OBJECTIVE:To establish the quality standard for Yiwei granule. METHODS:TLC was conducted for the qualitative identification of Coptis chinensis,Corydalis yanhusuo and Schisandra chinensis;HPLC was conduced for the content determination of paeoniflorin. The column was XDB C18 with mobile phase of methanol-water(25:75,V/V)at flow rate of 1.0 ml/min,detection wave-length was 230 nm and injection volume was 20μl. RESULTS:The TLC showed clear spots and good separation. The linear range of paeoniflorin was 7.575-60.6 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recov-ery was 96.97%-102.51%(RSD=1.92%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Yiwei granule.

Objective To establish the standard for quality control ofQingyan Granule. Methods The chief components of the preparation, Sophora Tonkinensis radix et rhizoma, Adenophorae radix, Lonicera japonica caulis, and Ophiopogonis radix were identified by TLC qualitatively. The contents of licorice glycosides and glycyrrhizic acid were determined by HPLC. The separation was performed on Thermo Syncronis C18 column (4.6 mm×250 mm, 5μm) with mobile phase consisted of acetonitrile with 0.05% phosphoric acid solution (A)-0.05% phosphoric acid solution (B), and gradient elution (0-8 min, 19%A;8-35 min, 19%→50%A). Detection wavelength was 237 nm, and flow rate was 1 mL/min.Results The spots in TLC were clear. There were spots with same color on the corresponding location of reference substance and reference herbal, negative control without interference. The linear range for licorice glycosides was 0.05-0.5μg (r=0.999 9). The average recovery was 99.97%, RSD=1.74% (n=9). The linear range for glycyrrhizic acid was 0.1-2μg (r=0.999 9). The average recovery was 99.74%, RSD=1.28% (n=9). Conclusion The method is simple, accurate, with high reproducibility, which can be used for quality control ofQingyan Granule.

Objective:To compare the steam distillation( SD) and supercritical CO2 ( SFE-CO2 ) method in the extraction of vola-tile constituents from such 3 traditional Chinese drugs as cnidium monnieri, radix sileris and atractylodes lancea in compound Kuhuang prescription. Methods:Essential oil was extracted from the 3 traditional Chinese drugs by SD and SFE-CO2 method, respectively. The components were identified by GC-MS, and their relative contents were calculated with peak area normalization method. Results:To-tally 36 components were isolated and identified using SD extraction and 31 components were identified using SFE-CO2 extraction, a-mong them, 22 components were the same and their relative molecular weight mainly concentrated within 200-230. Conclusion: SFE-CO2 method can extract effective components from the 3 traditional Chinese drugs in compound Kuhuang prescription with higher selec-tivity, which is the more suitable extraction method for essential oil from the prescription.

OBJECTIVE:To optimize the water-extraction technology of Compound kuhuang formulation. METHODS:The wa-ter-extraction technology of Compound kuhuang formulation was optimized by orthogonal design,with the total contents of matrine and oxymatrine,the total content of prim-O-glucosylcimifugin and 5-O-methylvisamminol,the content of berberine hydrochloride and extract yield,and with the extraction time,amount of water(3 reflux extractions)and soaking time as the factors;and verifi-cation tests were conducted. RESULTS:The extraction time had significant effect on comprehensive evaluation value (P<0.05). The optimal water-extraction technology was as follows as extraction time of 120 min,water amount of 8,6,and 6 times as the amount of herbs,and soaking time of 20 min. In verification tests,the average total content of matrine and oxymatrine was 3.152 6 mg/g(RSD=1.03%,n=3),that of prim-O-glucosylcimifugin and 5-O-methylvisamminol was 4.977 2 mg/g(RSD=2.27%,n=3),the average content of berberine hydrochloride was 3.345 0 mg/g (RSD=1.19%,n=3) and the average extract yield was 49.23%(RSD=2.43%,n=3). CONCLUSIONS:The optimal water-extraction technology of Compound kuhuang formulation is stable and feasible.

Objective To investigate the expression and significance of aromatase P450,COX-2,ER and PR in the patients with adenomysis.Methods The immunohistochemistry staining was used to detect the expression of aromatase P450,COX-2,ER and PR in ectopic endometrium and eutopic endometrium of 30 patients with adenomyosis and 30 cases in control group.Results The expression of aromatase P450,COX-2 in ectopic endometrium were significantly higher than those in eutopic endometrium (all P ＜ 0.05) ; The expression of ER,PR in ectopic endometrium were significantly lower than those in eutopic endometrium (all P ＜ 0.05),excepted the expression of PR in eutopic endometrium,the expressions of both ER and PR lose their periodical cycle.There was positive correlation between the expression of aromatase P450 and COX-2 in adenomysis group(P ＜ 0.05).In adenomyosis,the expression of aromatase P450,COX-2,ER and PR in dysmenorrhea subgroup were significantly higher than those in non-dys menorrhea subgroup(all P ＜ 0.05).Conclusion Aromatase P450,COX-2 and ER play important roles in the genesis and development of adenomyosis and dysmenorrheal,PR is not the main pathogenic factors of adenomyosis.The expressions of aromatase P450,COX-2 and ER in adenomyosis have nothing to do with endometrial cyclical change and are not subject to the regulation of ovarian hormones.

To review and analyze the pharmacological effect and underlying mechanism of Chinese lobelia. Methods:The references from home and abroad on the pharmacological effect and mechanism of Chinese lobelia in recent years were summarized and analyzed. Results:Chinese lobelia had wide range of pharmacological effects, such as anti-tumor, endothelial cells regulation, an-algesic and anti-inflammatory effect, inhibition against alpha glycosidase enzymes and myocardial ischemia reperfusion et al. Conclu-sion:A large number of effective compositions are in Chinese lobelia and the biological activity is extensive. The mechanism of biologi-cal activity should be further studied in-depth.

Objective:To re-establish the quality control standard for Weiling granules. Methods:The 4 chief herbs in the prepa-ration, radices paeoniae alba, licorice, rhizoma corydalis and hawthorn were identified by TLC qualitatively. The content of paeoniflor-in in radices paeoniae alba was determined by HPLC. The separation was performed on an Agilent XDB-C18 (250 mm × 4. 6 mm, 5μm)coulme with mobile phase consisting of acetonitrile-0. 1% phosphoric acid solution (10:90). The detection wavelength was 230 nm and the flow rate was 1. 0 ml·min-1 . Results:The spots in TLC were clear without any interference. The linear range for paeoni-florin was 0. 151-1. 212 μg(r=0. 999 9,n=5). The average recovery was 99. 63% and RSD was 2. 01%(n=9). Conclusion:The method is simple and accurate with high reproducibility, which can be used for the quality control of Weiling granules.

Objective:To evaluate the role of mitochondrial calcium homeostasis in hyperoxia-induced injury to alveolar epithelial cells typeⅡ(AECⅡ) in rats.Methods:Rat AECⅡcell lines (RLE-6TN) were randomly divided into 3 groups ( n=18 each) using a random number table method: nomoxia group (group C), hyperoxia group (group H), and hyperoxia plus ruthenium red group (group HR). The cell lines were cultured in a conventional incubator for 4 h in group C. The cell lines were incubated in an incubator with 90% O 2 for 4 h in group H. In group HR, ruthenium red 2 μmol/L was added first, and then the cell lines were incubated in an incubator with 90% O 2 for 4 h. After the treatment, the Ca 2+ concentrations in mitochondria were measured using the mitochondrial Ca 2+ specific fluorescent probe Rhod-2-AM, the ROS level was measured using the fluorescent probe DCFH-DA, and the cell morphology and mitochondrial ultrastructure were observed. Results:Compared with group C, the Ca 2+ concentrations in mitochondria and ROS level were significantly increased in H and HR groups ( P<0.05). Compared with group H, the Ca 2+ concentrations in mitochondria and ROS level were significantly decreased in group HR ( P<0.05). Compared group C, the cell volume was reduced, the morphology was shrunken round, the cell arrangement was loose, the mitochondrial double-layer membrane structure was broken, and the mitochondrial cristal fragments were observed in group H. Compared with group H, the cell morphology was significantly improved, and the slight damage to mitochondrial double-layer membrane structure was found, and the structure of mitochondrial cristae was normal in group HR. Conclusion:Mitochondrial calcium homeostasis is involved in the pathophysiological process of hyperoxia-induced injury to AECⅡ in rats.

Objective: To investigate the different efficacy of Yinchenhao Decoction (YCHD), a compound traditional Chinese herbal medicine, for liver cirrhosis induced by dimethylnitrosamine (DMN) or carbon tetrachloride (CCl(4)) in rats. Methods: To induce liver fibrosis, 0.5% DMN solution (2mL/kg body weight, i.p.) was given three consecutive days a week to male Wistar rats for 4 weeks. Cirrhotic rats were randomly divided into DMN group, YCHD group, Xiaochaihu decoction group by the end of the fourth week to accomplish a 2-week recipe treatment course. In CCl(4)-induced liver fibrosis model, 50% CCl(4)-olive solution was injected subcutaneously to rats at a dose of 2 mL/kg body weight twice a week to duplicate rat cirrhosis model. After 8 weeks, rats were divided into CCl(4) group, CCl(4) plus YCHD group and Xiaochaihu decoction group. For the YCHD group, YCHD was administered intragastrically once a day for 4 weeks. For DMN or CCl(4) model, by the end of 6 or 12 weeks respectively, rats were sacrificed for sampling to detect liver function, hepatic histological changes, hydroxyproline (Hyp) content and apoptosis-related gene expressions. Results: In DMN liver fibrosis model, hepatic fibrosis was obvious at week 2 and cirrhosis was evident at week 4 in DMN-treated rats. Compared to 6-week DMN group, hepatic pathological changes and liver function were improved significantly and content of Hyp decreased remarkably in YCHD group. In CCl(4)-induced liver fibrosis model, hepatic fibrosis was obvious at 8 weeks and cirrhosis was evident at 12 weeks in CCl(4)-treated rats. Compared to 12-week CCl(4) group, hepatic pathological changes and liver function were not obviously improvement in YCHD group. The results of gene chip showed that YCHD significantly decreased Fas, Bax and caspase-3 gene expressions, and increased Bcl-xL gene expression in the liver of DMN model. However, in the model induced by CCl(4), YCHD did not inhibit hepatocyte apoptosis induced by CCl(4), but increased tyrosine kinase receptor gene expression by 4.8 times. Conclusion: YCHD exerts more significant therapeutic effects on DMN-induced than CCl(4)-induced cirrhosis in rats in Hyp content and pathological change in liver tissue.

Objective:To establish the methods for the qualitative identification and content determination of aurantio-obtusin and chrysophanol in Zeju Jiangzhi tablets. Methods:A TLC method was adopted for the qualitative identification, and an HPLC method was used for the content determination. The determination was performed on a Wondasil C18 (250 mm × 4. 6 mm, 5 μm ) column with the mobile phase of acetonitrile -0. 1% phosphonic acid with gradient elution at the flow rate of 1. 0 ml?min-1 , the detection wave-length was 286 nm, the column temperature was 30℃ and the injection volume was 10μl. Results:The TLC spots of aurantio-obtusin and chrysophanol were clear and well-separated without any negative interference. The HPLC experiment results showed the good line-arity within the range of 1. 03-25. 72μg?ml-1(r=0. 999 9) for aurantio-obtusin, and 0. 48-11. 92μg?ml-1(r=0. 999 9) for chry-sophanol. The average recovery was 99. 21% and 98. 85%, and RSD was 0. 70% and 0. 73%, respectively (n=9). Conclusion:The method is simple, accurate and repeatable, which can be used for the qualitative identification and content determination of auran-tio-obtusin and chrysophanol in Zeju Jiangzhi tablets.