Objective; To establish a method for content determinate of Rutin in H ibiscus mutabilis L by RT- HLPC. Methods; The chromatographic conditions were as follows; Column; Ubondpak C18; the column temperature was 25℃and the detection wavelength was 370nm;Mobil phase consisted of 0.02mol/L KH2PO4 solution cotaining 1.0% glacial acetic acid -methanol (73:27, v:v). The flow rate was 1.0ml/min. SPD -2ASUV detector was used. The standard sample was used as control. Results; Good linearity was obtained for Rutin and other components in the range of (0.5-5) ?g/20?l. Peak area was favorable liner between the concentration of Rutin in range . The regression equation was Y =0.0001X +0. 1208,correlation coefficient r = 99.8% ,The recovery rate was 99. 88% , RSD was 1. 54% (n = 5 ). Conclusion; The method is convenient, precise, resulting in high separate efficacy, and suited for the content determination of Rutin in H ibiscus mutabilis L.

Objective To observe the effect of rutin on growth and proliferation of human hepatic cancer line(HepG2).Methods HepG2 cells were cultured in vitro,then cocultured with 50 to 250 ?mol/L rutin for 24 h.The inhibition rate of rutin on growth and proliferation of HepG2 was determined by MTT,~(3)H-TdR,and apoptotic cells were observed in fluorescent staining by Olympus fluorescent microscopy,and cell cycle was analysed by flow cytometry.Results Rutin inhibited HepG2 cells from growth and proliferation,and evoked apoptosis.Flow cytometry showed that 50 to 250 ?mol/L rutin caused an increase at G_(0)/G_(1) phase and a decrease at G_(2)/M phase and arrest at G_(0)/G_(1) phase in the cell cycle.Conclusion Rutin markedly inhibits the proliferation of HepG2 cells and induces apoptosis in a concentration-dependent manner.

Objective To explore the protective effects of quercetin on damage induced by oxidative stress and to clari?fy its molecular mechanism. Methods Chang liver cell cultures were randomly divided into control groups, H2O2 group and 3 doses of quercetin groups. Cell survival rate was detected with MTT. Cell apoptotic rate was measured by FACS(Fluores?cence-activated cell sorting). Intracellular reactive oxygen species (ROS) level in Chang liver cells were tested by flow cy?tometer. The DCF fluorescence intensity of DCFH-DA-stained intracellular ROS was observed by fluorescence microscope. The levels of malondialdehyde (MDA), superoxide dismutase(SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were determined in liver cells using commercial available kits. The expression of Nrf2 were detected by Western blot. Re?sults Compared with control, cell survival rate and levels of SOD, CAT and GSH-Px decreased significantly in H2O2 group (P < 0.05 ),while cell appotosis rate, content of MDA and mean fluorescence intensity(MFI) increased in H2O2 group (P <0.05). In comparison with H2O2, expression of Nrf2 protein was higher in all three quercetin treatment groups (P<0.05). Con?clusion Quercetin protected Chang liver cells from H2O2-induced oxidative stress, which may be caused by the increased ex?pressions of down stream antioxidant genes via activating the Nrf2-ARE signaling pathway.

The HBV X gene was amplified by PCR according to the pecob6 containing the whole fragment of adw subtype of HBV, then the fragment was inserted into the multiple clone site of pAdTrack-CMV. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 and the recombinant adenoviral plasmid pAd-X was generated. Then plasmid pAd-X was digested with Pac I and transfected into 293 cells for packaging and amplifying. Infection titer and rate were monitored by green fluorescent protein (GFP) expression. With restriction endonuclease analysis and PCR methods, it has been confirmed that HBV X gene was cloned into the adenovirus vector successfully. The expression of X protein in HepG2 cells was detected by Western-blot. The recombined adenovirus Ad-X was constructed successfully, which would contribute to the advanced functional study of HBV X protein.
Adenoviridae ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; metabolism ; Hep G2 Cells ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Trans-Activators ; biosynthesis ; genetics ; Transfection

OBJECTIVE: To determine the effect of exogenous hydrogen sulfide on experimental hepatic fibrosis in rats and its mechanism. METHODS: Wistar male rats were randomly divided into three groups: a normal control group (n=8), a model group (n=8), and a hydrogen sulfide prevention group (n=8). The rat model of hepatic fibrosis was reduced by intraperitoneal injection of carbon tetrachloride (CCl4). The prevention group, in addition to intraperitoneal injection of 40% CCl4, was intraperitoneally administered H2S once a day until 8th week. After the experiment, the liver function and liver fibrosis were assayed. Liver tissue samples were used for histopathological changes. The expression of TGF-β1 in liver tissue was detected by RT-PCR and Western blot. RESULTS: Compared with the model group, the levels of ALT, AST, HA, LN, and PC III in the sulfide group were significantly reduced (P<0.01 or P<0.05), ALB content was increased (P<0.05) in the serum, TGF-β1 expression was obviously reduced, and the degree of liver fibrosis was improved (P<0.05). CONCLUSION Exogenous hydrogen sulfide can effectively inhibit the development of hepatic fibrosis, reduce the expression of TGF-β1, and decrease the the sediment of extracellular matrix in the liver tissues.
Animals ; Carbon Tetrachloride ; Extracellular Matrix ; metabolism ; Hydrogen Sulfide ; pharmacology ; therapeutic use ; Liver ; pathology ; Liver Cirrhosis ; chemically induced ; pathology ; prevention & control ; Male ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; antagonists & inhibitors ; metabolism