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 establish a culture system in vitro of fetal and adult human retinal neural cells provide a model for the basic research of retinal neural cells and the medicinal exploitation. Methods Fetal human retinas(10~13 weeks after conception) and adult human retinas(20~40 years old) were dissected, dissociated, and put into culture plate which was coated with polylysine or rat tail gel. Specific growth factor EGF、FGF、BDNF or NT-4 were added to the culture medium. BrdU incorporation, Tunnel assessment and immuno-histochemistry and immuno-fluorescent staining were applied to determine cells proliferation, apoptosis and identify the component of cultured cells. Results Fetal human retinal cells and adult human retinal cells survived for up to 100 and 180 days in vitro. The addition of EGF、FGF、BDNF or NT-4 promoted the survival of both fetal and adult retinal neurons and stimultated proliferation of fetal retinal cells. The neurons or the rate of ganglion cells was observed with higher percentage in the group with growth factor adding than the group without. Conclusion Fetal and adult human retinal cells can be maintained in vitro and the fetal cells also can be expanded, which are helpful to generate retinal neurons for basic research and drug exploitation. The exogenous growth factors added to the culture medium can promote survival, proliferation and differentiation of retinal cells in culture. (Chin J Ocul Fundus Dis, 2002, 18: 279-282)
PURPOSE:To investigate the status and detailed structure of Rb gene in primary tumors and somatic cells of patients with retinoblastoma. To identify the character, origin and transmission of oncogenie point mutations. METHODS:DNA hybridization,SSCP analysis and PCR-associated direct sequencing. RESULTS:Among 108 RB patients examined 80 cases were found to have subtle alterations affecting Rb locus,including 44 cases with homozygous Rb point mutations, 20 cases with two independent heterozygous Rb point mutations, 16 cases with heterozygous mutations involved in one allele of Rb gene. Majority of bilateral RB patients and a small fraction of unilateral RB patients were detected to have a germline mutation. In addition the higher frequency of new germline mutation and parental origin of mutation were observed. CONCLUSION :Rb gene is closely associated with retinoblastoma. Two mutation events and resulting inaetivations of both Rb alleles are required for RB tumorigenesis. Based on our own data,the first event is exclusively point mutation. As for the second event,LOH accounts for two third of cases,point mutation for one third of cases. （Chin J Ocul Fundus Dis,1997,13: 12- 16）
OBJECTIVE:The hereditary form of retinoblastoma(RB)is a monogenic disorder which is due to germinal mutation of RB susceptibility gene located on 13q14.The majority of hereditary RB cases transmit as a Mendelian autosomal dominant inheritance that 50% of the offspring of a carrier will inherit the disorder susceptibility gene and all carriers will develop the disorder.The authors report 3 hereditary RB families with incompleted penetrance and irregular transmission of RB phenotype. METHOD:RFLPsamp;VNTRs for analysis of haplotype and SSCPamp;direct DNA sequencing for determination of RB point mutation. RESULTS:The mosaicism of Rb gene point mutation resulted in the incompleted penetrance and irregular transmission of RB phentype. CONCLUSION:DAN-based diagnosis can be used to differentiate the hereditary and nonhereditary forms of retinblastoma but only is the direct detection of disease-causing mutation reliable for determnation of carrier and estimation of th e risk for retinoblastoma. （Chin J Ocul Fundus Dis,1996,12: 37- 40）
Objective To assess the efficacy and safety of Chinese medicinal herbs for treating endometriosis. Methods We searched Cochrane Library, MEDLINE, EMBASE, CBM (from establishment to 2003). Randomized controlled trials (RCTs) and quasi-randomized controlled trials of patients with endometriosis were included. The quality of included studies such as randomization, blinding, allocation concealment and loss of follow up were evaluated and meta-analysis was performed by RevMan 4.3 software. Results Ten RCTs or quasi-RCTs involving 1 120 patients were included. Because of different therapies in the treatment and control groups, the results of outcome were described separately. Most of included studies suggested that the effects of traditional Chinese medicine (TCM) on general effect, pregnancy rate improvement and alleviating dysmenorrhoea were similar to Danazol or Tamoxifen, only a few studies showed better effects. There was no evidence to support that TCM was more effective than western medicine in reducing the size of endometriotic cysts. Only one study mentioned the recurrence rate and showed that TCM enema had lower recurrence rate than oral Tamoxifen with OR 0.17, 95%CI 0.04 to 0.67. Five studies mentioned adverse reactions and showed TCM had fewer adverse effects than western medicine. Conclusions Chinese medicinal herbs are effective in treating endometriosis with fewer adverse effects. The evidence is not b enough because of low quality of the included studies. Therefore, more high quality randomized controlled trials are required.