Applications of immobilization techniques in the screening of active constituents from traditional Chinese medicines / 药学学报

It has been an active approach to screen the active ingredients in traditional Chinese medicines (TCMs) according to the affinity property between small molecule compounds and biomaterials such as cells, bacteria and proteins. On the other hand, the biomaterials can be immobilized on a solid support before the screening procedure. The immobilization method not only can maintain the biological activities of biomaterials, but also have other advantages such as high efficiency, simple operation, easy to be continuous and automatic, etc. Carrier materials (solid supports) for the immobilization including silica gel, magnetic materials, hollow fiber, and the surface plasma resonance sensor chips have been used to immobilize biomaterials and successfully applied in the screening of active ingredients from TCMs. In this paper, applications of immobilization techniques in the screening of active components from TCMs were reviewed to provide a scientific reference to the future applications.

Expression of recombinant human IFNa-2b/IgG4 Fc fusion protein in a baculovirus insect cell system / 中华肝脏病杂志

To investigate a baculovirus insect cell system for expressing an interferon alpha 2b (IFNa2b)/immunoglobulin G-4 (IgG4) Fc fusion protein, which has long-acting antiviral effects. Human IFNa2b and IgG4 Fc cDNAs were generated by molecular cloning and inserted into a baculovirus shuttle vector, which was then transposed into the DH10 Bac strain to form recombinant Bacmid-IFN/Fc. The Bacmid-IFN/Fc was transfected into High five insect cells, and expression of the IFN/Fc fusion protein was detected by Western blotting and its biological activity was assessed by the cytopathic effect inhibition method. The IFNa2b and IgG4 Fc cDNA fragments were successfully amplified by RT-PCR using human peripheral lymphocytes. After cloning into the baculovirus shuttle vector, pFastBac1, and transforming into DH10 Bac competent cells, screening identified positive clones carrying the recombinant Bacmid-IFN/Fc. A Bacmid-IFN/Fc clone was successfully transfected into the High five insect cells and packaged into the baculovirus for expression of the IFN/Fc fusion protein. Western blotting revealed that the fusion protein expression was specific, and yielded a protein of 45 kD in size. The in vitro antiviral activity of the IFN/Fc fusion protein was 580 IU/mL. A novel IFN/Fc fusion protein was successfully generated using a baculovirus insect cell system, which may prove useful for providing future experimental data for development of a new long-acting interferon to treat chronic viral hepatitis.

Relationship between single nucleotide polymorphisms in the promoter of COX-2 gene and hereditariness to NAFLD / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the relationship between COX-2 gene and hereditariness to Nonalcoholic fatty liver disease by detecting single nucleotide polymorphisms in the promoter of COX-2 gene.</p><p><b>METHODS</b>Genotypes of 200 case patients with NAFLD and 206 control subjects were examined by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP). The DNA samples were extracted from the peripheral blood of all subjects.</p><p><b>RESULTS</b>Two SNPs, -1195G more than A and -765 G more than C, were identified with frequencies of variant alleles 54% and 5% in patients with NAFLD and 48% and 2% in control, respectively. A case-control analysis revealed a 1.13-fold (95% CI = 1.01-2.46) and a 2.35-fold (95% CI = 1.17-3.65) excess risk of developing NAFLD for -1195AA or -765CG genotype carriers compared with noncarriers. Compared with G-1195-G-765 containing haplotype, a greater risk of developing NAFLD was observed for A-1195-G-765 (OR =1.42; 95% CI =1.11-1.63) and A-1195-C-765 (OR = 4.24; 95% CI =1.72-14.22) containing haplotypes. A greater risk of developing NAFLD was observed for A-1195 and C-765 containing haplotype compared with other haplotype, suggesting an interaction between the -1195A and -765C in the context of haplotype.</p><p><b>CONCLUSIONS</b>These findings suggest that genetic variants in the COX-2 promoter may play an important role in mediating susceptibility to developing NAFLD in a Chinese population. -1195G more than A and -765G more than C in promoter region of Cyclooxygenase-2 gene, whose single nucleotide polymorphisms are related with development of NAFLD, are the significance factors of the susceptibility of NAFLD.</p>

Preliminary study on the effects of an artificial liver support system in the treatment of medicamentous liver insufficiency / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the effectiveness of an artificial liver support system.</p><p><b>METHODS</b>Thirty-two patients with medicamentous liver insufficiency were treated with an artificial liver support system in addition to the routine medicinal therapy. Thirty patients treated with routine medicinal therapy only served as controls.</p><p><b>RESULTS</b>The clinical symptoms (e.g. hepatic encephalopathy) and the laboratory indices (serum total bilirubin and prothrombin time) of the treatment group patients were obviously improved compared with those of the control group patients (P < 0.05). The cure rate and hospitalization days were 90.6% (26/32) and 47 days respectively in the treatment group, and 43.3% (13/30) and 72 days in the control group (P < 0.05).</p><p><b>CONCLUSION</b>Using an artificial liver support system combined with routine medicinal therapy is more effective than using medication alone.</p>