Age-related macular degeneration (AMD) is a multifactorial disease affected by environmental factors and genetic variation, which is a major cause of irreversible vision loss in the elderly. miRNA is a kind of endogenous non-coding RNA, which plays an important role in the pathogenesis of AMD, such as oxidative stress, pathological neovascularization and inflammation, by inhibiting or silencing the expression of transcription genes. miRNA has unique advantages in terms of ease synthesis, targeting and additive effect, a large number of experiments have proved the therapeutic potential of miRNA in AMD, which is expected to become a new method for the treatment of AMD in the future. Since the pathogenesis of AMD has not been fully elucidated, it is still necessary to continue to study the pathogenesis of AMD, the biological effects and mechanisms of various miRNA in the occurrence and development of AMD, and observe its therapeutic effects in AMD, so as to provide more effective options for the precise prevention and treatment of AMD.
ObjectiveTo observe the effects of NDRG1 on proliferation, migration and lumen formation of retinal vascular endothelial cells (RF/6A cells) in monkeys under high glucose condition. MethodsRF/6A cells were divided into normal group, mannitol group, high glucose group, small interfering RNA (siRNA) negative control group without target gene (siRNA group), 30 nmol/L siRNA down-regulated NDRG1 genome (siNDRG1 group) and 50 nmol/L siNDRG1 group. Normal group cells were cultured conventionally. The mannitol group was added with 25 mmol/L mannitol, and the high-glucose group was added with 25 mmol/L glucose. In the siRNA group, 25 mmol/L glucose was added, and then blank siRNA was added for induction. The 30 and 50 nmol/L siNDRG1 groups were added with 25 mmol/L glucose and induced with 30 and 50 nmol/L siRNDRG1, respectively. All cells were incubated for 24 h for follow-up experiments. Cell proliferation was observed by 4', 6-diaminidine 2-phenylindole staining. Cell counting kit-8 staining was used to detect cell activity. The expression level of NDRG1 mRNA and protein was detected by Western blot and real-time quantitative polymerase chain reaction. Cell migration was observed by cell scratch assay. Cell lumen formation assay was used to detect lumen formation. The two-tailed Student t test was used to compare the two groups. One-way analysis of variance was used to compare groups. ResultsThere were significant differences in cell proliferation rate (t=36.659, 57.645) mobility rate (t=24.745, 33.638) and lumen formation number (t=41.276, 22.867) between high glucose group and normal group and mannitol group (P＜0.01). Compared with normal group and mannitol group, the relative expression levels of NDRG1gene mRNA and protein in high glucose group were significantly decreased, with statistical significance (t=46.145, 21.541, 36.738, 32.976; P＜0.001). Compared with the siRNA negative group, the relative expression levels of NDRG1gene mRNA and protein in 30 nmol/L siNDRG1 group and 50 nmol/L siNDRG1 group were significantly decreased, and the differences were statistically significant (t=44.275, 40.7577, 57.167, 25.877; P＜0.01). Compared with normal group and siRNA group, cell mobility in 30 nmol/LsiNDRG1 group was increased, and the difference was statistically significant (t=57.562, 49.522; P＜0.01). Compared with normal group and siRNA group, the number of cell lumen formation in 30 nmol/LsiNDRG1 group was significantly increased in the same field of vision, and the difference was statistically significant (t=63.446, 42.742; P＜0.01). ConclusionDown-regulation of NDRG1 gene can improve the activity, migration and lumen formation of RF/6A cells under hyperglycemia.
Objective To evaluate the diagnostic value of Heidelberg retinal angiography(HRA) combined with optical coherence tomography (OCT) to detect neovascularization (CNV) in exudative agerelated macular degeneration (AMD) patients. Methods This is a cross-sectional study of a series of clinical cases. AMD diagnosis was established by international standard vision chart, Slit lamp microscope, direct or indirect ophthalmoscope examination. A total of 50 eyes (42 cases) of exudative AMD received HRA and frequency domain OCT scan. All 50 eyes received fundus fluorescein angiography (FFA) and frequency domain OCT simultaneously, and among them 15 eyes also received indocyanine green angiography (ICGA) at the same time. FFA and ICGA were carried out by conventional methods, CNV was localized by real-time localization technology of frequency domain OCT. In the radial and grid-like section from the areas with b fluorescence, image acquisition settings are 7 mu;m fault for each frame, 30 deg; intervals for radial section, 10 vertical and 10 horizontal scan lines for gridlike section. CNV can be divided into 4 types (typical CNV, partial typical CNV, occult CNV, CNV scarring) according to their boundaries demonstrated in FFA. Based on the features of the OCT images, there were 3 types of integrated image (sub-RPE type, sub-retinal type and mixed type). Results CNV was detected in all 50 eyes. There were 4 eyes (8%) of typical CNV, 11 eyes (22%) of partial typical CNV, 32 eyes (64%, including 27 eyes of RPE detachment and 5 eyes of passive late leakage) of occult CNV and 3 eyes (6%) of CNV scarring. There were 4 eyes (8%) of subRPE type (CNV under the RPE light band) , 16 eyes (32%) of subretinal type(interrupted light band of RPE and choroid capillary layer) and 30 eyes (60%) of mixed type of integrated image. Conclusion The image integration technology of the HRA and frequency domain OCT system provide a valuable tool to classify and measure CNV, which will benefit the clinical treatment of AMD patients.
ObjectiveTo observe the efficacy of intravitreal injection of conbercept (IVC) in the treatment of type 1 macular neovascularization (MNV) with different types of pigment epithelial detachment (PED) in neovascular age-related macular degeneration (nAMD). MethodsA retrospective clinical study. From June 2018 to June 2021, 42 patients with 42 eyes of nAMD type 1 MNV patients with different types of PED diagnosed in the ophthalmological examination of the Department of Ophthalmology, General Hospital of Central Theater Command were included in the study. All eyes underwent best corrected visual acuity (BCVA) and optical coherence tomography (OCT). The OCT examination was performed with a 3D-OCT 2000 instrument from Topcon Company in Japan. The fovea was scanned, and the PED height (PEDH), PED area (PEDA), PED volume (PEDV), and central foveal thickness (CFT) were measured. According to the OCT image features of PED, the affected eyes were divided into serous PED (sPED), fibrovascular PED (fPED), and hemorrhagic PED (hPED), and were grouped accordingly. Among the 42 eyes, 16 (38.1%, 16/42), 14 (33.3%, 14/42), and 12 (28.6%, 12/42) eyes were in the sPED group, fPED group, and hPED group, respectively. All patients received IVC treatment once a month for 3 consecutive months, and then on-demand treatment after assessment. BCVA and OCT were re-examined 3, 6, and 12 months after treatment, and the changes of BCVA, PEDH, PEDA, PEDV, and CFT in the affected eyes before and after treatment were compared, and repeated measures analysis of variance was used for statistical analysis. ResultsAt 12 months after treatment, the PEDH, PEDA and PEDV of the affected eyes in the sPED group, fPED group and hPED group were significantly lower than those before treatment, and the difference was statistically significant (P<0.05). The difference in the degree of improvement was －318.67±258.09 μm, －6.50±6.33 μm2, －1.95±1.78 μm3 in the hPED group; －119.31±224.13 μm, －0.86 ±5.00 μm2, －0.56±1.64 μm3 in the sPED group; fPED group were －53.93±92.51 μm, －0.76±2.54 μm2, －0.19±0.46 μm3. The improvement degree of the affected eyes in hPED group was significantly greater than that in sPED group and fPED group, and the difference was statistically significant (F=5.918, 6.029, 5.494; P＜0.05). Compared with the BCVA and CFT before treatment, 12 months after treatment, the difference was statistically significant in the fPED group and the hPED group (P＜0.05); there was no significant improvement in the sPED group (P＞0.05). There was no significant difference in the BCVA of the affected eyes in the three groups compared with those before treatment (F=0.817, 0.741, 0.848; P＞0.05). ConclusionConbercept can effectively improve or stabilize the visual function and anatomical morphology of eyes with type 1 MNV in nAMD with sPED, fPED and hPED, among which the anatomical effect is better for hPED.
Axon guidance molecules, slit glycoprotein (Slit) and Roundabout receptors (Robo) were firstly identified in the central neural system of Drosophila melanogaster. The Slit-Robo signal plays a crucial role in axon guidance, inflammation, tumor metastasis and angiogenesis, of which the role of Slit2-Robo pathway in angiogenesis has drawn a greater attention and still remains conflicting. Herein, we provide a review on the role of Slit2-Robo pathway in ocular angiogenesis and vascularization of other organs and systems. We hope this review will be the basis of further study on the mechanism of Slit2-Robo signaling on angiogenesis and provide new target for the therapy on ocular vascular disease
ObjectiveTo analyze the regulative rule of mRNA of vascular endothelial growth factor (VEGF) in mice with oxygen-induced retinopathy, and to elucidate the possible mechanism of occurrence of neovascularization in retinopathy of prematurity (ROP).MethodsSixty 7-day-old C57BL/6J mice were divided into oxygen-induced retinopathy group and control group. In oxygen-induced retinopathy group, 36 mice were exposed to 75% oxygen for 5 days and then to room air for 5 days; in control group, 24 mice were raised in room air. Vascular perfusion of fluorescein and retinal stretched preparation were used to observe the morphologic changes of retinal vessels. Reversal transcriptionpolymerase chain reaction (RT-PCR) was used to observe changes of VEGF mRNA in each group. ResultsIn oxygen-induced retinopathy group, the morphologic characteristics of retinal vessels were the unperfused area at the center of superficial and deepseated vessels, and the neovascularization appeared at mid-peripheral retina after 2 days in relative hypoxia condition. The results of RT-PCR showed space-time corresponding relation between expression of VEGF and neovascularization, which meant that the transcription of VEGF mRNA decreased in hyperxia conditionand increased in relative hypoxia condition. ConclusionHypoxia is the main reason of occurrence of retinal neovascularization; increased expression of VEGF caused by relative hypoxia after hyperxia might be effective in reducing the occurrence of neovascularization in ROP.(Chin J Ocul Fundus Dis, 2005,21:292-295)
Objective To determine the expression of the growth factors and the receptors related to angiogenesis in the intraocular tissues incarcerating in the sclerotomy sites. Methods Ten specimens from prolapsing intraocular tissues in sclerotomy sites during vitrectomy were obtained and serially sectioned in cryostate and were stained with a group of polyclonal antibodies against vascular endothelial growth factor(VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor-A(PDGF-A) and transforming growth factor-β1(TGF-β1) as well as their receptors by using a streptavidin peroxidase system. Results The tissues prolapsed from the sclerotomy sites were identified as retina(3 cases), vitreous tissues(3 cases), degenerated red blood cell components(2 cases), ciliary body(one case) and fibrous tissue(one case). All specimens expressed VEGF and bFGF as well as their receptors. PDGF-A, TGF-β1 and their receptors expressed in the most of specimens. The positive cells included retinal cells, ciliary non-pigmented epithelial cells and pigmented epithelial cells, fibrous cells and the cells in vitreous. Conclusions The intraocular tissues incarcerated in the sclerotomy entries express the growth factors and receptors related to angiogenesis. This might be one of the potential factors of developing anterior proliferative vitreoretinopathy. (Chin J Ocul Fundus Dis, 2002, 18: 34-37)
Purpose
To evaluate the efficacy of vitreous surgery and endolaser in a series of patients with retinal vein occlusion(RVO)with vitreous hemorrhage,neovascular membranes(NVM) and/or traction retinal detachment(TRD).
Methods
Clinical records were reviewed on 37 consecutive patients(38 eyes)who underwent vitreous surgery and endolaser for RVO with persistent vitreous hemorrhage,NVM and/or TRD.There were 19 patients(20 eyes)with retinal branch vein occlusion (BRVO)and 18 patients(18 eyes)with central retinal vein occlusion(CRVO).
Results
NVM and TRD were confirmed during operation in 27 and 23 eyes,respectively.Visual acuity improved postoperatively in 34 eyes(89.5%)including 22 eyes with 0.1 or better vision,and 4 eyes remained unchanged.CRVO group had longer history and less visual improvement after surgery.
Conclusions
Vitreous surgery and endolaser photocoagulation can improve the outcome in the majority of patients with RVO with vitreous hemorrage,NVM and/or TRD.
(Chin J Ocul Fundus Dis,1998,14:3-6)
Objective To review the research status of the neovascularization of adi pose tissue engineering in the past decade so as to provide theoretical references for the development of the rapid revascularization of tissue engineered adi pose. Methods The l iterature about the revascularization of adi pose tissue engineering was extensively reviewed andanalyzed, centering on 5 elements: specificity of histological structures and blood supply, revascularization mechanism, coculture of different seed cells, modification of scaffold, and microenvironment. Results Adi pose tissue engineering offers a new solution for soft tissue defects. However, there is still the unfulfilled need in the size of engineered adipose tissue (less than 1 mL), which was determined by the degree of neovascularization in engineered tissue. Overall, rapid neovascularization in engineering tissue is a key l ink of experimental study changing into cl inical appl ication. Conclusion Providing a sufficient supply with nutrients and oxygen by means of a sufficient and rapid neovascularization will be at the heart of any attempts to obtain bigger tissue engineered adipose to meet the demand of repairing large soft tissue defect.
Purpose
To estabalish a quantifying model of retinal neovascularization suitable for the study of pathogenesis and therapeutic intervention for the retinal neovascularization．
Methods
Sixteen one-week-old C57BL/6 mice were exposed to 75% oxygen for 5 days and then to room air and 16 mice of the same age kept in room air as controls.Ink-perfused retinal flatmount was examined to assess the oxygen-induced changes of retinal vessels.The proliferated neovascular response was quantitated by counting the nuclei of endothelial cells of new vessels extending from the retina into the vitreous in 6 mu;m sagittal cross sections． VEGF and bFGF were determined on the cross-sections after immunohistochemcal stain.
Results
Constriction and closure of the blood vessels were found under the hyperoxia condition,and dilation and proliferation were found under the relatively hypoxia status.There was a mean of 24 neovascular nuclei per cross-section in the oxygen-treated retina and less than 1 nucleus in the control group (P＜0.001).VEGF stain was found ber in the inner retinal layer of oxygen-treated mouse than in that of the controls.
Conclusion
The quantifying model of retinal neovascularization may fascilitate the further researches of medical intervention and pathogenesis of retinal neovacularization.
(Chin J Ocul Fundus Dis，2000，16：213-284)
