Objective To investigate the therapeutic effects of retinal angiomatosis in different clinical stages. To discuss the indication of vitrectomy for retinal hemangioblastoma. Methods The clinical data of 22 cases (33 eyes) were retrospectively analyzed. The retinal hemangiomas were divided into 5 stages according to their degrees of development from simple angioma without vessel dilation to feeder vessel dilation and intra-retinal exudates, local retinal detachment, massive retinal detachment and co mplication occurrence in proper order. The methods of treatment were laser photo coagulation, trans－scleral cryotherapy and vitrectomy. 13 eyes were treated with laser photocoagulation, 5 eyes with cryotherapy combined with laser and 11 eye s with vitrectomy. Tumor resection and silicone oil tamponade was performed in 3 eyes during vitrectomy. The patients were followed up for 46 months on average. Visual acuity (VA), the condition of the hemangioma and retina was compared pre- and post-operation respectively. Results In all 13 eyes treated with laser photocoagulation the hemangiomas regressed and the retina remained attached. VA improved in 2 eyes, and remained unchanged in 11 eyes. Cryother apy combined with laser photocoagulation was performed on 5 eyes. In this group, 4 eyesprime; hemangiomas regressed and no new hemangiomas occurred, proliferative vitreous retinopathy and vitreous hemorrhage was observed in 1 eye which vitrecto my was performed later. VA improved in 2 eyes, unchanged in 2 eyes and decreased in 1 eye. In the 11 eyes treated with vitreoretinal surgery, new hemangiomas wa s found in 1 eye, exudative retinal detachment was caused by hemangiomas in 2 eyes, proliferative vitreous retinopathy was observed in 2 eyes, and the retina re mained attached in 8 eyes. VA improved in 3 eyes, unimproved in 3 eyes, and decreased in 5 eyes. In the 3 eyes with surgical resection of retinal hemangioma during vitrectomy, 2 eyesprime; retina remained attached, 1 eye had exu dative retinal detachment and no new hemangiomas occurred. VA improved in 2 eyes and decreased in 1 eye. Conclusions Laser photocoagulation or combined with cryotherapy is effective in treating the hemangiomas in early stage. Vitrectomy is advisable for late stage of retinal angiomatosis, especially with vitreous hemorrhage, epiretinal membrane, proliferation and large scale of r etinal detachment. Surgical resection of isolated large retinal hemangioblastoma may be useful for selected patients. (Chin J Ocul Fundus Dis,2008,24:107-110)
Objective To observe the effect of tetramethypyrazine (TMP) on the expression of hypoxia-related factors in human umbilical vein endothelial cells (HUVECs). Methods The second to fifth passage cultured HUVECs were divided into five groups: control group, CoCl2induced hypoxic group and 50, 100, 200 mu;mol/L TMP treatment groups. HUVECs in control group were not treated. HUVECs inCoCl2induced hypoxic group were treated with 150 mu;mol/LCoCl2for four hours. HUVECs in 50, 100, 200 mu;mol/L TMP treated groups were pretreated with 150 mu;mol/LCoCl2 for four hours, followed by treatment with 50, 100, 200 mu;mol/L TMP for eight hours. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA levels of prolyl hydroxylase 2 (PHD2), hypoxia-induced factor-1alpha;(HIF-1alpha;) and vascular endothelial growth factor (VEGF). Protein levels of PHD2, HIF-1alpha;, and VEGF were detected using Western blot. Results Compared with the control group, theCoCl2 induced hypoxic group showed decreased mRNA and protein levels of PHD2 (t=3.734, 3.122;P＜0.05), while those of HIF-1alpha; and VEGF increased (HIF-1alpha; mRNA:t=4.589,P＜0.05; HIF-1alpha; protein:t=3.778,P＜0.05. VEGF mRNA:t=3.926,P＜0.05; VEGF protein:t=3.257,P＜0.05). Compared with theCoCl2 induced hypoxic group, 50, 100, 200 mu;mol/L TMP treated groups showed increased mRNA and protein levels of PHD2 (PHD2 mRNA: t=3.286, 3.617, 3.886;P＜0.05. PHD2 protein: t=2.813, 3.026, 3.078; P＜0.05); while those of VEGF decreased (VEGF mRNA: 50 mu;mol/L TMP: t=1.696,P＞0.05; 100 mu;mol/L TMP:t=2.974,P＜0.05; 200 mu;mol/L TMP: t=3.492,P＜0.05; VEGF protein: 50 mu;mol/L TMP: t=1.986,P＞0.05; 100 mu;mol/L TMP: t=2.976,P＜0.05; 200 mu;mol/L TMP:t=3.136,P＜0.05); although changes in HIF-1alpha;mRNA levels were not statistically significant (t=1.025, 0.726, -1.386;P＞0.05), showed a decrease in HIF-1alpha;protein levels (50 mu;mol/L TMP: t=2.056,P＞0.05; 100 mu;mol/L TMP:t=3.058,P＜0.05; 200 mu;mol/L TMP:t=3.828,P＜0.05). ConclusionIn HUVECs, TMP can upregulate the mRNA and protein expression of PHD2, while down regulating HIF-1alpha; protein expression and VEGF mRNA and protein expression under acute hypoxic conditions.
Objective To observe the clinical features of polypoidal choroidal vasculopathy (PCV) with retinal pigment epithelium (RPE) tears. Methods Twelve patients of PCV with RPE tears (12 eyes) were enrolled in this study. The patients included eight males and four females, with a mean age of 58.6 years (from 39 to 71 years old). All the patients were affected unilaterally, including eight right eyes and four left eyes. There were one eye with serous RPE detachment and 11 eyes with hemorhagic RPE detachment. All the patients were examined for fundus photography, fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA), three patients were examined for optical coherence tomography (OCT). The location of RPE tear was classified as within vascular arcade, on vascular arcade, and outside vascular arcade. The shape of tear was classified as crescent, semilunar, or irregular. The features of fundus, FFA, ICGA and OCT were observed. Results Fundus examination presents a gray lesion in all eyes. The location of tear were within vascular arcade in four eyes (33.3%), on vascular arcade in five eyes (41.7%) and outside vascular arcade in three eyes (25.0%). The shape of tears were crescent (one eye, 8.3%), semilunar (ten eyes, 83.3%) or irregular (one eye, 8.3%). The RPE tear region present transimitted fluorescence of at the early stage of FFA and hyperfluorescence with a clear border at late stage. There was no leakage, and at the border of hyperfluorescence, blockage fluorescence of rolled and contracted RPE was present. In ICGA manifestation, transimitted fluorescence was found in RPE tear region at early stage, and a clear border was seen in nine eyes at late stage. There was also blockage fluorescence in ICGA of contracted RPE. In OCT manifestation, the RPE reflections were disappeared, and at the margin of tear, the contracted RPE present a dense rolled b reflection. Conclusions In PCV patients, RPE tears are semilunar and usually located within or around the vascular arcade. Fundus angiography shows transimitted fluorescence at the RPE tear region, and curl blockage fluorescence at the edges. OCT shows RPE reflection is disappeared in the tear region and a b reflection at the edges.
Objective To observe the inhibitory effect of tetramethylpyrazine (TMP) on apoptosis of retinal neurons in oxygeninduced retinopathy (OIR). Methods 48 C57BL/6 mice were randomly divided into normal control group (n=18), OIR control group (n=18) and OIR TMP group (n=12). The mice of normal control group were raised in room air. From the postnatal day 7 (P7), mice of the other two groups were exposed to (75 3) % oxygen for 5 days and then returned to room air to establish OIR model. The mice of OIR TMP group received intraperitoneal injection of TMP (200 mg/kg) once a day from P12 to P16, meanwhile the mice of normal control group and OIR control group were injected with the same volume of normal saline containing of 0.1% DMSO. At P12, P14 and P17, the morphologic changes in retinal avascular zone and the number of retinal apoptotic cell were observed by HE staining and TUNEL assay. Results At P12, there were a few of chromatin condensation and pycnic nuclei in the inner nuclear layer (INL) of OIR control group. At P14, a great quantity (OIR control group) or some (OID TMP treated group) chromatin condensation and pycnic nuclei in the central INL were observed. At P17, the thickness of INL, inner plexiform layer (IPL) and outer plexiform layer (OPL) in the OIR control group were reduced; the thickness of INL, IPL and OPL in the OIR TMP group weas thinner than those in the normal control group and thicker than those in the OIR control group. At P12, the TUNEL-positive cells in the OIR control group was 6 times of the normal control group (F=587.217，P＜0.001). At P14, the difference of TUNEL-positive cells in three groups was significant (F=587.217，P＜0.001); the TUNEL-positive cells in the OIR control group was 28 times of the normal control group (t=49.813，P＜0.001); the TUNEL-positive cells in the OIR TMP group has reduced 50% compared with the OIR control group (t=42.434，P＜0.001). At P17, there was no significant difference in TUNEL-positive cells among the three groups (F=587.217，P＞0.05). Conclusions TMP can inhibit apoptosis of retinal cells in OIR significantly.
Objective To investigate the effects of recombinant adeno-associated virus type-2 (rAAV2) mediated delivery of pigment epitheliumderived factor (PEDF) on oxygen-induced retinal neovascularization (OIRNV) in mice. Methods A total of 22 C57/BL6 mice at the age of 3 days received intravitreal injections of 1 mu;l rAAV2-PEDF and rAAV2EGFP into the left eyes (experimental group) and the right eyes (control group). All mice were put into the oxygen box right after the injection to induce the OIRNV model.4 mice were sacrificed and PEDF protein in retina was measured by western blot at postnatal days 13 (P13). Twelve mice underwent retinal angiography with high molecular weight fluoresceindextran, and another 6 mice were sacrificed for retinal lectin immunohistochemistry staining at P17. Absolute and relative nonperfusion areas of retinal neovascularization were analyzed by Image-Pro Plus 5.1 software. Results The expression level of PEDF protein was higher in the experimental group than that in the control group.The absolute nonperfusion area was (0.96plusmn;0.22) mm2 in the experimental group and (1.96plusmn;0.34) mm2 in the control group; the difference between the two groups was significant（t=-8.554, P＜0.01）. The relative nonperfusion area was (8.64plusmn;1.52)% in the experimental group and (17.27plusmn;2.98)% in the control group with a significant difference between the two groups (t=-8.97, P＜0.01).The absolute area of retinal neovascularization was (0.37plusmn;0.11) mm2 in the experimental group which was obviously higher than (1.26plusmn;0.38) mm2 in the control group (t=-7.8, P＜0.01); the relative areas in experimental and control groups was (3.96plusmn;0.66)% and (11.45plusmn;2.06)%, respectively, whose difference is apparently(t=-8.51, P＜0.01).The areas of retina neovascularization were (0.11plusmn;0.003) mm2 and (0.41plusmn;0.02) mm2 in the experimental and control groups, respectively, and the difference between the two groups was significant（t=-5.14, P＜0.01）.Conclusions PEDF protein can stably express in the mice retina after rAAV2-PEDF transfetion. rAAV2-PEDF can decrease the retinal non-perfusion areas and inhibit the retinal neovascularization in OIRNV mice.
Objective To observe the influence of prolyl hydroxylase 2 (PHD2) expression on endothelial barrier dysfunction induced by high glucose in human retinal vascular endothelial cells (HRECs). Methods The HRECs were treated by different culture medium with various glucose concentrations (5 mmol/L glucose, 5 mmol/L glucose +25 mmol/L mannitol, 30 mmol/L glucose) as normal control group, mannitol control group and high glucose group, respectively. After the cells cultured for 24 and 48 hours, the protein levels of PHD2, hypoxia-inducible factor-1alpha; (HIF-1alpha;) and occludin was detected by Western blot; the expression of vascular endothelial growth factor (VEGF) in the supernatant was determined by enzymelinked immuno sorbent assay (ELISA); the transcription levels of PHD2, HIF-1alpha;, VEGF and occludin were determined by the reversetranscription polymerase chain reaction (RT-PCR); the paracellular permeability between endotheliums was detected by 7times;104 molecular weight FITCdextran. Results Compared with normal control group, the protein level of PHD2 in mannitol control group and high glucose group firstly decreased and then increased, the protein level of HIF-1alpha; increased while that of occludin decreased; the secretion of VEGF increased in high glucose group but not in mannitol control group (PHD2：F=7.618, 8.627;P＜0.05. HIF-1alpha;：chi;2=7.692, 7.652;P＜0.05. occludin：F=23.23, 7.317;P＜0.05. VEGF：F=10.768, 4.562; P＜0.05). Compared with normal control group, the mRNA levels of PHD2, HIF-1alpha;, VEGF and occludin in mannitol control group and high glucose group increased (PHD2：F=5.69, 14.27;P＜0.05. HIF-1alpha;：F=6.07, 10.47;P＜0.05. VEGF：F=12.31, 9.14;P＜0.05. occludin：F=8.77, 8.00;P＜0.05). Compared with normal control group, the paracellular permeability of mannitol control group and high glucose group increased (chi;2=20.57,F=56.09;P＜0.05). Conclusions High glucose induced altered expression of PHD2 which might play an important role in endothelial barrier dysfunction. The mechanism might be associated with HIF-1alpha; and VEGF.
Objective To investigate the expression and prolyl hydroxylase (PHD)2 in retina of diabetic rats．Methods Wistar rats were randomly divided into the control group (n=48) and the diabetes group (n=60). The rats in diabetes group were induced with streptozotocin (STZ) injection creating a diabetic retinopathy model. The same volume of citric acid buffer was injected into the rats in the control group. Fluorescence microscope was used to observe the retinal vasculature at one, three and six months after injection. Evans blue perfusion was used to detect the bloodretinal barrier (BRB) permeability. Immunohistochemical staining was used to observe the distribution of PHD-2 positive staining. Western blot was used to measure the protein expression of PHD-2, hypoxia inducible factor（HIF）-1alpha; and vascular endothelial growth factor (VEGF) every month from one to six months after injection. Results The vascularization was normal and form was clear in retina of rats in control group. The retinal blood vessels of rats in diabetes group showed significantly increased fluorescence. Compared with the control group, the BRB permeability was significantly increased in diabetes group (P＜0.05). Abundant expression of PHD-2 protein was detected in the inner layers of retina in control and diabetes group．Compared with the control group, the PHD-2 expression was decreased in diabetes group at one and two months after injection (t=16.230, 16.390;P＜0.05). The HIF-1alpha; expression was significantly increased in diabetes group at one, two and three months after injection (t=27.073, 36.709, 10.176; P＜0.05). The VEGF expression was significantly increased in diabetes group every month from one to six months after injection (t=13.547, 31.984, 21.897, 8.912, 9.019, 14.046; P＜0.05). Conclusions There is abundant expression of PHD-2 in the inner layer of retina in diabetic rats. PHD-2 may play an important role in diabetic retinopathy, which is correlated with VEGF．
Objective To observe the effect of intravitreal bevacizumab (IVB) for exudative age-related macular degeneration (eAMD), and analyze the reasons for treatment failure. Methods Eighteen eyes of 17 patients with eAMD who have been diagnosed by fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and optical coherence tomography (OCT) were enrolled in this openlabel, single treatment group and prospective study. The patients received the first IVB treatment after diagnosis, and received the 2nd, 3rd, 4th, 5th and 6th IVB treatment at 6, 12, 24, 36, 48 weeks after the first injection. The examinations of best corrected visual acuity (BCVA), intraocular pressure, slit lamp microscope, fundus photography, FFA, ICGA and OCT were performed before and after treatment. Non-responders were defined as patients who had BCVA loss more than 5 letters at week 52 compared with BCVA before treatment. The therapeutic effects of IVB for eAMD were observed. The central macular thickness (CMT), maximum retinal thickness (MRT) and morphological changes of FFA, ICGA and OCT before and after treatment were comparative analyzed. Results Of the 18 eyes, 12 eyes (66.67%) were effective to IVB, 6 eyes (33.33%) were non-responders. The average CMT and MRT of non-responders were 506.83, 635.33 mu;m before treatment, which decreased to 446.17, 563.67 mu;m at 52 weeks. They were reduced by 60.67 and 71.67 mu;m respectively, but there was no statistically significant differences (t=-1.572,-0.943; P=0.116, 0.345). FFA and ICGA examination found that the CNV lesions of 3 non-responder eyes located in the foveal region, and became scars after the 6th treatment. The other 3 non-responder eyes developed CNV repeatedly during the treatments, and after the 6th injections of bevacizumab the lesions were under control with less fluorescein leakage and macular retinal edema. ConclusionIVB treatment is effective for some eAMD patients, but is ineffective for patients with extensive expanded CNV and foveal CNV.
Objective To evaluate the safety and efficacy of dexamethasone intravitreal implant 0.7 mg (DEX) for treatment of macular edema associated with retinal vein occlusion (RVO). Methods This study was a six-month, randomized, double-masked, sham-controlled, multicenter, phase 3 clinical trial with a 2-month open-label study extension. Patients with branch or central RVO received DEX (n=129) or sham procedure (n=130) in the study eye at baseline; all patients who met re-treatment criteria received DEX at month 6. Efficacy measures included Early Treatment Diabetic Retinopathy Study (ETDRS), best-corrected visual acuity (BCVA), and central retinal thickness (CRT) on optical coherence tomography. Results Time to ≥15-letter BCVA improvement from baseline during the first 6 months (primary endpoint) was earlier with DEX than sham (P＜0.001). At month 2 (peak effect), the percentage of patients with ≥15-letter BCVA improvement from baseline was DEX: 34.9%, sham: 11.5%; mean BCVA change from baseline was DEX: 10.6±10.4 letters, sham: 1.7±12.3 letters; and mean CRT change from baseline was DEX: −407±212 μm, sham: −62±224 μm (all P＜0.001). Outcomes were better with DEX than sham in both branch and central RVO. The most common treatment-emergent adverse event was in-creased intraocular pressure (IOP). Increase sin IOP generally were controlled with topical medication. Mean IOP normalized by month 4, and no patient required incisional glaucoma surgery. Conclusions DEX had a favorable safety profile and provided clinically significant benefit in a Chinese patient population with RVO. Visual and anatomic outcomes were improved with DEX relative to sham for 3 - 4 months after a single implant.
ObjectiveTo observe the changes of macular morphology and blood flow after minimally invasive vitrectomy (PPV) in patients with severe non-proliferative diabetic retinopathy (sNPDR). MethodsA prospective clinical study. From January 2020 to April 2021, 17 consecutive sNPDR patients with 17 eyes who were diagnosed and received PPV treatment at the Zhongshan Ophthalmic Center of Sun Yat-sen University were included in the study. There were 12 males with 12 eyes and 5 females with 5 eyes; the average age was 55 years old; the average duration of diabetes was 11 years; the average glycosylated hemoglobin was 7.9%. Before the operation and 1, 3, and 6 months after the operation, all the affected eyes underwent best corrected visual acuity (BCVA), standard 7-field fundus color photography, and optical coherence tomography angiography (OCTA). An OCTA instrument was used to scan the macular area of the affected eye with in the range of 3 mm×3 mm to measure the central subfoveal thickness (CST), the thickness of the ganglion cell complex (GCC) in the macular area, the thickness of the retinal nerve fiber layer (RNFL), and the superficial capillary plexus (SCP) vessel density and perfusion density in the macular area, macular avascular zone (FAZ) area, a-circularity index (AI). Before the operation and 6 months after the operation, the least significant difference test was used for the pairwise comparison. ResultsBefore the operation, 1, 3, and 6 months after the operation, the FAZ area of the macular area were 0.34±0.14, 0.35±0.10, 0.37±0.10, 0.36±0.13 mm2, respectively; AI were 0.52±0.13, 0.54±0.11, 0.57±0.10, 0.60±0.11; CST was 282.6±66.7, 290.4±70.9, 287.2±67.5, 273.2±49.6 μm; GCC thickness were 77.1±15.5, 74.3±13.9, 72.6±16.2, 78.5±18.3 μm; the thickness of RNFL was 97.9±13.8, 101.3±14.6, 97.7±12.0, 96.1±11.4 μm, respectively. The overall blood flow density of SCP in the macula were (16.79±1.43)%, (16.71±1.82)%, (17.30±2.25)%, (17.35±1.22)%; the overall perfusion density were 0.32±0.02, 0.32±0.03, 0.33±0.03, 0.33±0.02, respectively. After the operation, the CST increased first and then decreased; the thickness of RNFL increased 1 month after the operation, and then gradually decreased. Comparison of the parameters before and 6 months after the operation showed that the AI improved, and the difference was statistically significant (P=0.049); the difference in FAZ area and the thickness of CST, GCC, and RNFL was not statistically significant (P=0.600, 0.694, 0.802, 0.712); There was no statistically significant difference in the retina SCP blood flow density and perfusion density in the macular area (P=0.347, 0.361). ConclusionCompared with before surgery, there is no significant change in macular structure and blood flow density in sNPDR patients within 6 months after minimally invasive PPV.