Objective To observe the retinal apoptosis of laser-induced retinal injury in mice after bone marrow mesenchymal stem cells transplantation. Methods Green fluorescent protein (GFP) labeled MSCs from C57BL/6 mice were cultured in vitro. A total of 135 C57BL/6 mice were divided into three groups including normal control group (15 mice), injured control group (60 mice) and MSCs treatment group (60 mice). Laser retinal injuries were induced by laser photocoagulation. One day after photocoagulation, 02 ml cell suspension, which contained 1times;106 GFP-MSCs, were injected into the mice in treatment group via tail vein, and the mice in injured control group were given equal volume of phosphate buffer solution. Animal were execute on three, seven, 14 and 21 days following laser damage. Hematoxylin and eosin (HE) staining was performed to assess the changes of injured retinas. The diameters of laser spots and areas with total loss of cells in outer nuclear layer (ONL) were analyzed by image processing software. The apoptosis of retinal cells was examined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. The migration of GFP-MSCs into the retina was observed by fluorescence microscope. Results HE staining showed that the retinal structures were integrated in normal control group. Retinal damages were observed both in injured control group and MSCs treatment group, but milder in the latter. Though the average diameter of area with total loss of cells in ONL of MSCs treatment group was less than the injured control group (t=5.769, P＜0.05), the diameters of laser spots show no difference (t=0.964,P＞0.05) on day three. Both the average diameter of laser spots (t=5.180, 5.417, 2.381) and area with total loss of cells in ONL (t=3.530, 3.224, 3.162) were less in the MSCs treatment group on day seven, 14 and 21 (P＜0.05). TUNEL staining shows that the apoptosis were decreased after MSCs transplantation on day three, seven, 14 and 21 (t=11.142, 7.479, 6.678, 3.953,P＜0.05). No apoptosis was observed in normal control group. Very few GFP-MSCs were observed in the retina at all time-points. They were only seen in the subretinal and choroidal neovascularization occasionally on day seven and 14. Conclusion MSCs transplantation can effectively limit the range of retinal laser damage and inhibit cell apoptosis.
Objective To observe the configuration and viability of full thickness human fetal retina after short-, mid- and long-term preservation. Methods Twenty-two full thickness human fetal retinae of gestational age of 12-24 weeks were coated by glutin and cut into 88 pieces, and then preserved in Ames' solution, DX solution, -80℃ refrigerator or under cryopreservation condition. The cell viability of retinal neuroepithelial layer was determined by trypan blue staining, retinal configuration was determined by light microscope and electromicroscope. Results The viability of neuroepithelial layer was (94.79plusmn;2.85) % in fresh fetal retina, gt;80% in Ames' solution within 4 hours, and gt;77% in DX solution within 2 days. There was no significant difference between those solution-preservations and the fresh fetal. In -80℃ refrigerator, the viability was (65.83plusmn;5.06)% after 7 days, and then dropped to (57.54plusmn;16.18)% at the end of the first month. Under the cryopreservation condition, the viability was (69.46plusmn;9.31)% at the end of first month. Light and transmission electron microscopy had not deteced any abnormals in the full thickness human fetal retina preserved in Ames' solution within 2 hours, but showed clear retinal layers with bigger intercellular space after preserved in DX solution for 2 days, in -80℃ refrigerator for 7 days and under cryopreservation condition for 1 month. Conclusion Ames' solution and DX solution can preserve good viability and configuration of full thickness human fetal retina in a certain time period.
Objective To investigate the features of optic disc formation and retinal nerve fiber layer(RNFL) changes in primary open-angle glaucoma with myopia (M-POAG). Methods On 63 eyes of 38 patients with M-POAG were imaged of the fundus,and were evaluated with the microcomputer image analyser,and were compared with the simple POAG (S-POAG) eyes. Results Variant features of the optic disc and RNFL atrophy were found in this M-POAG eyes.The shapes of the optic disc were revealed to be vertically or horizontally oval,obliquely inserted and irregular,the color of the most of optic disc was pallor.The pattern of glaucomatous cupping was saucer-like (28.6%),vertical (25.4%),oblique (23.8%),pot-like (9.5%),and focally or concentrically cupped.The quotient of the neuroretinal rim area and horizontal C/D ratio were significantly lower than those in S-POAG eyes (Plt;0.05,Plt;0.001).The focal point of the optic disc excavtions tended to be inferior.Most of the incidence in the focal atrophy of RNFL was located inferiorly,and the diffuse atrophy of RNFL was correlated positively with middle or late high-myopia POAG eyes (P＜0.005). Conclusion The variant features of the optic discs,glaucomatous cupping and RNFL atrophy formation in M-POAG eyes found in this series might be helpful in clinical diagnosis. (Chin J Ocul Fundus Dis,2000,16:81-84)
Purpose To study changes of cell cycle of vascular endothelial cell in non-proliferative diabetic retinopathy. Methods Alloxan induced Wistar-rats were employed and immunohistochemistry,Western blotting methods were used. Results The vascular endothelial cells of retinas of 8～20 weeks diabetic rats were observe to be cyclinD1,cyclinD3,cyclinB1,p21 and p27 positive stained with light and electronmicroscopies.CyclinE immuno-stained vascular endothelial cells was observed occasionally.Meanwhile,the evidences of morphologic changes of the vascular en dothelial cells were proved:less plasma,thinner cell,more bubble organelles than those of controls.But,the ultra-structures of pericytes and other type of retinal cells did not change and they also immunostain negative.Komas blue and Western blotting methods also proved that the vascular endothelial cells of retina of 20th week diabetic rats expressed cyclinD1,cyclinB1,p21 and p27 protein. Conclusion Glucose induced retinal vascular endothelial cells of 8～20th weeks diabetic rats enter cell cycle and were arrested at G1/S restriction point.This study also suggested that retinal vascular endothelial cells may possess the ability to resist glucose damage and mechanism of selfstability during very early stage of diabetes. (Chin J Ocul Fundus Dis,2000,16:173-176)
ObjectiveTo observe the effects of Rhodiola on the rat retinal tissue morphology and the hypoxia-inducible factor (HIF)-1α at simulated hypoxia at different altitudes. Methods Forty-eight adult female Sprague Dawley rats were randomly divided into the Rhodiola Intervention group (intervention group) and the control group, each group had 24 rats. The intervention group rats were treated with intraperitoneal injection of 10 ml/kg of large plants Rhodiola solution, and the control group rats were injected with same volume of saline. One hour after the injection, six rats were randomly selected from both of the two groups and reared in the plateau environment simulation laboratory modules with the oxygen partial pressure of 17.4, 14.6, 11.3 and 7.4 kPa, which simulated the altitudes of 1500, 3000, 5000 and 8000 meters indoor respectively. Six hours later the rat eyeballs were harvested for paraffin sections and analyzed by hematoxylin and eosin staining, and immunohistochemical staining to observe the expression of HIF-1α and p53. ResultsIn the control group, the rat retinal layers were edema and loose, the retinal thickness increased, the retinal structure was disorganized, the ganglion cells were swollen and degenerated, and some can observe the karyopyknosis, karyolysis and the reduced cells number. As the altitude increased, the pathological changes of retinal became more obvious. In the intervention group, the characteristics of rat retinal morphology were same with the control group, while the degree of morphology changes was lighter than the control group. HIF-1α and p53 expressed mainly in the ganglion cell layer and inner nuclear layer of rat retina in the control group. As altitude increased, the expression of HIF-1α and p53 were increased too, which was positive correlated (r=0.9846, P < 0.05). Compared with the control group, the rat retinal expression of HIF-1α increased, while expression of p53 decreased in the intervention group, and the differences were statistically significant (P < 0.05). ConclusionRhodiola can reduce the retinal tissue pathology damage caused by high altitude hypoxia, and its mechanism may be related to the increasing expression of HIF-1α and reducing expression of p53.
ObjectiveTo evaluate the effect of form deprivation myopia on optic nerve head and retinal morphology in guinea pigs using optical coherence tomography (OCT). MethodsTwenty guinea pigs aged from 4 to 5 weeks were chosen and randomly divided into the experimental group and control group, with 10 guinea pigs in each group. Form deprivation myopia was established for the right eyes of guinea pigs in experimental group for 4 weeks. The guinea pigs of control group were not intervened. Before and 4 weeks after form deprivation, refraction was measured by retinoscopy after cycloplegia; the axial length was measured by A-scan ultrasound; retinal nerve fiber layer (RNFL) thickness, optic nerve head and retinal morphology of guinea pigs were analyzed using OCT. ResultsBefore form deprivation, there were no statistically significant differences in spherical equivalent, axial length, RNFL thickness, disc edge area, optic disc area, average cup disc ratio, vertical cup disc ratio, cup volume, retinal thickness, or retinal volume between the experimental group and control group of guinea pig (P > 0.05). After 4 weeks of form deprivation, RNFL thickness of (64.9±17.7) μm in guinea pigs in experimental group was thinner compared to (97.9±25.1) μm in control group (t=-2.845, P=0.015). Retinal thickness of (142.7±3.4) μm in guinea pigs in experimental group was thicker compared to (138.4±3.5) μm in control group (t=2.338, P=0.038). There were no significant differences in disc edge area, optic disc area, average cup disc ratio, vertical cup disc ratio, cup volume or retinal volume between groups (P > 0.05). There were statistically significant differences in spherical equivalent, axial length, RNFL thickness, vertical cup to disc ratio cup volume, and retinal thickness between after and before form deprivation in the right eye of guinea pigs in the experimental groups (t=46.001, -50.119, 5.385, 3.447, -2.814, -8.911; P < 0.05), while there were no statistically significant differences in disc edge area, optic disc area, average cup disc ratio, or retinal volume (P > 0.05). ConclusionForm deprivation myopia has an effect on RNFL and retinal thickness.
Objective To determine the long-term changes in optic disc parameter and the thickness of circumpapillary retinal nerve fiber layer (CP-RNFL) and macular retina after acute primary angle closure. Methods Prospective clinical case-control study. A total of 26 patients (30 eyes) with acute primary angle-closure glaucoma (APACG) were in the APACG group, whose intraocular pressure were control after a single episode acute primary angle closure; 30 age-and sex-matched healthy subjects (30 eyes) in the control group. All subjects underwent three dimensional optical coherence tomography (3D-OCT) examination with 3D optic disk scanning or circle optic disk scanning and 6 mm×6 mm macular scanning. The parameters included average thickness of entire CP-RNFL, thickness of nasal, superior, temporal and inferior quadrant of CP-RNFL, disc area, disc cup area, rim area, cup/disc (C/D) area ratio, C/D horizontal diameter ratio and C/D vertical diameter ratio. The foveal retinal thickness, center retinal thickness (≤1 mm from the fovea), 4 quadrants of macular inner-ring ( > 1 mm but≤3 mm from the fovea) retinal thickness, 4 quadrants of macular outer-ring ( > 3 mm but≤6 mm from the fovea) retinal thickness, average thickness of macular retinal thickness and macular volume were measured and analyzed. Results The disc area, disc cup area, C/D area ratio, C/D horizontal diameter ratio and C/D vertical diameter ratio in APACG group were significantly bigger than the control group (t=3.22, 4.12, 3.90, 3.00, 3.23; P < 0.05), rim area was smaller than the control group (t=-2.63, P < 0.05). The average thickness (t=-6.68) and the thickness of superior (t=-5.90), temporal (t=-11.64) and inferior (t=-5.06) quadrants of CP-RNFL, center retinal thickness (t=-2.50), 4 quadrants of macular inner-ring retinal thickness (t=-4.91, -4.88, -2.83, -3.59), nasal (t=-2.13) and superior (t=-2.49) quadrants of macular outer-ring retinal thickness as well as average thickness of macular retinal thickness (t=-2.65) were significantly thinner than the control group (P < 0.05), and the macular volume (t=-2.69) was significantly smaller than the control group (P < 0.05). There was no statistically difference at nasal CP-RNFL (t=-0.11), foveal retinal thickness (t=-0.59), temporal (t=-0.67) and inferior (t=-1.02) quadrants of macular outer-ring retinal thickness between two groups (P > 0.05). Conclusions In comparison with the healthy subjects, the disc area, disc cup area, C/D area ratio, C/D horizontal diameter ratio, C/D vertical diameter ratio in APACG eyes were bigger, while rim area was smaller; the CP-RNFL and macular retinal thickness were thinner except nasal CP-RNFL, fovea, temporal and inferior quadrants of macular outer-ring retinal.
Objective To observe the macular morphology and circumpapillary retinal nerve fiber layer thickness (RNFL) in Parkinson's disease (PD) evaluated by spectral-domain optical coherence tomography (SD-OCT). Methods A total of 37 patients (74 eyes) with PD were in the PD group, 32 age- and sex-matched healthy subjects (64 eyes) in the control group. All subjects underwent SD-OCT examination with 5 line scanning, macular cube 512×128 scanning and optic disc volume 200×200 scanning. The retinal thickness, central foveal thickness (CFT), macular volume and thickness of circumpapillary, superior, inferior, nasal, and temporal of RNFL between two groups were comparatively analyzed. The relationship between SD-OCT parameters and age, disease duration, scores of Hoehn-Yahr and unified PD rating scale (UPDRS) Ⅲ in PD patients was analyzed by Pearson correlation analysis. Results Both of the retinal thickness and macular volume in PD group were significantly reduced than those in control group (t= −2.546, −3.410;P=0.012, 0.001). There was no difference of CFT (t= −0.463,P=0.644) and the thickness of circumpapillary (t= −1.645,P=0.102), superior (t= −0.775,P=0.439), inferior (t=−1.844,P=0.067), nasal (t= −0.344,P=0.732) and temporal (t= −0.541,P=0.590) of RNFL between two groups. The retinal thickness, macular volume, CFT and the thickness of circumpapillary, superior, inferior, nasal, temporal of RNFL had no relationship with age, disease duration and scores of Hoehn-Yahr and UPDRS Ⅲ in PD patients (P＞0.05). Conclusions In PD patients, the retinal thickness and macular volume are decreased, however, the circumpapillary RNFL have no obvious alterations.