflammatory mediators, including tumor necrosis factor-&#x3b1; (TNF-&#x3b1;) and interleukin-6 (IL-6)], raised activities of acetylcholinesterase (AChE), myeloperoxidase (MPO), beta-secretase 1 (BACE-1), caspase-1, and caspase-3, accompanied by a reduced number of CA1 pyramidal neurons and higher glial fibrillary acidic protein (GFAP) immunoreactivity. In contrast, microinjection of miR-220-3p reversed most of these alterations. The findings of this investigation imply that miR-220-3p may mitigate TMT-induced neurotoxicity, which is attributed to the suppression of hippocampal oxidative stress, neuroinflammation, and caspase-dependent apoptosis and pyroptosis, and part of its beneficial effect is associated with the upregulation of BDNF and Sirtuin-1.

Global contamination of agricultural products with aflatoxin (AF) is one of the most important concerns in the field of food safety and quality. Aflatoxin, when entering the food chain, can cause oxidative stress and hepatotoxicity. Black soldier fly larvae (BSFL) are an environmentally friendly insect whose extract is rich in valuable bioactive compounds with antioxidant properties. This study aimed to investigate the protective effect of the n-hexane extract of BSFL on oxidative stress, inflammation, and histopathological changes caused by Aflatoxin B1 (AFB1)-induced hepatotoxicity in rats. Thirty-five male Wistar rats with a weight range of 200 to 250 g were randomly divided into 5 equal groups including Control, AFB1 (75 &#x3bc;g/kg), BSFL (360 mg/kg), BSFL 180 mg/kg+AFB1, and BSFL 360 mg/kg+AFB1. At the end of the treatment on the twenty-eighth day, the animals were euthanized and samples were taken for liver enzymes, oxidative stress, inflammation, endoplasmic reticulum (ER) stresses, apoptosis marker, histopathology, and expression of ER stress-related proteins analyses. Data were analyzed by one-way ANOVA followed by Tukey's post-hoc test, with P < 0.05 considered statistically significant. According to the results of the study, AFB1 administration significantly increased the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), nitric oxide (NO), malondialdehyde (MDA), tumor necrosis factor alpha (TNF-&#x3b1;), interleukin 1 beta (IL-1&#x3b2;), and interleukin 6 (IL-6) (P < 0.001) and also decreased the levels of superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GPx), and catalase (CAT) compared to the control group (P < 0.001). These biochemical and inflammatory abnormalities caused by AFB1 were confirmed by histopathological observations of liver tissue. Administration of BSFL extract (at the higher dose of 360 mg/kg) resulted in significant reversal of biochemical, inflammatory, and hepatic markers and modulated GRP78/ATF6/IRE1 signaling in AF-poisoned rats. Our histological results showed that BSFL extract (360 mg/kg) could reduce steatosis, cellular swelling, and lobular inflammation induced after AF exposure. The results of this study indicate that BSFL extract, as a valuable bioactive substance with antioxidant properties, may attenuate biochemical indices, oxidative stress, and inflammation caused by AF-induced hepatotoxicity.

Depression is one of the most common neuropsychiatric disorders worldwide. Lipopolysaccharide (LPS), a bacterial endotoxin, induces depressive-like behaviors by activating microglia and promoting the release of pro-inflammatory mediators, including nitric oxide, eicosanoids, and various cytokines. Owing to its rich content of flavonoids, phenolic compounds, and terpenoids, Ferula aucheri possesses potent antioxidant and anti-inflammatory properties, which may contribute to its potential as an alternative treatment for depression. This study aimed to evaluate the antidepressant-like effects of the hydroalcoholic extract of F. aucheri on LPS-induced depression in mice. After preparation of the hydroalcoholic extract of F. aucheri, 30 mice were randomly divided into five groups: (1) Control; (2) LPS (1 mg/kg, i.p.); (3) LPS+fluoxetine (20 mg/kg, i.p.); (4) LPS+F. aucheri extract (100 mg/kg, i.p.); and (5) LPS+F. aucheri extract (200 mg/kg, i.p.). Lipopolysaccharide was administered to induce depressive-like behaviors, and after 24 hours, behavioral assessments were conducted using the forced swim test (FST), tail suspension test (TST), and open field test (OFT). Subsequently, immunohistochemical analysis was performed to assess the expression of toll-like receptor 4 (TLR4) and NF-&#x3ba;B in the brain tissue. The FST and TST results revealed that treatment with F. aucheri extract significantly reduced immobility time compared to the LPS group, particularly at the 200 mg/kg dose, which showed superior efficacy even compared to fluoxetine. The OFT confirmed that the observed behavioral changes were not due to alterations in locomotor activity. Immunohistochemical analysis revealed that LPS significantly increased the expression of TLR4 and NF-&#x3ba;B in the brain. Notably, treatment with F. aucheri (200 mg/kg) significantly attenuated the expression of both biomarkers compared to the LPS group (#P < 0.05 and ##P < 0.01, respectively). The findings suggest that F. aucheri exhibits antidepressant-like effects in an LPS-induced model of depression, potentially mediated through modulation of neuroinflammatory pathways involving TLR4 and NF-&#x3ba;B. Given its promising preclinical efficacy and mechanistic relevance, F. aucheri could be considered an appropriate candidate for future clinical investigations as an antidepressant agent.

Increased costs of prescription drugs impose tremendous economic burdens on patients globally, which usually result in low drug adherence and poor health outcomes. In Iran, out-of-pocket (OOP) expenditures are severe, particularly among vulnerable groups such as the elderly. This analysis reviews OOP expenditures of 51 medications in Iran to provide useful insights regarding healthcare policy. To analyze OOP payments for medications in Iran during a seven-year period, revealing trends, financial needs, and policy requirements that increase financial protection and drug access. We included 51 medicines, meeting the WHO/HAI minimum sample size requirement for reliability (n = 50), consisting of the universal core list (n = 14) and a supplementary list based on local disease prevalence (n = 37). Following RECORD guidelines for observational studies, data were sourced from the two largest national health insurance funds [Social Security Organization (SSO), Iran Health Insurance Organization (IHIO)], covering the majority of the Iranian population, and pharmacy records from 2016 to 2022. OOP costs, insurance coverage, and affordability were analyzed using impoverishing health expenditure (IHE) and catastrophic health expenditure (CHE) metrics. To control for economic confounders, costs were adjusted using purchasing power parity (PPP). Joinpoint regression assessed annual OOP changes, with significance set at P-value < 0.05. Over the seven-year period, PPP-OOP payments showed a non-significant decreasing trend [average annual percent change (AAPC): -1.2% for SSO; -1.38% for IHIO]. Despite this, OOP expenditure remained high, averaging approximately 40% of total medication costs. Significant heterogeneity was observed; for instance, Spironolactone saw the highest cost increase (AAPC +8.29%), while Insulin Glargine showed the largest decrease (AAPC -12.59%). High OOP payments were observed for medications like dexamethasone and chlorpromazine, while insulin and clozapine also carried high costs under insurance. The variability in OOP expenses across medicines highlights differences in pricing and insurance policies. Catastrophic expenses were prominent for certain medications, such as enoxaparin, especially at higher expenditure thresholds based on the 40% capacity-to-pay metric. Limitations included the use of administrative claims data subject to potential operational errors and the restriction of the sample to 51 medicines, which may limit generalizability to the broader pharmaceutical market. This research reveals high OOP spending for Iranian patients, with considerable drug-specific heterogeneity. The implications suggest that policy action in terms of price reform and wider insurance coverage is needed to minimize financial burden and ensure access to medicines. Further studies are required to determine the impact of high OOP spending on patient adherence and to inform specific health policies in Iran and beyond.

Endophytic fungi, which inhabit internal plant tissues without causing apparent disease, have emerged as rich sources of bioactive secondary metabolites. Hypericum species are well-known medicinal plants with diverse pharmacological properties; however, the endophytic fungi associated with these species remain largely unexplored. This study aimed to isolate and molecularly identify endophytic fungi associated with Hypericum helianthemoides, H. scabrum, and H. perforatum, and to evaluate the antibacterial activity of their extracts against selected pathogenic bacteria. Endophytic fungi were isolated from the root and stem tissues of three Hypericum species. Molecular identification was performed by amplifying and sequencing the internal transcribed spacer (ITS) and large subunit (LSU) regions of the rRNA gene. The antibacterial activity of the fungal extracts was assessed by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values against Staphylococcus aureus, S. epidermidis, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli. A total of 20 fungal isolates were obtained, mainly belonging to the phylum Ascomycota, with one isolate (Absidia sp.) belonging to Zygomycota. H. perforatum harbored the highest number of isolates (40%). Most isolates were newly reported endophytes of Hypericum species. The dominant genera were Fusarium and Alternaria. More than half of the isolates exhibited antibacterial activity, particularly against Gram-positive bacteria, whereas only three species were active against Gram-negative strains. Cladosporium subglobosum demonstrated the most potent antibacterial effect, with MIC and MBC values of 3.125 &#x3bc;g/mL. This study highlights the diversity and antimicrobial potential of endophytic fungi associated with Hypericum species. These findings suggest that such endophytes may represent promising sources of novel bioactive metabolites with potential pharmaceutical applications.

Building on our previous investigation of the total semipolar extract (SPE) of Artemisia kopetdaghensis in Plasmodium berghei-infected mice, the present study focuses specifically on the methoxylated flavonoid fraction (MFF). Flavonoids are increasingly recognized for their ability to suppress parasite growth and modulate host immunity. To clarify their role, we isolated and characterized the major flavonoid constituents of this plant and evaluated their antimalarial potential, both alone and in combination with chloroquine, in a mouse model. Aerial parts of A. kopetdaghensis were extracted using a chloroform:acetone (2:1) solvent mixture. The extract was then fractionated by column chromatography. Based primarily on ^1H-NMR spectra, the MFF was selected and further purified by preparative HPLC. Isolated compounds were identified by 1D and 2D NMR and mass spectrometry. In the in vivo antimalarial study, thirty-six infected female Balb/c mice were treated with MFF and evaluated for key parameters. Docking and molecular interaction studies were conducted using AutoDock v4.2.6 software to examine the interactions of constituents with cytokine protein targets: 1D9C [interferon-gamma (IFN-&#x3b3;)], 1B6C (TGF-&#x3b2;), 1BBN [(interleukin-4 (IL-4)], and 4HR9 (IL-17), separately. Phytochemical analysis of the MFF by HPLC revealed three flavones: 6-methoxytricin (20%), cirsilineol (10%), and cirsimaritin (70%), identified for the first time in A. kopetdaghensis, with cirsimaritin as the dominant constituent. In vivo, MFF treatment significantly reduced parasitemia and enhanced parasite suppression in P. berghei-infected mice. Cytokine profiling demonstrated suppression of TGF-&#x3b2; and IL-4 followed by their recovery, together with marked elevations of IFN-&#x3b3; and IL-17, indicating balanced modulation between pro-inflammatory and regulatory responses. These immunological findings were corroborated by molecular docking analyses, which confirmed binding interactions of cirsimaritin with cytokine receptor targets, providing mechanistic support for its immunomodulatory activity. This work extends our previous study on the SPE by moving from extract-level observations to constituent-specific insights. By combining phytochemical isolation with in silico receptor interaction analysis, we demonstrate that methoxylated flavonoids, particularly cirsimaritin, are key modulators of host immunity and promising candidates for further development as adjuncts or leads in antimalarial therapy.

Sepsis-related cardiomyopathy is mainly induced by uncontrolled inflammation and mitochondrial oxidative damage triggered by endotoxins, especially those of Gram-negative bacteria. Lipopolysaccharide (LPS), an endotoxin, activates the Toll-like receptor 4/nuclear factor kappa B (TLR4/NF-&#x3ba;B) signaling pathway and the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, which further activates caspase-1 to process the pro-inflammatory cytokines interleukin-1&#x3b2; (IL-1&#x3b2;) and interleukin-18 (IL-18). Chronic activation of this signaling pathway is responsible for mitochondrial damage and loss of membrane potential. Baicalein, a flavonoid derived from the herbal plant Scutellaria baicalensis is well recognized for its anti-inflammatory and antioxidant properties, although its full modulation of the NF-&#x3ba;B/NLRP3 mitochondrial signaling pathway is not thoroughly understood. In this study, we assessed whether baicalein has a protective effect on H9c2 cardiomyoblasts against inflammatory and mitochondrial injury induced by LPS through regulation of inflammatory pathways involving NF-&#x3ba;B, the inflammasome, caspase-1 activity, cytokine expression, and oxidation. H9c2 cells were pre-exposed to different baicalein concentrations (5 - 20 &#xb5;M) and, after 1 hour, exposed to 1 &#xb5;g/mL of LPS. Analysis included cell viability measurements by MTT assay, lactate dehydrogenase (LDH) release, and microscopic evaluation by phase-contrast microscopy. Expression levels of premature Nlrp3, Il1B, Il18, and Casp1 (0 - 4 h) were analyzed by real-time polymerase chain reaction (PCR). Protein levels of inflammasome and NF-&#x3ba;B pathway proteins were evaluated by immunoblot analysis. Activity of caspase-1 and cytokine secretion of IL-1&#x3b2; and IL-18 were evaluated by colorimetry and enzyme-linked immunosorbent assay (ELISA), respectively. Mitochondrial membrane potential (&#x394;&#x3a8;m) was evaluated by staining with JC1 dye, and reactive oxygen species (ROS) levels were evaluated by fluorescence with DCFH2DA; N-acetylcysteine (NAC) was used as an antioxidant positive control. LPS strongly elevated the transcription of Nlrp3 and Il1b at 2 hours, in addition to enhancing protein expression of NLRP3, activity of caspase-1, and secretion of IL-1&#x3b2; and IL-18. Underlying these pro-inflammatory responses were mitochondrial depolarization, augmented ROS production, diminished cell survival, and cytotoxicity. Prior administration of baicalein mitigated NF-&#x3ba;B activation, diminished priming of the inflammasome response at both transcriptional and protein levels, and decreased caspase-1 activity and secretion of the cytokines. Moreover, baicalein maintained mitochondrial membrane potential and diminished ROS within the cells, comparable to NAC, and improved cellular viability and morphology. Baicalein confers substantial protection against LPS-induced inflammatory and oxidative damage in H9c2 cardiomyoblasts by concurrently suppressing NF-&#x3ba;B activation, mitigating NLRP3 inflammasome signaling, curbing caspase-1-driven cytokine maturation, and stabilizing mitochondrial function through reduction of ROS. These findings identify baicalein as a promising candidate for targeting the ROS-inflammasome axis in sepsis-associated cardiac dysfunction and related inflammatory cardiac disorders.

The continuous emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants necessitates the rapid development of novel therapeutics, particularly those targeting conserved viral proteins. Peptide-based drugs offer high specificity and low toxicity, making them ideal candidates. This study employed an integrated computational approach, combining structural biology, molecular docking, and molecular dynamics (MD) simulations, to design and evaluate novel peptide analogs targeting three key proteins of SARS-CoV-2: the Spike (S) protein, RNA-dependent RNA polymerase (RdRp), and nucleocapsid (N) protein. The first step involved preparing a dataset containing anti-SARS-CoV-2 peptides using the DRAVP database and a literature survey. Then, the best inhibitory peptides were screened using the AVPPred tool, and analogous peptides were designed based on the selected lead peptide. The designed peptides were then investigated in terms of their structure, physicochemical properties, and antiviral potency. Additionally, molecular docking, performed using the specialized nCoVDock2 server, showed that all designed analogs exhibited highly favorable binding. Specifically, the best-performing analogs achieved remarkable docking scores in the range of -200 to -300 a.u. (arbitrary units), indicating a strong predicted relative binding affinity for their respective targets. The top-ranked complexes were then subjected to 100 ns explicit solvent MD simulations. Our findings suggest that peptide W is the most effective analogue for inhibiting S protein, achieving a relative docking score of -303.41 a.u., in contrast to the -284.12 a.u. relative docking score of the EK1 lead peptide. Regarding the inhibition of RdRp protein, the top newly designed analogue is peptide A5, which has a relative docking score of -187.36 a.u., compared to the score of -121.3 a.u. for lead peptide 5, respectively. The leading novel analogue for inhibiting the N protein is A7, which has a relative docking score of -317.69 a.u., surpassing the relative docking score of -255.48 a.u. for Plectasin. The MD results confirmed the high dynamic stability of the W (targeting S protein) and A5 (targeting RdRp) complexes, demonstrating low Root Mean Square Deviation (RMSD) and maintaining critical hydrogen bonds and hydrophobic interactions throughout the trajectory. The use of bioinformatics algorithms to develop engineered peptides with high affinity for SARS-CoV-2 virulence proteins offers a promising outlook for peptide-based therapies against SARS-CoV-2. It also presents a promising approach for developing therapeutic methods against other viral diseases. Furthermore, these computational insights lay the groundwork for subsequent in vitro and in vivo validation studies to ascertain the therapeutic efficacy and safety profiles of the identified peptide candidates.

Alzheimer's disease (AD) is a chronic neurodegenerative disorder characterized by downregulation of potassium voltage-gated channel subfamily a member 2 (KCNA2) proteins. Potassium voltage-gated channel subfamily a member 2 is involved in the regulation of neuronal excitability by restoring neuronal potassium-chloride co-transporter 2 (KCC2) functions. Coumarin derivatives exert neuroprotective effects via upregulation of KCC2 proteins. Daphnetin (DPN; 7,8-dihydroxy coumarin) is a polyphenolic compound known to attenuate cognitive dysfunction. However, the role of DPN in the attenuation of AD-associated cognitive dysfunctions through regulation of KCC2 functions has not yet been investigated. The present study was designed to investigate the role of DPN against amyloid-&#x3b2; oligomer-induced AD in mice. In this study, a total of six groups with eight male Swiss albino mice per group were used. The simple randomization method was adopted for unbiased assignment of animals based on age, sex, and weight variations. Alzheimer's disease in mice was induced by intracerebroventricular (i.c.v.) injection of amyloid-&#x3b2; oligomer (A&#x3b2;; 4 &#x3bc;g/4 &#x3bc;L). The test compounds, i.e., DPN (40, 80, and 120 mg/kg of body weight), and donepezil (DP, 2 mg/kg), were administered orally (p.o.) for 21 consecutive days. Behavioral changes, including the Morris water maze (MWM) test, water Y-maze alternation test (WYMA), and novel object recognition test (NORT), were assessed according to the experimental protocol. Furthermore, hippocampal brain tissue biomarkers, namely acetylcholinesterase (AChE) activity, thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH), and KCC2 levels, were also estimated. In addition, A&#x3b2;-associated brain histopathological changes were evaluated using the eosin and hematoxylin staining method. Six mouse hippocampus tissue samples were used for the assessment of tissue biomarkers, and the remaining two brain tissues were used for histological observations. Behavioral data were statistically analyzed by two-way analysis of variance (ANOVA), and biomarkers were analyzed by one-way ANOVA. The 95% confidence level (P < 0.05) was set for confirmation of statistical significance. The results revealed that administration of A&#x3b2; enhanced escape latency time (ELT) and reduced time spent in the target quadrant (TSTQ) values in the MWM test; increased transfer latency (TL) values in the WYMA test; and reduced percentage location preference (%LP) while increasing percentage Recognition Index (%RI) in the NORT test. Furthermore, A&#x3b2; induced increases in AChE activity and TBARS levels, along with reductions in GSH and KCC2 levels. It also caused neurodegeneration in the CA3 hippocampus region. However, DPN ameliorated the above A&#x3b2;-induced changes in cognitive behaviors, biomarkers, and histopathological levels. Daphnetin attenuates A&#x3b2;-associated AD progression via inhibition of AChE activity, scavenging of free radicals, reduction of inflammation, and restoration of neuronal KCC2 channels. Hence, it may be a potential therapeutic agent for the treatment of AD. However, more extensive studies are required to confirm this therapeutic potency in different AD conditions and various animal species.

Hepatocellular carcinoma (HCC) is a lethal liver malignancy associated with substantial morbidity and mortality and remains a major socioeconomic burden worldwide. Given the high risk associated with HCC, novel therapeutic approaches are urgently needed. Angelica sinensis is a traditional Chinese herbal remedy that contains bioactive phytochemicals with promising anticancer properties, making it a potential candidate for evaluation in HCC. This study integrated computational network pharmacology, molecular docking, and in vitro cellular assays to elucidate the mechanisms by which A. sinensis exerts anti-HCC effects in Hep-G2 cells. Phytochemicals were identified using the Traditional Chinese Medicine Systems Pharmacology database and filtered according to drug-likeness and oral bioavailability criteria. Key compounds (&#x3b2;-sitosterol, alpha-cephalin, sitogluside, and stigmasterol) were further analyzed using the SuperPred Target Prediction tool to predict potential targets. HCC-related targets were curated from GeneCards and refined using the GeneCards Inferred Functionality Score. The overlap between compound-related and disease-specific targets was used to construct a protein-protein interaction network using STRING, which was subsequently visualized in Cytoscape to identify hub genes. Molecular docking between stigmasterol and the top 3 hub genes was evaluated using the CB-Dock2 online server, and all preparations were performed in BIOVIA Discovery Studio. In vitro, Hep-G2 cells were treated with varying concentrations (0, 25, 50, 100, and 200 &#xb5;g/mL) of A. sinensis extract. Cell viability, clonogenic potential, apoptosis induction, and migratory capacity were evaluated using the MTT assay, clonogenic assay, Annexin V/PI staining, and Transwell migration assay, respectively. Western blotting was used to assess the expression of key hub proteins in Hep-G2 cells. In silico analysis identified an initial pool of 126 phytochemicals, which was refined to 4 key compounds yielding 139 unique targets. Intersection with HCC-related targets produced 123 common targets, generating a network comprising 119 nodes and 520 edges. STAT3, NFKB1, and TLR4 were identified as pivotal hub genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the hub genes demonstrated significant enrichment in HCC-related pathways and biological processes. Molecular docking indicated a strong binding affinity of stigmasterol to STAT3, NFKB1, and TLR4, with binding energies of -7.8, -6.9, and -6.9 kcal/mol, respectively. In vitro assays showed a dose-dependent reduction in Hep-G2 cell viability and colony formation, significant induction of apoptosis, and marked inhibition of migration. Western blotting confirmed significant downregulation of STAT3, NFKB1, and TLR4 expression at higher extract concentrations. This study concluded that A. sinensis herbal extract was predicted to exert potent antiproliferative effects against HCC through the possible modulation of key signaling pathways and the targeting of hub genes, supporting its potential for further therapeutic development.

The use of antioxidants has been shown to mitigate the symptoms of acute pancreatitis (AP) and improve associated biomarkers. In light of the antioxidant properties of mesna, this pilot study was undertaken to determine whether the addition of mesna to standard treatment for AP offers superior outcomes compared with standard treatment alone. In a randomized open-label pilot trial, patients diagnosed with AP were enrolled in the study. Intravenous mesna at a dose of 400 mg/day was administered for 7 days. Clinical indicators, including the Harmless Acute Pancreatitis Score (HAPS), amylase, lipase, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and malondialdehyde (MDA), were measured at baseline and after the 7-day treatment period. Although MDA levels significantly decreased in the group receiving mesna combined with standard therapy compared with the group receiving standard therapy alone after 7 days (P = 0.01), no significant differences were observed between the 2 groups in terms of HAPS, amylase, lipase, CRP, or ESR. These findings suggest that the addition of mesna at a daily dose of 400 mg did not influence the overall outcome of AP after 7 days. Further large-scale studies with varying doses and durations of mesna administration are recommended.