Objective To investigate the expression and characteristics distribution of ciliary neurotrophic factor (CNTF) and its receptor during the development of retina of healthy Sprague-Dawley（SD）and Royal College of Surgeons (RCS) rats with hereditary retinal degeneration. Methods The expression and distribution of ciliary neurotrophic factor and its receptor were detected by immunohistochemical staining in the retinal paraffin sections of SD and RCS rats from newborn to 12 moths old. Results In the normal retina of SD rats 0-7 days after birth, positive CNTF staining was found in all of the retinal layers and the staining of ganglion cells strengthened and other cells weakened as the age of rats increased; the staining of ganglion cells reached the peak at the 4th week and lasted till the agedness. The same results of the CNTF staining were also found in RCS rats retina. Weak positive staining of CNTFR in all of the retinal layers was seen in the 0-3-day-old SD rats; the ganglion cells were darkly stained and incontinuous positive staining at the site which would develop to be the external segment was found; as the age increased, the positive staining of external segment of photoreceptor enhanced and reached the peak at the 14-28th day after birth. At the 56th day, the staining of ganglion cells in retina of SD rats was strengthened while the staining of external segment weakened till the agedness. The expression of CNTFR in retina of 3-14-day-old RCS rats was the same as which of normal SD rats basically, but the staining of external segment weakened obviously from the 21st day on, and negative staining of external and positive ganglion cells were detected at the 28th day till the agedness. Conclusions Expression of CNTF in normal SD rats and RCS rats with hereditary retinal degeneration is almost the same. The presence of significant difference of expression of CNTFR between normal SD rats retina and RCS rats retina may provide the experimental gist for the CNTF treatment to retinal degeneration. (Chin J Ocul Fundus Dis, 2006, 22: 120-123)
Objective To observe the morphological changes and gene expression during the transdifferentiation of adult retinal pigment epith elial(RPE) cells into neuronal phenotype in vitro induced by retrovirus and ciliary neurotrophic factor (CNTF). Meothds The adult RPE cells derived from CRL 2302 were infected by retrovirus with green fluoresence protein(GFP)and then were transfected further by liposome mediated CNTF expressing plasmid.The cellular ability of producing CNTG,and the expression of CNTF, CNTF receptor (CNTFR), and signal transduction molecule janus tyrosine kinases (JAK) were detected by enzyme linked immunosorbent assay, immunohistochemical stainin gand Western blotting method. Results After infected by retrovirus, the configuration of adult RPE cells didnrsquo;t change much, but expressions of neurons and some glial cells markers likeneurofilament (NF) protein and glial fibraillary acidic protein (GFAP) were detected. After further transfected by CNTF expressing plasmid, RPE cells which expressed CNTF highly and continuously had differential neurocytes; the expression of CNTFR didnrsquo;t change, but the distribution position changed to the cell membrane; expression of signal transduction molecule JAK increased obviously. Conclusion The adult RPE cells may transdifferentiate into neurons induced by retrvirus and CNTF. The transdifferentiation may relate to CNTF-CNTFR-JAK signal transduction pathway. (Chin J Ocul Fundus Dis, 2006, 22: 400-403)
Objective To observe the effects of immunologic cytokines or anti-angiogenesis gene transfer mediated by electroporation for choroidal melanoma cells.Methods The human embryo kidney cells and malignant choroidal melanoma cells were transfected with plasmids pNGVL-mIL2, pNGVL-mIL12, pCI-sFLK-1, pCR3.1-antiVEGF121,pCI-ExTek. Then the expression of mIL2, mIL12, sFLK-1, VEGF and ExTek were detected by enzymelinked immunosorbentassay (ELISA) and Western blot. Nude mice models of malignant choroidal melanoma were established and they were divided into four groups randomly. Each group was treated with 30 mu;l of 0.9% NaCl, 30 mu;g pNGVL, 30 mu;g antiVEGF121+sFLK-1+ExTek and 30 mu;g mIL2+mIL12 respectively by electroporation. Seven，14, 21, 28, 35 and 42 days after treatment, the tumor volumes were measured to calculate the tumor inhibition rate. Results ELISA and Western blot showed that mIL2，mIL12，sFLK-1 and ExTek were expressed after electroporation，VEGF expression was decreased remarkably. After treatment，the tumors of mIL2+mIL12 group were greatly inhibited with a tumor inhibition rate of 97.33%，while the tumors of antiVEGF121+sFLK-1+ExTek and pNGVL group were partially inhibited with tumor inhibition rates of 53.33% and 36.33% respectively.Conclusions Immunologic cytokines transfer mediated by electroporation can inhibit the growth of melanoma，but anti-angiogenesis only have a mild effects.