Background and Objectives:For the development of an arteriogenic gene therapy in peripheral artery occlusive disease, we developed a novel angiogenesis assay, with electroporation-mediated naked DNA delivery to the skeletal muscle. Materials and Methods:The levels of the expression CAT were compared between pJDK and pc-DNA3.1, in HeLa and C 2 C 12 cell lines, and skeletal muscle. The well known angiogenic gene, pJDK-hVEGF 165 , was injected, intramuscularly, into the tibialis anterior muscle of Balb/C mice, which was followed by electroporation. Two days later, the anterior tibialis muscles were divided into halves, embedded, and cultured in growth factor-reduced Matrigel. The capillary network area formed by the newly sprouting tube-like structures was calculated. For validation of this ex vivo assay, the connective tissue growth factor gene (pJDK-CTGF) was tested both by this new assay, and by the mice-hind limb ischemia model, with Laser Doppler imaging. Results:The pJDK showed a significantly higher level of CAT expression than the pcDNA3.1. From the pJDK-hVEGF 165 injected explants, endothelial cell migration and tube-like formation occurred on day 2, and the capillary network formation peaked on day 7. The capillary network formation in the pJDK-hVEGF 165 group was markedly increased to that in the pJDK group. From the skeletal muscle assay, the pJDK-CTGF showed no angiogenic activity or attenuated VEGF-induced capillary network formation. The LDI flux ratio, on day 10 in the mice-hind limb ischemia model, for the mice treated with the pJDK-CTGF and pJDK-hVEGF 165 was significantly lower than that of the mice treated with the pJDK-hVEGF 165 alone. Conclusion:The skeletal muscle ex vivo assay, using an electroporationmediated naked DNA delivery, is a simple, quantitative and reproducible method for assessing angiogenic genes. CTGF could be an anti-angiogenic factor due to its inhibition of VEGF. (Korean Circulation J 2003 ; 33 (4) : 321-332)
Gyupyo (圭表, Gnomon) consists of Gyu (圭, Measuring Scale) and Pyo (表, Column), and was one of the traditional astronomical instruments in East Asia. Daegyupyo (Large Gnomon) was manufactured in the Joseon dynasty around 1434 ~ 1435. To increase the measurement accuracy, it was equipped with a Hoengyang (橫梁, Cross-bar) and used a Youngbu (影符, Shadow-Definer) which was invented during the Yuan dynasty (1271 ~ 1368). The cross-bar was installed on the top of the column and this structure was called Eol (臬). In addition, three plumbs hanging from the cross-bar was employed to vertically built Eol on the measuring scale. This method was also used to not only check the vertical of Eol but also diagnose the horizontal of the cross-bar. Throughout this study, we found that a cross-bar in a gnomon has played three important roles; measurement of the shadow length made by the central part of the Sun, increase of the measurement precision using the shadow-definer, and diagnosis of the vertical of Eol and the horizontal of the cross-bar itself using the three plumbs. Hence, it can be evaluated that the employment of a cross-bar and a shadow-definer in a gnomon was a high technology in the contemporary times. In conclusion, we think that this study is helpful for understanding the Large Gnomon of the Joseon dynasty.
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