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.

Aims Tocomparethepharmacokineticsof ephedra alkaloids,cinnamic acid and cinnamic alcohol in Ephedrae, Cinnamomi and Ephedrae-Cinnamomi herb couple through UPLC-MS/MS in rats respective-ly,and to investigate the effect of combination on phys-iologicaldisposition.Methods Plasmasampleswere collected at different times after oral administration of Ephedrae,Cinnamomi and Ephedrae-Cinnamomi herb couple extracts.The concentrations of ephedra alka-loids,cinnamic acid and cinnamic alcohol in plasma samples were determined by UPLC-MS/MS.DAS 3. 0 was used to calculate pharmacokinetic parameters.The differences of samples in two groups were conducted with univariate statistical analysis using SPSS 13. 0. Results ComparedwithEphedraegroup,theCmaxof norephedrine hydrochloride, norpseudoephedrine hydrochloride,ephedrine hydrochloride,pseudoephed-rine hydrochloride,and methylephedrine hydrochloride in Ephedrae-Cinnamomi herb couple group were signif-icantly greater (P<0. 05 );the AUC0-t of norpseudo-ephedrine hydrochloride was significantly greater (P<0. 05 );the MRT0-t of norephedrine hydrochloride, phedrine hydrochloride, pseudoephedrine hydrochlo-ride,and methylephedrine hydrochloride were signifi-cantly less (P <0. 05 );the T1/2z of norephedrine hydrochloride,phedrine hydrochloride,and methyl-ephedrine hydrochloride were significantly less (P <0. 05 ).The AUC0-t and MRT0-t of cinnamic acid and cinnamic alcohol were significantly greater than those inCinnamomigroup(P<0.05).Conclusion The combination of Ephedrae and Cinnamomi improves the absorption concentration of five ephedra alkaloids, Slows down the elimination of norephedrine hydrochlo-ride,phedrine hydrochloride,pseudoephedrine hydro-chloride,and methylephedrine hydrochloride,and in-creases the bioavailability of cinnamic acid and cin-namic alcohol.

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

ObjectiveThe human angiotensin converting enzyme2 （hACE2） transgenic mouse model was used to clarify the antiviral efficacy of BD-77 against a novel coronavirus SARS-CoV-2 and explore the action mechanism of BD-77 against SARS-CoV-2. MethodSARS-CoV-2 Omicron and Delta variant strains-infected VeroE6 cell models were established and administered with BD-77 to observe the antiviral effect of BD-77 in vitro. A kit was used to detect the effect of BD-77 in vitro on the binding of spike S protein of SARS-CoV-2 virus （Delta/Omicron） to angiotensin converting enzyme2 （ACE2）. Chromatography was adopted to detect the binding of BD-77 to the S protein and N protein of the novel coronavirus. hACE2 transgenic C57BL/6 mice were divided into a blank control group， SARS-CoV-2 infection group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1， with eight mice in each group. The pneumonia model of SARS-CoV-2-infected hACE2 transgenic mice was built to observe the survival of the mice， detect the virus titer of the lung tissue of the mice， and observe the lesions in the lung tissue. ResultBD-77 had a certain inhibitory effect on Omicron and Delta variant strains in vitro， with median inhibitory concentration （IC₅₀） of 526.3 mg·L^-1 and 653.0 mg·L^-1， respectively. BD-77 had no significant inhibitory effect on the binding of the S protein of WT， Omicron， and Delta variant strains of SARS-CoV-2 to ACE2 and had no binding effect with the S protein and N protein of the novel coronavirus. No mice in the blank group died， while the mortality rate of SARS-CoV-2-infected mice was 75%. There was a large amount of virus replication in the lung tissue of the mice and large areas of inflammatory infiltration in the lung tissue and interstitium. Compared with the model group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1 could reduce the mortality of mice， significantly lower the virus titer in the lung tissue of mice （P<0.05）， and improve lung lesions. ConclusionBD-77 demonstrated significant inhibitory effects against SARS-CoV-2 virus in vitro and in vivo. However， its mechanism of action did not involve direct inhibition of the virus itself or intervention in the virus-host binding process. This finding suggests that the mechanism of action of BD-77 needs to be thoroughly investigated and elucidated by further experiments.