From the chloroform-soluble fraction of the ethanol extracts of the whole plant of Taxus chinensis var. mairei (Lemee et Levl), four compounds were isolated by using repeated column chromatography on silica gel and Sephadex LH-20. Based on spectroscopic data (UV, IR, ESI-MS, 1H NMR and 13C NMR), the compounds were identified as taxamairin K (1), 2α, 4α-dideacetoxy-7β-benzoyloxy-5β,20-epoxy-9α,10β,13α,15-tetrahydroxy-11(15→1)abeotaxa-11-ene (2), 7β-xylosyl-taxol (3), 10-deacetoxy-7-xylosyl-taxol (4). Among them, taxamairin K is a new compound.

A sensitive LC-MS/MS method to determine cocaine and its major metabolite benzoylecgonine in guinea pig's hair has been established. About 20 mg of decontaminated hair sample was hydrolyzed with 0.1 mol·L-1 HCl at 50 ℃ overnight, in the presence of cocaine-d3 and benzoylecgonine-d8 used as internal standards, and then extracted with dichlormethane. The analysis was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Positive electrospray ionization (ESI+) and multiple reactions monitoring (MRM) mode were used. The limit of detection (LOD) for cocaine and benzoylecgonine was 1 pg·mg-1. The calibration curves of extracted standards were linear over the range from 5 pg·mg-1 to 250 pg·mg-1 (r2≥0.999 7). The method was validated and applied to the analysis of guinea pig's hair after a single dose administration of cocaine hydrochloride. Cocaine and benzoylecgonine were not only detected, but also quantified in guinea pigs hair.

Five compounds, huyouyisu (1), vomifoliol (2), isoferulic acid (3), 1,2,3-trihydroxy-phenol (4) and p-hydroxy-phenol (5), were isolated from the peels of Citrus changshan-huyou Y.B.Chang for the first time by utilizing repeated column chromatography on silica gel. Among them, huyouyisu (1) is a new compound. The structure was elucidated by using 1D and 2D spectra.

The present study is to investigate the protective actions of guggulsterone against the cytotoxicity produced by exposure to hydrogen peroxide (H2O2) in PC12 cells. It was evaluated by MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide] reduction assay, lactate dehydrogenase (LDH) release assay, and the release of nitric oxide (NO). ROS and Ca2+ in cells were evaluated by DCFH and Fura 2-AM, respectively. Mitochondrial membrane potential (MMP) was assessed by the retention of rhodamine 123 (Rh 123). Apoptosis and morphological alteration in PC12 cells were monitored with flow cytometry and electric microscope. Vitamin E, a potent antioxidant, was employed as a comparative agent. The results showed that preincubation of PC12 cells with guggulsterone (0.1-10 μmol·L-1) prevented cytotoxicity induced by H2O2. Extracellular accumulation of LDH, NO and intracellular accumulation of ROS, Ca2+ resulting from H2O2 were significantly reduced by guggulsterone. Incubation of cells with H2O2 caused a marked decrease in MMP, which was significantly inhibited by guggulsterone. The percentage of H2O2-induced apoptosis in PC12 cells was 24.3%, and decreased in the presence of guggulsterone (0.1-10 μmol·L-1) by .8.4%, 15 .9%, 11.8%, respectively. Guggulsterone exhibited comparable potency against oxidative stress induced by H2O2 in PC12 cells as that of vitamin E. The present findings showed that guggulsterone attenuated H2O2-induced cytotoxicity, extracellular accumulation of LDH and NO, intracellular accumulation of ROS and Ca2+, loss of MMP, and apoptosis, which may represent the cellular mechanisms for its neuroprotective action.

A highly sensitive, rapid and selective liquid chromatography-tandem mass spectrometry(LC-MS/MS) method for the quantitative determination of retinamido-ester in rat plasma was developed and validated. A simplified protein precipitation with acetonitrile was employed for the sample preparation.The separation was carried out on an Agilent TC C18 column ( 150 mm×4. 6 mm ID, 5 μm particle size)with the mobile phase consisted of methanol-water-formic acid (93 : 7 : 0.1). Simvastatin was used as internal standard. The detection was performed on a trap-quadrupele tandem mass spectrometer by selected reaction monitoring (SRM) scan mode via atmospheric pressure chemical ionization (APCI). The range of and inter-day precision values were between 95.97% and 104. 43%, and RSD was between 4. 63% and 10. 69%, respectively. This method was applied to determine the pharmacokinetic parameters. The main pharmacokinetic parameters of retinamido-ester after oral administration via gastric gavage of 2. 5, 5, 10mg·kg-1 were as follows,T1/2;(11.28±7.23),(8.90±3.82),(8.01±5.56)h; AUC0-∞:(103.41±61.46),(190.23±74.99),(421.66±299.20)ng·h·mL-1;MRT:(6.31±0.75),(5.98±0.71), (6.18±0.97) h; CL/F: (30. 10 ± 13.67), (29.58±10.59), (31.18 ±17.51)L·h-1·kg-1;Vd/F:(414.94±159.82),(356.16±139.85),(369.28±322.73)L·kg-1,respectively.

The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide(NO) secretion of mouse macrophages stimulated by lipopelysaccharide (LPS) for 24 h was assayed by Griess reagent system. We found that RCE had potent anti-inflammatory effects on mice. RCE had littl ecell toxic effect on mouse lymphocytes and macrophages. RCE strongly inhibited the excessive production of inflammatory mediators ( NO, CD69, CD25, CD71 ), in a dose-dependent manner, like cyclosporine A injection. RCE could inhibit proliferation of CD3<'+> T lymphocytes. These data suggested that RCE migh texhibit anti-inflammatory effect by inhibiting the activation and proliferation of mouse lymphocytes and the NO secretion of mouse macrophages.

Effect of new calcium antagonist m-nisoldipine (m-Nis) on MCT-induced PH in rats and its mechanisms were investigated. Rats were injected with a single doxe(60mg·kg-1)of MCT subcutaneously to induce PH. Pulmonary haemedynamic measurement and lung tissue morphological investigations were undertaken. The MDA production and SOD activity in the serum were tested. PCNA,ERK1 and p-ERK expressions were analyzed by Western blotting. The expressions of 5-HT and PCNA were observed with immunohistochemistry. Results suggested that the PAP, right ventricular index and the degree of muscularization of small pulmonary artery were elevated markedly in MCT group, which was attenuated by m-Nis treatment. A significant reduction in MDA production and an increase in the SOD activity in the serum were also observed in all three m-Nis groups. The number of PCNA and 5-HT positive smooth muscle cells increased significantly in MCT group, and m-Nis treatment attenuated the expression obviously. Western blotting results suggested that the protein expression of PCNA and the ratio of p-ERK/ERK1 increased markedly in MCT group and decreased by m-Nis. In conclusion, m-Nis protected against MCT-induced PH by decreasing PAP, right ventricular index, PAMSCs proliferation and pulmonary artery remodelling, which may be related to the reduction of 5-HT and the suppression of the ERK/MAPK signal pathway.

To study xanthones from the barks of Garcinia xanthochymus, the constituents were isolated by normal-phase and reverse-phase silica gel column chromatography from the EtOAc extract. Their structures were elucidated by spectral analysis. Three new xanthones were purified and identified as 1,2,5-trihydroxy-6-methoxyxanthone (1), 1,4,6-trihydroxy-5-methoxyxanthone (2), 1,2,7-trihydroxy-4-( 1,1-dimethylallyl) xanthone (3).

To investigate the chemical constituents of the roots of Aconitum taipaicum, silica gel column chromatography was used for the isolation and purification of compounds. A new norditerpenoid alkaloid, isodelelatine (1), along with five known alkaloids, atisine (2), delfissinol (3), liangshanine(4), hypaconitine (5) and delelatine (6) were isolated and identified. The structure of the new compound was elucidated on the basis of spectral data.

One new quinoline alkaloid and seven known bisabolane sesquiterpenes: 2-(2'-methyl-1'-propenyl)-4, 6-dimethyl-7-hydroxyquinoline(1), 2, 5-dihydroxybisabola-3, 10-diene(2), 4, 5-dihydroxybisabola-2, 10-diene(3), turmeronol A(4), bisacurone(5), bisacurone A(6), bisacurone B(7), bisacurone C(8), as well as dehydrozingerone(9) and zingerone(10) were isolated from the root tuber of Curcuma longa. Their structures were identified by spectral evidence. Compound 1 is a new compound, compounds 6-8 were isolated from this plant for the first time and compounds 9-10 from Curcuma for the first time.