Establishment and identification of HUVEC cell strains with over-expression and low expression of RACK 1 / 基础医学与临床

Objective To establish several human umbilical vein endothelial cell ( HUVEC ) strains with over-ex-pression or low expression of receptor for activated C kinase 1 ( RACK1 ) , which will provide an effective tool for future studying the function of RACK1 in arrhythmia.Methods The full-length cDNA sequence of RACK1 gene was amplified and inserted into pIRES2-EGFP.At the same time, designed and synthesised complementary DNA sequences of 3 pairs of short hairpin structure and a pair of negative control sequence , then subcloned into the plas-mid pGenesil-1 .The HUVEC cells were transfected with these plasmids and screened by using G 418 .And the expression of RACK1 mRNA and protein in the cells were assayed by qRT-PCR and Western blot , respectively . Results RACK1 eukaryotic expression vector and siRNA expression vectors of RACK 1 were constructed success-fully.After a 48 h transfection of HUVEC cells with the recombinant vectors and G 418 selection, the positive cell clones were obtained .qRT-PCR and Western blot showed that over-expression vector and interference vectors could effectively enhanced and knocked-down RACK1 expression in HUVEC strains .Conclusions HUVEC cell strains with over-expression and low expression of RACK 1 have been successfully established .

Effective antiviral therapy with entecavir in chronic hepatitis B virus carriers / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the short-term effect and safety of entecavir for the treatment of chronic hepatitis B (CHB) virus carriers.</p><p><b>METHODS</b>Ninety-three cases of CHB virus infection (hepatitis B surface antigen (HBsAg)-positive, hepatitis B e antigen (HBeAg)-positive, hepatitis B core antibody (HBcAb)-positive, HBV DNA≥1x10(5) copies/mL) were divided into two groups: CHB virus carrier (47 cases) and CHB (46 cases). All of the 93 cases were given 0.5 mg entecavir orally once a day for 48 weeks. Virology, serology and biochemistry tests were perrmed at treatment weeks 0, 4, 12, 24 and 48. Side effects of entecavir and the incidence of liver cirrhosis and hepatocellular carcinoma were recorded.</p><p><b>RESULTS</b>The CHB virus carrier and CHB group had complete virological response rates of 14.9% and 17.4% at week 4, 51.1% and 63.0% at week 12, 76.6% and 89.1% at week 24, and 97.9% and 100% at week 48, respectively; there was no significant difference between the two groups (P>0.05). The CHB virus carrier and CHB group had partial virological response rates of 42.6% and 47.8% at week 4, 57.44% and 65.2% at week 12, 85.0% and 89.1% at week 24, and 100% and 100% at week 48, respectively; there was no significant difference between the two groups (P>0.05). No cases in either group experienced virologic breakthrough during the treatment course. The CHB virus carrier and CHB group had serological response (HBeAg-negative) rates of 0 and 4.3% at week 4, 2.1% and 8.7% at week 12, 4.3% and 13.0% at week 24, and 8.5% and 21.7% at week 48, respectively; there was no significant difference between the two groups (P>0.05). The CHB virus carrier and CHB group had HBeAg seroconversion rates of 0 and 0 at week 4, 0 and 4.4% at week 12, 2.1% and 10.9% at week 24, and 6.4% and 17.4% at week 48, respectively; there was no significant difference between the two groups (P>0.05). No case in either group showed HBsAg-negativity and seroconversion during the treatment course. The CHB group had a biochemical response (alanine aminotransferase normalization) rate of 26.1% at week 4, 65.2% at week 12, 91.3% at week 24, and 97.8% at week 48.No case in either group showed biochemical breakthrough during the treatment course. There were no cases of liver cirrhosis or hepatocellular carcinoma in either group. There were no side effects of the entecavir treatment experienced in either group.</p><p><b>CONCLUSION</b>Antiviral therapy with entecavir is effective, safe and well tolerated in CHB virus carriers.</p>

A analysis of the proteome of HL60 cell line induced by a new steroidal drug as a monocytic inducer / 中国药理学通报

Aim The potentiality as a differentiation inducer of the new steroidal drug(NSC67657) had been studied.Then the expression differences of protein between treated and untreated HL60 cell line could be analysed.Method Firstly,the expression patterns of C/EBP alpha gene and protein were observed between treated and untreated HL60 cell line.Then the proliferation of HL60 cell line could be investigated by MTT.At the same time cellular chemical staining could be employed to investigate which direction HL60 cell line would be induced by NSC67657.Then the flow cytometry(FCM) could be employed to detect the profile of differentiation of HL60 cell line induced in different time and at different drug concentrations,by which the most suitable drug concentration and inducing time could be found. Following that,the information of cellular cycle and ultramicrostructure could be analysed by FCM and electronmicro scope,by which whether the apoptosis had happened or not under the drug treatment could also be found. Finally,the protein of these two group HL60 cell lines could be separated by modified two-dimensional electrophoresis (2-DE).Results The expression of C/EBP alpha gene and protein could be promoted under the treatment of NSC67657.Then the proliferation of HL60 cell line was inhibited significantly.From cellular chemical staining,the monocytic differentiation could be easily found and the perfect inducing time and drug concentration were defined as 10 ?mol?L-1NSC67657 and constantly inducing HL60 cell line within 5 days.The cellular number of G0~G1 was increased and hardly any apoptosis which might be happened during drug inducing ability could be seen.The protein of HL60 cell lines were separated by modified 2-DE technology.Then there were 14 protein spots which could only be found in the differentiated gels,on the other hand,20 protein spots could only be found in the undifferentiated gels,which would have been analyzed by MALDI-TOF.Conclusions HL60 cell line could be induced to monocyte by NSC67657 which could also stimulate the C/EBP? in the early stage.2-DE could separate the protein directly which expressed differentially,from which some proteins essential in cellular differentiation might be found.