The expression of COX-2 in VEGF-treated endothelial cells is mediated through protein tyrosine kinase.

Cyclooxygenase (COX), existing as the COX-1 and COX-2 isoforms, converts arachidonic acid to prostaglandin H2, which is then further metabolized to various prostaglandins. Vascular endothelial growth factor (VEGF) has been shown to play important roles in inflammation and is upregulated by the prostaglandin E series through COX-2 in several cell types. Here, we have investigated the effects of VEGF on the COX isoform expressed in human umbilical vein endothelial cells (HUVEC). The signalling mechanism of the COX isoform expressed in endothelial cells activated with VEGF will be also investigated using the tyrosine kinase inhibitor, genistein, and protein kinase C inhibitor, staurosporine. The activity of COX-2 was assessed by measuring the production of 6-keto-prostaglandin F1alpha in the presence of exogenous arachidonic acids (10 microM, 10 min) by enzyme immunoassay. The expression of COX isoform protein was detected by immunoblot using specific antibodies. Untreated HUVEC contained no COX-2 protein. In HUVEC treated with VEGF (0.01-50 ng/ml), COX-2 protein, but not COX-1, and COX activity were increased in a dose-dependent manner. Interestingly, the increased COX-2 protein and activity in response to VEGF (10 ng/ml) was inhibited by the tyrosine kinase inhibitor, genistein (0.05-5 microg/ml), but not by the protein kinase C inhibitor, staurosporine (0.1-10 ng/ml). Thus, the induction of COX-2 by VEGF in endothelial cells was mediated through protein tyrosine kinase, and the uses of specific COX-2 inhibitors in these conditions, in which VEGF was involved, might have a role

Untargeted Metabolomics Analysis using LC-MSQTOF for Metabolite Profile Comparison between Patients with Myofascial Pain of Upper Trapezius Muscle versus Controls

Objective: This study aims to identify different biomarkers of Myofascial pain syndrome (MPS) using untargeted metabolomics screening. Materials and Methods: In a case-control study, serum samples from MPS patients (n = 19) and healthy controls (n = 10) were analyzed using reverse-phase liquid chromatography and mass spectrometry quadrupole time-of-flight (MS-QTOF). The resulted raw data was processed with Progenesis QI data analysis software. The HMBD database was used to identify the metabolites based on their fold change (>1.2), variable importance plot (>1) with P < 0.05. MetaboAnalyst 5.0 was used to generate metabolic network analysis for all identified metabolites. Results: The MPS group reported significantly higher pain on visual analog scale when compared with control while most of the other routine blood chemical profiles were not different. Twenty-seven metabolites were analyzed and identified with untargeted metabolomics analysis which could distinguish MPS patients from healthy controls. Inosine and chenodeoxycholic acid were abundant in the MPS group, whereas the others were low. Metabolites were divided into three categories: lipids, nucleotides, and organic compounds. Possible MPS metabolites included lysoSM (sphingomyelin), lysoPC (lysophosphatidylcholine), lysoPE (lysophosphatidylethanolamine), triglyceride, and inosine. Conclusion: These metabolite profiles, including glycerophospholipids mechanism and purine metabolism, indicate that the inflammatory process might be related to the mechanisms of MPS. A larger sample size, a different trigger point location, and modifications in therapy afterward should all be further explored

Effects of Different Durations of 9-Square Dance Exercise Versus Treadmill Exercise on the Physical Fitness and Quality of Life of Healthy Volunteers: A Pilot Randomized Controlled Trial

Objective: To evaluate the impact of 9-square dance exercise (9SDE) on physical fitness and quality of life compared to traditional treadmill exercise (TME). Materials and Methods: In total, 33 healthy volunteers (10 men, 23 women) were recruited and randomly assigned into three groups: 9 square dance exercise for 8 minutes (9SDE-8), 9 square dance exercise for 30 minutes (9SDE-30), or treadmill exercise (TME). Exercises were done three times a week for 12 weeks and physical fitness tests were performed for all the groups at weeks 0, 6, and 12. Participants were assessed using the European Quality of Life Measure 5 Domains and 5 Levels questionnaire (EQ-5D-5L). Results: Significant improvements in cardiorespiratory endurance, leg strength, and flexibility were demonstrated in the 9SDE-30 group (p<0.05). There was no significant difference in physical fitness between the 9SDE-30 and TME groups. The 9SDE-8 group showed a significant improvement in utility in the EQ-5D-5L questionnaire (p<0.05), while the TME group showed a significant improvement in directly evaluated health status (p<0.05). 9SDE-30 and TME showed similar improvements in cardiorespiratory endurance and leg strength. Conclusion: Considering its low-resource requirement and overall utility, coupled with its effectiveness in promoting cardiovascular fitness and leg strength, 9SDE represents a viable exercise alternative for those with limited time and resources

Identification of Apigenin and Luteolin in Artemisia annua L. for the Quality Control

Objective: To identify active compounds and establish the chemical fingerprint of Artemisia annua L. for the quality control. Methods: Thin-layer chromatography (TLC) conditions were developed to screen for 2 common flavonoids (apigenin and luteolin). Three mobile phases were used to isolate these flavonoids in 80% ethanolic extract of A. annua. Hexane : ethyl acetate : acetic acid (31:14:5, v/v) and toluene : 1,4-dioxane : acetic acid (90:25:4, v/v) were used in normal phase TLC (NP-TLC), and 5.5% formic acid in water : methanol (50:50, v/v) were used in reverse phase TLC (RP-TLC). Chromatograms were visualized under visible light after spraying with Fast Blue B Salt. Apigenin and luteolin bands were checked by comparing their Rf values and UV-Vis absorption spectra with reference markers. Results: Apigenin and luteolin were simultaneously detected with good specificity in RP-TLC condition, while only apigenin was detected in NP-TLC condition. Apigenin band intensity was higher than luteolin band intensity in both conditions. Conclusion: This knowledge can be applied to the development of quality control assessments to ensure product efficacy and consistency

Identification of Apigenin and Luteolin in Artemisia annua L. for the Quality Control

Objective: To identify active compounds and establish the chemical fingerprint of Artemisia annua L. for the quality control. Methods: Thin-layer chromatography (TLC) conditions were developed to screen for 2 common flavonoids (apigenin and luteolin). Three mobile phases were used to isolate these flavonoids in 80% ethanolic extract of A. annua.Hexane : ethyl acetate : acetic acid (31:14:5, v/v) and toluene : 1,4-dioxane : acetic acid (90:25:4, v/v) were used in normal phase TLC (NP-TLC), and 5.5% formic acid in water : methanol (50:50, v/v) were used in reverse phase TLC (RP-TLC). Chromatograms were visualized under visible light after spraying with Fast Blue B Salt. Apigenin and luteolin bands were checked by comparing their Rf values and UV-Vis absorption spectra with reference markers. Results: Apigenin and luteolin were simultaneously detected with good specificity in RP-TLC condition, while only apigenin was detected in NP-TLC condition. Apigenin band intensity was higher than luteolin band intensity in both conditions. Conclusion: This knowledge can be applied to the development of quality control assessments to ensure product efficacy and consistenc

Effect of Kao-Ta (9-Square Step Exercise) and Kao-Ten (9-Square Dance Exercise) on Balance Rehabilitation in Patients with Balance Disorders

Objective: To study the effect of Kao-ta (9-square step exercise) and Kao-ten (9-square dance exercise) on balance improvement in patients with balance disorders. Methods: This prospective pilot study in patients with balance disorders was conducted at the outpatient clinic, Department of Otorhinolaryngology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand from December 2015 to December 2016. Patients diagnosed by clinical symptoms and at least one abnormal condition on posturography were taught how to perform Kao-ta and Kao-ten exercise. Participants were provided with the equipment necessary to create a nine square grid at home. They were instructed to perform 3 minutes of Kao-ta followed by 2 minutes of Kao-ten twice per day for at least 45 days in an 8-week period. Posturography and visual analogue scale (VAS) of balance symptom severity were compared between before and after exercise program.  Results: Eleven patients with balance disorders were included. The mean age was 57.2±12.9 years (range: 33-70), and all patients were women. The average composite equilibrium score at baseline was 64.4±8.1. After 8 weeks of Kao-ta and Kao-ten, the average composite equilibrium score increased to 73.8±10.2 (p<0.01). The median (P25, P75) of the abnormal equilibrium score condition decreased from 2 (1, 3) at baseline to 1 (0, 2) after 8 weeks (p=0.016). The median VAS of balance symptom severity decreased from 4 (3, 6) at baseline to 2 (0.2, 5.5) after 8 weeks (p=0.028). Conclusion: Kao-ta and Kao-ten exercise can improve symptoms in patients with balance disorders after 8 weeks of exercise

Efficacy of 0.0125% Capsaicin Patch at Acupuncture Point for Pain Relief in Knee Osteoarthritis: A Randomized Controlled Trial

Objective: To study the efficacy of 0.0125% capsaicin patch at acupuncture point for pain relief in knee osteoarthritis (OA). Methods: This randomized, double-blind, placebo-controlled clinical trial was conducted during September 2014to March 2015 Sixty-two ambulatory patients over 50 years of age with a diagnosis of knee OA were included. All enrolled patients had a pain score of 4-7 out of 10. Participants were randomized into either the treatment (capsaicin, n=31) or control (placebo, n=31) group. Interventions: Capsaicin vs. placebo patch at ST34 (1), SP10 (2), ST35 (3), EX-LE4 (Neixiyan) (4), ST36 (5), and SP9 (6) acupuncture points for 4 weeks. Main outcome measure: Pain subscale of modified Thai version of Western Ontario and McMaster (WOMAC) Osteoarthritis Index was assessed at baseline, 2, 4, 6, and 8 weeks after treatment. Results: Pain subscale of the modified Thai version of WOMAC showed no significant difference between groups at baseline, 2, 4, 6, and 8 weeks after treatment. After 2 weeks of treatment, the pain subscale score in the treatment group was significantly decreased from baseline (p<0.001). After 4 weeks of treatment, the pain subscale score in the placebo group was significantly decreased from baseline (p<0.006). Conclusion: Capsaicin patch at acupuncture point for pain relief in knee OA yielded no significant difference between groups. However, significant pain relief from baseline was observed in the treatment group after 2 weeks of treatment and in the placebo group after 4 weeks of treatment. Further study with higher capsaicin concentration and/or larger size patch should be considered

Upregulation of CYP 450s expression of immortalized hepatocyte-like cells derived from mesenchymal stem cells by enzyme inducers

<p>Abstract</p> <p>Background</p> <p>The strenuous procurement of cultured human hepatocytes and their short lives have constrained the cell culture model of cytochrome P450 (CYP450) induction, xenobiotic biotransformation, and hepatotoxicity. The development of continuous non-tumorous cell line steadily containing hepatocyte phenotypes would substitute the primary hepatocytes for these studies.</p> <p>Results</p> <p>The hepatocyte-like cells have been developed from hTERT plus Bmi-1-immortalized human mesenchymal stem cells to substitute the primary hepatocytes. The hepatocyte-like cells had polygonal morphology and steadily produced albumin, glycogen, urea and UGT1A1 beyond 6 months while maintaining proliferative capacity. Although these hepatocyte-like cells had low basal expression of CYP450 isotypes, their expressions could be extensively up regulated to 80 folds upon the exposure to enzyme inducers. Their inducibility outperformed the classical HepG2 cells.</p> <p>Conclusion</p> <p>The hepatocyte-like cells contained the markers of hepatocytes including CYP450 isotypes. The high inducibility of CYP450 transcripts could serve as a sensitive model for profiling xenobiotic-induced expression of CYP450.</p

The Effects of Thai Herbal Ha-Rak Formula on COX Isoform Expression in Human Umbilical Vein Endothelial Cells Induced by IL-1 β

Objective. To investigate the modulated effects of HRF on cyclooxygenase isoform expression and its activity, using the human umbilical vein endothelial cell (HUVEC) model induced by interleukin-1 beta (IL-1β). Methods. Cells were treated with indomethacin (positive control), HRF, and its components at various concentrations prior to treatment with IL-1β at 24 h. Cell viability was determined by MTT assay. Moreover, the anti-inflammatory effects of HRF and its components through mRNA and protein expression were established using real-time quantitative PCR and Western blot, respectively. COX activity was identified via exogenous and endogenous PGE2 productions using the EIA. Result. There was no cytotoxicity in HUVECs treated with HRF. None of the experimental conditions used in the study affected the expression of COX-1, but COX-2 protein expression was inhibited at concentrations under 10 µg/mL. Despite the significantly increased levels of exogenous PGE2, HRF had no effect on COX-2 mRNA expression. However, the production of PGE2 was lower at a concentration of 100 µg/mL HRF than at a concentration below 10 µg/mL. Interestingly, each component of HRF revealed different effects of the Ha-Rak formula. Conclusion. Our preliminary findings suggest that HRF and its components provide diverse modulation of COX-2 and PGE2 at the in vitro level