Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (contained 0. 2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0. 8 mL/min. Results Seventeen char-acteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is repro-ducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex PheUodendri.

BACKGROUND: Angiogenesis is an important prognostic indicator for malignant tumors. Breast cancer overexpressing oncogene HER-2/neu often denotes a poor prognosis. Many studies have demonstrated the antitumor effect of genistein against breast cancer.OBJECTIVE: To study the relationship between HER-2/neu expression and angiogenesis in breast cancer as well as the effect of genistein on the angiogenesis in HER-2/neu-overexpressing breast cancer.DESIGN: A randomized controlled observatory experiment with nude mice.SETTING: Department of nutrition and food hygiene of a military medical university.MATERIALS: Twenty specific pathogen-free(SPF) normal female BALB/c nude mice weighing (10 ± 2) g, aged 3 to 4 weeks, were purchased from the Experimental Animal Center of the Third Military Medical University.METHODS: This study was carried out in the Department of Nutrition and Food Hygiene, Third Military Medical University from June 2001 to March 2002. HER-2/neu-overexpressing breast cancer cell line MCF-7/HER-2 was generated by transfecting MCF-7 cells with human HER-2/neu cDNA. MCF-7/HER-2 and MCF-7 cells were inoculated in female BALB/c nude mice to establish tumor-bearing mouse models. Four weeks after the inoculation, the mice with MCF-7/HER-2 xenografts were randomly divided into control,genistein treatment, and anti-HER-2/neu antibody treatment groups to receive corresponding treatments every other day for two weeks, at the end of which the tumor volume, microvessel density(MVD) and vascular endothelial growth factor(VEGF) expression in the xenografts were measured.MAIN OUTCOME MEASURES: MVD and VEGF expression in the xenograft tumor. Secondary outcome measures: Identification of HER-2/neu-transfected from MCF-7-transfected cells and the tumor volume.RESULTS: The MVD was 16 ±6, 98 ±21, 56± 18, and 52 ± 19 in each visual field in the MCF-7 xenografts group, control group, genstein treatment group and anti-HER-2/neu antibody treatment group recpectively. MVD and VEGF expression in MCF-7/HER-2 xenografts were higher than that in MCF-7 xenografts, and was reduced after treatment with genistein or anti-HER-2/neu antibody. The changes of tumor volume in these xenografts were consistent with the changes of MVD and VEGF.CONCLUSION: HER-2/neu overexpression in breast cancer promotes angiogenesis, and genistein can inhibit angiogenesis and growth of HER-2/neu-overexpressing breast cancer to improve the prognosis.

Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0.1 % phosphoric acid (contained 0.2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0.8 mL/min. Results Seventeen characteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is reproducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex Phellodendri.

Objective Previous studies have demonstrated that LPS can induce endothelial cell activation and the expression of E-selectin. In this study, we examined whether A20 gene could inhibit the expression of E-selectin in endothelial cells induced by LPS. Methods With the help of DOTAP, endothelial cells were transfected with pCDNA3.1 EHA20. The postive cell clones were selected with G418.The stable transfection and expression of A20 in the endothelial cells were determined by immunofluorescence analysis. The E-selectin expression was checked by immunofluorescence, Western blot and in situ hybridization. Results Abundant A20 stable expression in endothelial cells transfected with pCDNA3.1 EHA20 was confirmed by immunofluorescence analysis. E-selectin expression increased in LPS-inducible endothelial cells. A20 gene inhibited 90% LPS-inducible E-selectin expression(P

BACKGROUND:The cytokine has an effect of immunoregulation and immediate induction in the reconstruction of periodontal tissue. At present, the role and mechanism of hepatocyte growth factor involving the reconstruction of periodontal tissue under orthodontic force are stil unclear.
OBJECTIVE:To explore the mechanism underlying hepatocyte growth factor in the tooth movement and periodontal tissues remodeling under the inflammation periodontal tissue condition.
METHODS:Thirty Sprague-Dawley rats, aged 8 weeks, were used to establish periodontitis model. The obtained model was randomly divided into two groups:inflammatory control group and inflammatory force group. In the force group, rats were treated with the fixed orthodontic appliance by 50 g forces in the maxil ary first molars. After 1, 3, 5, 7 and 14 days of tooth movement, five rats were sacrificed respectively. Hematoxylin-eosin staining and immunohistochemistry methods were used to analyze the expression and distribution of hepatocyte growth factor in the periodontium for rats at different tooth movement stages.
RESULTS AND CONCLUSION:Hematoxylin-eosin staining results showed that, remodeling of periodontal tissues existed in al groups. (2) The immunohistochemical results showed that hepatocyte growth factor had positive expression in periodontal tissue, and the distribution was even in the control group. In the force group, hepatocyte growth factor expression was increased and reached the peak on day 5, then began to decline. Osteoblast, fibroblast and osteoclast were strongly expressed. The findings indicate that, hepatocyte growth factor is involved in the periodontal tissues remodeling under orthodontic force, and inflammation can increase the expression of hepatocyte growth factor in periodontal tissue.

Objective To investigate the effects of zinc finger protein gene A20 on the inhibition of lipopolysaccharide (LPS) induced interleukin 8 (IL 8) expression in endothelial cells. Methods Plasmid pcDNA3.1EHA20 was transfected into human umbilical vein endothelial cells (HUVECs) by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of A20 in HUVECs were determined by immunofluorescent analysis. IL 8 expression was detected by sandwich ELISA with double monoclonal antibodies. Results High expression of A20 gene in HUVECs transfected with pCDNA3.1EHA20 was confirmed by immunofluorescent analysis. IL 8 expression increased in LPS induced endothelial cells. A20 gene could inhibit more than 70% LPS induced IL 8 expression ( P

Objective To investigate the mechanisms of genistein in the growth inhibition in the tumor angiogenesis. Methods The effects of genistein and vascular endothelial growth factor receptor (VEGFR) on the proliferation of ECV304 cells were observed by cell growth curves and flow cytometry. The level of the phosphorylation of VEGFR and the activity of protein tyrosine kinase (PTK) in ECV304 cells were detected by immunoprecipitation and kinase activity analysis. Results Vascular endothelial growth factor (VEGF) alone could facilitate the proliferation of ECV304 cells, up regulate the phosphorylation and PTK activity of VEGFR, but genistein alone could inhibit the growth of ECV304 cells, stop the cell cycle mainly at the phase of G 2/M, induce apoptosis, and down regulate the phosphorylation and PTK activity of VEGFR ( P

0.05).After that,both systolic and diastolic function began to decrease.In 4 weeks after the operation,both the the maximum rate of left ventricular isovolumic systolic pressure + dp/dtmax and the maximum rate of left ventricular isovolumic diastolic pressure-dp/dtmax decreased to the lowest levels [(1249.89 ? 95.82) mm Hg/s and(-1316.40 ? 58.31) mm Hg/s,respectively];and then in 6 weeks after the operation,echocardiography showed that the left ventricular short axis fractional shortening FS reached the lowest level [(18.70 ? 3.83)%].Moreover,we found that the FS was highly related with the + dp/dtmax(r=0.864,P

Objective: To study the effects of octylpheno(lOP)and genistein(GEN)on the expressions of estrogen receptors(ER),amplified in breast cancer 1(AIB1) and proliferating cell nuclear antigen(PCNA) in rat breast cancer.Method:Female SD rats were randomly divided into control,model,GEN treated,OP treated,GEN and OP combined treated groups.RT-PCR and immunohistochemical methods were used to detect the expressions of AIB1,ER,PCNA on normal mammary gland and mammary cancer.Results: Compared with control mammary gland,AIB1 mRNA,ER mRNA,PCNA and ER expressions were up-regulated in mammary cancer.Compared with mammary cancer in model group,the level of AIB1mRNA,ER mRNA and ER expressions were significantly decreased in GEN treated group,while they were significantly increased in OP treated group and PCNA expression was significantly increased too.Compared with OP treated group,the level of AIB1mRNA,ER mRNA,ER and PCNA expressions were partly decreased in GEN and OP combined treated group.Conclusion: Octylphenol can up-regulate the levels of AIB1,ER and PCNA expressions in mammary cancer and may increase the incidence of 7,12-dinmethylbenz[a]anthracene(DMBA)-induced mammary cancer.Genistin or genistein combined with octylphenol can downregulate the levels of AIB1,ER and PCNA expressions in mammary cancer,and inhibit theiincidences.

Objective: To study the effects of genistein on the expression of urokinase-type plasminogen activator (uPA) and the activity of protein tyrosine kinase (PTK) in MDA-MB-453 cells, and explore the molecular mechanism of anti-angiogenesis in HER-2/neu-overexpressing breast cancer by genistein. Methods:Western blot, immunoprecipitate, reverse transcription-polymerase chain reaction (RT-PCR) and kinase activity analysis technics were used to observe the expression of uPA and the protein phosphorylation of HER-2/neu receptor and the activity of protein tyrosine kinase (PTK) in MDA-MB-453 cells treated by genistein for 24, 48, 72 h. Results: The expression of uPA and the protein phosphorylation of HER-2/neu receptor and the activity of PTK were significantly decreased after treated with 5?10-5mol/L genistein, which had a time-dependence. Conclusion: Genistein could inhibit the activity of PTK and the protein phosphorylation of HER-2/neu receptor, and down-regulate the exprssion of uPA at transcription and translation levels in breast cancer cells. This might be a part of molecular mechanism of genistein anti-angiogenesis in HER-2/neu-overexpressing breast cancer.