Pakistan Journal of Pharmaceutical Sciences

Abstract: Background: Moderate hamstring strain is a common sports injury impairing daily and athletic function. While topical nonsteroidal anti-inflammatory drugs (NSAIDs) offer localized pain relief with reduced systemic effects, high-quality evidence specific to hamstring injury is limited. Objectives: To evaluate the effect of a short-course topical diclofenac epolamine patch combined with rehabilitation on pain and functional recovery in moderate hamstring strain. Methods: In this single-center, double-blind, placebo-controlled trial, 120 patients were randomized to an experimental group (diclofenac patch twice daily for 14 days plus rehabilitation) or a control group (placebo patch plus identical rehabilitation). Outcomes assessed included pain measured by the Visual Analogue Scale (VAS), as the primary endpoint, muscle strength, knee range of motion (ROM), functional back-kick test, time to rehabilitation criteria and safety (adverse events, liver/renal function). Results: The diclofenac group demonstrated significantly greater pain reduction at days 14, 21 and 28 post-intervention (all p < 0.01). Muscle strength recovery was superior at days 21 and 28, knee ROM improved more at day 28 and functional back-kick performance was better at day 28 (all p < 0.05). Time to meet rehabilitation criteria was significantly shorter in the experimental group (p = 0.002). Adverse event incidence and changes in liver/renal function were similar between groups (all p > 0.05). Conclusion: A short-course topical diclofenac epolamine patch, combined with rehabilitation training, effectively alleviates pain, accelerates functional recovery, and is safe for treating moderate hamstring strain, supporting its use in clinical and sports rehabilitation.

Abstract: Background: Pseudomonas aeruginosa is widely recognized as a ubiquitous, drug-resistant, biofilm-forming bacterium that produces numerous virulence factors and plays a prominent role in hospital-acquired infections. Objectives: This study aimed to determine the prevalence of multidrug resistance (MDR), biofilm formation and biofilm-associated genes among P. aeruginosa isolates recovered from burn wound infections. Methods: A prospective cross-sectional observational study was conducted in which total of 176 burn wound samples were collected and processed for bacterial isolation. Identification of isolates was confirmed by polymerase chain reaction (PCR) targeting the oprL gene. Antimicrobial susceptibility testing was performed using the Kirby–Bauer disc diffusion method. Biofilm-forming ability was assessed using the microtiter plate assay. Furthermore, PCR was used to detect biofilm-associated genes, including pelA, pelD, cupA, algD, pelF, pslA, pslB and pslD, in biofilm-producing isolates. Results: Out of 176 burn wound samples, 74 (42%) were positive for P. aeruginosa, among which 58 isolates were identified as MDR while all isolates remained susceptible to polymyxin B. Biofilm formation was observed in all MDR isolates, with 13.79%, 44.82% and 41.37% categorized as weak, moderate and strong biofilm producers, respectively, whereas only a limited proportion of non-MDR isolates demonstrated biofilm-forming ability (P = 0.001). Among MDR biofilm-forming isolates, biofilm-associated genes such as pslA and algD were each detected in 96.55% of isolates while cupA were identified in 94.82% of isolates. Notably, cupA showed a statistically significant association with biofilm-forming intensity (P = 0.020), suggesting its potential role in enhanced biofilm establishment among MDR isolates. Conclusion: Overall, the high prevalence of multidrug-resistant, biofilm-forming P. aeruginosa isolates recovered from burn wounds is an important microbiological concern. The frequent coexistence of multidrug resistance and biofilm-forming capacity observed in this study suggests a possible association that may complicate infection management and emphasizes the importance of continuous surveillance and effective infection control strategies.

Abstract: Background: Hepatocyte nuclear factor 4α (HNF-4α) is a liver-enriched transcription factor that is downregulated in hepatocellular carcinoma (HCC). However, the molecular mechanisms by which it regulates the malignant phenotype of HCC cells remain unclear. Objectives: To investigate whether HNF-4α affects HCC cell proliferation, invasion and apoptosis by regulating the IGFBP-2/Notch1 signaling pathway. Methods: HNF-4α was overexpressed and silenced in HepG2 cells using a pcDNA3.1-HNF-4α plasmid and HNF-4α-specific siRNA, respectively. RT-qPCR and Western blotting verified transfection efficiency. Cell proliferation and invasion were assessed by MTT and Transwell assays, respectively. Apoptosis was evaluated by Caspase 3 activity assay. IGFBP-2 secretion was measured by ELISA. Protein expression of Bcl-2, Bax, IGFBP-2 and Notch1 was detected by Western blotting. Results: HNF-4α overexpression significantly inhibited HepG2 cell proliferation (by approximately 35% at 48 h and 45% at 72 h) and invasion (by approximately 55%), while markedly increasing Caspase 3 activity (approximately 2.8-fold) and the Bax/Bcl-2 ratio (from 0.42 to 2.18). Moreover, HNF-4α overexpression significantly suppressed IGFBP-2 secretion (by approximately 60%) and downregulated IGFBP-2 (by approximately 55%) and Notch1 NICD (by approximately 50%) protein expression (all P < 0.001). Conversely, HNF-4α knockdown produced opposite effects, significantly promoting proliferation (by approximately 40% at 48 h and 55% at 72 h) and invasion (by approximately 70%), decreasing Caspase 3 activity (by approximately 55%) and the Bax/Bcl-2 ratio (from 0.45 to 0.19) and upregulating IGFBP-2 (approximately 2.2-fold) and Notch1 NICD (approximately 1.8-fold) expression (all P < 0.01). Conclusion: HNF-4α inhibits HCC cell proliferation and invasion and promotes apoptosis through negative regulation of the IGFBP-2/Notch1 signaling pathway, providing new experimental evidence for the tumor-suppressive mechanism of HNF-4α in HCC.

Abstract: Background: Transdermal fentanyl is widely used for pain management; however, whether high systemic doses exert direct hepatotoxic effects or trigger intrinsic liver apoptotic pathways independently of systemic hypoxia remains unclear. Objectives: This study aimed to investigate the dose-dependent effects of transdermal fentanyl on hepatic structural integrity, biochemical function and markers of intrinsic mitochondrial apoptosis (Bcl-2 and caspase-3) in rats. Methods: Thirty-two Wistar albino rats were randomly assigned to four groups (n = 8): a control group and three experimental groups treated with transdermal fentanyl at doses of 3.125 µg/kg/h (Group I), 6.25 µg/kg/h (Group II) and 12.5 µg/kg/h (Group III). Patches were applied to the abdominal region and replaced every 72 hours for nine days. At the end of the experiment, liver tissues were collected for biochemical and histopathological and molecular analyses of apoptotic markers. Apoptotic changes were assessed by measuring caspase-3 and Bcl-2 levels. Results: Serum alanine aminotransferase (ALT) and total cholesterol levels were significantly higher in Group II and III than in the control group (p<0.05). Glucose levels were significantly increased in all fentanyl-treated groups (p<0.05). Total protein levels were significantly elevated in Group III compared to the control group (p<0.05). Lactate dehydrogenase (LDH) levels were significantly lower in Groups II and III (p<0.05). The anti-apoptotic marker Bcl-2 was significantly decreased in Group III compared with the other groups (p<0.05), whereas no statistically significant differences were observed in caspase-3 levels among any of the groups. No evident histopathological alterations were detected in liver tissues. Conclusion: High-dose transdermal fentanyl induces significant biochemical alterations and reduces Bcl-2 levels in rat liver tissue, suggesting a potential shift toward apoptotic signaling despite the absence of significant caspase-3 changes. However, short-term administration does not appear to cause evident histopathological damage.

Abstract: Background: Oral fixed-dose combinations of Amlodipine Besylate and Valsartan are widely prescribed for hypertension but are associated with first-pass metabolism and variable bioavailability. Buccal drug delivery provides a favorable substitute to improve systemic availability and provide sustained release. Objectives: This study aimed to develop and assess mucoadhesive buccal patches containing Amlodipine Besylate and Valsartan to enhance bioavailability and achieve controlled drug release over 24 hours. Methods: Buccal patches were developed via the solvent-casting method using Hydroxypropyl Methylcellulose, Polyvinylpyrrolidone, Methylcellulose, Sodium Alginate and Metolose as backing membrane polymers. All the prepared films were evaluated for physicochemical parameters by means of X-ray Diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM) and in vitro and ex vivo drug release. In-vitro drug release was studied with USP Apparatus II (paddle method) in 500 ml 6.8 pH phosphate buffer at 37 ± 0.5°C and 50 rpm for 24 hours. Ex vivo permeation studies were performed using a Franz diffusion cell fitted with rabbit buccal mucosa under similar conditions. Results: The formulations generally exhibited acceptable physicochemical characteristics within the evaluated parameters. The developed films appeared transparent with a uniform distribution of components. FTIR analysis did not indicate any detectable incompatibility or significant interaction between the drugs and excipients under the study conditions. Among all formulations, F1 contained the highest drug release and permeation. After 24 hours, cumulative release reached 84.45% for amlodipine and 88.03% for valsartan. Ex-vivo permeation was 76.31% and 78.42% for amlodipine and valsartan, respectively. Furthermore, buccal films' histopathological and stability studies were found to be harmless and functioned as an efficient dosage system. Conclusion: The developed mucoadhesive buccal patches successfully delivered both drugs in a sustained manner. Formulation F1 showed superior release and permeation profiles, suggesting its potential as substitute to conventional oral dosage forms for improved hypertension management.