Retinoblastoma susceptibility gene(Rb gene)is a tumor suppressor,which often deleted,mutated or inactivated in many malignant tumors.We construct an antisense Rb gene expression vector DOLRBAS and transfected it into human embryonic lung fibroblast(HEL cells)by electroporation technique.With the transient expression of antisense Rb gene the Rb protein synthesis is reduced and the growth rate of HEL cells in increased,but no coloney of HEL cells formed in soft agar.This result indicates that Rb gene can block cell division of HEL cells,but inactivation of Rb gene only is not sufficient for malignancy transformation of HEl cells. （Chin J Ocul Fundus Dis,1993,9:198-201）
Objective To investigate the correlation of expression of Fas/Fas ligand (FasL) and apoptosis in retinoblastoma (RB)． Methods The expression and distribution of Fas/FasL were detected by using immunohistochemical staining in 32 cases of RB． Light microsc opy (32 cases), electron microscopy (4 cases) and TdT mediated biotin-d UTP nick-end labeling (TUNEL) (12 cases) were used to study apoptosis in RB． Results Apoptotic RB cells mostly located at RB regress area． Chromatin margination and apoptotic bodies were found in RB． TUNEL posi tive labeling cells especially located in tumor regress area． Positive immunola beling for Fas and FasL was found in all RB specimens． There was a highly signi ficant and positive correlation between the expression of Fas/FasL and apoptotic indices (AI) (Plt;0.01 or 0.001)． Conclusion The results suggest that apoptotic cell death is prevalent in RB and it may be one type of the most dominant cell death． Fas system may play an important role in oncogenesis and progression of RB, and the up-regulation of Fas system expression might induce RB cell apoptosis． (Chin J Ocul Fundus Dis, 2001,17:21-23)
Objective To observe the retinoblastoma (RB) reexamination of children with new and recurrence retinoblastoma under special circumstances.MethodsFrom January 2, 2020 to March 15, 2020, 30 children with RB who had fundus examination in Henan Children's Hospital were enrolled in this study. Among them, 14 were male, 16 were female; 18 were monocular and 12 were binocular. The average age was 37.07±18.15 months. The mean age of initial diagnosis was 20.23±13.77 months. Two patients had a family history (6.67%). In 42 eyes, stage B, C, D and E were 7, 8, 20 and 7 eyes, respectively. Twenty-one eyes had finished the treatment course and 21 eyes were during treatment. All the children underwent RetCam fundus examination, orbital MRI, ocular B-ultrasound and so on. Whether the children had new tumor or recurrence at different treatment stages was observed.ResultsAmong 7 eyes in stage B, there was no recurrence or new tumor at the end of treatment or in the process of treatment. Among 8 eyes in stage C, there were 1 eye with new tumor and 1 eye with activity tumor at the end of treatment. Among 20 eyes in stage D, there were 1 eye with recurrence tumor at the end of treatment, 3 eyes with new tumor and 7 eyes with activity tumor at the end of treatment. Among 7 eyes in stage E, 5 eyes had eyeball enucleation and 2 eyes were receiving treatment; there were 1 eye with activity tumor at the end of treatment, 1 eye with recurrence tumor, 1 eye with activity tumor. Among 18 monocular eyes, there were 11 eyes in the treatment process, 2 eyes with new tumor, 1 eye with recurrence tumor and 3 eyes with activity tumor. Of the 24 binocular eyes, 10 were receiving treatment and there were 3 eyes with new tumor, 6 eyes with activity tumor. Twenty-one eyes had finished the treatment course, the average time required for follow-up was 3.71±0.31 months, and the average time delayed for follow-up was 6.43±1.66 weeks. There was a recurrence of tumor in 1 patient who had finished the whole treatment, the incidence was 4.76%. In the course of treatment, 21 eyes were required to have a follow-up time of 3 weeks, and the average delayed follow-up time was 6.00 ± 1.89 weeks. There were 5 eyes with new tumors, with a incidence of 21.74%. Nine eyes still had activity and needed to be treated in time.ConclusionsThe higher the risk of tumor staging, the more relapses and new tumors. The patients who are being treated, the time of delayed follow-up, the higher the recurrence or new tumor than the children who have finished the treatment course and delayed the follow-up. The children who have relapsed or new tumor in the treatment course of binocular are higher than the children who have monocular.
Objective To observe the effect of resveratrol on multidrug resistance (MDR) in human retinoblastoma cells treated. Methods RB cells in logarithmic growth phase were divided into experimental group and control group. RB cells in experimental group were cultured with different concentrations of resveratrol (6.25, 12.50, 25.00, 50.00, 100.00 mu;mol/L) for 24 and 48 hours. The proliferation (absorbance value) was assayed using methyl thiazolyl tetrazolium (MTT). RB cells were cultured with 50.00 mu;mol/L resveratrol for 48 hours. The expressions of MDR-1, cyclooxygenase-2 (COX-2)、multidrug resistance-associated protein-1 (MRP-1), glutathione-S-transferases-pi; (GST-pi;) were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The RB cells of the control group were cultured with 0.5% dimethyl sulfoxide. Results Compared with the control group, the absorbance value decreased in experimental groups (6.25, 12.50, 25.00, 50.00 mu;mol/L) in a dose dependent manner (F=4.782，P＜0.05). The difference of absorbance value between 50.00 and 100.00 mu;mol/L experimental groups was not significant (F=6.351,P＞0.05). Compared with the control group, the mRNA (t=9.170, 5.758, 4.152, 4.638) and protein (t=3.848, 5.955, 4.541, 3.514) expression levels of MDR-1, MRP1, COX-2, and GST-pi; decreased in the experimental group (P＜0.05). Conclusion Resveratrol can down-regulate the expression of MDR in RB cells.
Purpose To establish orthotopic heterotransplanted model of retinoblastoma(RB)into the vitreous cavity of nude mice for experimental studies of gene therapies in vivo. Methods To prepare the concentrated cell suspensions of human RB cell line HXO-Rb44 and inject them into the vitreous cavities of 10 nude mice(20 eyes).The experimental mice were randomly divided into 2 groups:the first group,5 mice (10 eyes) were injected with 5 mu;l of the cell suspensions,and the second group,5 mice (10 eyes)with 10 mu;l．The transplanted RB was observed by inspection and ophthalmoscopy,and graded semi-quantitatively in vivo.Light and electrone microscopes were used for histopathological examination,ABC immunohistochemical methods for NSE and GFAP,and flow cytometric investigations for DNA index(DI) and s-phase fraction (SPF). Results All of the models of RB were successfully established by injecting the cell suspensions of human RB cell line into the mice vitreous cavities.The immunohistochemical reaction of the transplanted tumor cells to NSE was positive,and negative to GFAP．The DNA index was found to be more than 1.1,and the determination of SPF indicated the presence of proliferative ability of the transplanted RB cells. Conclusion The above findings reveal that the transplanted tumor cells were similar to those of the parent cells,hence the orthotopic heterotransplantation of RB via nude mice vitreous cavity injection is a superior method in establishing the animal model for the further experimental studies of gene therapies in vivo. (Chin J Ocul Fundus Dis,1998,14:144-148)
OBJECTIVE：Observing the clinical and pathological features of Coats disease. METHODS:Reviewing the clinical data and pathologic slides duly confirmed by pathology of 19 cases of Coats disease,which belonging to our college's Laboratory of Ophthalmologic Pathology from 1959 to 1994. RESULTS: 14 males,5 females,aged 1-18 years. More boys were affected than girls in the age group under 10 and that difference between both sexes became gradually less as they grew older. The main pathologic changes were the vascular dilatation and congestion of the outer layer of the retina,the uneven thickness of the vascular walls and the proliferation of the connective tissue. Retinal protuberance was seen in most of the advanced cases.with bleeding and vascular changes on its surfaces. The main pathologic changes were the detachment of retina and the appearance of many foam cells and crystals of cholesterol in the subretnal fluid,and calcification and ossification of the outer layer of the retina were found in some cases. CONCLUSION :Cytological examination of the subretinal fluid might be the liable method in differentiating between the Coats disease and retinoblatstoma. （Chin J Ocul Fundus Dis,1996,12: 157-159）
ObjectiveTo identify the pathogenic mutation in a three generation Chinese family with low penetrance retinoblastoma (RB). Methods8 from 9 family members received complete ophthalmic examinations. DNA was extracted from 6 family members. Using exon combined target region capture sequencing chip to screen the candidate disease-causing mutations. Sanger sequencing were used to confirm the disease-causing mutation. ResultsAmong 9 family members, the proband (Ⅲ2) was bilateral RB, Ⅲ1 was unilateral RB, Ⅲ3 was dead for bilateral RB. Normal fundus were observed in the left eye ofⅢ1 and the eyes of other family members except the proband. Sequence analysis of RB1 gene revealed a missense mutation c.1981C > T (p.Arg661Trp) in the proband and two carriers (Ⅱ2, Ⅱ3), but not in the two normal subjects (Ⅱ1, Ⅱ4). We suspect that the RB penetrance in the family was 50%. ConclusionsThere is a missense mutation c.1981C > T in a Chinese family with low penetrance RB. The RB penetrance is 50%.
Objective A short-term observation study of ocular surgery with or without systemic chemotherapy to treat retinoblastoma (RB). Method The clinical data of 66 RB cases treated at Beijing Tongren Hospital from August 2006 to September 2007 were retrospective reviewed.All patients received ultrasound (B scan/CDI), radiology examination (CT scan/MRI) and RetCam retina examination, and classified according to the International Intraocular Retinoblastoma Classification (IIRC) method. Focal therapies (include laser and cryotherapy) were applied to Group A, systemic chemotherapy (CCTV protocol, Cyclosporine, Carboplatin, Teneposide and Vincrinstine) plus ocular surgery (include focal therapy, enucleation and orbital exenteration) to Group B, C, D and E. Patients were followedup for more than 6 months. Result Of 66 cases (36 male/30 female), 16/66 cases were bilateral RB and 50/66 were unilateral. Follow up period was from 6 to 13 months (mean time 8.6 month). According to IIRC, there were 5 eyes in Group A, 6 eyes in Group B, 5 eyes in Group C, 15 eyes in Group D and 51 eyes in Group E( include extraocular disease). 22/82 eyes were conserved at the end of follow up, including 5/5(100%) of group A, 6/6(100%) of group B, 5/5(100%) of Group C, 4/15(26.7%) of Group D and 2/51(3.9%) of Group E. 5/66 (7.6%) patients died during this period, they all were in Group E. Conclusion Ocular surgery combined with systemic chemotherapy is an effective method to treat retinoblastoma. The consequences depends on the classification with higher conservative rate of eyeballs in early and middle stage (Group A, B, C), and lower rate in late stage (Group D and E).
Objective To study hyperthermia induced apoptosis and the effect of aspirin on hyperthermia induced apoptosis in retinoblastoma cells. Methods Retinoblastoma cells (Y79) were divided into two groups:hyperthermia groups,hyperthermia+aspirin (0.18～18mu;g/ml) groups.Heat shock condition:44℃,heat shock time:10,20,30, and 40 minutes respectively.The following events were studied after heat shock by using FAC Scan: ①cell apoptosis; ②heat shock protein 70 (HSP70) expression；③bcl-2 expression. Results Apoptosis was induced by the treatment of hyperthermia (44℃) in Y79 cells in a heat dose dependent fashion.Longer time heating (44℃，40 minutes) induced necrosis rather than apoptosis.Aspirin could rescue Y79 cells from hyperthermia induced apoptosis in a dose dependent manner．HSP70 was induced in Y79 cells after heat shock,it was further enhanced by the treatment of aspirin(＞1.8mu;g/ml).Heat shock itself showed no effect on bcl-2 expression in Y79 cells,aspirin,on the other hand,could enhance bcl-2 expression in a modest level in heat treated Y79 cells. Conclusions Hyperthermia may induce apoptosis in Y79 cells which can be protected by use of aspirin．The enhancement of HSP70 and bcl-2 expression in Y79 cells by the treatment of aspirin in heating condition may be responsible for the protective function. (Chin J Ocul Fundus Dis, 1999, 15: 143-145)
Intraocular tumors is a serious blinding eye disease, which has a serious impact on patients' vision and even life. At present, the main treatments include surgical treatment, radiation therapy, chemotherapy, laser therapy and combination therapy. In recent years, with the wide application of anti-vascular endothelial growth factor (VEGF) in the treatment of ocular diseases, many studies have confirmed that anti-VEGF drugs play an important auxiliary role in the treatment of intraocular tumors and its complications. In terms of the therapeutic effect, intravitreal anti-VEGF combined with other methods have a good prognosis in the treatment of choroidal metastatic carcinoma and retinoblastoma, while the therapeutic effect of uveal melanoma is still controversial. In the treatment of intraocular tumor complications, intravitreal anti-VEGF also has a good effect on the secondary lesions of choroidal osteoma and radiation retinopathy. As for drug safety, intravitreal anti-VEGF can significantly reduce the toxic and side effects of systemic chemotherapeutic therapy. However, the dosage and medication regimen of anti-VEGF drugs in the treatment of intraocular tumors and their complications have not been unified in current studies, and further basic and clinical trials are still needed to explore in the future.