Objective To investigate the development and metastasis of malignant choroidal melanoma cell strain OCM-1-gfp modified with green fluorescent protein(GFP) and the factors which affected the tumor biological behaviors. Methods GFP was transfected into malignant melanoma cell strain OCM-1.Melanoma cells with high and stable expression of GFP were injected into subretinal space and the subcutaneous space of hind leg of Balb/c nude mouse respectively in order to establish orthotopic and heterotopic transplanted tumor models.The development and metastasis process of orthotopic tumor models was observed directly by fluorescence microscope，and the size of the hypodermal tumor was measured by vernier.The expressions of 13 genes in melanoma were detected by means of immunohistochemistry staining. Results Malignant choroidal melanoma cell strain OCM-1 stably expressed GFP and preserved the characteristics of parental generation，OCM-1-gfp may develop melanoma and continue to metastasize in nude mouse.Positive expression of most of the antibodies，including Rb，p53，p21，E2F，NFkappa;B，cyclin D1，proliferation cellular nuclear antigen（PCNA），bcl2、bclXL/S，bax，and epithelial growth factor（EGF）and its receptor（EGFR），was found.While the staining of inhibition gene p16 was negative. Conclusions GFP is the marker for observing the development and metastasis of malignant choroidal melanoma in vivo.The rate of tumor formation and development process in orthotopic models does not differs much from which in heterotopic models of malignant choroidal melanoma.The expressions of lots of genes in malignant choroidal melanoma developed from OCM-1-gfp including p16、p53、NFkappa;B，cyclin D，PCNA，EGF，and EGFR are abnormal. (Chin J Ocul Fundus Dis, 2006, 22: 170-173)
Objective To evaluate the application value of scanning laser angiography with a wide-field contact lens system in the diagnosis of choroidal melanoma. Methods Twenty-four patients with choroidal melanoma were randomly divided into two groups, who underwent fundus fluorescein angiography and indocyanine green angiography scanning with the wide-field contact and non-contact lens system respectively in order to acquire the 150deg;wide-field and 30deg;view image data. The quality of the images was comprehensively evaluated. Results Satisfying images were acquired from all of the 24 patients. Widefield contact lens system indicated the accurate adjacent relation between the lesion position and the other dissection mechanisms, and also provided the general information about the size of the tumor and the perfusion of fluorescien or indocyanine green in the blood vessels. At the same time, it enlarged the view scope 3-5 times, which make for the screening of the peripheral lesions. Conclusions Scanning laser angiography with a wide-field contact lens system has important application value in the diagnosis of choroidal melanoma. (Chin J Ocul Fundus Dis, 2006, 22: 166-169)
ObjectiveTo observe the effectiveness of radiotherapy for refractory choroidal hemangioma. MethodsEight patients (8 eyes) with choroidal hemangioma were enrolled in this retrospective study. All the patients had received laser or photodynamic therapy before without effectiveness. The patients included 7 males and 1 females. The age was ranged from 11 to 54 years old, with an average of (27.50±15.18) years. All the patients were affected unilaterally, including 3 right eyes and 5 left eyes. There were 5 eyes with circumscribed choroidal hemangioma, 3 eyes with diffused choroidal hemangioma. All eyes had extensively exudative retinal detachment. The vision was from light sensation to 0.01. The volume of the tumors was ranged from 1.96 to 5.35 cm3, with a mean of (3.37±1.06) cm3. All the patients were treated with X rays by conventional fractional radiotherapy. Four of 8 patients were applied 24Gy totally in 8 fractions, while the other 4 patients were applied 46Gy in 23 fractions. Follow-up period ranged from 7 to 95 months, with medium of 42 months. ResultsRetinas reattached in all the eyes while exudation being absorbed. No retinal detachment happened again. To the last follow-up, the vision was from light sensation to 0.6. Visual activity improved in 6 eyes while 2 eyes improved obviously. Visual acuity was stable in remaining 2 eyes. The volume of the tumors decreased to 1.24-2.16 cm3, with a mean of (1.68±0.30) cm3. The percentage of the tumor decreased by 14.6-72.7, with an average of (44.89±21.30)%. No radiotherapy-associated complication occurred. ConclusionRadiotherapy is an efficient and safe treatment for refractory choroidal hemangioma.
Objective To observe the macular morphological changes of choroidal melanoma with coherence tomography (OCT) after plaque radiotherapy (PRT). Methods A total of 48 patients (48 eyes) with choroidal melanoma who underwent125I PRT were enrolled in this study. All the patients were examined documenting OCT to get the image of macula. The macula of all the patients was not involved. The median visual acuity was 0.4plusmn;0.2, which ranged from 0.02 to 1.0. There were 18 eyes (37.5%) with retinal detachment, 12 eyes (25.0%) with retinal pigment epithelium (RPE) changes, seven eyes (14.6%) with macular edema, epimacular membrane, detachment combined with edema, exudation and RPE changes, 11 eyes (22.9%) with normal macular structure. The median follow-up time was (10.4plusmn;5.9) months, which ranged from one to 24 months. The tumor control situation and visual acuity were observed in follow-up period. The same equipment and methods of OCT were used to return visit in follow-up period. The macular morphological changes at the final visit and its relationship with PRT and visual acuity were contrastively analyzed. Results All the patients had good control of tumor. The vision acuity improved in two eyes (4.2%), unchanged in 10 eyes (20.8%), and decreased in 36 eyes (75.0%). The differences of the visual acuity was statistically significant between before and after treatment (Z=-3.778,P＜0.05). There were 13 eyes (27.1%) with retinal detachment; nine eyes (18.8%) with RPE changes; 17 eyes (35.4%) with macular edema, detachment combined with edema, exudation and RPE changes; six eyes (12.5%) with proliferation, atrophy, detachment combined with edema, exudation and epimacular membrane;three eyes (6.3%) with normal macular structure. There were 15 patients (31.3%) with two or more abnormal macular morphology after PRT. Conclusions Retinal detachment, RPE changes, macular edema and exudation are common abnormal macular morphology after PRT. The incidence rate of abnormal macular morphology is increased. There are 31.3% patients with two or more abnormal macular morphology.
Objective To isolate and purify the melanoma stem cells (MSC) in choroidal melanoma OCM-1 cells. Methods OCM-1 cells were resuscitated, and after cultured in standard Dubecco's modifided Eagle's medium (DMEM)/F12, they were cultured in serum-free medium (SFM). The cultured MSC were isolated and purified, and the positive rate of CD133, the specific markers of neurostem cells, was observed by flow cytometry (FCM). The 6th generation of the cells were stained by musashi-1 immunocytochemistry, and the rate of the positive cells was observed under the microscope. Results After the Adherent OCM-1 cells cultured in SFM, the number of the adherent number decreased obviously. The cells at the 6th generation grew as the suspended gobbets, which represented the typical grow manner of the stem cells. Positive CD133 could be found in the cells of different generations, which was 2.5％, 21.7％, and 57.8％ in the non-isolated OCM-1 cells, the 1st generation of isolated cells, and the 2nd generation cells, respectively. The positive rate of CD133 in the cells at the sixth generation was 79.8% with b positive expression of musashi-1. Conclusion MSC is in the human choroidal melanoma OCM-1 cells. The suspended stem cells may be purified by limited differentiation and serial passage in SFM. (Chin J Ocul Fundus Dis, 2007, 23: 87-90)
Objective To evaluate the clinical and histopathological features of diffuse choroidal melanoma. Methods The clinical and histopathological data of 11 patients with diffuse choroidal melanoma were reviewed retrospectively. Those patients were referred to Tianjin Eye Hospital because of visual loss or ophthalmalgia (10 cases), or Coats disease with secondary glaucoma and atrophy bulbi (1 case). The clinical disgnosis included choroidal tumor or melanoma (8 cases), absolutestage glaucoma (2 cases) and atrop hy bulbi with Coats disease (1 case). Nine patients received enucleation, and 2 patients received enucleation combined with orbital exenteration. The cellular proliferation was assessed by Ki-67staining. Results All 11 tumors had grown flatly with a wide base ranged from 12 to 20 mm, and tumor thickness ranged from 2 to 4 mm. There were 9 cases of mixed cell type, 1 case of epithelioid cell type and 1 case of necrotic cell type. The tumors invaded into the sclera in 7 cases and orbital cavity in 3 cases. Secondary glaucoma was found in 7 cases. On average, 9% (7%13%) of tumor cells were Ki67 positive and most of them located at the tumor base. There were more Ki67 positive epithelioid tumor cells than Ki67 positive spindle-shaped cells. Conclusions Diffuse choroidal melanoma had a special growth pattern and is difficult to be recognized, sometimes could be misdiagnosed as glaucoma or other choroidal tumors. With its wide base, this tumor could easily invade the orbit and metastate, and its prognosis is very poor.
Objective To explore the clinicopathological features and misdiagnosed causes of choroidal hemangioma. Methods Seven misdiagnosed cases(7eyes) of choroidal hemangioma,which were enucleated,were analysed retrospectively. Results One of the 7 cases was misdiagnosed as absolute phase of the secondary glaucoma,and 6 of them as choroidal melanoma before the enucleation.The majority of cases in this series manifested themselves clinically and pathologically in progressive loss of visual acuity and a flat elevated tumor located at the posterior ocular fundus near the optic disc and associated with exudative retinal detachment.And also there were occasionally small focal or linear pigmentary deposites obser ved on the surface of the neoplasm. Conclusion A flat elevated discoid tumor in the posterior fundus with extensive exudative retinal detachment might be a clinicopathological feature of the choroidal hema ngioma. (Chin J Ocul Fundus Dis,1999,15:91-93)
ObjectiveTo observe the clinical effect of prolonged photodynamic therapy (PDT) irradiation time combined with intravitreal injection of ranibizumab in the treatment of circumscribed choroidal hemangioma (CCH).MethodsA retrospective clinical study. From March 2012 to March 2018, 51 eyes of 51 patients diagnosed in Shenzhen Eye Hospital were included in the study. Among the patients, the tumor of 36 eyes were located in macular area, of 15 eyes were located outside macular area (near center or around optic disc). All patients underwent BCVA, color fundus photography, FFA, ocular B-scan ultrasonography and OCT examinations. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. OCT showed 48 eyes with macular serous retinal detachment. of 36 eyes with tumor located in macular area, the logMAR BCVA was 0.05±0.05, the tumor thickness was 4.5±2.2 mm, the diameter of tumor was 9.7±3.6 mm. Of 15 eyes with tumor located outside macular area, the logMAR BCVA was 0.32±0.15, the tumor thickness was 3.8±1.4 mm, the diameter of tumor was 7.7±1.9 mm. PDT was performed for all eyes with the irradiation time of 123 s. After 48 h, all patients received intravitreal injections of 0.5 mg ranibizumab (0.05 ml). At 1, 3 and 6 months after treatment, the same equipment and methods before treatment were used for related examination. BCVA, subretinal effusion (SRF), tumor leakage and size changes were observed. BCVA, tumor thickness and diameter before and after treatment were compared by t test.ResultsAt 6 months after treatment, the tumor was becoming smaller without scar formation. FFA showed that the blood vessels in the tumor were sparse compared with those before treatment, and the fluorescence leakage domain was reduced. OCT showed 43 eyes of macular serous detachment were treated after the combined treatment. The logMAR BCVA were 0.16±0.15 and 0.55±0.21 of the eyes with tumor located in or outside macular area, respectively. The difference of logMAR BCVA between before and after treatment was significant (t=－2.511, －2.676; P=0.036, 0.040). Both the tumor thickness (t=3.416, 3.055; P=0.011, 0.028) and diameter (t=4.385, 4.171; P=0.002, 0.009) of CCH patients were significantly reduced compared with that before treatment.ConclusionThe tumor of CCH can be reduced by prolonged PDT irradiation time combined with intravitreal injection of ranibizumab.
ObjectiveTo comparatively observe features of choroidal osteoma by multimodal fundus imaging methods. MethodsThis is a retrospective case study. Sixteen patients (16 eyes) with choroidal osteoma were enrolled in this study. The patients included 6 males (6 eyes) and 10 females (10 eyes), with an average age of (30.5±2.4) years. All patients received examination of best-corrected visual acuity, slit lamp microscope, indirect ophthalmoscopy, fundus color photography, fundus autofluorescence (AF), fundus fluorescein angiography (FFA) and spectral domain optical coherence tomography (SD-OCT). The tumors were classified as fresh lesion (clear boundary and rosy tumor with smooth surface) and obsolete lesions (pale and flat tumor with obvious patches). The tumor features of color fundus photography, AF, FFA and SD-OCT were comparatively observed. ResultsThere were 5 fresh lesions and 11 obsolete lesions. Color fundus photography showed the tumor color was orange-red or yellow-white with clear boundary and retinal blood vessels on the surface of the tumor. The color of fresh lesion was rosy. In general, choroidal osteoma shown weak AF, however AF of fresh tumor was slightly stronger than the obsolete tumor, and retinal detachment region showed relatively stronger AF. FFA of fresh tumor indicated uniform intense fluorescence with clear boundary at late stage, much stronger than obsolete tumor. SD-OCT showed mesh-like reflected signal in the choroidal layer, but different from the surrounding choroidal vascular structures. ConclusionsThe tumor color is orange-red or yellow-white in color funds photography, which shown weak AF. FFA showed mottled hyperfluorescence in the early stage and tissue staining at the late stage. SD-OCT showed mesh-like reflected signal in the choroidal layer.