腸病毒71 型( EV 7 1)好發在嬰幼兒及學齡兒童，在重症患者當中，超過半數的人，會出現肢體殘障，或是智能發展遲緩，也因此成為國內公共衛生及流行病學重要的議題。目前仍無有效對抗腸病毒之疫苗或藥物，而根據文獻報導指出，小兒麻痺病毒之5 端非轉譯區(5' -UTR) 具有一個intemal ribosome enter site (IRES) ，此區之序列發生突變時會影響小兒麻痺病毒之轉譯能力，進而影響病毒之複製能力;在腸病毒71 型之5 端非轉譯區亦被報導具有一個IRES 序列，故本實驗希望藉由建立一帶有腸病毒71 型之IRES 及報導基因(Luciferase) 核酸序列的質體DNA 轉染人類神經母細胞瘤細胞，以建立in vitro 的快速藥物篩選平台，藉由從傳統中藥中篩選出可能有效影響腸病毒71 型IRES 活性之抑制劑，以作為探討中醫藥方防治腸病毒71 型感染之可行性。結果顯示，我們成功建立帶有腸病毒71 型之IRES 報導基因之質體，並建立一快速藥物篩檢平台，以多種中草藥驗證，發現白虎湯與升麻葛根湯可促進腸病毒EV71-IRES 轉譯之活性，而大黃、升麻、金銀花、甘露消毒丹、甘露飲、柴葛解肌湯等對EV71-IRES 轉譯活性無影響。藉由本實驗所建立之EV71 -IRES 快速藥篩平台可用以分析中草藥對腸病毒71 型IRES 之活性影響，並具潛力用以篩選出具有抑制腸病毒71 型之中藥。

Enterovirus71 (EV71) belongs to the Picornaviridae, a family of RNAviruses. EV71 infection was usually found in the infant and child and more thanhalf of the severely symptomatic patients wil1 experience sequelae involving limbweakness /atrophy or slow intellectual development. It is a big problem in thepublic health and epidemiology. There are stil1 no effective drugs or vaccines caninhibit EV71. Because both of EV71 and poliovirus are belonged toPicomaviridae, and some of the clinical symptoms of these viruses are similar.Therefore, we propose that some ofthe research models of poliovirus can be usedin the EV71' s studies. The 5' -UTR of Poliovirus has been reported to involvea region of intemal ribosome enter site (IRES). Moreover, the single-nucleotidemutation of IRES has been reported to regulate the translation of poliovirus andviral replication. Furthermore, EV71 has been reported to involve an IRES regionin 5' -UTR. The aim of this study wi1l construct the plasmid with EV71-IRESregion and luciferase gene and transfect into human neuroblastoma cells as themodel to mimic the process of translation of EV71 in the cells. We will also usethis model as in vitro platform to screen the traditional Chinese herbal medicinesand to estimate the feasibility of EV71 treatment by TCM. The result shows thatEV71 IRES reporter gene had been established as a high throughput screeningsystem for anti-EV71-IRES activity of TCMs in this study. Among TCMs selectedin this study, BOR HU TONG and SHENG MA GER GEN TONG were foundenhanced the translational activity of EV71-IRES. Other formulae, such as RheiRhizoma 、Cimicifugae Rhizoma 、Lonicera japonica 、GAN LUH SHIAUDWUDAN 、GANLUHYIIN 、CHAIR GER JIEE JI TONG, have no effect onEV71-IRES translation. We demonstrate the EV71-IRES reporter construct couldbe used as a high throughput screening system for TCM regulated EV7I-IRESactivity analysis. This screening platform has the potential for finding out theTCMs that with EV71 inhibition activity.