[Effects of Eupolyphaga Sinensis Walker on erythrocyte's CR1 activity and anti-cardiolipin antibody level in rats with stagnation of blood].

Aim: To study the effect of Eupolyphaga Sinensis Walker(ESW) on red blood cell immune adherence (RCIA) and serum anticardiolipin antibody level in rat model of Yin-deficiency Huo-excess with chronic blood stasis.

Methods: The model was made by i.m.

Expression of Wild-type and Mutant p16 in H460 Cell Line.

Two p16 mutants, P48L and D74N, were obtained by site-directed mutagenesis using two step PCR method. Mutant p16 cDNA and wild-type p16 cDNA were colned into the mammalian expression vector pcDNA3 to construct p16 expression vector pCMV-p16, pCMV-p16P48L and pCMV-p16D74N, respectively. After the introduction of these expression vectors into human lung cancer cell line H460 in which endogenous p16 gene was homozygously deleted, exogenous p16 expression was detected in G418 resistant cells by Northern blotting and immunocytochemistry staining.

Production of Anti-P16 Sera by Genetic Immunization and Protein Immunization.

Genetic immmunization is a new method of producing antibody, which is different from protein immunization. In order to produce anti-P16 sera and to find out the difference between genetic immunization and protein immunization, fusion protein GST-P16 and eukaryotic expression vector pCMV-p16 were injected in rabbit respectively. Western blot analysis showed that the titer of sera from protein immunization was 1:625, which was much higher than the titer from genetic immunization sera.