Objective To construct the expression short hairpin RNA （shRNA） targeting gene kir2. 1 in rat myocardial cells, named pEGFP6 kir2. 1, and to observe the effects on the expression of messenger RNA（mRNA） and protein of gene kir2. 1 as well as the changes of myocardial beating rates. Methods Five RNA interference （RNAi） sites targeting the rat kir2. 1 gene was selected, designed and synthesized five pairs of oligonucleotides fragments ,annealed them to double-strand, then cloned them into the vectors containing U6 promoter,obtained the vector expressing five aim genes. Rat myocardial cells were divided into three groups： Experimental group, negative plasmid control group and normal control group. Reverse transcription-polymerase chain reaction（RT PCR） and Western-blot were carried out to detect the expression of the mRNA and protein of gene kir2.1 and the beating rates of myocardial cells were observed after 72 h. Results The expression of mRNA and protein of gene kir2. 1 of experimental group were markedly lower than that of other two control groups after 72 h（P〈0.01）. There was no statistically significant between two control groups. The beating rate in experimental group was much faster than other two control groups （P〈0.01）, remained unchanged in both negative plasmid control group and normal control group. Conclusion Plasmid pEGFP6-kir2.1 could suppress the expression of the mRNA and protein of kir2.1 and increase the rat cardiac muscle cell beats.
Objective To review the current status and problems in developing cardiac biological pacemaker(CBP) by cell transplantation. Methods The l iterature over the past decade concerning CBP constructed through celltransplantation was reviewed and summarized. Results Experiments in vivo testified that the cell transplantation was feasible for CBP construction, and the transplantation of sinus atrial node cell and stem cell was still the predominant method for constructing CBP. However, such problems as difficult ampl ification of transferred cardio muscle cell, low success rate of CBP construction as well as unstable function of CBP make it lag behind the tremendous cl inical demands. The gene transfection technology might be one of the approaches to resolve these issues. Conclusion As one feasible method for CBP construction, the cell transplantation has a bright future in the cl inical appl ication and is worthy of further study.