Brain protection and recirculation therapy with non-viral vector gene

1)将人b-内啡肽编码序列与部分NL1基因连接，构建新的融合基因，构建具有控制和特异性分泌b-内啡肽的细胞系。携带Tet-on系统的HEK293细胞在多西环素（Dox）作用下以剂量依赖的方式分泌b-内啡肽。该细胞系被包裹并移植到猴子的脑脊液空间，但对给予的Dox （i.m）没有反应。另一方面，pomc上的neuro2a - tet细胞被包裹并移植到猴子的脑脊液空间。他们在给药后以剂量依赖的方式在猴子的脑脊液中分泌ACTH。从这些结果来看，脑脊液空间中被包裹细胞的肽分泌可以通过Tet-on系统进行控制。HEK293细胞可能不适合本实验。2)将含有lacZ、GFP-ODN或DsRed的HVJ-E注射到大鼠和猴子的大池中，并灌注4%多聚甲醛。取脑和脑膜，切片50 μm厚。检测了猴子和大鼠的大脑和脑膜lacZ、GFP-ODN或DsRed基因的转染情况。基因转染的效果在两种动物中都不稳定。原因不明，未做生理检查。

1)To create a cell line with controlled and specific secretion of b-endorphin, a new fusion gene was constructed by joining human b-endorphin coding sequence to part of NL1 gene. HEK293 cells carrying Tet-on system transfected with this fusion gene secreted b-endorphin in a dose-dependent manner upon doxycycline(Dox) administration. This cell line was encapsulated and transplanted in the monkey's CSF space, but failed to respond the administered Dox (i.m.). On the other hand, Neuro2A-Tet-on-POMC cells were encapsulated and transplanted in the monkey's CSF space. They secreted ACTH in the monkey's CSF in a dose-dependent manner upon Dox administration. From these results, peptide secretion from encapsulated cells in the CSF space can be controlled through Tet-on system. HEK293 cells may not fit for this experiment.2)HVJ-E containing the lacZ, GFP-ODN or DsRed was injected into the cisterna magna of rats and monkeys, and they were perfused with 4% paraformaldehyde. Their brains and meninges were removed and 50-μm-thick sections were made. The brains and meninges of both monkeys and rats were examined about the gene transfection of lacZ, GFP-ODN or DsRed. The efficacy of gene transfection was not stable in both animals. It's reason wasnot clear and the physiological examination has not been done.