ABSTRACT
<p><b>OBJECTIVE</b>To explore the role of microRNA on the myocardial microvascular endothelial cells (CMECs) of ischemic heart rats in the process of angiogenesis and related regulation mechanism.</p><p><b>METHODS</b>Myocardial ischemic rats model was established by coronary ligation.Seven days after operation, the ischemic CMECs were cultured by the method of planting myocardium tissue and identified by immunocytochemistry to observe the biological characteristics of ischemic CMECs angiogenesis, to determine the window period of migration, proliferation, tube formation in the process of its angiogenesis. Dynamic expression changes of microRNA in the process of ischemic CMECs angiogenesis was detected using microRNA chip and further verified by real-time PCR, the core microRNA of the ischemic CMECs was defined and the predicted target genes of core microRNA were determined by bioinformatics methods and real-time PCR. At the same time, the protein expression of target gene and angiogenesis related genes of p38MAPK, PI3K,Akt,VEGF were measured by Western blot.</p><p><b>RESULTS</b>The CMECs of rats presented typical characteristics of microvascular endothelial cells, and factor VIII, CD31 related antigens were all positively stained by immunocytochemical analysis. The migration window period was on the first day, and the tube formation window period was on the second day of both control and ischemic groups, while the proliferation window period was on the third day for the normal group, and the sixth day for ischemic group. According to the expressional difference and their relationship with angiogenesis, miRNA-223-3p was ultimately determined as the core microRNA in the process of ischemic CMECs angiogenesis, real-time PCR verified this hypothesis. Bioinformatics methods predicted that Rps6kb1 is the target genes of miRNA-223-3p, the pathway analysis showed that Rps6kb1 could regulate angiogenesis via HIF-1α signal pathway. Moreover, the mRNA and protein expression of VEGF, p38MAPK, PI3K,Akt, which were the downstream molecules of Rps6kb1/HIF-1α signal pathway, were also significantly downregulated in ischemic CMECs from migration and proliferation stage.</p><p><b>CONCLUSION</b>Our results show that the miRNA-223-3p is the core microRNA of ischemic CMECs angiogenesis. MiRNA-223-3p could regulate Rps6kb1/HIF-1α signal pathway, inhibit the process of migration and proliferation of ischemic CMECs angiogenesis. MiRNA-223-3p is thus likely to be a core target for enhancing angiogenesis of ischemic heart disease.</p>
Subject(s)
Animals , Rats , Blotting, Western , Endothelial Cells , Physiology , Endothelium, Vascular , Hypoxia-Inducible Factor 1, alpha Subunit , MicroRNAs , Pharmacology , Myocardial Ischemia , Myocardium , Myocytes, Cardiac , Neovascularization, Pathologic , Phosphatidylinositol 3-Kinases , Platelet Endothelial Cell Adhesion Molecule-1 , RNA, Messenger , Ribosomal Protein S6 Kinases, 70-kDa , Signal TransductionABSTRACT
Case recruitment of large-scale clinical trials should be strictly checked in quality and quantity for it is the key to clinical trial. This study discusses the main difficulties and countermeasures in the case recruitment of large sample, multi-center clinical trials according to the national research project "Myocardial Infarction Secondary Prevention Study in Traditional Chinese Medicine".
ABSTRACT
Abstract: Large-scale clinical trial is an important measure of clinical evaluation on drugs. This paper introduces the concept and features of large-scale clinical trial, the possibility and necessity of large-scale clinical trial of traditional Chinese medicine, as well as its administration and quality control, with Myocardial Infarction Secondary Prevention Study in Traditional Chinese Medicine (MISPS-TCM), a National Program Subject, as an example.
ABSTRACT
Background Traditional Chinese medicine (TCM), especially herbal medicine, has been widely used in China and now is also being increasingly used in other countries for the treatment of cardiovascular diseases. Although many studies have demonstrated that certain Chinese herbal products are effective and safe for the treatment of cardiovascular diseases, most of these lack sufficient quality. Therefore, large randomized clinical trials and further scientific research to determine its safety, effectiveness are necessary.QiShen YiQi Dripping Pills (QSYQDP) is a herbal preparation clinically used in the treatment and prevention of coronary artery disease. Preliminary observations have shown its safety and effectiveness. Methods/Design This randomized, controlled trial will recruit 3600 patients with a history of myocardial infarction. Patients will be randomized into two groups by a Centr-Randomized System. One group receives QSYQDP, the other group receive aspirin. This trial protocol will describe eligibility criteria, detailed information on the treatment definition, blinding, endpoints, statistical methods, sample size determination, data management, legal aspects, and the current status of the trial. Discussion This trial is one of the first randomized, controlled clinical trial to evaluate the efficacy and safety of traditional Chinese herbal medicine in the treatment and secondary prevention of coronary artery disease. The results of this study should help to define the role of TCM in modern medical care, as well as to provide the management strategy for CAD patients in China and other countries.
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AIM： To investigate the effects of 6， 7－ dimethoxycoumarins on the proliferation and the cell cycle of human lung carcinoma cell（ PA） in vitro METHODS： The effects of 6， 7－ dimethoxycoumarins on the cytomorphology， growth and cell cycle of PA cells were evaluated by MTT assay and flow cytometry in vitro RESULTS： 6， 7－ dimethoxycoumarins could inhibit the proliferation of PA cells in a dose－ dependent manner The inhibition action of 6， 7－ dimethoxycoumarins plateaued at 160μ g/ml(52 4％ inhibition) The agent in dose of 80μ g/ml led to a decline in the proportion of cells to those in G2/M phase， made the cells remain in G0/G1 phase and reduced the proliferation index(PI) of PA cells CONCLUSION： In vitro， 6， 7－ dimethoxycoumarins can inhibit the proliferation of PA cells， which leads to arrest of the cells in G0/G1 phase by way of inhibiting DNA synthesis
ABSTRACT
AIM:To investigate the effects of 6,7-dimethoxycoumarins on the proliferation and the cell cycle of human lung carcinoma cell(PA)in vitro METHODS:The effects of 6,7-dimethoxycoumarins on the cytomorphology,growth and cell cycle of PA cells were evaluated by MTT assay and flow cytometry in vitro RESULTS:6,7-dimethoxycoumarins could inhibit the proliferation of PA cells in a dose-dependent manner The inhibition action of 6,7-dimethoxycoumarins plateaued at 160?g/ml(52 4% inhibition) The agent in dose of 80?g/ml led to a decline in the proportion of cells to those in G2/M phase,made the cells remain in G0/G1 phase and reduced the proliferation index(PI) of PA cells CONCLUSION:In vitro,6,7-dimethoxycoumarins can inhibit the proliferation of PA cells,which leads to arrest of the cells in G0/G1 phase by way of inhibiting DNA synthesis