Objective To investigate the related factors of effects on distant visual acuity after photodynamic therapy (PDT) for choroidal neovascularization (CNV). Methods One hundred and thirty-five cases (135 eyes ) of CNV treated with PDT were observed. The gender, preoperative distant and near visual acuity, disease course, pathogeny, area of CNV, types of CNV ascertained by fundus fluorescein angiography (FFA), and changes of CNV in FFA were recorded. Multi-factor regression analysis of visual acuity within 1 month and 3 months after PDT was performed with SPSS statistics software. Results The distant visual acuity within 1 month postoperatively was related to the preoperative distant visual acuity, the area of CNV and the changes in the FFA（P=0.000,0.030,0.062）, and 3 months after PDT, it was related to the distant and near visual acuity preoperatively and the changes in the FFA（P=0.000,0.054,0.034）. The condition of distant visual acuity within 1 month postoperatively was related to the FFA type of CNV and the disease course（P=0.018,0.08）. Conclusion The smaller the area of CNV is, the better postoperative distant visual acuity would be. The proportion of improvement of visual acuity is relatively higher in patients with classic CNV. Early treatment for the patients with the indicatio may improve the visual acuity effectively. (Chin J Ocul Fundus Dis，2004，20：292-294)
ObjectiveTo observe the expression of Rap1, guanosine triphosphate-Rap1 (GTP-Rap1), vascular endothelial growth factor (VEGF) and β-catenin in experimental choroidal neovascularization (CNV).MethodsForty-two brown Norwegian rats were randomly divided into a blank control group (7 rats) and a model group (35 rats). Both eyes were enrolled. The CNV model was established by holmium ion laser photocoagulation in the model group. At 3, 7, 14, 21, and 28 days after photocoagulation, fluorescein fundus angiography (FFA) and choroidal vascular smear were performed to observe the degree of fluorescein leakage and CNV area in rats; Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to detect the expression of Rap1, GTP-Rap1, VEGF, β-catenin and mRNA in CNV.ResultsThe results of FFA examination showed that a large disc-shaped fluorescein leaked in the photo-condensation spot 14 days after photocoagulation. Laser confocal microscopy showed that compared with 7 days after photocoagulation, CNV area increased at 14, 21, 28 days after photocoagulation, and the difference were statistically significant (t=3.725, 5.532, 3.605;P＜0.05). Western blot showed that there was no significant difference in the relative expression of Rap1 protein in CNV at different time points after photocoagulation between the two groups (P=0.156). Compared with the blank control group, the relative expression of GTP-Rap1 protein was significantly decreased, the relative expression of VEGF and β-catenin protein were significantly increased in the model group (P=0.000). The results of RT-PCR showed that there was no significant difference in the relative expression of Rap1 mRNA at different time points after photocoagulation between the two groups (P=0.645), but there were significant difference in the relative expression of β-catenin mRNA (P=0.000). At 7, 14, 21 and 28 days after photocoagulation, there were significant difference in the relative expression of GTP-Rap1 and VEGF mRNA between the two groups (P=0.000).ConclusionsThe expression of GTP-Rap1 in experimental CNV is significantly lower than that in normal rats.
ObjectiveTo investigate whether desferrioxamine (DFO) can enhance the homing of bone marrow mesenchymal stem cells (BMSCs) and improve neovascularization in random flaps of rats.MethodsBMSCs and fibroblasts (FB) of luciferase transgenic Lewis rats were isolated and cultured. Forty 4-week-old Lewis male rats were used to form a 10 cm×3 cm rectangular flap on their back. The experimental animals were randomly divided into 4 groups with 10 rats in each group: in group A, 200 μL PBS were injected through retrobulbar venous plexus; in group B, 200 μL FB with a concentration of 1×106 cells/mL were injected; in group C, 200 μL BMSCs with a concentration of 1×106 cells/mL were injected; in group D, cells transplantation was the same as that in group C, after cells transplantation, DFO [100 mg/(kg·d)] were injected intraperitoneally for 7 days. On the 7th day after operation, the survival rate of flaps in each group was observed and calculated; the blood perfusion was observed by laser speckle imaging. Bioluminescence imaging was used to detect the distribution of transplanted cells in rats at 30 minutes and 1, 4, 7, and 14 days after operation. Immunofluorescence staining was performed at 7 days after operation to observe CD31 staining and count capillary density under 200-fold visual field and to detect the expressions of stromal cell derived factor 1 (SDF-1), epidermal growth factor (EGF), fibroblast growth factor (FGF), and Ki67. Transplanted BMSCs were labeled with luciferase antibody and observed by immunofluorescence staining whether they participated in the repair of injured tissues.ResultsThe necrosis boundary of ischemic flaps in each group was clear at 7 days after operation. The survival rate of flaps in groups C and D was significantly higher than that in groups A and B, and in group D than in group C (P<0.05). Laser speckle imaging showed that the blood perfusion units of flaps in groups C and D was significantly higher than that in groups A and B, and in group D than in group C (P<0.05). Bioluminescence imaging showed that BMSCs gradually migrated to the ischemia and hypoxia area and eventually distributed to the ischemic tissues. The photon signal of group D was significantly stronger than that of other groups at 14 days after operation (P<0.05). CD31 immunofluorescence staining showed that capillary density in groups C and D was significantly higher than that in groups A and B, and in group D than in group C (P<0.05). The expressions of SDF-1, EGF, FGF, and Ki67 in groups C and D were significantly stronger than those in groups A and B, and in group D than in group C. Luciferase-labeled BMSCs were expressed in the elastic layer of arteries, capillaries, and hair follicles at 7 days after transplantation.ConclusionDFO can enhance the migration and homing of BMSCs to the hypoxic area of random flap, accelerate the differentiation of BMSCs in ischemic tissue, and improve the neovascularization of ischemic tissue.
ObjectiveTo study the inhibitory effects of pigment epithelium derived factor (PEDF) on oxygen-induced retinal neovascularization in mice, and to investigate the possible involvement of interleukin-1β (IL-1β) in the neovascular-inhibitory function of PEDF.
Methods A total of 140 postnatal day (P)7 C57BL/6 mice were randomly divided into normal control group, oxygen-induced retinopathy (OIR) model group, PEDF treatment group and PBS treatment control group. All mice except normal control group with their mothers were exposed to (75±2)% oxygen environment for 5 days and then kept in room air for another 5 days to establish the OIR model. Mice in normal control group were kept in room air only. At P12 and P14, respectively, mice in PEDF treatment group received intravitreous injections of 1 μl PEDF (2 μg/μl), while PBS treatment control group received the same volume of PBS (10 mmol/L, pH7.4).All mice were euthanized at P17 and eyes were isolated. The changes of retinal vessels were observed on retinal flat mounts and cryosections by fluorescence microscopy. Retinal specimens were prepared for IL-1β protein and mRNA analysis by Western blot and real time fluorescence quantitative reverse transcription-polymerase chain reaction (Real-time RT-PCR).
ResultsChanges of retinal vessels had been viewed by fluorescence microscopy on flat-mounted retina, the relative retinal neovascularization areas were significantly increased in OIR model group compared with normal control group (t=15.02, P < 0.01), and the relative retinal neovascularization areas were obviously smaller in PEDF treatment group than those in PBS treatment control group (t=5.96, P < 0.01). Fluorescence staining revealed that retinal vascular tufts were extending from outer plexiform layer (OPL) to ganglion cell layer (GCL) of the retina along with multiple interconnections; Neovascular tufts in OIR model group and PBS treatment control group were presenting distinctly more than those of normal control group and PEDF treatment group. The specific expression levels of IL-1β protein in retinas of OIR mice by Western-blot analysis were higher than those of normal control group(t=3.35, P < 0.05), While these of PEDF treatment group showed a considerable decline in comparison with PBS treatment control group (P < 0.01), and there were no difference in normal control group and PEDF-treated group (F=11.764, P > 0.05). Similarly, expression levels of IL-1β mRNA tested by Real-time RT-PCR were obviously increased in the OIR model group when compared to normal control group(t=4.43, P < 0.01). After treated with PEDF, expression levels of IL-1β mRNA showed a considerable decrease when compared to PBS treatment control group (P < 0.01), and there were no difference in normal control group and PEDF-treated group (F=11.15, P > 0.05).
ConclusionsPEDF can inhibit oxygen-induced retinal neovascularization. The mechanism may be related to that PEDF can downregulate the expression of IL-1β in retina.
Two-hundred and forty-five eyes of 240 cases of branch retinal vein occlusion (BRVO) were analysed to determine the risk factors influencing the development of retinal neovascularization (NV).There were 208 eyes with major BRVO, 37 eyes with macular BRVO, 79 eyes with major BRVO developed NV, and the incidence of NV in this series was 37.9%.The incidence of vitreous hemorrhage in these eyes was 15.9%(39 eyes). The risk factors influencing the development of retinal NV in BRVO seem as follows: (]) the extent of retinal capillary nonperfusion area, (2)inefficiency of arterial infusion, (3) the extent of venous block at the arteriovenous crossing, (4) the duration of follow-up since onset of BRVO, and (5) the lack of collateral formation, Because BRVO has a long natural history, it is recommended that the patients should be followed-up for a long time If the vessels at peripheral retina closed, fluorescein angiography should be performed without hesitation and if the nonperfusion area is greater than 20-30 disc area, one should follow the patient carefully.As soon as the new vessels appear, laser photocoagulation should be applied without delay.
（Chin J Ocul Fundus Dis,1994,10:67-70）
ObjectiveTo evaluate the inhibitory effect of small interfering RNA (siRNA) targeting peroxisome-proliferator-activated receptor-γcoactivator-1α(PGC-1α) on retinal neovascularization in the mouse.
MethodsEighty seven-day-old C57BL/6J mice were divided into normal group, model blank group, model control group and PGC-1αsiRNA group, twenty mice in each group. Mice in the normal group were kept in normal room air. Mice in the model blank group, model control group and PGC-1αsiRNA group were induced for retinal neovascularization by hypoxia. Liposome with PGC-1αsiRNA (1 μl) and liposome with negative control siRNA (1 μl) were injected into the vitreous in the PGC-1αsiRNA group and model control group respectively when mice were moved out to room air from the cabin (Postnatal 12). No injection were performed in the model blank group. At postnatal 17, fluorescein angiography was used to assess the vascular pattern.The proliferative neovascular response was quantified by counting the nuclei of new vessels extending from the retina into the vitreous in cross-sections. PGC-1αand vascular endothelial growth factor (VEGF) level in retina were measured by real-time polymerase chain reaction (real-time PCR) and Western blot. Inhibition efficiency of PGC-1αsiRNA on PGC-1αand VEGF was calculated.
ResultsMice in the normal group showed reticular distribution of retinal blood vessels. Central nonperfused retina, neovascular tufts and fluorescein leakage were seen in the model blank group and model control group. Neovascular tuft and fluorescein leakage were decreased in the PGC-1αsiRNA group compared to the model blank group and model control group. The neovascular nuclei were increased in the model blank group and model control group compared to the normal group (P < 0.05). The neovascular nuclei were decreased in the PGC-1αsiRNA group compared to the model blank group and model control group (P < 0.05). The expression of PGC-1αmRNA and protein in retina was increased significantly in the model blank group and model control group as compared with normal group, while decreased 54% and 53% respectively in the PGC-1αsiRNA group as compared with model blank group and model control group (P < 0.05). The expression of VEGF mRNA and protein in retina was increased significantly in the model blank group and model control group as compared with normal group, while decreased significantly in the PGC-1αsiRNA group (decreased 48% and 40% respectively) as compared with model blank group and model control group (P < 0.05).
ConclusionsIntravitreal injection of PGC-1αsiRNA mediated by liposome can inhibit retinal neovascularization in the mouse effectively.
Objective To investigate the possible relationship between choroidal neovascularization (CNV) and macular choroidal watershed zones (CWZ) in patients with age-related macular degeneration (AMD). Methods Fifty-seven selected indocyanine green video angiograms (ICGA) of 57 patients (57 eyes) with AMD were evaluated, and 35 ones of the healthy fellow eyes of 35 patients with unilateral non-AMD fundus diseases were selected as age-matched control. The video angiograms were evaluated for investigating the relationship between CNV and macular CWZ. Results In 57 eyes with AMD, 35 (61.4%) had macular CWZ, while in 35 control patients only 3 (8.57% ) had. The difference was significant (Plt;0.05). In 43 eyes with exudative AMD and CNV, 32 (74.4%) had macular CWZ, including 29 eyes (90.6%) with CNV caused by macular CWZ. Conclusion Macular CWZ could be a predilection site of CNV in exudative AMD. (Chin J Ocul Fundus Dis，2003，19：76-78)
Objective To detect expression of NF-κB in the inner retina and in vestigate the inhibitoryeffect of pyrrolidine dithiocarbamate on retinal neovascularization in rats. Methods The rat models with retinopathy were set up un der the hypoxia condition, and fluorescein fundus angiography (FFA) was used to observe the retinal neovascularization. The expressions of NF-κB in the inner retina in rats with and without neovascularization were detected by immunohisto chemical method. PDTC was intraperitoneally injected in rats with neovascularization to observe the expression of NF-κB in the inner retina and the effect on retinal neovascularization. Results Hypoxia induced NF-κB activation in the retinal glial cells and endothelial cells. But immuno-staining intensity for NF-κB and adhesion molecules were reduced by PDTC intraperitoneal injection. Retin al angiogenesis in rats were suppressed effectively (P＜0.05). Conclusions NF-κB activation correlates with retinal neovascularization closely. PDTC may inhibit the NF-κB activation and prove beneficial in the treatment of ischemic neovascularization. (Chin J Ocul Fundus Dis，2003，19：201-268)
Objective To observe the clinical characteristics of diabetic neovascularization on the disc (DNVD).Methods The clinical data of 526 patients (1052 eyes) who were diagnosed as diabetes in Department of intern medicine, as diabetic retinopathy by ophthalmoscope and fundus fluorescein angiograph (FFA) was retrospectively reviewed. All patients were carried out with best corrected visual acuity（BCVA）, slitlamp microscope,ophthalmoscope and FFA after mydriasis. In which, who has neovascularization on the optic disc with ophthalmoscopy and FFA examination were included in this study.The relationship between the occurrence and development of DNVD and phase of DR, disease duration, the level of blood glucose and panretinal photocoagulation were analyzed. Results DNVD was found in167/1052eyes (15.87%). There were 91 eyes (54.49%) with BCVA＜0.1, 58 eyes (34.73%) with BCVA＜0.4 but ge;0.1，and 18 eyes(19.78%) with BCVAge;0.4. Retinal neovascularization was located in the surface of disc surface or within 1PD from the optic disc；Those vessels filled early and rapidly, and with local b fluorescence due to fluorescence leakage at middle and late stage of FFA examination.All 167 DNVD eyes are proliferative diabetic retinopathy (PDR) with 43 eyes (25.75%) in stage IV,52 eyes (31.14%) in stage V and 72 eyes (43.11%) in stage VI.Of those DNVD eyes,there were 5 eyes (2.99%) with course of diabetes ＜3 years,12 eyes (7.19%) s＜5 years but ge;3 years, 21 eyes (12.57%)＜10 butge;5 years, 56 eyes (33.53%)＜15 but ge;10 years and 73 eyes (43.71%) ge;15 years. There were 15 eyes (8.98%) with fasting blood glucose (FBG)＜7.0 mmol/L,26 eyes (15.57%) with FBG＜9.0 but ge;7.0 mmol/L,50 eyes (29.94%) with FBG＜12.0 but ge;9.0 mmol/L and 76 eyes (45.51%) with FBG ge;12.0 mmol/L;there were 28 eyes (16.77%) with 2 hour postprandial blood glucose（2hPBG）＜10.0 mmol/L, 35 eyes (20.96%) with 2hPBG＜12.0 but ge;10.0 mmol/L,42 eyes (25.15%) with 2hPBG ＜16.0 butge;12.0 mmol/L and 62 eyes (50.30%) with 2hPBG ge;16.0 mmol/L. The occurrence of DNVD and duration of diabetes, FBG and 2hPBG were all positively correlated (r=0.991,0.984,0.960, P=0.001, 0.016, 0.040) by the Person correlation analysis. 15 eyes (5.84%) of DNVD happened in 257 eyes who treated with PRP in severe nonproliferative diabetic retinopathy (NPDR),152 eyes (19.12%) DNVD happened in 795 eyes who untreated with PRP in severe NPDR，the differences were statistically significant (chi;2=25.659,P＜0.01) between them.Conclusion DNVD happened commonly in DR, the occurrence of DNVD is intensive related with diabetic retinopathy stage,duration of diabetes,FBG and PBG.
