Background: Chronic obstructive pulmonary disease is a progressive lung disease that involves airway inflammation, chronic bronchitis, and emphysema. Yukgunja-tang, one of the traditional Asian herbal medicines, has been used widely in treating patients with gastrointestinal diseases in Korea. Objective: Here, we investigated its efficacy on the inflammatory response using a mouse model of cigarette smoke (CS) exposure together with lipopolysaccharide (LPS) treatment. Materials and Methods: Over 4 weeks, mice were exposed to CS on 5 days/week and instilled intranasally with LPS on days 8 and 23. Yukgunja-tang water extract (YTWE) was administered to mice on the same 5 days. Results: YTWE administration significantly reduced the numbers of inflammatory cells and levels of pro-inflammatory cytokines in bronchoalveolar lavage fluid compared with CS plus LPS-exposed mice. Moreover, YTWE inhibited the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and IκBα proteins induced by CS plus LPS treatment. Histologically, YTWE attenuated the infiltration of inflammatory cells into peribronchial lesions, thickening of alveolar walls and accumulation of collagen in the lung tissues. Conclusion: Our findings suggest that YTWE prevents CS plus LPS-induced lung inflammation by inhibiting p38 MAPK and IκBα signaling. Therefore, YTWE might be a potential drug for the treatment of lung inflammation induced by CS exposure. Abbreviation used: BALF: Bronchoalveolar lavage fluid; COPD: Chronic obstructive pulmonary disease; CS: Cigarette smoke; HPLC: High-performance liquid chromatography; IL: Interleukin; LPS: Lipopolysaccharide; MAPK: Mitogen-activated protein Kinase; NF-κB: Nuclear factor kappa-B; ROF: Roflumilast; TNF-α: Tumour necrosis factor-alpha; YTWE: Yukgunja-tang water extract.

Background: Cordyceps sinensis (CS), a traditional Chinese herbal medicine, has many pharmacological effects. Doxorubicin (DOX) is a spectrum antitumor drug, but it has toxicity to multiple organs. Objective: The objective of this study was to evaluate the effect of fermented CS on regulating hepatic energy metabolism against DOX-treated hepatic toxicity in rats. Materials and Methods: Male Sprague-Dawley rats were randomly divided into six groups and administrated orally for 23 days as follows: normal control group, CS (1.50 g/kg/d) control group, DOX control group, and DOX + CS (0.75 g/kg/d, 1.5 g/kg/d, 3.00 g/kg/d) groups. Rats in DOX-treated groups were intraperitoneally injected with DOX at a dose of 2.5 mg/kg at every 48 h, and repeated for six times. At the end of the experiment, the mortality and liver index, blood biomarkers about the hepatic injury, hepatic histopathological changes, hepatic energy metabolism, and hepatic cyclic adenosine monophosphate (AMP) were measured. Results: DOX-induced a higher mortality, the damage of the liver manifests itself in the increase of liver weight index and the activity of serum aspartate aminotransferase, the decrease of serum contents of total protein and albumin, and histopathological changes, and the disorders of energy metabolism including the decrease of phosphocreatine, adenosine-triphosphate, adenosine diphosphate, AMP, and total adenine nucleotides in hepatic tissues. Fermented CS not only could attenuate those changes but also could increase the content of hepatic cAMP. Conclusion: DOX induces hepatic injury accompanying hepatic energy metabolism disorders. Fermented CS regulates the disorders of hepatic energy metabolism, thereby attenuating liver injury caused by DOX. Abbreviation used: CS: Cordyceps sinensis; DOX: Doxorubicin; cAMP: Cyclic adenosine monophosphate; AST: Aspartate aminotransferase; TP: Total protein; ALB: Albumin; ATP: Adenosine triphosphate; ADP: Adenosine diphosphate; AMP: Adenosine monophosphate; TAN: Total adenine nucleotides.

Background: Capsicum annuum L. and Capsicum frutescens L., of family Solanaceae, contain capsaicinoids with several pharmacological effects. However, their pungency limits the long-term use through the gastrointestinal tract. Objective: To obtain phytoformulations of microparticles containing capsaicinoids for reducing their pungency by oral route to achieve an anti-ulcer effect. Materials and Methods: Microparticles of poly(ε-caprolactone) containing capsaicinoids were prepared by simple emulsion/solvent evaporation. An optimized reverse-phase high-performance liquid chromatography method with ultraviolet (UV) detection for quantifying capsaicinoids into microparticle phytoformulations was then developed and validated. Chromatographic conditions consisted of a C18 reverse-phase analytical column (250 mm × 4.60 mm, 5 μm) using a mixture of acetonitrile and water (70:30 v/v) adjusted to pH 4.5 as mobile phase at a flow rate of 0.75 mL/min with UV detection at 280 nm. The developed method was validated as per the ICH guidelines with respect to specificity, linearity, limit of quantification, limit of detection, accuracy, precision, and robustness. The gastroprotective activity of pure capsaicinoids and loaded microparticles against ethanol-induced gastric ulcer in rats was performed. Results: Microparticles were successfully obtained. Analytical validation provided suitable results regarding all parameters investigated. These phytoformulations presented suitable encapsulation efficiency higher than 90%. Regarding the ulcerative lesion index (ULI) scores, poly(ε-caprolactone) (PCL) microparticles containing 5% of capsaicinoids (ULI = 16.3 ± 1.8 points) was statistically similar (P > 0.05) to ranitidine (ULI = 15.3 ± 1.4 points) and omeprazole (ULI = 8.0 ± 1.2 points). Conclusion: Capsaicinoids-loaded PCL microparticles reveal the potential to be suitable candidates as controlled drug delivery system for the therapeutic management of ulcer. Abbreviations used: CAP: Pure capsaicin; EE: Encapsulation efficiency; HPLC: High-performance liquid chromatography; LOD: Limit of detection; LOQ: Limit of quantitation; MC0: Unloaded microparticles; MC3: Capsaicinoids-loaded poly(ε-caprolactone) microparticles (3%); MC5: Capsaicinoids-loaded poly(ε-caprolactone) microparticles (5%); MC10: Capsaicinoids-loaded poly(ε-caprolactone) microparticles (10%); OME: Omeprazole; PCL: Poly(ε-caprolactone); PVA: Poly (vinyl alcohol); PTFE: Polytetrafluoroethylene; RAN: Ranitidine; RSD: Relative standard deviation; SD: Standard deviation; SEM: Standard error of mean; TCM: Caprylic/capric triglycerides; TRPV: Transient receptor potential vanilloid; TRPV1: Transient receptor potential vanilloid 1; ULI: Ulcerative lesion index

Background: Saponins from Rhizoma Panacis Majoris (SRPM) are confirmed to have cardioprotective effect against myocardial ischemia injury by reducing oxidative stress, while its underlying mechanism has not been elucidated until recently. Objective: The objective of this study was to investigate the SRPM's cardioprotection and elucidate its underlying mechanisms. Materials and Methods: Adult male rats were received SRPM treatment in the presence or the absence of the silent information regulator 1 (Sirt1) inhibitor Ex-527 or nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor ATRA for 14 days and subjected to myocardial ischemia for 0.5 h and then 2 h reperfusion. Cardiac function, infarct size, antioxidant enzyme activities, ROS level and the related mRNA and protein expressions of antiapoptosis protein Bcl-2, proapoptosis protein Bax, caspase-3, caspase-9, Sirt1 and Nrf2-relatived signaling pathways were assessed. Results: SRPM was confirmed to have cardioprotective effects by ameliorating cardiac function, decreasing infarct size, reducing serum creatine kinase (CK), creatine kinase isoenzyme, lactate dehydrogenase and ROS releases and malondialdehyde level, raising total antioxidant capacity, superoxide dismutase, glutathione peroxidase, catalase activities, upregulating myocardial Sirt1, Nrf2, Bcl-2 protein and manganese superoxide dismutase, heme oxygenase-1, NAD(P)H-quinone oxidoreductase 1 and γ-glutamylcysteine synthetase mRNA expressions and downregulating acetylated forkhead box O 1, acetylated peroxisome proliferator-activated receptor γ coactivator 1α, Bax, cleaved caspase-3 and cleaved caspase-9 protein expressions; histopathological observations provided supportive evidence for the aforementioned results. Interestedly, its protective effects were significantly blocked for its combination with Ex-527 or ATRA. Conclusion: The studies demonstrated that SRPM exerted beneficially protective effects on myocardial ischemia/reperfusion injury. It was possibly related to reducing oxidative stress damage by activations of Sirt1 and Nrf2-related antioxidant signaling pathways. Abbreviations used: Ac-FoxO1: Acetylated FoxO1, Ac-Pgc-1α: Acetylated Pgc-1α, ARE: Antioxidant-response element, CK: Creatine kinase, CK-MB: Creatine kinase isoenzyme, DEPPD: N, N-diethyl-paraphenylendiamine, ELSD: Evaporative light scattering detection, FoxO: Forkhead box O, GCLC: Glutamate-cysteine ligase catalytic subunit, GSH-Px: Glutathione peroxidase, HO-1: Heme oxygenase-1, H₂O₂: Hydrogen peroxide, Keap-1: Kelch-like ECH-associated protein 1, HPLC: High-performance liquid chromatography, LDH: Lactate dehydrogenase, MDA: Malondialdehyde, NAD+: Nicotinamide adenine dinucleotide, MnSOD: Manganese superoxide dismutase, NF-κB: Nuclear factor-kappa B, NQO1: Nicotinamide quinone oxidoreductase 1, NAD(P) H: NAD(P)H-quinone oxidoreductase 1, Nrf2: Nuclear factor erythroid 2-related factor 2, Pgc-1α: Peroxisome proliferator-activated receptor γ coactivator 1α, Sirt1: Silent information regulator 1, SOD: Superoxide dismutase, SRPM: Saponins from Rhizoma Panacis Majoris, I/R: Ischemia/reperfusion, T-AOC: Total antioxidant capacity, TTC: 2,3,5-triphenyltetrazolium chloride.

Background: Endophytic fungi are of a growing interest as prominent sources of structurally unique bioactive natural products. Objective: This study aims to isolate and characterize bio-metabolites from the endophytic fungus Fusarium sp. isolated from Mentha longifolia L. roots as well as to assess the antimicrobial and cytotoxic potential of these metabolites. Materials and Methods: The endophytic fungi Fusarium sp. was cultured on a rice medium. The rice cultures' ethyl acetate extract was separated using various chromatographic techniques (SiO₂, RP-18, and sephadex LH-20) to afford four metabolites. Their structural characterization was achieved by various spectroscopic analyses, as well as comparing with the published data. Results: A new ergosterol derivative namely, fusaristerol A (22E,24R-5β,8β-epidioxyergosta-22-en-3β-yl decanoate) (1), along with ergosta-7,22-diene-3β,5α,6β-triol (2), ergosta-5,7,22-triene-3β-ol (3), and (22E,24R)-ergosta-7,22-dien-3β-ol (4), was separated. Fusaristerol A (1) possessed a significant antifungal activity toward Candida albicans with minimum inhibitory concentration (MIC) value of 8.3 μg/disc compared to clotrimazole (MIC 5.1 μg/disc). Moreover, it displayed a potent cytotoxic potential toward HCT-116 cell line with an half maximal inhibitory concentration (IC₅₀) value of 0.21 μM, compared to doxorubicin (IC₅₀ 0.06 μM). Conclusion: This is the first report for separation of a 5,8-epidioxy ergostane derivative from Fusarium sp. Fusaristerol A may provide a new promising candidate for the development of a potential anti-candida and cytotoxic agent. Abbreviations used: BT-549: Ductal carcinoma; CC: Column chromatography; COSY: Correlations spectroscopy; CDCl₃: Deuterated chloroform; EtOAc: Ethyl acetate; DMSO: Dimethyl sulfoxide; EIMS; Electron-impact mass spectrometry; EP: Ergosterol peroxide; GCMS; Gas chromatography mass spectrometry; HCl: Hydrochloric acid; H₂O: Water; H₂SO₄: Sulfuric acid; HMBC: Heteronuclear multiple bond correlation experiment; HRMS: High-resolution mass spectrometry; HRESIMS: High-resolution electrospray ionization mass spectrometry; HCT-116: Colorectal adenocarcinoma; HSQC: Heteronuclear single-quantum correlation; IC₅₀: Half maximal inhibitory concentration; IR: Infrared; KBr: Potassium bromide; KOH; Potassium hydroxide; LTQ: Linear trap quadruple; MeOH; Methanol; MIC: Minimum inhibitory concentration; MCF-7: Human breast adenocarcinoma; NMR: Nuclear magnetic resonance; PDA: Potato dextrose agar; RP: Reversed phase; SiO₂: Silica gel; TLC: Thin-layer chromatography; VLC: Vacuum liquid chromatography.

Background: Phaleria macrocarpa (PM) has been used conventionally to cure hypertension. Objective: The objective of the study is to determine the active fraction and its chemical composition responsible for antihypertensive activity of PM fruit using a bioactivity-guided investigation on different extracts of the fruit. Materials and Methods: Among the extracts of PM, water extract (WE) showed prominent effect in screening test by inhibiting noninvasive blood pressure (BP) in spontaneously hypertensive rats (SHRs). WE was investigated further using hypertensive and normotensive experimental models. Results: WE caused dose-dependent hypotensive effect in normotensive rats with adrenergic and cholinergic effects by inhibiting the elevated levels of norepinephrine hydrochloride and acetylcholine hydrochloride induced mean arterial pressure (MAP) and heart rate (HR). In addition, WE enhanced the activity of nonselective β-agonist isoprenaline on MAP and inhibited the increased HR. Similarly, WE demonstrated significant inhibition on pulse wave velocity (PWV), MAP, and HR in SHRs. Fractions of WE were tested for vasorelaxation effect on rat aortic explant. Among the fractions, water fraction-4 (PF-4) showed pronounced effects. Column chromatography of PF-4 yielded two subfractions; among them, sub-fraction-2 (SF-2) displayed significant vasorelaxation effect in endothelium-denuded and endothelium-intact aortas. Further, SF-2 revealed significant inhibition in calcium influx and mobilization from intracellular stores. Gas chromatography-mass spectroscopy analysis of SF-2 revealed abundance of kaempferol 3-O-β glucuronide, mangiferin, gallic acid, and rutin. Conclusions: This study demonstrates that polar phytochemical fraction of PM fruit has a promising potential of reducing PWV, BP, and HR. This antihypertensive effect is probably due to the inhibition of arterial tone and extracellular calcium influx. Abbreviations used: WE: Water extract; SHRs: Spontaneously hypertensive rats; NE-HCl: Norepinephrine hydrochloride; MAP: Mean arterial pressure; HR: Heart rate.

Background: Anti-aging health drinks currently are one of the largest consumer drinks. People are more concerned about their health and quality of health life in order to increase longevity. We have investigated that many anti-aging health drink products in Thailand are developed below the standard and do not meet the Thai FDA requirements in terms of bioactivities and toxicity analysis. Objective: This study aimed to evaluate the bioactivity quantities, active ingredients, total phenolic contents, and different extraction methods essential to obtain definite value of bioactivities, quantified antioxidant activity, and also acute toxicity analysis of Clitoria ternatea Linn flower extract drink. Materials and Methods: We performed ultrasonication extraction and maceration with 40% and 50% ethyl alcohol in different periods of time and also analysis of antioxidant activities with 2,2-diphenyl-1-picrylhydrazyl, ferric-reducing antioxidant power, and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid methods, total phenolic compound, active in gradients by high-performance liquid chromatography (HPLC). Results: We found that C. ternatea Linn flower extract drink consisted of high contents of gallic acid and rutin by HPLC method. There was no acute toxicity of C. ternatea Linn flower extract health drink. Abbreviations used: DPPH: 2,2-diphenyl-1-picrylhydrazyl; FRAP: Ferric-reducing antioxidant power; ABTS: 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid; HPLC: High-performance liquid chromatography; EtOH: Ethanol; PTZ: (2,4, 6—tripyridyl—striazine); CH₃OH: Acetic acid; CH₃OH: Methyl alcohol.

Background: Bergenia species contain various therapeutically important compounds such as phenolic compounds-arbutin, bergenin, tannins, gallic acid, flavonoids, minerals, and many others. Bergenia plants show antibacterial, antiviral, anticancer, antidiabetic, diuretic and immunostimulant activities. Materials and Methods: Bergenia leaf extracts from Bergenia crassifolia, Bergenia ciliata, and Bergenia x ornata on lymphocyte activation using flow cytometry method were investigated. Their activation was then monitored with the help of the increasing fluorescence intensity. Results: By activating of cells, there has been an increase in the amount of the established antibodies. Bergenia extracts significantly stimulated the expression of CD69 on lymphocytes in the final concentration of 3.13 and 6.25 mg/ml. The values of stimulation indices of B. crassifolia and B. x ornata extract significantly statistically did not differ. Conclusion: The values of stimulation indices of B. crassifolia and B. x ornata extract significantly statistically did not differ.

Background: Ferula vesceritensis is an indigenous plant of Algerian Sahara riche in sesquiterpene coumarins. Objective: In this study, we investigated the biological activity of sesquiterpene coumarins; coladin, ferulenol, and lapiferin (10_-acetoxy-6_-angeloyloxy-8_,9_-epoxy-trans-caxotan-4_-ol) isolated for the first time from the crude extract CH₂Cl₂—MeOH (1:1) of the roots of F. vesceritensis Coss. et Dur. Materials and Methods: Structures of coladin, ferulenol, and lapiferin were determined by extensive nuclear magnetic resonance (NMR) analyses, including 1D-(¹H and¹³C) and 2D-NMR experiments (correlation spectroscopy, heteronuclear single-quantum coherence, heteronuclear multiple bond correlation (HMBC), and nuclear overhauser effect spectroscopy) as well as high-resolution electron ionization mass spectra and mass spectroscopy analyses. Results: Tested on mouse B16F1 melanoma cells, ferulenol, coladin, and lapiferin exhibited a significant decrease in cell proliferation and a decrease of mitochondrial dehydrogenase activity evaluated on living FAO cells and B16F1 melanoma cells with the WST-1 throughout an apoptotic pathway. They displayed pro-apoptotic effects observed by a decrease in mitochondrial membrane potential and the mitochondrial respiratory rate on isolated liver mitochondria in a dose-dependent manner. Conclusion: Our results highlight the importance of the sesquiterpene coumarins extracted from F. vesceritensis as new biologically active natural products against cancer cells. Abbreviations used: FAO: Hepatoma cell line; B16F1: Pulmonary metastasis melanoma cell line.

Background: Narcissin is well known for their various biological activities. Objective: To recover narcissin from the Opuntia ficus-indica fruits (OFIF), an efficient method was developed by a combination of response surface methodology (RSM) and high-speed countercurrent chromatography (HSCCC). Materials and Methods: Optimization of extraction conditions of narcissin from OFIF was determined using RSM with three-level-three-factor Box-Behnken design (BBD). Then, a rapid and efficient method for the isolation of narcissin from the rich narcissin extracts was developed using HSCCC. Results: Regression analysis showed a good fit of the experimental data and the optimal condition was obtained as extraction time (X₁), 6.02 h; solvent to material ratio (X₂), 8.16 mL/g; and ethanol concentration (X₃), 93.48%. Then, the rich narcissin extracts were separated by HSCCC with a two-phase solvent system composed of n-hexane: ethyl acetate: methanol: water (1.5:5:5:1.5, v/v/v/v) in one step within 60 min. As a result, 12 mg of narcissin was isolated from 100 mg of crude extract with purities of 98.5%, as determined by high-performance liquid chromatography (HPLC). Conclusion: This study can be useful to the development of industrial extraction processes, including further studies concerning the optimal number of sequential steps to enhance the efficacy of a large-scale extraction system. Abbreviations used: BBD: Box-Behnken design; HPLC: High-performance liquid chromatography; HSCCC: High-speed countercurrent chromatography; OFIF: Opuntia ficus-indica fruits; RSM: Response surface methodology

Background: Emodin can ameliorate insulin resistance in diabetes mellitus (DM), but the molecular mechanisms are still uncertain. Objective: The objective of this study is to identify the potential molecular mechanisms of emodin-mediated type 2 DM treatment. Methods: We treated the type 2 diabetic KKAy mice with emodin in different doses. Biochemistry data were collected, and the expression of peroxisome proliferator activated receptor γ (PPARγ) and Glucose transporter (GLUT)-2/4 were examined in liver, muscle, and adipose tissues using immunohistochemistry and reverse transcriptase polymerase chain reaction. The expression of IRS-1, PI3K, pAkt-ser473, and FoxO1 were also tested in these tissues. Results: Our data demonstrated that the levels of cholesterol, higher fasting plasma glucose, total cholesterol, total triacylglycerol, low-density lipoprotein cholesterol, free fatty acid, C creative protein, and tumor necrosis factor-α (P < 0.05), lower high-density lipoprotein, and insulin sensitivity index (P < 0.05) were ameliorated by emodin in a dose-dependent manner (P < 0.05). In addition, emodin was also identified to improve insulin sensitivity in KKAy diabetic mice (P < 0.05). In DM, the expression of PPARγ and GLUT-2 was down-regulated in liver (P < 0.05) as well as in muscle and adipose tissues (P < 0.05) when compared with the controls. However, the decreased levels were subject to emodin treatment in a dose-dependent manner. Meantime, emodin was identified to up-regulate the expression of IRS-1, PI3K, Akt-ser473 (P < 0.05), while FoxO1 (P < 0.05) was down-regulated. Conclusion: These results suggest that emodin represents a promising target to improve insulin sensitivity by enhancing liver glucose utilization, glucose uptake of muscle, and fat through IRS/PI3K/Akt/FoxO1 pathway. Abbreviations used: DM: Diabetes mellitus; PPARγ: Peroxisome proliferator activated receptor γ; GLUT: Glucose transporter; IRS: Insulin receptor substrate; PI3K: Phosphatidylinositide 3 kinase; Akt(PKB): Protein kinase; FoXO1: Fork head box protein O1.

Background: Previous studies reported that Momordica charantia (MC) improves several metabolic parameters, yet outcomes from numerous trials are contradictory. Objectives: This study aimed to assess MC efficacy for improving glycemic status, lipid profile, and body weight. Materials and Methods: The databases included PubMed, Cochrane Register of Controlled Trials, Scopus, CINALH, AMED, ThaiLIS, and Thai Medical Index, from inception to June 2016. References from retrieved articles were also evaluated. For this analysis, we selected randomized placebo versus controlled intervention trials conducted in humans dosed with various forms of MC, excluding studies where patients coadministered other medications. We performed a quality assessment of the retrieved studies using Jadad's scoring and Cochrane's risk of bias assessment. Results: Eight studies (507 participants) met inclusion criteria, which included six randomized controlled trials (RCTs). Meta-analysis revealed a reduction in fasting blood sugar (FBS) (weight mean difference [WMD] −25.03 mg/dL; 95% confidence interval [CI] −41.17,-8.89) and hemoglobin A1C (HbA1C), favoring MC (WMD −0.20%; 95% CI −0.36, −0.04). Similar results were observed for LDL-C (WMD −5.86 mg/dL; 95% CI: −10.83, −0.89), total cholesterol (WMD −6.29 mg/dL; 95% CI: −10.64, −1.93), and triglyceride (WMD −16.22 mg/dL; 95% CI: −26.40, −6.04). Moreover, patients administering MC experienced a significant reduction in body weight (WMD v3.45 kg; 95% CI −6.73, −0.16). Conclusions: MC may improve fasting blood glucose levels, lipid profile, or body weight. A large, well-designed RCT and head-to-head comparison using a standardized preparation of MC will provide definitive data on specific participants. Abbreviations used: ACROBAT: A Cochrane risk of bias assessment tool, WMD: Weight mean difference, CI: Confidence interval, SDs: Standard deviations, FBS: Fasting blood sugar, OGTT: Oral glucose tolerance test level; HDL: High-density lipoprotein, TC: Total cholesterol, TG: Triglyceride, BMI: Body mass index.

Background: The screening of Indonesian medicinal plant extracts against T47D cell line and identifying active chemical constituents of Alpinia galanga rhizome were conducted. Materials and Methods: Thirty methanol extracts of Indonesian medicinal plants were screened for possible anti-breast cancer. The T47D cell line was used as a target model. Chromatography techniques were used to purify the chemical constituents of A. galanga rhizome. Structural elucidation of isolated compounds was conducted by means of 1D and 2 D NMR (nuclear magnetic resonance) spectrometry. Their activities were also examined on MCF7, WiDr, and HeLa cell lines. Results: Five samples (A. galanga, Clonorchis sinensis, Piper cubeba, Santalum album, and Vitex trifolia) showed a strong activity with 96.4%, 91.9%, 87.6%, 82.6%, and 88.7% inhibition, respectively. Since A. galanga exhibited the most potent activity, its IC₅₀value was determined with a dose-dependent effect with the IC₅₀value of 21.2 μg/mL. Its methanol extract was also separated based on chromatography techniques producing the following four compounds: trans-p-coumaryl alcohol (1); p-coumaryl acetate (2); [1'S]-1'-acetoxy chavicol (3); and trans-p-coumaryl diacetate (4). Compounds 3 and 4 showed significant activity with the IC₅₀values of 17.3 and 25.4 μg/mL, respectively. On the other hand, compounds 1 and 2 showed weak activity. All compounds exhibited a similar feature against MCF7, WiDr, and HeLa cell lines. Conclusions: On the structure-activity relationship observation, acetoxy group at a para position could be a key contributor to the effect. Thus, an acetoxy phenylpropanoid could be a good model for future anti-breast cancer lead compound development. Furthermore, extract plants should have demonstrated their potential to inhibit cancer cells. Abbreviations used: TLC: Thin layer chromatography, RPMI: Roswell Park Memorial Institute, NMR: Nuclear magnetic resonance, HMBC: Heteronuclear multiple bond correlations.

Background: Phillyrin and (+)-pinoresinol 4-O-β-D-glucoside are the major active furofuran-type lignans in Fructus Forsythiae. The metabolic routes and metabolites of these two lignans are not well understood yet. Objective: In this study, we attempted to identify the human-intestine bacterial metabolites of lignans from Fructus Forsythiae. Materials and Methods: Two natural compounds, phillyrin and (+)-pinoresinol 4-O-β-D-glucoside were incubated with human fecal microflora in an anaerobic incubator for 72 h and the metabolites with highly sensitive ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) were analyzed. Results: As a result, nine metabolites were determined using a Syncronis^™ C₁₈column (particle size 1.7 mm) in a gradient elution system. These metabolites were then identified according to the mass fragmental mechanism, MS/MS fragment ions, and previous publications. The results of this study indicated that the major metabolites of furofuran-type lignans are through the processes of hydrolysis, demethylation, reduction, dehydroxylation, and oxidation. Conclusions: Lignans can be metabolized by intestinal microbiota and the intestinal bacteria play a critical role in the metabolism of components administered orally. Abbreviations used: GAM: General anaerobic medium; UPLC/Q-TOF-MS: Ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry; ESI: Negative ion electrospray; RT: Retention time; TCM: Traditional Chinese Medicine.

Background: Studies have reported that depression is associated with increased level of oxidative stress and accompanied with decreased Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1-nuclear factor (erythroid-derived 2)-like 2 (Keap1-Nrf2) signaling pathway. Berberine has been shown to possess properties on protection against neurodegenerative diseases, but its effects on chronic unpredictable mild stress (CUMS) induced depressive such as behaviors and Keap1-Nrf2 signaling pathway are unknown. Objective: This study was designed to evaluate the effect of berberine on CUMS induced depression animal model and investigate the underlying mechanisms. Materials and Methods: We established CUMS depressant rats model and treated CUMS rats with berberine. Sucrose preference test, forced swim test , and tail suspension test were used to measure behavioral changes. We used quantitative polymerase chain reaction and western blot to test the levels of cytokines in the hippocampus. Results: We found that CUMS rats displayed obvious depressive-like behaviors. Moreover, berberine treatment prevented depressive behaviors in CUMS rats accompanied with suppression of oxidative stress markers. Further experiments showed that berberine treatment up-regulated the expression of Keap1-Nrf2 antioxidant signaling pathway and its downstream neuroprotective factors. Conclusion: The present results suggested that treatment of berberine significantly ameliorated depressive-like behaviors in CUMS rats and enhanced Keap1-Nrf2 antioxidant signaling pathways in hippocampus. Abbreviations used: MDD: Major depressive disorder; CUMS: Chronic unpredictable mild stress; FST: Forced swim test; TST: Tail suspension test; Keap1: Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1; Nrf2: nuclear factor (erythroid-derived 2)-like 2; ARE: Antioxidant response elements; SOD: Superoxide Dismutase; MDA: Malonic dialdehyde; IMI: Imipramine.

Background: Hypericin is an anthraquinone derivative of Hypericum hookerianum, a shrub from Palni hills of Southern India which possess various medicinal values. Aim: The present study was aimed to evaluate the anti-inflammatory activity of hypericin using RAW 264.7 macrophage cell line. Materials and Methods: The anti inflammatory property was determined by assessing the inhibitory action on lipopolysaccharide (LPS) stimulated nitric oxide (NO) production and pro inflammatory mediators/cytokines. RAW 264.7 macrophages cell line was used as in vitro inflammatory model. Cytotoxicity was evaluated using methylthiazolyldiphenyl tetrazolium bromide assay and NO estimation was carried out using Griess method. Gene expression of pro—inflammatory cytokines such as inducible nitric oxide synthase, cyclooxygenase 2, tumor necrosis factor α, interleukin—1 β (IL—1 β), and IL—6 was also evaluated. Results: The results exhibited that hypericin significantly suppressed LPS induced NO production with a concomitant decrease in the levels of pro—inflammatory cytokines. Conclusion: The study demonstrates the potential of hypericin as an effective anti—inflammatory agent. Abbreviations used: NSAID: Nonsteroidal anti-inflammatory drug; NO: Nitric oxide; IL — 6: Interleukin — 6; IL — 1β: Interleukin — 1β; TNF — α: Tumour necrosis factor — α; iNOS: inducible Nitric oxide synthase; COX — 2: Cyclooxygenase - 2; NF-κB: nuclear factor kappa B; PGE2: prostaglandin E2; LPS: Lipopolysaccharide; DMEM: Phenol free Dulbecco's modified Eagle medium; MTT: methyl thiazolyl diphenyl tetrazolium bromide; DMSO: Dimethyl sulfoxide; PBS: phosphate buffer saline; RNA: ribonucleic acid; q-PCR: quantitative polymerase chain reaction.

Background: The seed of Cuscuta chinensis Lam. (Cuscuta, Convolvulaceae), is widely used in traditional Chinese medicine, with a variety of biological activity. However, the efficacy of C. chinensis has not been investigated in renal injury caused by sepsis. Objective: Our research was carried out to evaluate the effect of ethanol extract of C. chinensis seeds (ECCS) on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) in Kunming mice, and explore its underlying mechanisms. Materials and Methods: ECCS was administered orally (10, 30, and 100 mg/kg bodyweight, once daily) to the AKI mice for 14 consecutive days. On the 14^th day, LPS (7 mg/kg) was administered to induce AKI. Blood urea nitrogen (BUN), creatinine (Cr), cytokines, and antioxidant index were estimated. The protein phosphorylation was tested by Western blot. Results: Results indicated that ECCS (100 mg/kg) decreased LPS-induced augmented BUN by 58.54% (P < 0.001) and reduced Cr by 61.57% (P < 0.01), respectively. In addition, the evident reversion of renal pathological damage was observed in ECCS-pretreated mice. ECCS (100 mg/kg) also exhibited a reduction of cytokines (tumor necrosis factor-α, interleukin [IL]-1 β, and IL-6, P < 0.05 for all) in AKI mice. Furthermore, ECCS blocked the activation of the nuclear factor-κB (NF-κB) (p65 and inhibitor kappa B subunit) and reduced the toll-like receptors 4 (TLR4) expression. Conclusions: The protective effect of ECCS against LPS-induced AKI was at least partially associated with suppressing of a TLR4-NF-κB signaling pathway, which provides evidence of the renal protective function of C. chinensis extract. Abbreviations used: AKI: Acute kidney injury; LPS: Lipopolysaccharide; NF κB: Nuclear factor κB; IκB: Inhibitor kappa B; IL: Interleukin; ECCS: Ethanol extract of C. chinensis seed; SOD: Superoxide dismutase; TLR4: Toll like receptor 4; BUN: Blood urea nitrogen; Cr: Creatinine; MDA: Malondialdehyde; PVDF: Polyvinylidene fluoride; TBST: TBS-Tween solution; iNOS: Inducible NOS; ROS: Reactive oxygen species; TH: T Helper; TBHB: Tert-butyl hydroperoxide; IFN-γ: Interferon-γ.

Background: Excessive osteoclast formation and over-activated function lead to a series of osteoclast-related diseases. Suppression of osteoclastogenesis is likely to be an effective means for the treatment of these diseases. Objective: In this study, we investigated the effects of alpha-mangostin (α-MAG), a natural compound derived from Garcinia mangostana, on osteoclast formation and function in RAW264.7 cells. Materials and Methods: Different concentrations of α-MAG were used to explore its effects on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis and further, we investigated the mechanism by Western Blotting. Results: The study revealed that α-MAG attenuated RANKL-induced osteoclastogenesis. Moreover, the effects were confirmed to be caused by the suppression of the phosphorylation of the extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) signaling and this inhibitory effect could be rescued by the administration of the JNK and p38 agonist anisomycin. Conclusion: The study results demonstrated that α-MAG could impair RANKL-induced osteoclastogenesis by inhibiting the ERK and JNK signaling pathways in RAW264.7 cells. Abbreviations used: α-MAG: α-mangostin, RANKL: Receptor activator of nuclear factor-κB ligand, ERK: Extracellular signal-regulated kinase, JNK: c-Jun N-terminal kinase, ANI: Anisomycin, M-CSF: Macrophage colony-stimulating factor, MAPK: Mitogen-activated protein kinase, IκBα: Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha, NFATc1: Nuclear factor of activated T cells c1, TRAP: Tartrate-resistant acid phosphatase, SEM: Scanning electron microscope, CTSK: Cathepsin K, CTR: Calcitonin receptor, DC-STAMP: Dendritic cell-specific transmembrane protein, TRAF6: Tumor necrosis factor receptor-associated factor 6, AP-1: Activator protein-1

Background: Hypecoum leptocarpum Hook. f. et Thoms., which is used in traditional Tibetan medicine as an antipyretic, antitussive, analgesic, and anti-inflammatory agent, contains a variety of alkaloids that could be responsible for its analgesic and anti-inflammatory properties. Objective: The present study was designed to investigate the anti-inflammatory activity of the total alkaloids from H. leptocarpum (AHL) in vitro and to elucidate the chemical structure of the anti-inflammatory components in AHL. Materials and Methods: Chemical characterization was performed using liquid chromatography/quadrupole-time-of-flight mass and diode-array detector-high performance liquid chromatography. The anti-inflammatory effects of AHL were investigated by measuring the production of inflammatory cytokines using enzyme-linked immunosorbent assay and mRNA expression by real-time polymerase chain reaction in lipopolysaccharide-induced RAW 264.7 macrophages. Results: Chemical analysis of AHL revealed the presence of seven alkaloids, protopine (13.3%), cryptopine (1.5%), leptopidinine, leptocarpine, corydamine, dihydroleptopine, and oxohydrastinine. AHL significantly suppressed the production of nitric oxide (NO), interleukin-1 beta (IL-1 β), IL-6, and tumor necrosis factor-alpha (TNF-α) in LPS-induced RAW 264.7 cells. The maximum levels of suppression of NO, IL-1 β, IL-6, and TNF-α were 86.8% ± 2.2%, 70.1% ± 1.5%, 100.1% ± 2.5%, and 50.8% ± 3.6%, respectively. IC₅₀values of suppression of cytokine production by AHL were 7.47 ± 2.81 μg/mL (NO), 0.12 ± 0.28 μg/mL (IL-1 β), 0.56 ± 0.37 μg/mL (IL-6), and 18.95 ± 5.23 μg/mL (TNF-α). AHL was also shown to downregulate mRNA expression of inducible NO synthase, IL-1 β, IL-6, and TNF-α in vitro. Conclusion: The study provides convincing evidence that AHL has strong anti-inflammatory activity. The potent activity is likely a result of synergy between the different alkaloids. Abbreviations used: The total alkaloids from H. leptocarpum: AHL; Nitric oxide: NO; Interleukin-1 beta IL-1β; Interleukin-6: IL-6; Tumor necrosis factor-alpha: TNF-α; Prostaglandin E2: PGE2; Inducible nitric oxide synthase: iNOS; Nonsteroidal anti-inflammatory drugs: NSAIDs; lipopolysaccharide: LPS; The total ion chromatograms: TIC; The liquid chromatography/quadrupole-time of flight: LC/Q-TOF; Nuclear factor-kappa B: NF-κB; Janus kinase-signal transducers and activators of transcription: JAK-STAT.

Background: Cordyceps militaris and Cordyceps sinensis belong to the same genus, but the different species, with immunity improvement, antibacterial, and antihypertensive effect, and the studies on the functions of C. militaris mainly primarily focus on those of its polysaccharides and polypeptides currently. The latest studies have found that some of the polypeptides with immunomodulatory effect can widely regulate immune functions at multiple levels, improve immunity, and enhance immune functions to ensure the healthy body, showing an important significance. Materials and Methods: C. militaris polypeptide prepared with the enzymolysis method was taken as the research object in this study. The differentially expressed genes and the related cell signal transduction pathway were screened by mRNA expression microarray. STEM software V1.3.6 (short time-series expression miner, http://www.cs.cmu.edu/jernst/stem/) was used for the clustering of the gene functions, and David and KEGG database were applied for the analysis of the related functions. Results: One thousand seven hundred and forty-eight differentially expressed genes were selected finally and three of them were validated by quantitative polymerase chain reaction. The results showed that gene Hist1h2bp, Ctsg, and Elane were involved in the regulation of C. militaris on the immune activity of mice. Conclusion: Gene Hist1h2bp, Ctsg, and Elane may be the potential targets of C. militaris polypeptide, which may provide an important theory basis for the further research and development of C. militaris polypeptide. Abbreviations used: C. militaris: Cordyceps militaris; ConA: Concanavalin A; SRBC: Sheep red blood cells; PCR: Polymerase chain reaction; RT PCR: Real time quantitative; HC50: Half values of hemolysin; CMP: Cordyceps militaris polypeptide; SLE: Systemic lupus erythematosus; GO: Gene Ontology.

Background: The plant Euphorbia esula has been used by ancient Chinese people to treat cancer and other ailments and is used by present doctors or folks as an assistant treatment for some kind of cancers, but the mode of action is unclear. Objective: To investigate the influence of E. esula full extract on chemoresistance, cell migration/invasion, and apoptosis of multidrug-resistant human stomach cancer cells. Materials and Methods: After treating multidrug-resistant human stomach cancer SGC7901/VCR cells with E. esula extract at varying concentrations, the inhibition of cell proliferation was detected using thiazolyl blue. Sensitivity to the chemotherapeutic drugs, adriamycin and paclitaxel, was evaluated using half maximal inhibitory concentration. Cell cycle progression was analyzed by flow cytometry. The inhibitions of cell migration and invasion were examined by Transwell method. The induction of apoptosis and the apoptotic rate were studied by electron microscopy and flow cytometry, respectively. Activation of caspase 3 enzyme was inspected by ultraviolet spectrophotometry. Results: E. esula extract could increase the sensitivity of SGC7901/VCR cells to the chemotherapeutic drugs, adriamycin and paclitaxel. The proliferation, migration, and invasion of SGC7901/VCR cells were significantly inhibited by E. esula extract (P < 0.01 compared with negative control), which showed time- and dose-dependent manners (P < 0.05 and P < 0.01, respectively). Cell cycle was arrest at the S phase. E. esula extract also induced apoptosis in SGC7901/VCR cells, and the apoptotic rate was increased significantly with drug concentration and with treatment time (P < 0.01 compared with negative control, P < 0.05 between concentrations and time points). E. esula extract upregulated enzymatic activity of caspase 3. Conclusion: E. esula extract could reverse SGC7901/VCR cell's resistance to chemotherapeutic drugs and could inhibit cell proliferation, migration, and invasion, interfere with cell cycle progression, and induce apoptosis in SGC7901/VCR cells. Abbreviations used: E. esula: Euphorbia Esula; FCS: fetal calf serum; IC50: half maximal inhibitory concentration; OD: optical density.

Background: Puerariae Lobatae Radix is widely used in the pharmaceutical, food, and cosmetic industries. For these applications, when grading roots according to diameter, thick roots are always considered to be of better quality. Objective: The objective of this study is to qualitative and quantitative analysis of isoflavonoids profiling in various tissues of P. lobata roots. Materials and Methods: Ultraperformance liquid chromatography quadrupole/time-of-flight-mass spectrometry (UPLC-QTOF/MS) and high-performance liquid chromatography separation and ultraviolet-visible detection were used to identify and profile the isoflavonoids detected in various tissues of P. lobata roots. Results: Consequently, 82 peaks were detected by UPLC-QTOF/MS, and 28 isoflavonoid compounds were identified in the various tissue samples. According to the results of this study, the thick roots of P. lobata are of better quality for medical use than the thin ones. The fine roots and the periderm of roots contained slightly lower isoflavonoid abundances. The results indicated that isoflavonoids, particularly puerarin, mainly accumulated in the xylem and phloem. Conclusions: This study provides a new and practical method for evaluating the quality of P. lobatae Radix. Abbreviations used: ANOVA: One-way analysis of variance; CNKI: Chinese National Knowledge Infrastructure; HPLC-UV: High performance liquid chromatography separation and ultraviolet-visible detection; PPRC: Pharmacopoeia of the People's Republic of China; RSD: Relative standard deviation; UPLC-QTOF/MS: Ultraperformance liquid chromatography quadrupole/time-of-flight-mass spectrometry.

Background: Previous phytochemical studies showed that extracts from leaves and seed oil of Catalpa plants showed antioxidant and antitumor activities. However, the active components and their roles and molecular mechanisms were still incompletely identified. Objective: We aimed to extract and identify the active fraction from Catalpa bungei leaves and investigate its underlying mechanism in suppressing cervical cancer cell survival. Materials and Methods: Extracts from C. bungei leaves with 80% methanol were separated into different fractions. The component with optimal inhibitory effect on HeLa cell growth was isolated, identified, and examined. Results: Extracts from C. bungei leaves with methanol were separated into petroleum ether, ethyl acetate (EA), n-butanol, and water fractions. The chemicals in EA fraction exhibited optimal inhibition effects, which were further separated into 29 fractions by the silica column chromatography. The compound with optimal antitumor activity was eventually determined to be ursolic acid (UA) based on the results of Sephadex column chromatography and¹H nuclear magnetic resonance analysis. UA at 5.0 and 10.0 μg/mL substantially inhibited the growth and migration of HeLa cells. UA also retarded cell cycle at G0/G1 stage and promoted cell apoptosis through activating death receptor and mitochondria-associated pathways. Conclusion: UA isolated from C. bungei leaves could inhibit growth and migration and induce apoptosis in HeLa cells. Abbreviations used: UA: Ursolic acid; EA: Ethyl acetate; HPV: Human papillomavirus, PARP: Poly ADP-ribose polymeras; ROS: Reactive oxygen species; PBS: Phosphate buffered saline; SRB: Sulfonyl rhodamine B; DMSO: Dimethyl sulfoxide; TLC: Thin layer chromatography; NMR: Nuclear magnetic resonance; CAS: Chemical abstracts service.

Background: Catha edulis is an evergreen plant belonging to the Celastraceae family, used by the people of Africa, Yemen, and some part of Saudi Arabia as a recreational plant. They chew it alone or in combination with alcohol for attaining euphoria. Objective: There are contradictory studies in the genotoxic potential of the leaves of this plant. So far, the accomplished studies were conducted with much high doses. Hence, the current study has been designed to check the genotoxicity of khat-leaves extract with subcytotoxic concentration in human peripheral blood lymphocytes. Materials and Methods: We have employed comet assay, micronuclei (MN) analysis, and Ames test to evaluate the objectives. Results: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay has revealed that there is a dose-dependent reduction in cell viability with an IC₅₀value of 76.5 μg/ml at 24-h treatment. The results of the comet assay have showed a nonsignificant difference between control and treatment up to 40 μg/ml extract. There was no dose-dependent increase in the frequency of binucleated cells with MN and reduction of cell proliferation in all the doses. The extract showed no mutagenic effect, in both TA98 and TA100 strains even with S9 activation. Conclusion: Under the experimental conditions employed in the present study, khat has been found to be nongenotoxic at subcytotoxic level. Abbreviations used: DMSO: Dimethyl sulfoxide; GIT: gastrointestinal tract; HPBL: Human peripheral blood lymphocytes; KCL: Potassium chloride; LCQ: Low-content quantification; MMC: Mitomycin C; RPMI: Roswell park memorial institute; SWGDRUG: Scientific working group for seized drugs.

Background: Fingerprint analysis plays a key role in quality control of herbal medicines due to its technical capacity to represent the chemical diversity of these complex matrices. Several traditional Brazilian species showed very little data in their chemical profiles. Objective: Thus, the purpose of this study was to evaluate the chemical profiles of Brazilian herbal species by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Materials and Methods: The herbal materials of 7 species (Anacardium occidentale, Annona muricata, Guazuma ulmifolia, Phyllanthus niruri, Psidium guajava, Punica granatum, and Spondias mombin) were collected from three different locations in Northeast Brazil, botanically authenticated and their chemical profile analyzed by TLC (cinnamics, flavonoids, and tannins) and by HPLC (polyphenols). Results: The chromatographic data showed the similarities between chemical profiles of the sample fingerprints, confirming the presence of several classes of secondary metabolites as well as the identification of different chemical standards (catechin, chlorogenic acid, caffeic acid, ellagic acid, gallic acid, quercetin, or rutin). Conclusion: The chromatographic profiling of the herbal drugs by TLC and HPLC were successfully characterized and allowed for the identification of promising chemical markers, improving the state of art in the quality control of the herbal species investigated in this study. Abbreviations used: HPLC: High-performance liquid chromatography; PVDF: Polyvinylidene fluoride; RP-LC: Reverse-phase liquid chromatography; TLC: Thin-layer chromatography; UV-spectrum: Ultraviolet spectrum.

Background: Pouzolzia zeylanica (L.) Benn. (Family: Urticaceae), which is widely distributed in China, is consumed as food and as a traditional herbal medicine for the treatment of numerous diseases. Objective: The aim of the present study was to explore the antioxidant compounds of P. zeylanica. Materials and Methods: A petroleum ether extract, an ethyl acetate extract (EAE), an n-butanol extract, and the remaining part were separated by liquid—liquid extraction from a 90% aqueous ethanol extract and a water extract of P. zeylanica and were evaluated using two complementary antioxidant assays, namely 2,2-diphenyl-1-picrylhydrazyl-free radical scavenging and ferric-reducing antioxidant power assays. Results: The EAE, which possessed the highest antioxidant activity, contained the highest total phenolic content (263.5 ± 4.8 mg/g) and total flavonoid content (388.3 ± 5.6 mg/g). In addition, three compounds with antioxidant activity, quercetin, kaempferol, and N-[2-(3-hydroxy-4-methoxyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl) prop-2-enamide, were isolated from the EAE. Conclusions: These results demonstrate that the flavonoids and phenolic compounds from P. zeylanica could be used as natural antioxidants. Abbreviations used: P. zeylanica: Pouzolzia zeylanica (L.) Benn., PE: Petroleum ether extract, EAE: Ethyl acetate extract, BE: n-butanol extract, RE: Remaining part, WE: Water extract, DPPH: 2,2-Diphenyl-1-picrylhydrazyl, FRAP: Ferric-reducing antioxidant power, TPC: Total phenolic content, TFC: Total flavonoid content, BHA: Butylated hydroxyanisole, BHT: Butylated hydroxytoluene, TPTZ: 2,4,6-Tris (2-pyridyl)-s-triazine, HPLC: High-performance liquid chromatography, CC: Column chromatography.

Background: Prosopis glandulosa Torr. (Fabaceae) is native to Africa, America, and Asia. In India, it is abundantly found in southern parts. The plant is reported to contain polyphenols, flavonoids, mesquitol-catechin dimers, indolizidine alkaloids, and triterpenes, imparting antitumor, antimicrobial, anti-infective, and anti-parasitic activities. Aim: The present study deals with the isolation and characterization of flavonoid C-glycosides from P. glandulosa Torr. leaves. Materials and Methods: P. glandulosa leaves were subjected to hot aqueous extraction at 90°C—100°C to yield crude aqueous extract (PGA). PGA was fractionated and then subjected to chromatography over Diaion^® HP-20 column and semi-preparative high-performance liquid chromatography to provide flavonoid C-glycosides. Results: We have identified and characterized vicenin-2/isomer and schaftoside (s) from P. glandulosa leaves through liquid chromatography-electron spray ionization-mass spectrometry/mass spectrometry (LC-MS/MS) and nuclear magnetic resonance. LC-MS/MS chromatogram displayed peaks at retention time of 9.69 and 10.37 with m/z 594 and major fragments at m/z 473 [M-H-120]⁻ (loss of glucose unit) for “vicenin-2/isomer.” Interestingly, five major peaks were observed with retention times of 10.75, 11.17, 11.92 (major), 12.92, and 13.32 corresponding to the same mass of m/z 564 with major fragments at m/z 473 [M-H-90]⁻ (loss of arabinose unit) and 443 [M-H-120]⁻ (loss of glucosyl unit), together termed as “schaftosides.” Conclusion: To the best of our knowledge, this is the first report on flavonoid C-glycosides (vicenin-2/isomer and Schaftosides) from P. glandulosa leaves which may act as chemomarker(s) for the standardization and quality control of this plant. Abbreviations used: PGA: Aqueous extract of Prosopis glandulosa leaves; PGM: Methanol fraction of PGA; PGAM: Aqueous-methanolic fraction of PGA; LC-ESI-MS/MS: Liquid chromatography-electron spray ionization-mass spectrometry/mass spectrometry

Background: Atractylodes japonica has been commonly used to treat gastrointestinal (GI) disorders in Korean traditional medicine. Interstitial cells of Cajal (ICCs) are pacemaker cells in the GI tract and can regulate GI motility. Objective: To investigate the effects of the extract of Atractylodes japonica (AJE) on pacemaker potentials generated by ICCs from murine small intestine. Materials and Methods: Enzymatic digestion was performed to dissociate ICCs. All experiments on ICCs were performed after 12 h of culture. The whole-cell patch-clamp configuration was used to record pacemaker potentials generated by ICC. Results: AJE (0.1—1 mg/mL) depolarized pacemaker potentials in a concentration-dependent manner and decreased the amplitudes of pacemaker potentials at all concentrations in the current-clamp mode. Pretreatment with Y25130 (a 5-HT₃receptor antagonist), RS39604 (a 5-HT₄receptor antagonist), or SB269970 (a 5-HT₇receptor antagonist) had no effects on depolarization of pacemaker potentials induced by AJE. In addition, pretreatment with 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide (a muscarinic M₃receptor antagonist) inhibited AJE-induced depolarization of pacemaker potential; however, pretreatment with methoctramine (a muscarinic M₂receptor antagonist) did not affect depolarization of pacemaker potentials induced by AJE. In the presence of an external Na⁺-free solution, the pacemaker potentials decreased, and under this condition, AJE did not depolarize the pacemaker potentials. Flufenamic acid, a nonselective cation channel (NSCC) blocker, decreased the pacemaker potential, which in turn inhibited AJE-induced depolarization of pacemaker potential. Conclusion: The results of this study suggest that AJE depolarized the pacemaker potentials generated by ICC by stimulating muscarinic M₃receptors, but not 5-HT receptors, through NSCCs. Therefore, AJE can be a novel prokinetic agent. Abbreviations used: ICCs:Interstitial cells of Cajal, GI:Gastrointestinal, NSCC:Non-selective cation channel, TRP:Transient receptor potential.

Objective: The capitulum of Chrysanthemum indicum (Compositae) is the source of a crude drug used in Japanese traditional Kampo and traditional Chinese medicine. Previous research showed that C. indicum flowers promote adipocyte differentiation through the activation of peroxisome proliferator-activated receptor (PPAR)-γ that may be an important mechanism for controlling systemic insulin resistance or other biological functions. Materials and Methods: The capitulum of C. indicum was extracted with methanol, and its PPAR-γ agonistic activity was measured using luciferase assay. The active ingredients were isolated by the activity-guided fractionations. Results: We isolated (Z)-tonghaosu and (E)-tonghaosu, and (E)-tonghaosu had PPAR-γ agonistic constituents by the activity-guided fractionation from the capitulum of C. indicum. The isomer (Z)-tonghaosu did not show PPAR-γ-agonistic activity. Conclusion: (E)-tonghaosu is one of the active ingredients as PPAR-γ agonist in the capitulum of C. indicum. Abbreviations used: ANOVA: One-way analysis of variance; DMEM: Dulbecco's modified minimum essential medium; FBS: Fetal bovine serum; HPLC: High-performance liquid chromatography; MCI: Methanol extract of the capitulum of Chrysanthemum indicum; NMR: Nuclear magnetic resonance; PPAR: Peroxisome proliferator-activated receptor.

Background: Boeravinone B (BB), a therapeutic biomarker isolated from roots of Boerhavia diffusa, possesses immunomodulatory properties. Objective: In the present study, we investigated cytotoxicity and immunomodulatory effect of BB on human dendritic cells (DCs). Materials and Methods: In this study, buffy coat of human healthy volunteers was collected and the monocytes were separated. Effect of various concentrations of BB on DCs from induced differentiation of monocytes was studied on day 7 using ELISA microplate reader. The effect of BB on the immature DCs generated in the presence and absence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin4 (IL4) was also noted. The binding of BB with the DC maturation markers CD80, CD83, and CD86 including antigen uptake were evaluated using fluorescent-activated cell sorter (FACS). Results: Viability of DCs was not affected at BB concentration of 100 μg/ml. Phytohemagglutinin (PHA), BB showed increase in DC expression of CD80 expression up to 6.7% (P < 0.001) and 7.27% (P < 0.001) and increase in antigen uptake up to 7.67% (P < 0.001) and 8.87% (P < 0.001) when compared with dimethyl sulfoxide control. PHA and Boeravinone B supplemented with GM-CSF and IL4 showed increase of CD83 expression up to 6.86% (P < 0.01) and 4.63% (P < 0.05). However, PHA and BB showed increase of CD86 expression up to 6.6% (P < 0.01) and 6.73% (P < 0.01) when compared with control. Conclusion: The results justify that BB induced differentiation of monocytes into mature DCs. Abbreviations used: DCs: Dendritic cells, ELISA: Enzyme-linked immunosorbent assay, GMCSF: Granulocyte-macrophage colony-stimulating factor, IL4: Interleukin4, PHA: Phytohemagglutinin, CD: Cluster of differentiation, DMSO: Dimethyl sulfoxide, FACS: Fluorescent activated cell sorter.