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Hepatitis B virus (HBV) that causes Hepatitis B with a high chronic rate, could lead to hepatic cirrhosis and hepatocellular carcinoma. The IL28B gene belongs to a new interferon family λ and its genetic polymorphisms are identified to be associated with treatment effect and viral clearance. The purpose of this review is to discuss the role of the IL28B gene in treatment response and viral clearance of HBV-infected individuals, and further to reveal the mechanisms. This review could provide a theoretical basis for personalized medicines of HBV patients.
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OBJECTIVE To investigate the inhibitory effect of lead acetate on transient receptor potential A1(TRPA1)channel. METHODS TRPA1-mediated calcium influx in mice dorsal root ganglion(DRG) neurons and HEK293 cells expressing nouse TRP1 (mTRPA1) and human TRPA1 (hTRPA1) was recorded by intracellular calcium imaging. TRPA1-mediated currents were detected by two-electrode voltage clamp. RESULTS Lead acetate 3.0 and 10.0μmol·L-1 inhibited external calcium influx in DRG neurons by(36.7 ± 4.1)% and(79.4 ± 3.1)%(n=5),respectively. The inhibitory effect of lead acetate on hTRPA1-mediated current was concentration-dependent. Lead acetate 0.3, 1.0, 3.0, 10.0 and 30.0μmol · L-1 inhibited the amplitudes of currents by(1.0 ± 0.7)%,(11.6 ± 0.8)%,(57.7 ± 3.2)%,(93.6 ± 2.6)%and(91.2±2.0)%(n≥4),respectively,with the IC50 2.4μmol·L-1. CONCLUSION TRPA1 channel may be an endogenous target of lead. Lead acetate inhibits TRPA1 channel at a very low concentration.
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Background: rabies virus [RABV] is a deadly neurotropic virus that causes the disease of rabies in humans and animals. L protein is one of the large structural protein of rabies virus, which displays multiple enzymatic activities, and is required for viral transcription and replication
Objectives: a truncated L protein of Rabies virus is being cloned, expressed and purified to produce relevant polyclonal Antibody
Materials and Methods: the gene fragment of L protein of RABV was subcloned into prokaryotic expression vector pET- 28a and transformed into E. coli Rosetta DE3 host strain. The recombinant L protein of RABV was expressed and characterized by SDS-PAGE and western blot analysis using anti-his tag antibody. Mice were immunized with the purified recombinant L protein, the reaction of the anti-serum was checked by immunofluorescence and dot-blot, respectively
Results: the results of PCR and sequencing confirmed that the fragment of L gene of RABV was successfully cloned into the expression vector. The expression of recombinant L protein fragment induced by IPTG was confirmed by the band of 43 kDa in SDS-PAGE and western blot. The antiserum of purified L protein immunized mice was reacted with RABV infected N2a cells and suckling mouse brain tissue lysates
Conclusions: our data showed that the recombinant L protein produced by pET-28a vector was very successful, and the purified L protein could efficiently induce the antibody response in mice. The antiserum could recognize the virus in RABV infected cells and tissue very well
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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 .