Background: Baihe Zhimu Tang (BZT) is a widely used traditional Chinese medicinal formula in treating various diseases; however, its active components have remained unknown. Materials and Methods: Based on the metabolic fingerprinting technique and liquid chromatography/diode array detection-electrospray ionization-mass spectrometry (LC/DAD-ESI-MS), a method for rapid screening and analysis of the multiple absorbed bioactive components and metabolites of an oral solution of Baihe Zhimu Tang (BZT) in rabbit plasma, urine and feces after oral administration of BZT was developed. Results: The results obtained from a comprehensive comparative analysis of the fingerprints of the BZT and its metabolic fingerprints in rabbit biological samples indicated that 19 components in the BZT were absorbed into the rabbit's body. Both of them were tentatively identified from their MS and UV spectra and retention behaviors by comparing the results with the reported literature. In addition, only six components were found in the metabolic fingerprints, which suggested that they might be metabolites of some components in the BZT. Conclusion: The findings demonstrated that the proposed method could be used to rapidly and simultaneously analyze and screen the multiple absorbed bioactive constituents and metabolites in a formula of traditional Chinese medicines (TCMs) by comparing and contrasting the chromatographic fingerprints with its metabolic fingerprints. This is very important not only for the pharmaceutical discovery process and the quality control of crude drugs, but also for explaining the curative mechanism of TCMs.

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Background: A decoction prepared with barks of Adenanthera pavonina and Thespesia populnea is a herbal formulation which has been prescribed in Sri Lanka in the treatment of cancer patients for many years. This study was designed to investigate its phytochemical and antioxidant properties. Materials and Methods: The total phenolics and flavonoids were determined using Folin-Ciocalteau and aluminum chloride colorimetric methods, respectively. Gallic acid content in the decoction was determined using high-performance liquid chromatography. The antioxidant properties were evaluated by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical assay, nitric oxide scavenging assay, deoxyribose method, and the reducing power of the decoction. Results: The concentration of total phenols, flavonoids, and gallic acid of the decoction were 34.13 ± 3.54 w/w % gallic acid equivalents, 41.37 ± 0.57 w/w % of (-)-Epigallocatechin gallate equivalents, and 0.58 ± 0.24 mg/g, respectively. The EC50 for DPPH, nitric oxide scavenging, and deoxyribose assays were 7.24 ± 0.50, 14.02 ± 0.66, and 53.21 ± 2.82 μg/ml, respectively. Reducing power of the decoction increased with the concentration. Conclusion: These investigations suggested that the decoction prepared with A. pavonina and T. populnea can be a potential source of nutraceuticals with antioxidant activity.

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Background: In this study, the antidepressant activity of ethanolic extract of Pinus eldarica Medw needles was assessed using forced swimming test (FST) in rats. Materials and Methods: Male Wistar rats were randomly divided into six Groups and treated as follows: first group was received only reserpine (6 mg/kg, i.p.), second group was received reserpine (6 mg/kg, i.p.) and imipramine (10 mg/kg, i.p.), three experimental groups received reserpine (6 mg/kg, i.p.) and three doses of pine needle extract (100, 300, and 500 mg/kg, p.o.) respectively and the final group (control group) received only vehicle (5% DMSO, i.p.). Results: Acute oral administration of ethanolic extract of P. eldarica Medw needles at a dosage of 300 mg/kg reduced reserpine-induced increase in immobility time in the FST, demonstrating an antidepressant effect in the FST. Additionally, extract treatment did not modify the ambulation and rearing evaluated in open field test, indicating that antidepressant effect found in the forced swimming test was not based on the stimulation of locomotor activity. Conclusion: These results indicate that ethanolic extract of Pinus eldarica needles possesses an antidepressant activity.

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Context: In irritable bowel syndrome (IBS), disturbance of bowel motility is associated with infiltration of inflammatory mediators and cytokines into the intestine, such as neutrophils, myeloperoxidase (MPO), tumor necrosis factor alfa (TNF-α), and lipid peroxide. Aims: Regarding promising anti-inflammatory and anti-oxidative effects of Hypericum perforatum (HP) extract, besides its anti-depressant effect, this study was designed to evaluate the effects of HP in an experimental model of IBS. Settings and Design: IBS was induced by a 5-day restraint stress in rats. The HP extract was administered by gavage in doses of 150, 300, and 450 mg/kg for 26 days. Fluoxetine and loperamide were used as positive controls. Gastric emptying and small bowel and colon transit, besides the levels of TNF-α, MPO, lipid peroxidation, and antioxidant power, were determined in colon homogenates. Statistical Analysis Used: Data were analyzed by one-way ANOVA followed by Tukey's post hoc test for multiple comparisons. Results: A significant reduction in small bowel and colonic transit (450 mg/kg), TNF-α, MPO, and lipid peroxidation and an increase in antioxidant power in all HP-treated groups (150, 300, and 450 mg/kg) were seen as compared with the control group. Gastric emptying did not alter significantly when compared with the control group. Treatment with loperamide (10 mg/kg) significantly inhibited gastric emptying and small bowel and colonic transit, while flouxetine (10 mg/kg) decreased gastric emptying, TNF-α, MPO, and lipid peroxidation and increased the antioxidant power of the samples in comparison with the control group. Conclusions: HP diminished the recruitment of inflammatory cells and TNF-α following restraint stress not in a dose-dependent manner, possibly via inhibition of MPO activity and increasing colon antioxidant power, without any difference with fluoxetine. The HP extract inhibits small bowel and colonic transit acceleration like loperamide but has minimal effect on gastric emptying.

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Background: Aconitum napellus (Acn) is used topically to relieve pain, itching and inflammation, and internally to reduce febrile states, among others. Any circadian time-related consequences of Acn administration are unknown. The objective of this study was to explore the effects of two doses of Acn on body temperature (BT) of mice treated at six different times over 24 hours. Materials and Methods: BALB/c female mice were housed in six chambers (six mice each) with air temperature 24 ± 3°C, humidity 60 ± 4%, and a 12-hours light (L)/12-hours dark cycle, but with L-onset staggered by 4 hours between chambers so that study at one external test time resulted in six test times (02, 06, 10, 14, 18 and 22 hours [h] after light onset). Rectal temperature (RT; in °C) was measured at baseline (B) and 1 hour after oral treatment with placebo (P) or two doses of Acn (6C and 30C, two studies each) in six studies over an 8 day span. The difference in RT for each mouse from the respective B + P timepoint mean RT was computed following each Acn treatment, and data from each of the six studies (original RT and difference from B + P) were analyzed for time-effect by analysis of variance (ANOVA) and for circadian rhythm by 24-hour cosine fitting. Results: A circadian rhythm in RT was found at B and after P (mean: 35.58°C vs. 35.69°C; peak: 15:31 h vs. 15:40 h) and after each Acn dose (30C or 6C). Acn induced hyperthermia and the overall change in BT was rhythmically significant for each dose (mean = +1.95°C vs. +1.70°C), with greatest hyperthermia observed during the L-span for each dose (peak = 08:56 h vs. 05:17 h). Conclusion: Acn administered around the clock induced hyperthermia overall and in a time-dependent manner, with greatest effects during the resting (L) span. Thus, time of day may significantly impact the outcome of Acn and other homeopathic treatments and should be considered in determining optimal dosing and treatment time(s) in order to increase the desired outcome and decrease undesired effects.

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Background: Phyllanthus (Euphorbiaceae) species have long been used in folk medicine to treat various pathological conditions including liver diseases. Some species of Phyllanthus were found to exhibit hepatoprotective activity against drugs or toxins and this property was majorly attributed to phyllanthin and hypophyllanthin. In this study, we examined the hepatoprotective activity of five different species of Phyllanthus, namely, Phyllanthus amarus, Phyllanthus fraternus, Phyllanthus maderaspatensis, Phyllanthus urinaria, and Phyllanthus Rotundifolius. The extracts were also evaluated for the presence of key phytoconstituents, phyllanthin and hypophyllanthin. Materials and Methods: The extracts were evaluated for hepatoprotective activity against tert-butyl hydroxide (t-BH)-induced cytotoxicity using human hepatocarcinoma cells (HepG2 cell line). Results: Only P. urinaria and P. maderaspatensis exhibited significant hepatoprotective activity as evident from increased cell viability. The HPLC profile revealed that except P. amarus, the other extracts did not contain phyllanthin and hypophyllanthin. Conclusion: P. urinaria and P. maderaspatensis demonstrated dose-dependent hepatoprotective activity and hence, can provide promising therapeutic interventions against chemical-induced liver damage.

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Background: Zingiber officinale Rosc. is not only an important medical plant in China, but also one of the most commonly used plant spices around the world. Early researches in Z. officinale Rosc. were focused on rapid propagation, germplasm preservation, and somatic embryogenesis, only a few reports focused on the generation of tetraploid ginger plants with colchicines treatment in vitro. Materials and Methods: The adventitious buds were submerged into different concentrations of colchicine water solution for different time to induce polyploid plants, and the induced buds were identified by root-tip chromosome determination and stomatal apparatus observation. Eighteen selected tetraploid lines were transferred to the field, and the leaf characteristics, rhizome yield, contents of volatile oil and gingerol were respectively evaluated to provide evidence of high-yield and good qualities of tetraploid ginger. Results: The induction rate reached as high as 33.3% of treated buds. More than 48 lines of autotetraploid plants were obtained. All tetraploid plants showed typical polyploidy characteristics. All of the 18 selected tetraploid lines possessed higher rhizome yield and overall productivity of volatile oil and gingerol than those of the control. Conclusion: Five elite lines have been selected for further selection and breeding new varieties for commercial production in agricultural production.

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Objective: To optimize the extraction technology for polysaccharides from the root of Limonium sinense (Girard) Kuntze, Plumbaginaceae and evaluate the antioxidant capacity of polysaccharides from L. sinense (LSEP) Materials and Methods : One-singer factor and response surface methodology(RSM) were established to extract the polysaccharides from L. sinense. Then, the 1,1-diphenyl-2-picrylhydrazyl free radical, hydroxyl radical(.OH), and 2,2'-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt free radical assays were established to measure the antioxidant capacity of the LSEP in vitro. Results: According to analysis, extraction temperature significantly affected extraction yield. The optimum extraction conditions for LSEP were as follows: extraction temperature, 95°C; ultrasonic time 50 minutes; and dosage liquor ratio, 1: 12. Under these conditions, the experimental yield of crude LSEP was 12.80±0.19% which was well matched with the predicted models. The antioxidant capacity data suggested that LSEP has strong antioxidant activity. Conclusion: One-singer factor and RSM were used to extract of LSEP are simple and feasible and LSEP could be developed as a nutraceutical agent for itsstrong antioxidant activity.

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Background: Hwangryunhaedok-tang is a traditional herbal prescription that has sedative activity, hypotensive and anti-bacterial effects. Objective: In this study, we investigated the alteration of contents of components in Hwangryunhaedok-tang, antioxidant activity and neuroprotective activity by fermentation with Lactobacillus acidophilus KFRI 128. Materials and Methods: Contents of three marker compounds (geniposide, berberine and palmatine) and unknown compounds in the Hwangryunhaedok-tang (HR) and the fermented Hwangryunhaedok-tang (FHR) were measured and compared using the established high-performance liqued chromatograph coupled with a photodiode (HPLC-DAD) method. The antioxidant activity of HR and FHR were determined by DPPH free radical and hydrogen peroxide (H₂ O₂ ) scavenging assay. Also, the neuroprotective activities of HR and FHR against glutamate-induced oxidative stress in a mouse hippocampal cell line (HT22) were evaluated by MTT assay. Results: The contents of geniposide and palmatine were decreased but the content of berberine was increased in the FHR. And the contents of unknown compounds (1), (2), (3), (4) and (5) in the HR were altered by fermentation. Electron donating activity (EDA, %) value of FHR was higher than HR for DPPH radical scavenging activity and H2O2 scavenging activity, respectively. In the MTT assay, FHR showed more potent neuroprotective activity than HR by 513.90%. Conclusion: The FHR using microorganism could convert compounds in HR and enhance the antioxidant and neuroprotective activity.

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Background: The hypoglycemic effects of hexane, chloroform and methanol extracts of leaves of Azadirachta indica (AI) were evaluated by oral administration in streptozotocin-induced severe diabetic rats (SD). Materials and Methods: The effect of chronic oral administration of the extract for 28 days was evaluated in streptozotozin diabetic rats. Lipid peroxidation, glycogen content of liver and skeletal muscles, insulin, superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), oxidized glutathione (GSSG) levels were determined. In addition, advanced glycation end product formation (AGEs) was evaluated. Results: The most active extracts were obtained with chloroform. Chloroform extract from AI shows increased levels of SOD, GSH, GSSG and CAT, hepatic glycogen content, glucose-6-phosphatase and insulin plasma levels, which also decreased the glucokinase (GK), lipid peroxidation and insulin resistance. The chloroform extract exhibited significant inhibitory activity against advanced glycation end product formation with an IC₅₀ average range of 79.1 mg/ml. Conclusion: Azadirachta indica can improve hyperlipidemia and hyperinsulinema in streptozocin-induced diabetic rats and, therefore, AI can be potentially considered to be an antidiabetic-safe agent.

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Objective: The aim was to study the in vitro and in silico interactions of cleistanthin A and B on the adrenergic and cholinergic receptors using isolated animal tissues and bioinformatics tools. Materials and Methods: The alpha adrenergic receptor activities of cleistanthin A and B were studied in vitro using a guinea pig vas deferens preparation. The beta adrenergic receptor activities of cleistanthin A and B on an isolated rat heart were studied in vitro using a modified Langendorff apparatus. The effects of cleistanthin A and B on the nicotinic and muscarinic cholinergic receptors were studied in vitro using rabbit vas deferens and rabbit jejunum, respectively. All the drug responses were recorded using a data acquisition system through a variable force transducer. The receptor-ligand interactions of cleistanthin A and B with adrenergic and cholinergic receptor proteins were determined using the ArgusLab molecular modeling and drug docking program. Results: Cleistanthin A and B significantly inhibited the actions of the alpha adrenergic receptor and the nicotinic cholinergic receptor. Cleistanthin A and B shifted the dose-response curve to the right with an increased EC₅₀ value of phenylephrine and acetylcholine. Both cleistanthin A and B did not have any significant effect on the beta adrenergic and muscarinic cholinergic receptors. Conclusion: Cleistanthin A and B block the alpha adrenergic and nicotinic cholinergic receptors, but these compounds do not interact at all with the beta adrenergic and muscarinic cholinergic receptors.

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Background: Teucrium polium L. (TP) have been used in herbal medicine for different purposes such as antispasmodic, antidiabetic and lowering blood lipid. In the present study, the impact of aqueous-ethanol extract of TP on blood pressure, heart rate and intraventricular pressure was investigated in rabbit. Materials and Methods: Twenty-four NWZ rabbits weighed (2-3 kg) were randomly divided into four groups. In each experiment, two groups of six rabbits received jugular injection of either TP extract (20, 40 and 80 mg/kg) or normal saline for blood pressure effects and two groups for intraventricular pressure. Then, blood pressure, heart rate and intraventricular pressure were measured via carotid cannula using pressure transducer connected to a power lab system, and the data were pooled from independent, single-blinded experiments for each group. Results: Treatment with 80 mg/kg of TP extract significantly depressed the mean arterial blood pressure (12.5%, P< 0.05). However, there was no significant decrease in the 20 or 40 mg/kg dose or normal saline treatment group. Moreover, the extract increased (dp/dt)max (P<0.05), maximum left ventricular pressure (LVP_max) (P<0.05) and decreased (dp/dt)min significantly (P<0.05), there was no meaningful effect on left ventricular end-diastolic pressure (LVEDP). Conclusion : The present results demonstrated the extract had no effect on the heart rate, but showed a positive inotropic on the heart and hypotensive effects. These data suggested that hypotensive effect may counterbalance by the inotropic effect of the extract.

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Background: The aim of this study was to investigate the possible protective role of antioxidant treatment with ellagic acid (EA) on lung injury after intestinal ischemia-reperfusion (I/R) injury using biochemical and histopatological approaches. Materials and Methods: Forty rats were divided into four groups as control, control + EA, I/R, and I/R + EA. The control and control + EA groups were also anesthetized and subjected to laparotomy, but without clamp application. The control + EA and I/R + EA groups were given EA (85 mg/kg) orally prior to experiment. The I/R and I/R + EA groups underwent 30 minutes of intestinal ischemia and 1 hour of reperfusion. In all groups, serum total antioxidant capacity (TAC) and malondialdehyde (MDA) levels were determined. TAC, total oxidative status (TOS), and oxidative stress index (OSI) in lung tissue were measured. Lung tissue histopathology was also evaluated by light microscopy. Results: TAC levels were higher in control, EA, and I/R + EA groups while TOS, OSI, and MDA levels were lower in these groups compared with I/R group. Serum MDA levels were significantly higher in I/R + EA group than that of control group. Lung tissue TAC levels were lower in I/R + EA group while OSI values were higher in that groups compared with EA group. Histological tissue damage was milder in the EA treatment group than in the I/R group. Conclusion: These results suggest that EA treatment protected the rats lung tissue against intestinal I/R injury.

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