Objective To explore the role of genistein (genistcin, GEN) on human breast cancer cell proliferation and its mecha-nism. Method GEN MTT method was used to detect the proliferation of GEN on estrogen-dependent breast cancer cells MCF-7 and non-es-trogen-dependent breast cancer cell proliferation of MDA-MB231. Estrogen receptor.antagonist IC1182780 was used to observe the effect of GEN. FACS was used to analyze the cell cycle of breast cancer cells. Results GEN promote MCF-7 cell proliferation, and make G1/S transition. Estrogen receptor antagonist can block the proliferation effect on MCF-7 cells but no effect on MDA MB231. Conclusion GEN has estrogen-like activity, which was mediated by estrogen receptor (ER).

AIM: To study the feasibility, security a nd validity of the intrapericardial injection with a trans-diaphragmatic approach in chronic heart failure rats and its value in the study of gene therapy for hea rt diseases, and further investigate whether adeno-associated virual gene transf er of sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) gene can improve ven tricular function in chronic heart failure (HF) rats. METHODS: An animal model of heart failure was obtained by creati ng descending aortic constriction in rats. Recombinant adeno-associated virus, c arrying enhanced green fluorescent protein (EGFP) gene and recombinant adeno-ass ociated virus carrying SERCA2a gene, were respectively injected into pericardium of heart failure rats in different groups (group HF+EGFP and group HF+SERCA2a) by intrapericardial injection with a trans-diaphragmatic approach. After 30 days , hemodynamic parameters were measured and analyzed. Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein, and Western blotting was performed to detect the expression of SERCA2a. RESULTS: Green fluorescence was detected in cryosection of the h earts in all rats in group HF+EGFP and the expression of green fluorescent prote in was ubiquitously. The expression of SERCA2a in all rats in group HF+SERCA2a w as more than those in group HF and group HF+EGFP. And overexpression of SERCA2a improved the systolic and diastolic function of heart failure rats significantly and the hemodynamic parameters were similar with those of the controls. CONCLUSION: The intrapericardial injection with a trans-diaphrag matic approach suggests a simple, safe, efficient and cheap technique for the ge ne therapy of chronic heart failure. Gene thransfer of SERCA2a may be a new appr oach for the treatment of chronic heart failure.

BACKGROUND:In chronic congestive heart failure (CHF),both protein expression and activity of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) are decreased, which results in abnormal regulation of calcium ion and systole and diastole dysfunction.OBJECTIVE:To observe the short-and middle-term therapeutic effects of adeno-associated viral gene transfer of SERCA2a in treatment of CHF in rats,and to investigate the possible mechanisms.DESIGN:Randomized-controlled design.SETTING:Department of Geriatric Cardiology and Department of Pathophysiology,General Hospital of Chinese PLA.METHODS:Male Sprague-Dawiey rats,were randomized into 4 groups:sham-operation group,CHF group,CHF+ green fluorescent protein (GFP) group and CHF+SERCA2a group. Gene transfer 10 and 30 days two time points were set in each group.CHF rat models were established by coarctation of abdominal aorta.At postoperative 18 to 20 weeks,the successful rats in CHF group were intraperitoneally injected with 500 μL aseptic normal saline,and those in the CHF+ GFP group and CHF+ SERCA2a group were injected with the same dose of Adeno-associated viral gene with GFP and SERCA2a gene respectively.MAIN OUTCOME MEASURES:①At 10 and 30 days after gene transfer,hemodynamic parameters,the SERCA2a protein expression was analyzed by Western Blot,and SERCA2a activity was carded out by modified Jones method.②The difference of proteome of hearts was detected by expression proteomics in between CHF+ SERCA2a group and CHF group at 30 days.③Electrophoretic separation and quantitation of cardiac myosin heavy chain(MHC) isoforrns were done at 30 days.RESULTS: ①In CHF group and CHF+GFP group, SERCA2a expression and activity were lower than those in sham-operation group.At 10 days after gene transfer.SERCA2a expression was increased by 80.7%and still lower than that of sham-operation group,and activity was also increased by 89.9%,but still lower than that of sham-operation group.And at 30 days,the SERCA2a protein expression and activity in the CHF+SERCA2a group reached to levels of sham-operation group. ②At day 10,LVSP and LV +dp/dtmax and LV-dp/dtmax were increased and LVEDP was decreased dramatically,but they still did not reach to the levels of sham-operation group.At day 30,the parameters were normalized back to control levels.There were no significant differences in each index between CHF+SERCA2a group and sham-operation group.③Overexpression of SERCA2a increased some energy metabolic enzymes of hearts at 30 days.④There were a downregulation of α-MHC and an upregulation of 3-MHC in failing hearts.Overexpression of SERCA2a increased the expression of α-MHC and decreased the expression of β-MHC to the levels of sham-operation group at 30 days.CONCLUSION: Adeno-associated viral gene transfer of SERCA2a can enhance SERCA2a functions, maintain calcium homeostasis,improve cardiac energy metabolism,and normalize the expression of MHC isoforms in CHF rats.As a result,the heart functions can be improved. So adeno-associated viral gene transfer of SERCA2a has good therapy effects on CHF rats in short and relatively long periods.