Tropical Journal of Pharmaceutical Research

Mir-204 regulates the biological behavior of childhood leukemia cells by binding to 3'UTR end of target gene and reducing the level of the gene

2024-06-25T09:24:39+00:00

Purpose: To study the regulatory impact of mir-204 on the biological behavior of childhood leukemia cells and to elucidate its mechanism of action.

Methods: A total of 112 children with leukemia who were treated in Three Gorges University Yichang Central People's Hospital, Yichang, China from July 2018 to June 2020 were randomly divided into study and control groups (n = 56 each). The expression levels of mir-204 and hepatocyte growth factor (HGF) were determined using real-time polymerase chain chain reaction (RT-PCR) while the cell proliferation potential was determined using the CCK-8 method. Apoptosis and cell cycle progression of each group were evaluated using flow cytometry while changes in cell invasion and migration were assessed by Transwell assay. Luciferase reporter assay was used to determine the binding of mir-204 and HGF to 3'UTR.

Results: The expression level of mir-204 in study group was significantly lower than that in control group, while expression level of HGF was significantly higher (p < 0.01) compared to control group. There was no significant change in cell proliferation capacity at 24 h. The expression level of mir-204 was significantly decreased, while the expression level of HGF was significantly increased (p < 0.05). Luciferase activity of wild-type HGF 3'UTR in mir-204 overexpression group was significantly decreased, relative to mir-204 negative control group (p < 0.01). There was no significant difference in luciferase activity in mutant HGF gene between the two groups (p > 0.05).

Conclusion: Mir-204 reduces HGF expression level via HGF 3'UTR end, thereby inhibiting cell proliferation, invasion and migration, while promoting apoptosis, blocking cell cycle and regulating biological behavior of childhood leukemia cells.

Effect of anti-CD3-EGFR bispecific antibody-labeled DCCIK immune cells on proliferation of lung cancer cells

2024-06-25T09:40:49+00:00

Purpose: To investigate the effect of DC-CIK immune cells labeled with anti-CD3-EGFR bispecific antibody (BSAB) on the proliferation of lung cancer (LC) cells in order to generate data useful for enhancing the therapy of LC.

Methods: The anti-CD3 and anti-EGFR monoclonal antibodies were labeled on the surface of DC-CIK cells using chemical coupling method. Successful preparation of anti-CD3-EGFR BSAB was determined using SDS-PAGE. Two groups of cells were used: labeled group and a DC-CIK group. The cytotoxic effect on A549 LC cells was determined in vitro, and the tumor inhibition capacity was determined in 30 nude mice with LC.

Results: The killing rate of EGFR-positive A549 cells in labeled group was more severe than the killing rate in DC-CIK cells (p < 0.05). Cellular growth rate was significantly lesser in labelled A549 LC cells than in DC-CIK group. After 2 months of treatment, nude mouse tumor size in labeled group was smaller than the tumor volume in DC-CIK group. No obvious adverse reactions (ARs) were observed in both groups.

Conclusion: In vitro, CD3-EGFR BSAB-labeled DC-CIK immune cells produce cytotoxic effect and inhibit the proliferation of LC cells, while in vivo studies reveal that the cells produce good therapeutic effect on LC.

Complex assessment of in vitro anti-melanoma action of medicinal plant Cotinus coggygria Scop

2024-06-25T09:46:58+00:00

Purpose: To elucidate the anticancer mechanism and molecular targets of the aqueous fraction from Cotinus coggygria leaf extract in A375 malignant melanoma cells.

Methods: Aqueous ethanol extract from the leaves of C. coggygria was fractionated into the aqueous and chloroform fractions by solvent–solvent partition. A375 cells were treated with the aqueous fraction at various concentrations and incubated for 72 h. The effect of the aqueous fraction on programmed cell death by staining with Annexin V/Propidium iodide ias well as on cell viability and adhesion through labeling with fluorescein diacetate were determined under fluorescence microscopy. In addition, the clonogenicity assessment, comet assay evaluation of genotoxic properties, characterization of thermodynamic behavior of cells by differential scanning calorimetry, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) on the expression of genes involved in processes of apoptosis and cell cycle control were determined.

Results: The number of viable adherent cells was reduced by 38.00 and 85.06 % at 20 and 80 µg/mL, respectively compared to untreated cells. In addition, a significant, dose-dependent reduction in the clonogenicity of melanoma cells was observed, culminating in an almost complete inhibition of cancer cells colony-forming ability at 40 and 60 µg/mL of the fraction. Treatment with the fraction increased comets of Classes 3 and 4 to 35 %, while comets of Class 5, which corresponds to the highest level of DNA fragmentation, increased to 2.25 %. Also, the gene damage index (GDI) was highest (1.2) at the concentration of 40 μg/mL. Furthermore, the expression levels of BAX, CASP8 and CDKN1A were upregulated and MYC expression decreased after treatment with the aqueous fraction.

Conclusion: Cotinus coggygria possesses significant in vitro anti-melanoma potential. Future studies will be focused on the in vivo assessment of its therapeutic qualities in experimental animals.

Inhibitory effect of Asparagus officinalis extract on allergic asthma in Swiss albino mice

2024-06-25T11:32:08+00:00

Purpose: To determine the inhibitory effect of Asparagus officinalis extract (AOE) on allergic asthma induced by carrageenan (CGN).

Methods: Forty male mice were divided into 4 groups: the 1st group was untreated control; 2nd group was treated orally and daily with 500 mg/kg AOE for one week; the 3rd group was treated with single dose of CGN 2 %w/v (200 µL/mice) intraperitoneally and left for one week; while the 4th group was treated first with CGN as in 3rd group and treated with AOE as 2nd group after CGN injection for one week. After treatment, the animals were sacrificed and blood samples were subjected to white blood cell count. IL6 and TNF-α were measured in the lung homogenate while histopathological analysis was performed for lung samples.

Results: A single dose of CGN resulted in significant increase in white blood cell (WBC) count and proinflammatory cytokines (IL6 and TNF-α; p < 0.05). Histopathological analysis showed severe lung alterations such as accumulation of infiltration cells that blocked alveolar sacs, over- secretion of collagenous fibers, extracellular matrix and hyaline membranes. Moreover, treatment with AOE after CGN injection significantly reduced WBC count (p = 0) and pro-inflammatory cytokines (IL-6 and TNF-α) that were raised by CGN (p = 0). Furthermore, AOE reduced the pathological signs that were induced by CGN leading to improvement of lung function and reduction of collagenous fibers and hyaline membranes.

Conclusion: Asparagus officinalis extract reduces the allergic effect and lung pathological signs induced by CGN. Since AOE inhibits the production of inflammatory cytokines, it has the potential to be developed as a source of active pharmaceutical ingredients for the management of lung and airway injury.

Supercritical fluid carbon dioxide extract of Alpinia oxyphylla ameliorates dextran sulfate sodium-induced intestinal barrier damage in mice

2024-06-25T11:41:35+00:00

Purpose: Inflammation and oxidative stress are the leading causes of intestinal barrier dysfunction and a wide range of diseases. The purpose of this study was to explore the protective effect and elucidate potential mechanisms of supercritical carbon dioxide extract from Alpinia oxyphylla Miq. (AOE) on intestinal inflammation.

Methods: The AOE was extracted by supercritical carbon dioxide extraction and its components were determined. Cytotoxicity of the extract (0.05 to 20μg/mL) was determined on Caco-2 cells. In vitro assessment of whether AOE protected against LPS-induced intestinal barrier dysfunction was done on Caco-2 cells pretreated with non-cytotoxic concentrations (0.5μg/mL and 1 μg/mL) of the extract. Experimental colitis of mouse model was induced by drinking water containing 3% dextran sulfate sodium (DSS). The intestinal permeability was assessed by TEER and fluorescein isothiocyanate conjugated dextran (FITC). Pathological evidence and possible mechanism were verified by tissue staining and molecular methods including quantitative real-time polymerase chain reaction (qRT-PCR) and western blot.

Results: TEER and FITC testing indicated that AOE pretreatment significantly improved intestinal barrier hemostasis (p<0.05). Furthermore, AOE pretreatment counteracted DSS-induced upregulation of pro-inflammatory cytokines and oxidative stress (p<0.05). Lastly, two crucial signal pathways regarding hemostasis of intestinal barrier, NF-κB and NOX1-LCN2, were significantly attenuated upon AOE pretreatment (p<0.05).

Conclusion: This study sheds lights on the protective effect of AOE for intestinal hemostasis and supports the development of AOE-based intestinal protective agents.

Antioxidant, anti-inflammatory and antiulcer effects of Moroccan Ceratonia siliqua pulp in animal models

2024-06-25T11:49:56+00:00

Purpose: To investigate the potential antioxidant, anti-inflammatory and antiulcer effects of methanol extract of Ceratonia siliqua (Fabaceae).

Methods: The methanol extract of Ceratonia siliqua pulp was obtained using Soxhlet apparatus. Xylene (20 µL) induced ear inflammation administered intraperitoneally and HCl/ethanol solution (1 mL) induced gastric ulceration administered intra-gastrically were used to induce inflammation and gastric ulcer, respectively. Twenty-four mice were randomly divided into 4 groups (n = 6). Furthermore, eighteen rats (Sprague Dawley) were divided into 4 groups. Control group received (10 mL/kg of saline), Ceratonia siliqua group received (250 and 500 mg/kg), and other groups received diclofenac sodium (10 mg/kg; p,o), and omeprazole (30 mg/kg; p,o). Phytochemical screening was carried out by following standard procedures. The percentage of inhibition, pH, and ulceration index were measured using pH metric titration and degree of ulceration. The antioxidant capacity was determined in vitro using radical DPPH scavenging and FRAP assays.

Results: Phytochemical screening reveals the presence of flavonoids, tannins, quinones and sterols. Acute toxicity assessment showed that methanolic extract of Ceratonia siliqua has a DL₅₀ higher than 5 g/kg. Oral administration of methanolic extract of Ceratonia siliqua (500 mg/kg) reduced significantly (p < 0.001) the percentage inflammation and ulceration index in both assays in comparison to control groups, these effects were comparable to those observed in reference drugs. The extract possessed a strong antioxidant capacity with low IC₅₀.

Conclusion: Ceratonia siliqua extract has antioxidant, anti-inflammatory and anti- ulcer properties. Standardization of Ceratonia siliqua extract will be required for the development of lead molecules against ulcers and inflammation.

Molecular characterization of methanol extract of Artemisia annua leaf and its antifungal activity on clinical isolates of Candida albicans

2024-06-25T09:57:11+00:00

Purpose: To molecularly characterize and determine the antifungal properties of methanol-leaf extract of Artemisia annua (A. annua) on C. albicans isolates collected from three tertiary hospitals within Enugu State, Nigeria.

Methods: Five hundred (500) isolates of Candida albicans were collected from medical microbiology laboratories in Bishop Shanahan Hospital (BSH), University of Nigerian Teaching Hospital (UNTH), and Enugu State University Teaching Hospital (ESUTH). Anti-fungal sensitivity profiles of different azoles were determined using the disk diffusion technique. Thirty of the collected isolates were evaluated using the polymerase chain reaction (PCR) which targeted both 18S messenger RNA (rRNA) and Ergosterol II (ERGII) resistance gene sets for C. albicans. Anti-fungal activity of different concentrations (25, 50, 100, 200, 400, and 800 µg/mL) of the extract of Artemisia annua on resistant isolates was investigated using the agar well diffusion technique.

Results: All the isolates were resistant to azole drugs (ketoconazole (30 µg), miconazole (30 µg), clotrimazole (10 µg), itraconazole (30 µg), voriconazole (1 µg), and fluconazole (25 µg)). The PCR revealed that thirteen isolates (43 %) were positive for 18S rRNA gene and nine (30 %) possessed ERGII-resistant gene. Primers specific for C. albicans genotypes generated a PCR product of approximately 665 bp. Primers specific for ERGII resistant gene generated PCR product of approximately 1.4 kb. Extract of A. annua had antifungal activity against resistant C. albicans isolates with minimum inhibitory concentration ranging from 76.03 to 40.74 µg/mL.

Conclusion: Resistant C. albicans species have been genotypically characterized and the antifungal activity of A. annua has been reported. Methanol extract of A. annua inhibits C. albicans better than azole drugs.

Effect of casein supplementation on HBD-1, Th-17 and IL22 in a preterm Rattus norvegicus chorioamnionitis model

2024-06-25T10:12:02+00:00

Purpose: To investigate casein's role in the expressions of human beta-defensin (HBD)-1, T helper subset (Th)-17, and interleukin (IL)-22 in preterm Rattus norvegicus (RNs).

Methods: Twenty-four healthy, first-time pregnant RNs were divided into three groups (G). Group 1 (G1) received regular casein (R) 0.4 g/RN/day, while G2 and G3 received casein supplementation (S) 2 g/RN/ day. Each RN was given 10 g of food and drank 10 mL. On day 10, a single intraperitoneal lipopolysaccharide (LPS) was injected with 100 g/kg body weight. Lipopolysaccharide was injected into G3 as a premature model with chorioamnionitis. Histopathological examination was carried out to measure the expression of HBD-1, Th-17, and IL-22 using methyl green counterstaining. Chorioamnion specimens were taken and fixed in 4 % paraformaldehyde, and then evaluated by immunohistochemical (IHC) procedures to obtain immunoreactive score (IMS).

Results: The IMS expression of HBD-1 in G1 was 1.187 ± 0.372, while G3 was 9.562 ± 2.351 (p < 0.05). Th-17 expression were G1: 2.062 ± 0.140, G2: 6.122 ± 1.347, and G3: 9.112 ± 2.019 (p < 0.05). Furthermore, IL-22 expressions were G1: 1.825 ± 0.468, G2: 4.700 ± 1.614, and G3: 7.800 ± 1.669 (p < 0.05).

Conclusion: Casein supplementation increases HBD-1, Th-17, and IL-22 expression in preterm RN and in the LPS- induced RN chorioamnionitis model. The increase in HBD-1, Th-17, and IL-22 indicate that casein plays a role in the protection and control of chorioamnionitis tissue inflammation.

Validation of UV-visible spectrophotometric method for niclosamide in different media

2024-06-25T10:23:31+00:00

Purpose: To validate a UV-visible spectrophotometric technique for evaluating niclosamide (NIC) concentration in different media across various values of pH.

Methods: NIC was investigated using a UV-visible spectrophotometer in acidic buffer solution (ABS) of pH 1.2, deionized water (DW), and phosphate buffer solution (PBS), pH 7.4. The characterization of NIC was done with differential scanning calorimeter (DSC), powder X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The UV analysis was validated for accuracy, precision, linearity, and robustness.

Results: The DSC spectra showed a single endothermic peak at 228.43 °C (corresponding to the melting point of NIC), while XRD and FTIR analysis confirmed the identity, crystallinity and purity of NIC. In all media, the measured concentration of NIC was within ± 5 % of the actual value, which confirmed accuracy. The percentage relative standard deviation values were < 1 %, reflecting the precision of the method. The range of concentration measured was between 2 and 24 µg/mL, and all coefficient of determination (R2 ) values were > 0.99, indicating the linearity of the established analytical method. The limit of detection (LOD) and limit of quantification (LOQ) values were 0.122 and 0.407 µg/mL in ethanol, 0.530 and 1.766 µg/mL in ABS (pH 1.2), 0.224 and 0.747 µg/mL in DW, and 0.798 and 2.662 µg/mL in PBS, pH 7.4. The robustness was confirmed as the measured concentration under slight changes in temperatures and wavelengths were insignificant (p > 0.05).

Conclusion: Based on the results above, the UV- visible spectrophotometric method under investigation was validated to be accurate, precise, linear, and robust in all the different media for the determination of NIC.

Efficacy of diterpene ginkgolide meglumine injection in convalescent cerebral infarction and hemorheology

2024-06-25T10:36:53+00:00

Purpose: To investigate the effect of diterpene ginkgolide meglumine injection (DGMI) compared to betahistine in convalescent cerebral infarction (CI).

Methods: 80 convalescent patients with CI from March 2021 and March 2022 were randomly grouped into control and study groups comprising 40 participants each. Control and study groups received 20 mg betahistine and 25 mg DGMI respectively in 250 mL normal saline for 1 week. The efficacy, National Institute of Health Stroke Scale (NIHSS) score, hemorheology, and adverse effects were investigated in control and study groups

Results: Total effective rate was significantly higher in study group compared to control group (p < 0.05). Also, the National Institute of Health Stroke Scale (NIHSS) score, plasma viscosity (PV), whole blood reduced viscosity, fibrinogen (FIB), erythrocyte aggregation index (EAI), and incidence of adverse effect were significantly lower in study group compared to control group (p < 0.05).

Conclusion: Administration of DGMI to patients in the recovery period of CI improves neurological deficit, and average blood flow velocity in the brain, and reduces the incidence of adverse effects. Further studies with larger sample sizes and multi-center approaches will be required to validate these findings.

Efficacy of Lactobacillus live capsules in combination with metronidazole for the treatment of bacterial vaginosis

2024-06-25T10:44:16+00:00

Purpose: To evaluate the effectiveness of metronidazole in combination with Lactobacillus live capsules for the treatment of bacterial vaginosis (BV).

Methods: A retrospective study of 100 BV patients was conducted between June 2020 and February 2022 at Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Jinhua, China. Patients were assigned to control (48 patients received metronidazole only) and study groups (52 patients received metronidazole and Lactobacillus live capsules). Short-term efficacy, changes in vaginal flora distribution, recurrence rates, and occurrence of adverse effects were investigated.

Results: The study group showed significantly higher overall efficacy compared to control group (p < 0.05). After treatment, vaginal flora analysis revealed a more balanced microbiota in the study group, indicating a positive effect of combined therapy. However, recurrence rates at one and three months after treatment did not significantly differ between the two groups (p > 0.05). Both treatment regimens were generally well-tolerated, with only a single report of mild adverse reactions in the study group.

Conclusions: The combined therapy of metronidazole and Lactobacillus live capsules enhances shortterm therapeutic effectiveness and promotes a healthier vaginal microbiota. Although, the combined therapy does not significantly impact recurrence rates, it demonstrates a favorable safety profile. The lack of attention to non-bacterial constituents of the vaginal flora and potential long-term adverse effects of Lactobacillus live capsules represent an area for future study.

Establishment of a failure mode and effects analysis for high-risk breviscapine-based traditional Chinese medicine injection

2024-06-25T10:49:15+00:00

Purpose: To assess the failure modes and effects of clinical application of breviscapine-based traditional Chinese medicine injection (TCMI).

Methods: 229 reports on clinical application errors, medication errors or management measures relating to breviscapine injection were collected by searching various databases. A clinical application and safety evaluation questionnaire was formulated for use in the failure mode and effects analysis (FMEA) of breviscapine injection. The questionnaire survey was then distributed to 100 doctoral, nursing and pharmaceutical personnel in Xinxiang Central Hospital who were randomly chosen to participate.

Results: A total of 81 (83.5 %) valid questionnaires were retrieved. A total of 29 potential failure types, failure causes and failure effects were identified. Mean values of all risk priority numbers (RPN) of the 29 failure modes were ranked comprehensively. The failure modes identified as top 10 risk factors include; detailed information regarding the patient’s medical, allergy and family disease history not being provided to physicians (75.22); incorrect choice of drug manufacturer and lot number (74.95), drugs not being dispensed on the spot (72.16); inappropriate choice of solvent (71.31); drugs not suitable for combination therapy (70.81); inappropriate choice of solvent dosage (69.14); individual patient differences not taken into consideration (67.07); infusion rate too fast (67.00); off-label drug use (65.32); and age of the patient not taken into consideration (64.96).

Conclusion: As a risk management tool, the FMEA conducted in this study reduces potentially dangerous clinical application of high-risk TCMI, standardizes the medication process and significantly reduces occurrence of adverse reactions to TCMI. Future studies are required to validate these claims.

Clinical effect of suspension training and Mulligan technique in combination with celecoxib in the treatment of chronic non-specific lower back pain

2024-06-25T11:22:19+00:00

Purpose: To investigate the clinical efficacy of suspension training and Mulligan technique in combination with celecoxib in the management of chronic lower back ache of unknown causes.

Methods: A total of 100 subjects with persistent lower back pain of unknown etiology treated in the Sports Medical Rehabilitation Center, Shijiazhuang, China from June 2019 to December 2020, were assigned at random to control and study cohorts (n = 50/group). Celecoxib (100 to 200 mg/day) was taken orally, once or twice daily by the control cohort, while the other cohort received suspension training, Mulligan technology (once a day, 8 weeks) and celecoxib. The treatment effect, pain and dysfunction scores, inflammatory indicators, quality of life, and complications were compared on Visual Analogue Scale (VAS) and Oswestry Disability Index (ODI).

Results: The effect of Macnab standard in study group was significantly better than that in control group (p < 0.05). After treatment, VAS and ODI scores in both cohorts were significantly lower than pretreatment values, but were significantly lesser in the study cohort. Furthermore, TNF-α level, and levels of CRP and IL-6 were significantly reduced when compared with pre-treatment levels, and were significantly lower in study cohort. The GQOL-74 scores in both groups were significantly increased after treatment, but were significantly lower in the control cohort (p < 0.05). No significant variations were seen in cases of complications between both cohorts.

Conclusion: The use of suspension training and Mulligan technology in combination with celecoxib in treating persistent lower back pain of unknown cause is beneficial in reducing lower back pain, mitigating dysfunction, and improving patients' quality of life. There is however a need for more studies to validate these findings.