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%.
ObjectiveTo observe the application value of RetCam Ⅲ in the diagnosis of retinoblastoma (RB). MethodsThe clinical data of 125 eyes from 94 patients with RB were retrospectively analyzed. According to the International Intraocular Retinoblastoma Classification, the results of binocular indirect ophthalmoscope and RetCam Ⅲ was compared. The consistency between RetCam Ⅲ and binocular indirect ophthalmoscope was analyzed by Kappa test. The detectable rates of tumor between binocular indirect ophthalmoscope, RetCam Ⅲand B scan ultrasound were comparatively analyzed. ResultsBy binocular indirect ophthalmoscope, there were 18 eyes (14.4%) in stage A, 15 eyes (12.0%) in stage B, 10 eyes (8.0%) in stage C, 28 eyes (22.4%) in stage D, and 54 eyes (43.2%) in stage E. Using RetCam Ⅲ, the classification was 18 eyes (14.4%) in stage A, 16 eyes (12.8%) in stage B, 11 eyes (8.8%) in stage C, 27 eyes (21.6%) in stage D, and 53 eyes (42.4%) in stage E. The RB classification of RetCam Ⅲ was quite consistent with binocular indirect ophthalmoscope (Weighted Kappa Coefficient=0.979 8, 95% confidence interval=0.960 1-0.999 5). A total of 150 tumors from stage A-D were observed by binocular indirect ophthalmoscope. In which, 147 tumors (98.0%) were found by RetCam Ⅲ, 122 tumors (81.3%) were found by B scan ultrasound. No serious complication was found in these patients. ConclusionsRetCam Ⅲ can detect RB of different stage in clinic. It is quite consistent with binocular indirect ophthalmoscope.
ObjectiveTo analyze the clinical features, treatment and prognosis of asymptomatic patients with retinoblastoma.MethodsA retrospective series of case study. Eight asymptomatic patients (11 eyes) with the diagnosis of retinoblastoma by screening enrolled in Department of Ophthalmology of The Eye-ENT Hospital of Fudan University from January 2006 to March 2019 were included. There were 6 males and 3 females ranging from 2 days to 20 months, with a median age of 6 months. Five patients were unilateral retinoblastoma while 3 patients were bilateral. Based on the International Classification of Intraocular Retinoblastoma, 4 eyes were stage A, 3 eyes were stage B and 4 eyes were stage C. One patient had family history. Four patients were evaluated the Rb1 mutation. Routine ophthalmic examinations and ultra-wide field fundus imaging were performed on the 16 parents and 3 siblings of the 8 patients. Systemic intravenous chemotherapy was performed using the Carboplatin, Vincristine, Etoposide protocol, intra-arterial chemotherapy using Carboplatin and Melphalan, and local treatment involved cryotherapy and transpupillary thermotherapy. The mean follow-up time is 47.25 months.ResultsNone of the 8 children had any ocular symptoms. Six patients received intravenous chemotherapy (5-6 times), 1 patient received intra-arterial chemotherapy (3 times), and 1 patient just received local treatment. Among the 11 eyes, 9 eyes were treated with local cryotherapy and 8 eyes were treated with transpupillary thermotherapy. During the follow-up period, 2 patients had new tumor, and the average time was 6.3 months after the last chemotherapy. At the last follow-up, the tumor disappeared in 11 eyes, remained stable in 11 eyes. The eye protection rate was 100% (8/8) for patients without eyeball excision. The best corrected visual acuity was 0.1 for 3 eyes and 1.0 for 5 eyes. Three eyes were not found. One heterozygous mutation of Rb1 gene [1c.35_69del (p.T12fs)] was identified in 1 patient, and the other 3 patients were not detected. One had bilateral bulbar tuberculosis of the 16 parents, 1 had bilateral RB of the 3 siblings. They were the mother and brother of a child with bilateral RB.ConclusionsFundus screening is helpful for the detection of early RB. The eye protection rate is high and the long-term vision prognosis is good after systemic or topical chemical drugs (IVC, IAC) and ocular topical treatment (cryopreservation and transpupillary thermotherapy).
Objective To observe the microvessel density（MVD）and expression of vascular endothelial growth factor (VEGF) in retinoblastoma（RB）before and after comprehensive treatment and explore its clinical correlations with tumor infiltration and recurrence. Methods Sixty-one paraffin specimens of RB were divided into enucleation without comprehensive treatment group (untreated group，52 cases), planned enucleation before comprehensive treatment group (planned enucleation group，six cases) and tumor recurrence after comprehensive treatment group（recurrence group，three cases）. There were optic nerve invasion in 19 cases，no optic nerve infiltration in 33 cases. The MVD and VEGF expressions of 61 paraffin specimens were detected by streptavidin biotin-peroxidase (SP) immunohistochemistry with monoclonal antibody of CD34 and VEGF. Real time PCR was performed for the VEGF expression of planned enucleation group and recurrence group.Results In the untreated group，the MVD and VEGF expression of optic nerve infiltration cases were significantly higher than those of cases without optic nerve infiltration（t=-2.4685, P=0.017; chi;2=8.416 6，P=0.032 8）.Tumor microvessel regression, decreased MVD, occlusion and hyaline changes of blood vessels were observed in the planned enucleation group in the course of systemic chemotherapy. Many neovascularized capillaries and increased MVD were observed in tumor tissues of the recurrence group. The VEGF expression of planned enucleation group was lower than that in the recurrence group.Conclusions There was no significant difference on VEGF expression in RB between with and without comprehensive treatment. The increasing MVD and VEGF expression of cases without comprehensive treatment were related to the optic nerve infiltration. And the increasing MVD may also play an important role in RB recurrence after comprehensive treatment.
Objective To observe the regional expression of hypoxia inducible factor-1alpha; (HIF-1alpha;) in retinoblastoma and its relationships with vascular endothelial growth factor (VEGF), Bax and Ki-67. Methods Immunohistochemical study for HIF-1alpha;, VEGF, Bax and Ki-67 was performed in 39 paraffinaceous examples of retinoblastoma. Each pathological section was divided into five regions: the surface region, the central part, the bottom part, the choroidal region and seeding tumors. The expressions, correlations and distributional differences of these factors were all invested both integrally and regionally. Results In the 39 cases of retinoblastoma, 10 cases (25.6%) were negative for HIF-1alpha;; 29 cases (74.4%) were positive for HIF-1alpha;, including 17 cases (43.6%) (+), 12 cases (30.8%) (++). Regionally, HIF-1alpha; was positive in 71.1%, 36.8%, 84.2%, 54.5% and 82.1% of the cases in the surface region, the central part, the bottom part, the choroidal region and seeding tumors, respectively, which was statistically reliable (chi;2=24.55，P＜0.001). The positive rate of VEGF, Bax and Ki-67 was 53.8%, 66.7% and 59.0%, respectively. In different regions, the positive rates of VEGF and Bax were different (chi;2=26.77, 22.79; P＜0.001), but there was no regional distinctions in the expression of Ki-67 (chi;2=0.47, P=0.976). Both the expression of VEGF and Bax had a positive correlation with that of HIF-1alpha;(rs=0.48, 0.39; P=0.002, 0.021), but there was no relationship between the expressions of Ki-67 and that of HIF-1alpha; (rs=0.09, P=0.606). Regionally, the expressions of VEGF, Bax and HIF-1alpha; shared similar distributional features: positive rates were higher in the surface region, bottom part and seeding tumors, and were lower in the central part and choroidal region, which was different from the expression of Ki-67. Conclusion The anoxic zones are more likely to be located in the marginal parts in retinoblastoma, and the expressions of VEGF and Bax had a positive correlation with that of HIF-1alpha; in different regions in retinoblastoma.