The therapeutic effects of panretinal cryotherapy(PRC)on proliferative diabetic retinopathy(PDR)were prospectively investigated in 80 eyes with PDR of 44 patients.Forty eyes with PDR(20 of them stage Ⅳ and 20 stage Ⅴ)received PRC operation.Of the 80 eyes,the other 40 ones with stage Ⅳ and Ⅴ similar to the formers were observed conservatively as controls.The follow up duration was 2 years.We found that in the cases of stage Ⅳ,no more remarkable visual loss was found after operation.There was a significant difference comparing with the control（P＜0.002）,and the retinal neovascularization regressed more noticeably than the control group（P＜0.001）.In the cases of stage Ⅴ,the incidence of the traction retinal detachment was 55% in the operated group,and was 20% in the control group.There was a statistic difference between them（P＜0.05）.The clearance of the vitreous hemorrhage was more rapid in the operated group than the control（P＜0.025）.The above results suggest that cryotherapy is suitable for the cases of earlier stage which cannot be performed with photocoagulation for any reasons,but not for the patients with advanced retinal proliferation.Photocoagulation for any reasons,but not for the patients with advanced retinal proliferation. （Chin J Ocul Fundus Dis,1993,9:148-151）
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)
Objective To observe the effect of Twist gene interference on the migration and pAkt protein expression of Rhesus retinal vascular endothelial cell line. Methods The Rhesus retinal vascular endothelial cells (RF/6A) were divided into Twist interference plasmid group, negative control group, and phosphate buffered solution (PBS) group; plasmid vectors were transfected via liposome gene transfection method. Migrated endothelial cells was detected and counted by Transwell chamber assay. Matrigel was used in endothelialcell tube formation; the inhibitory effect of Twist gene interference on endothelial cell tube formation was observed.The effect of Twist gene interference on the expression of pAkt protein in RF/6Acells was measured by Western blot. Results The number of migrated endothelial cells in Twist interference plasmid group was lower than that in the negative control and PBS group (F=23.786，P=0.000).The number of endothelial cell tubes in Twist interference plasmid group was apparently less than that in the negative control and PBS gorup (F=7.159，P=0.014). The expression of pAkt protein in Twist interference plasmid group decreased markedly.Conclusion Twist gene interference can suppress the migration of retinal endothelial cells via inhibiting the expression of pAkt protein.
Objective To compare two kinds of myofascial flap encapsulating adi pose-derived stromal cells (ADSCs) in adi pogenic efficacy in vivo, and to provide experimental basis for the efficient transplantation of free adi pose tissue. Methods ADSCs were isolated from the subcutaneous adipose tissue in the neck of 10 New Zealand rabbits (aged 3-4 months old, male and female, weighing 2.0-2.5 kg), and primary culture and subculture of ADSCs were conducted. When the cells at passage 3 covered 70%-80% of the bottom of the culture flask, BrdU (10 μg/mL) was appl ied to label the cells for 48 hours before performing immunofluorescence staining. Oil red O staining observation was conducted to thosecells 2 weeks after being induced towards adi pocyte, al izarin red staining observation was performed 3 weeks after being induced towards osteoblast, and alcian blue staining was conducted 2 weeks after being induced towards chondrocyte. Besides, after being induced towards adipocyte for 2 weeks, 1 × 107 ADSCs/piece at passage 3 labeled by BrdU was seeded into Col I (10 mm × 10 mm × 5 mm/piece) to prepare cell carrier complex. The experiment was divided into two groups: group A in which vascular pedicled dextral latissimus dorsi fascial flap was adopted to encapsulate the complex; group B in which dextral gluteus maximus fascial flap with no specific vessel pedicle was appl ied to encapsulate the complex. Rabbits in each group went through autogenous ADSCs transplant and self control. The implants were dislodged 8 weeks after operation, HE staining and immunohistochemistry staining were performed to testify cambium, the wet weight and micro vessel count of the cambium in each group were tested, immunofluorescence staining was performed to determine the origin of cambium and microvascular endothel ium. Results The nucleus of ADSCs positive for BrdU label ing showed green fluorescence under fluorescence microscope, with the positive label ing ratio of ADSCs above 90%. For ADSCs at passage 3, the formation of red l ipid droplets within cells was observed 2 weeks after being induced towards adipocyte, red calcium nodules were evident 3 weeks after being induced towards osteoblast, and highly congregated cell mass positive for alcian blue staining appeared 2 weeks after being induced towards chondrocyte. Eight weeks after operation, neogenetic blood vessel grew into scaffolds and no obvious fibreencapsulation was observed in group A, while few blood vessel grew into scaffolds in group B. The wet weight of cambium in group A and B was (0.149 5 ± 0.017 3) g and (0.095 3 ± 0.012 7) g, respectively, indicating there was a significant difference between two groups (P lt; 0.01). HE staining showed the formation of neogenetic adipose tissue and the growth of micrangium in the implant, and the degradation and absorption of scaffold. The micro vessel count of group A and B was 31.2 ± 4.5 and 19.3 ± 2.6, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Eight weeks after operation, the immunofluorescence staining of cambium showed that the cell nucleus of regenerated adi pocytes and partial capillary endothel ium in groups A and B presented green fluorescence. Conclusion ADSCs encapsulated by vascular pedicled latissimus dorsi fascial flap and collagen protein scaffold complex has a higher adi pogenic efficacy in vivo than the gluteus maximus fascial flap with no specific vessel pedicle.
Objective To investigate auto-cortex of crystalline lens-induced neovascular epiretinal membrane（NVERM）by micro-injuring posterior c apsule of crystalline lens. Methods twenty four C57BL/6 mouse between 4-6 weeks were selected, and divided into two groups randomly: auto-cortex of crystalline group and the control group. The auto-cortex of crystalline group was treated by penetrating the posterior capsule of lens and washing out the lens cortex into the mouse vitreous using PBS (phosphate buffered solution), while the control group were injected PBS into vitreous merely. Clinical change s were followed by slit-lamp examination and photograph. The eye balls were enu cleated at the day of 3, 7, 14 and 28 after operation. Both HE and immunohistoch emistry were used to detect the pathological changes. Results postoperative one to three days, 11 of 12 mouse in autocortex of crystalline g roup, lens appear to alba turbid at different levels one after another, and then develop into highdensity chinaware white. Postoperative (po) three days, HE s taining shows cortex of lens debris transmigrated in vitreous cavity, and some o f which approached to internal limiting membrane and lead it to rough and discon tinue; Po7-14 days, the capillary in retina expanded, migrated and broke though t internal limiting membrane which got to the pro retina and became the new ves sels. And typical NVERM were observed. Po28 days, some vascularslike structure formed in vitreous cavity. None of mouse in control group developed NVERM. Conclusion Auto-cortex of crystalline lens can induced neovascular epiretinal membrane in C57BL/6 mouse. (Chin J Ocul Fundus Dis,2008,24:118-121)
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.
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 investigate the effect of Nodal protein on retinal neovascularization under hypoxia. MethodsIn vivo animal experiment: 48 healthy C57BL/6J mice were randomly divided into normal group, oxygen-induced retinopathy (OIR) group, OIR+dimethyl sulfoxide (DMSO) group and OIR+SB431542 group, with 12 mice in each group. Retinal neovascularization was observed in mice at 17 days of age by retina flat mount. Counts exceeded the number of vascular endothelial nuclei in the retinal inner boundary membrane (ILM) by hematoxylin eosin staining. In vivo cell experiment: human retinal microvascular endothelial cells (hRMEC) were divided into normal group, hypoxia group, hypoxia+DMSO group and hypoxia +SB431542 group. The cell proliferation was detected by thiazolyl blue colorimetry (MTT). The effect of SB431542 on hRMEC lumen formation was detected by Matrigel three-dimensional in vitro molding method. Cell migration in hRMEC was detected by cell scratch assay. The Seahorse XFe96 Cell Energy Metabolism analyzer measured extracellular acidification rate (ECAR) of intracellular glycolysis, glycolysis reserve, and glycolysis capacity. One-way analysis of variance was used to compare groups. ResultsIn vivo animal experiment: compared with normal group, the neovascularization increased in OIR group (t=41.621, P＜0.001). Compared with OIR group, the number of vascular endothelial nuclei breaking through ILM in OIR+SB431542 group was significantly reduced, and the difference was statistically significant (F=36.183, P＜0.001). MTT test results showed that compared with normal group and hypoxia+SB431542 group, the cell proliferation of hypoxia group and hypoxia+DMSO group was significantly increased, and the difference was statistically significant (F=39.316, P＜0.01). The cell proliferation of hypoxia+SB431542 group was significantly lower than that of hypoxia+DMSO group, and the difference was statistically significant (t=26.182, P＜0.001). The number of intact lumen formation and migration cells in normal group, hypoxia group, hypoxia+DMSO group and hypoxia+SB431542 group were statistically significant (F=34.513, 41.862; P＜0.001, ＜0.01). Compared with the hypoxia+DMSO group, the number of intact lumen formation and migrating cells in the hypoxia+SB431542 group decreased significantly, and the differences were statistically significant (t=44.723, 31.178; P＜0.001, ＜0.01). The results of cell energy metabolism showed that compared with the hypoxia +DMSO group, the ECAR of intracellular glycolysis and glycolysis reserve in the hypoxia +SB431542 group was decreased, and the ECAR of glycolysis capacity was increased, with statistical significance (t=26.175, 33.623, 37.276; P＜0.05). ConclusionSB431542 can inhibit the proliferation, migration and the ability to form lumens, reduce the level of glycolysis of hRMECs cells induced by hypoxia.
With the rapid development of fundus imaging technology, it is of great significance to establish a new naming system for neovascular age-related macular degeneration (nAMD) based on the multi-mode imaging. In 2020, an international panel of retina specialists, imaging and image reading center experts, and ocular pathologists reached a consensus after repeated discussions, a new name for nAMD subtype and related lesions was established based on the previous knowledge of fundus fluorescein angiography and pathology, combining indocyanine green angiography, optical coherence tomography and optical coherence tomography angiography with current pathological knowledge, in order to help ophthalmologists to study nAMD. The consensus proposed the term "macular neovascularization" and classified it into type 1, type 2 and type 3. Many lesions related to macular neovascularization, such as pigment epithelial detachment, hemorrhage, fibrosis, rip of retinal pigment epithelium and so on, were named. The new designation will help improve clinical communication between different studies, establish standard definitions and terms between reading centers and researchers, and further promote the understanding and communication of nAMD among ophthalmologists.
Purpose To investigate the roles of vascular endothelial growth factor (VEGF),glutamate and gamma;- aminobutyric acid (GABA) in neovascularization of proliferative diabetic retinopathy (PDR). Methods Vitreous samples were collected from 25 patients (27 eyes)with PDR and 14 patients (14 eyes) with idiopathic macular hole.VEGF levels were determined by ELISA.Amino acids analyses were performed by high-performance liquid chromatography. Results Patients with PDR had significantly higher concentrations of VEGF (median 0.41 ng/ml,quartile 0.54 ng/ml) than controls (median 0.017 ng/ml,quartile 0.01 ng/ml)(Plt;0.001).The levels of glutamate [(11.7plusmn;3.0) mu;mol/L] and GABA [(7.2plusmn;3.9mu;mol/L)] were also higher in patients with PDR than glutamate [(5.8plusmn;0.7) mu;mol/L] and GABA [(3.3plusmn;2.9) mu;mol/L) in controls (Plt;0.05).The glutamate level and GABA level had a significantly positive correlation with VEGF level. Conclusions The increased levels of glutamate and GABA indicate retinal ischemia.The correlations of glutamate and GABA levels with an elevated VEGF level provide biochemical support for ischemi induced neovascularization in patients with PDR.The findings present novel modalities in treatment of PDR. (Chin J Ocul Fundus Dis,2000,16:162-165)