Objective To study the chemical constituents of the EtOAc soluble fraction obtained from the stems and branches of Euonymus alatus with anti-myocardial ischemic effect. Methods Chemical constituents were isolated and purified by solvent extraction together with various chromatographic techniques, and the structures were identified by physicochemical properties and spectral analyses. ResultsFive compounds, 1-[3-?-D-glucopyranosyloxy)-4, 5-dihydroxyphenyl]-ethanone (Ⅰ), phenethyl alcohol 8-O-?-D-glucopyranosyl-(1→2)-?-D-glucopyranoside (Ⅱ), eugenyl-O-?-D-glucopyranoside (Ⅲ), hyperin (Ⅳ), and hesperidin (Ⅴ) were isolated. Conclusion Compound Ⅰ is determined as a new compound named guijianyuside. Compounds Ⅱ and Ⅲ are isolated from this plant for the first time.

Objective To observe the protective effects of various extracts from Flos Daturae (FD), including active fraction (AF-FD), withanolides constituents (WC-FD), and flavonoids constituents (FC-FD) on Chinese hamster ovary (CHO) cells against cytotoxicity induced by DMSO. Methods The survival rate of CHO cells was examined by MTT assay and LDH leakage assays. Results Using MTT assay, coincubation of CHO cells with 3% DMSO for 24 h resulted in a significant reduction of survival rate of CHO cells. AF-FD was tested in a range of 10—80 ?g/mL to improve the survival rate of CHO cells in a dose-dependent manner. FC-FD (2.5—20 ?g/mL), but not WC-FD (30—120 ?g/mL), could significantly relieve the injury induced by 3% DMSO in CHO cells. In the measurement of LDH leakage, coincubation of CHO cells with 4.5% DMSO for 24 h obviously increased LDH release. However, all the compounds tested, including AF-FD (10—80 ?g/mL), WC-FD (30—120 ?g/mL), and FC-FD (2.5—20 ?g/mL) had no effect on LDH leakage induced by 4.5% DMSO. Conclusion The findings suggest that the FC-FD may protect CHO cells from DMSO cytotoxicity assessed by MTT assay, which may be associated with improving mitochondrial function, but not protecting the membrane injury of CHO cells.

AIMTo assess the sex-difference on flutamide metabolism in rat liver microsomes useing rat cytochrome P450, 1A2, inhibitory monoclonal antibody.

METHODSLiver microsomes were prepared from male or female rats. Protein concentration and total cytochrome P450 content were determined. Incubation mixture included liver microsomes (1.0 nmol.L-1), reduced form of nicotinamide adenine dinucleotide phosphate (NADPH, 0.1 nmol.L-1), CYP1A2 (1:400) and flutamide (2 mg.L-1). The incubation time was 30 min. The concentration of flutamide and its major metabolite 2-hydroxyflutamide were analyzed by reverse high-performance liquid chromatography. The mobile phase was a mixture of methanol-acetonitrile-water-diethylether (40:20:35:1) with methyltestosterone as internal standard. The detection wavelength was 234 nm. The reaction mixture was extracted with acetic ether 4 mL. Sex-difference on flutamide metabolism was expressed as the difference between the concentration ratio of 2-hydroxyflutamide to flutamide in male and female rat liver microsomes.

RESULTSThe recoveries of flutamide and 2-hydroxyflutamide for the proposed method were more than 75%. The formation of 2-hydroxyflutamide from flutamide was inhibited by CYP1A2 antibodies (1:400) in male and female rat liver microsome for 30 min of incubation time, but the inhibition of flutamide metabolism in female rat was stronger than that in male. The concentration ratios of 2-hydroxyflutamide to flutamide were (1.5 +/- 0.6) and (0.9 +/- 0.4) in male and female rat liver microsomes, respectively (P < 0.01).

CONCLUSIONThe results indicate that the activity of male rat CYP1A2 is higher than that of the female rat. There is difference in sex-related rate of flutamide metabolism in rat liver microsomes.

Androgen Antagonists ; metabolism ; Animals ; Antineoplastic Agents, Hormonal ; metabolism ; Cell Separation ; Cytochrome P-450 CYP1A2 ; metabolism ; Female ; Flutamide ; analogs & derivatives ; analysis ; metabolism ; Male ; Microsomes, Liver ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sex Factors

Objective To explore the clinical features and genetic etiology of children with cystinuria with onset of kidney stone. Methods The clinical data of 3 children with cystinuria with onset of kidney stone and the gene analysis results of SLC3A1 and SLC7A9 by PCR sequencing were retrospectively analyzed.Results Three male children were from three unrelated families, kidney stone were presented in 2 cases at 1 year old and 1 case at 14 years old. The blood amino acid spectrum was normal in all 3 cases, while the free carnitine were decreased. The urinary amino acid spectrum indicated that cystine, ornithine, arginine,and threonine increased.Gene analysis confirmed that 1 case had homozygous mutations of SLC7A9 gene c.325G>A, and his parents were carriers of c.325G>A heterozygous mutation;other 2 cases had heterozygous mutations of SLC3A1 gene, c.1365delG and c.1113C>A heterozygous mutation in one case, and c.1897_1898insTA and c.1093C>T heterozygous mutation in one case, and their parents were heterozygous mutation carriers. After treatment with potassium citrate and L-carnitine, the conditions were improved in all cases. Conclusions Inherited metabolic disease should be considered for children with kidney stone. Urine amino acid analysis and gene detection are important methods for the diagnosis of cystinuria.