Triterpenic Acids from Potentilla parvifolia and Their Protective Effects against Okadaic Acid Induced Neurotoxicity in Differentiated SH-SY5Y Cells.

Six triterpenic acids were separated and purified from the ethyl acetate extractive fraction of ethanol extracts of Potentilla parvifolia FISCH. using a variety of chromatographic methods. The neuroprotective effects of these triterpenoids were investigated in the present study, in which the okadaic acid induced neurotoxicity in human neuroblastoma SH-SY5Y cells were used as an Alzheimer's disease cell model in vitro. The cell model was established with all trans-retinoic acid (5 µmol/L, 4 d) and okadaic acid (40 nmol/L, 6 h) treatments to induce tau phosphorylation and synaptic atrophy. Subsequently, the neuroprotective effects of these triterpenic acids were evaluated in vitro by this cell model. Results from the Western blot and morphology analysis suggested that compounds 3-6 had the better neuroprotective effects. Furthermore, we tested the level of mitochondrial reactive oxygen species and mitochondrial membrane potential of these compounds in SH-SY5Y cells by flow cytometry technology to investigate the potential neuroprotective mechanism of these compounds. All of the results indicated that maybe the mechanism of compounds 5 and 6 is to protect the cell from mitochondrial oxidative stress injuries.

Triterpenoids with antioxidant activities from Myricaria squamosa.

A new triterpenoid, namely myricarin C (compound 1), together with three known compounds myricarin A (compound 2) and myricarin B (compound 3), 3α-hydroxy-D-friedoolean-14-en-28-oic acid (compound 4), was isolated from the overground part of Myricaria squamosa. Compound 2 and compound 3 existed in the solution by the form of cis-trans isomers. Their structures were elucidated by means of extensive spectroscopic methods, including 1D-NMR, 2D-NMR, and HR-ESI-MS. The antioxidant properties of all compounds were calculated based on the DPPH radical scavenging activities. Results showed that myricarin A and myricarin C had general antioxidant activities with EC50 values of 40.90 µg/ml, 42.22 µg/ml, respectively, compared to the control, rutin (5.17 µg/ml). The EC50 values of myricarin B was 195.81 µg/ml. Compound 4 had no antioxidant activities.

[Determination and analysis of trace elements in Lycium barbarum L. from different regions of Qinghai province].

For comparison of the quality of Lycium barbarum L., the authors determined 11 trace elements in the fruits of Lycium barbarum L. from 12 different regions of Qinghai province by ICP-MS and ICP-AES. Meanwhile, 7 trace elements essential for human body were selected to the object of principal component analysis by SPSS statistic software. Three principal component equations were obtained, and the regression equation related to principal component was also set up. The research is very important to quality analysis and to obtaining high quality Lycium barbarum L. , and provided science basis for the development and utilization of Lycium barbarum L. in Qinghai province.

[Rapid determination of fatty acids in soybean oils by transmission reflection-near infrared spectroscopy].

In the present research, a novel method was established for determination of five fatty acids in soybean oil by transmission reflection-near infrared spectroscopy. The optimum conditions of mathematics model of five components (C16:0, C18:0, C18:1, C18:2 and C18:3) were studied, including the sample set selection, chemical value analysis, the detection methods and condition. Chemical value was analyzed by gas chromatography. One hundred fifty eight samples were selected, 138 for modeling set, 10 for testing set and 10 for unknown sample set. All samples were placed in sample pools and scanned by transmission reflection-near infrared spectrum after sonicleaning for 10 minute. The 1100-2500 nm spectral region was analyzed. The acquisition interval was 2 nm. Modified partial least square method was chosen for calibration mode creating. Result demonstrated that the 1-VR of five fatty acids between the reference value of the modeling sample set and the near infrared spectrum predictive value were 0.8839, 0.5830, 0.9001, 0.9776 and 0.9596, respectively. And the SECV of five fatty acids between the reference value of the modeling sample set and the near infrared spectrum predictive value were 0.42, 0.29, 0.83, 0.46 and 0.21, respectively. The standard error of the calibration (SECV) of five fatty acids between the reference value of testing sample set and the near infrared spectrum predictive value were 0.891, 0.790, 0.900, 0.976 and 0.942, respectively. It was proved that the near infrared spectrum predictive value was linear with chemical value and the mathematical model established for fatty acids of soybean oil was feasible. For validation, 10 unknown samples were selected for analysis by near infrared spectrum. The result demonstrated that the relative standard deviation between predict value and chemical value was less than 5.50%. That was to say that transmission reflection-near infrared spectroscopy had a good veracity in analysis of fatty acids of soybean oil.

The glycosides from Lomatogonium rotatum.

A new phenyl glycoside, 2-(3'-O-beta-D-glucopyranosyl)benzoyloxygentisic acid (1), along with seven known glycosides 2-8 was isolated from Tibetan herbal medicine Lomatogonium rotatum. The structures of the compounds were elucidated by spectroscopic methods including 1D and 2D NMR techniques and MS data.

[Preparation of Prussian Blue@Yeast Catalyst and Its Heterogeneous Fenton Performance].

A novel heterogeneous Fenton catalyst, Prussian blue@yeast (PB@yeast), was prepared via facile self-assembly synthesis. The as-synthesized composite was characterized by field emission scanning electron microscopy (FE-SEM), energy-dispersive spectroscopy (EDS), powder X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). All of the results indicated that PB nanoparticles were uniformly dispersed on the surface of yeasts with stable core-shell morphology. Degradation of the model anionic fluorescent whitening agent CXT indicated that the PB@yeast catalyst presented a synergistic effect of adsorption and heterogeneous Fenton performance. Owing to the high adsorption capacity of yeast, the CXT molecules were easy to move to the active site of the catalyst, promoting the electron transfer between Fe(Ⅲ) and Fe(Ⅱ) and then enhancing the catalytic activity of the Fenton reaction effectively. Furthermore, the yeast support could improve the dispersity and stability of PB nanoparticles, which maintained excellent catalytic activity and stability after being recycled four times.

Raspberry pulp polysaccharides inhibit tumor growth via immunopotentiation and enhance docetaxel chemotherapy against malignant melanoma in vivo.

It has been reported previously that the systemic efficacy of chemotherapeutic agents is substantially restricted for some cancer types, including malignant melanoma. Therefore, the development of more effective treatment modalities remains a critical, albeit elusive, goal in anticancer therapy. The study presented here evaluates the antitumor activity of raspberry pulp polysaccharides (RPPs) against malignant melanoma using a murine tumor-bearing model. Furthermore, the underlying mechanism of this antitumor activity has also been investigated. The results show that while RPP exhibits no direct cytotoxic effect on HT-29, MGC-803, HeLa, Bel-7402, L02 and B16F10 cells in vitro, it does demonstrate a dose-dependent growth inhibition of melanoma in vivo with an inhibition ratio of 59.95% at a dose of 400 mg kg(-1). Besides this, the body weight and spleen index in tumor-bearing mice have also been improved in RPP-treated groups. RPP is also found to induce splenocyte proliferation and is able to upregulate the activity of immune-related enzymes, including acid phosphatase (ACP), alkaline phosphatase (AKP), lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in the spleen of tumor-bearing mice. The levels of tumor necrosis factor α (TNF-α), interferon γ (IFN-γ) and interleukin 2 (IL-2) in the serum of tumor-bearing mice show to be effectively increased upon RPP treatment. Histopathological analyses show that RPP induces tumor tissue necrosis by increasing inflammatory cell infiltration and causes no lesions to liver and kidney tissues. Remarkably, RPP further enhances the antitumor effect of the chemotherapeutic drug docetaxel and alleviates docetaxel-induced liver and kidney lesions in tumor-bearing mice. These findings indicate that RPP exhibits antitumor activity in vivo against malignant melanoma, partly by enhancing the cellular immune response of the host organism. In summary, RPP features critical properties to potentially find use as an immunopotentiating agent or as a chemotherapy adjuvant agent for the treatment of malignant melanoma.

[Adsorption of the TiO2 @ yeast composite microspheres for adsorbing Fluorescent Whitening Agent-VBL in fixed bed].

In this work, the adsorption potential of TiO2@ yeast composite microspheres to remove Fluorescent Whitening Agent-VBL (FWA-VBL) from aqueous solution was investigated using fixed-bed adsorption column. The effects of pH(2.0-8.0), bed height (1-3 cm), inlet concentration (20-80 mg x L(-1)) and feed flow rate (5-11 mL x min(-1)) on the breakthrough characteristics of the adsorption system were determined. The results showed that the highest bed capacity of 223.80 mg x g(-1) was obtained under the condition of pH 2.0, 80 mg x L(-1) inlet dye concentration, 1.0 cm bed height and 5 mL x min(-1) flow rate. The adsorption data were fitted to three well-established fixed-bed adsorption models, namely, BDST model, Thomas model and Yoon-Nelson model. The results fitted well to the three models with coefficients of correlation R2 > 0.980 in different conditions. The TiO2 @ yeast composite microspheres have desired regeneration ability and could be reused for four times.

[Determination of trace arsenic and mercury in medicine animal horns by hydride atomic fluorescence spectrometry].

A hydride atomic fluorescence spectrometry has been developed for the determination of trace arsenic and mercury in medicinal animal horns. This method was simple, rapid and sensitive. The samples were digested with HNO3-HClO4(5:1), under the optimum conditions, the allowable amounts of 26 coexist elements were determined. The detection limits of this method were as follows: As 1.50 ng.mL-1, Hg 0.80 ng.mL-1. The relative standard deviations of 11 determinations were 1.2%-2.3% for As and 2.5%-4.7% for Hg. The recoveries were 98.9%-103.8% for As and 95.4%-105.0% for Hg.

Protective effect of seed oil of Herpetospermum pedunculosum against carbon tetrachloride-induced liver injury in rats.

OBJECTIVES: To investigate the protective effect of Herpetospermum pedunculosum (H. pedunculosum) seed oil against carbon tetrachloride (CCl4)-induced liver damage. METHODS: This experimental study was conducted at the Northwest Institute of Plateau Biology, Chinese Academy of Sciences, and Yantai University, China from November 2012 to May 2013. The H. pedunculosum seed oil was extracted using supercritical carbon dioxide. The antioxidant activities of H. pedunculosum seed oil were assayed in vitro by 2,2-diphenyl-1-picrylhydrazyl assay, lipid peroxidation assay, and antihemolytic assay. Adult Sprague Dawley rats were randomly divided into 6 groups (10 rats/group) including control, CCl4, CCl4+bifendate, and CCl4+H. pedunculosum seed oil (3 different doses) groups. RESULTS: The CCl4-induced liver lesions include hepatocyte necrosis, ballooning degeneration, calcification, and fibrosis. Moreover, CCl4 damage results in an obvious increase of serum triglycerides, high-density lipoprotein, low-density lipoprotein, malondialdehyde, total bilirubin, alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase activity. In addition, CCl4 also significantly decreased the activities of superoxide dismutase (SOD). By contrast, H. pedunculosum seed oil administration significantly ameliorated the CCl4-induced liver lesions, lowered the serum levels of hepatic enzyme markers, and increased the activities of SOD. CONCLUSION: The results of this study show that H. pedunculosum seed oil can be proposed to protect the liver against CCl4-induced oxidative damage in rats, and the hepatoprotective effect might be correlated with its potent antioxidant and free radical scavenging effect.

Analysis of estrogenic compounds in environmental and biological samples by liquid chromatography-tandem mass spectrometry with stable isotope-coded ionization-enhancing reagent.

A sensitive and reliable stable isotope labeling technology was developed for the determination of estrogenic compounds in environmental and biological samples based on the derivatization of estrogenic compounds with 10-methyl-acridone-2-sulfonyl chloride (d(0)-MASC) and its deuterated counterpart d(3)-MASC. The labeling reaction of MASC with estrogenic compounds is simple and robust and can be carried out under mild conditions within 5 min. Internal standard-based quantification was achieved by this labeling strategy without the need of using expensive internal standard analogy to every analyte of interest. Meanwhile, the sensitivity obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was enhanced by 2-3 orders of magnitude compared to the underivatized counterparts. Application of the stable isotope labeling technology in relative and absolute quantification of estrogenic compounds in complicated samples indicated that the labeling strategy was effective in overcoming matrix effects. The proposed method was successfully applied to the analysis estrogenic compounds in different environmental and biological samples with high sensitivity and accuracy.

Preparative separation and purification of four cis-trans isomers of coumaroylspermidine analogs from safflower by high-speed counter-current chromatography.

High-speed counter-current chromatography (HSCCC) was successfully applied for the first time to isolate and purify four cis-trans isomers of coumaroylspermidine analogs from Safflower. HSCCC separation was achieved with a two-phase solvent system composed of chloroform-methanol-water (1:1:1, v/v/v) with the upper phase as the mobile phase. In a single run, a total of 1.3mg of N(1), N(5), N(10)-(E)-tri-p-coumaroylspermidine (EEE), 4.4mg of N(1)(E)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (EZE), 7.2mg of N(1)(Z)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (ZZE), and 11.5mg of N(1),N(5),N(10)-(Z)-tri-p-coumaroylspermidine (ZZZ) were obtained from 100mg of crude sample. High Performance Liquid Chromatography (HPLC) analysis showed that the purities of these four components are 95.5%, 98.1%, 97.5% and 96.2%, respectively. The chemical structures were identified by ESI-MS, (1)H NMR and (13)C NMR.

MMPT as a reactive oxygen species generator induces apoptosis via the depletion of intracellular GSH contents in A549 cells.

MMPT, (5-[(4-methylphenyl)methylene]-2-(phenylamino)-4(5H)-thiazolone), a thiazolidin compound, was identified in our laboratory as a novel antineoplastic agent with a broad spectrum of antitumor activity against many human cancer cells. A previous study showed that MMPT inhibited cell growth, and induced apoptosis in H1792 cells. In this study, the antiproliferative activity of MMPT was investigated. The results showed that MMPT was able to inhibit A549 cell growth in a time- and dose-dependent manner by blocking cell cycle progression in the G2 phase and inducing apoptosis. MMPT induced DNA fragmentation and caspase activation in A549 cells, both of which are hallmarks of apoptosis. The apoptotic process was accompanied by the generation of reactive oxygen species, depletion of glutathione (GSH), and reduction the GSH/GSSG ratio, suggesting that MMPT may induce apoptosis in A549 cells through a reactive oxygen species dependent pathway. Treatment with a thiol antioxidant, NAC, showed the recovery of GSH depletion and the reduction of reactive oxygen species levels in MMPT-treated cells, which were accompanied by the inhibition of apoptosis. In contrast, L-buthionine sulfoximine (BSO), a well-known inhibitor of GSH synthesis, aggravated GSH depletion and cell death in MMPT-treated cells. In conclusion, we have demonstrated that MMPT inhibits the growth of A549 cells by inducing a G2 arrest of the cell cycle and by triggering apoptosis accompanied with the depletion of GSH.

Development of a new HPLC method with precolumn fluorescent derivatization for rapid, selective and sensitive detection of triterpenic acids in fruits.

Triterpenic acids are widespread in plants and have multiplicity of biological properties. Unfortunately the method for accurate analysis of these compounds remains poorly investigated. This study proposed a highly sensitive and selective precolumn derivatization method for accurate determination of five triterpenic acids (betulinic acid, betulonic acid, maslinic acid, ursolic acid and oleanolic acid) in fruits using acridone-9-ethyl-p-toluenesulfonate (AETS) as fluorescent labeling reagent by HPLC with fluorescence detection (FLD). Response surface methodology was employed to optimize the derivatization reaction, ensuring the sufficient labeling of the analyzed components. The rapid separation of five triterpenic acids could be achieved in as little as 16 min. This developed method offered the exciting detection limits of 1.68-2.04 ng/mL. When applied to several popular fruits in China, it revealed satisfactory applicability and reproducibility. This developed method also exhibits powerful potential for accurate detection of triterpenic acids from other foodstuffs and nature products.

Simultaneous determination of monoamine and amino acid neurotransmitters in rat endbrain tissues by pre-column derivatization with high-performance liquid chromatographic fluorescence detection and mass spectrometric identification.

A sensitive and efficient method for simultaneous determination of glutamic acid (Glu), gamma-amino-butyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) in rat endbrains was developed by high-performance liquid chromatography (HPLC) with fluorescence detection and on-line mass spectrometric identification following derivatization with 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC). Different parameters which influenced derivatization and separation were optimized. The complete separation of five neurotransmitter (NT) derivatives was performed on a reversed-phase Hypersil BDS-C(18) column with a gradient elution. The rapid structure identification of five neurotransmitter derivatives was carried out by on-line mass spectrometry with electrospray ionization (ESI) source in positive ion mode, and the BCEOC-labeled derivatives were characterized by easy-to-interpret mass spectra. Stability of derivatives, repeatability, precision and accuracy were evaluated and the results were excellent for efficient HPLC analysis. The quantitative linear range of five neurotransmitters were 2.441-2x10(4) nM, and limits of detection were in the range of 0.398-1.258 nM (S/N=3:1). The changes of their concentrations in endbrains of three rat groups were also studied using this HPLC fluorescence detection method. The results indicated that exhausting exercise could obviously influence the concentrations of neurotransmitters in rat endbrains. The established method exhibited excellent validity, high sensitivity and convenience, and provided a new technique for simultaneous analysis of monoamine and amino acid neurotransmitters in rat brain.