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Background Retinal neovascularization is pathological basis of a variety of fundus diseases,but its pathogenesis is unclear.Studies showed that the expression level of radixin in retina is remarkably increased in retinal neovascularization-related diseases.It is presumed that silencing or down-regulating the abnormal expression of radixin is helpful for curing retinal neovascularization-related diseases.Objective This study was to investigate the inhibitory effect of radixin short hairpin RNA (shRNA) plasmid on expression of radixin gene in retina of oxygen-induced retinopathy (OIR) mice.Methods Sixty-four 7-day-old C57BL/6J mice were randomly divided into normal control group, model control group, radixin shRNA plasmid group and shRNA plasmid group by random number table.There were 16 mice in every group.OIR models were established by exposing the mice in an environment of (75±2) ％ oxygen for 5 days and then returned to the normal air in the model control group,radixin shRNA plasmid group and shRNA plasmid group,while the mice of the normal control group were fed in the normal air environment.Radixin shRNA plasmid or control shRNA plasmid at the dose of 1 μg was intravitreally injected in 12-day-old mice of the radixin shRNA plasmid group or shRNA plasmid group, respectively.Five days later, FD-2000S angiography was performed on the mice of each group and then retinal flatmounts were prepared for the observation of retinal vessels.The mice from various groups were sacrificed and retinal sections were prepared.The vascular endothelial nucleus and new blood vessels extending inner limiting membrane (ILM) were examined by hematoxylin and eosin staining;the expression of radixin in the retinas was detected using immunochemistry;the relative expression levels of radixin mRNA and protein were quantitative assayed by real-time quantitative RCR and Western blot, respectively.The use and care of the animals adhered to the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.Results The distribution of retinal vessels was normal in the normal control group.Non-perfusion zone at the posterior pole of retina, circuity of blood vessels,leakage of vessel wall and new blood vessels were found in the mice of the model control group.Non-perfusion zone and microaneurysms were also exhibited in the shRNA plasmid group.However,these findings were slight in the radixin shRNA plasmid group.The surface of ILM was in discontinuity in the model mice and shRNA-injected mice with more vascular endothelial cell nucleus and more tubes extending ILM than that in the radixin shRNA plasmid group.The immunochemistry results showed that the expressions of radixin in the normal control group and radixin shRNA plasmid group were weaker than those in the model control group and control shRNA plasmid group.The relative expression levels of radixin mRNA were 1.002±0.043,2.236-±0.093,0.556±0.015 and 2.272±0.096 in the normal control group, model control group,radixin shRNA plasmid group and control shRNA plasmid group,and those in the radixin shRNA plasmid group were significantly reduced in comparison with the normal control group, model control group and the shRNA plasmid group (all at P＜0.01).The relative expression levels were 1.000±0.082,1.193±0.021,0.263± 0.016 and 1.235±0.005 in the normal control group,model control group,radixin shRNA plasmid group and shRNA plasmid,with the lowest expression level in the radixin shRNA plasmid group (all at P＜0.01).Conclusions Radixin shRNA can downregulate the expression of radixin gene in the retinas of OIR mice and further inhibit pathological retinal neovascularization.
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Background Hypoxia is the main factor of retinal neovascularization and is closely associated with retinal ganglial cells (RGCs) degeneration.However, the study of retinal neural tissue lesions is rare.Objective This study was to investigate the influence of hypoxia environment on the expression of annexin A2 (ANXA2) in mouse RGC-5 cells and explore the mechanism of RGCs damage induced by hypoxia.Methods Immortalized mouse RGC-5 cells were cultured in high glucose DMEM with 10％ fetal bovine serum.The cells were identified by detecting the expression of Thy-1 ,a specific biomarker of RGCs.CoCl2 was added into the medium at the final concentrations of 50,100,200 and 300 μmol/L, and the cells without CoCl2 served as the control group.Cell viability (absorbance) was assayed by cell counting kit-8 (CCK-8) method in 12,24 and 48 hours after addition of CoCl2.The hypoxic cell models were established in DMEM with 100 μmol/L CoCl2 and divided into the hypoxic 3-hour group, hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group,with the normal cultured cells as the normal control group.Apoptotic cells were determined by using hoechst 33342 stain.The expression levels of ANXA2 mRNA and protein in the cells were detected by real-time quantification PCR and Western blot,respectively.The expression and location of ANXA2 in the cells were examined by using immunofluorescence technique.Results The cultured cells grew well and showed the fusiform and polygonal shape,with positive expression of Thy-1 protein.Compared with the normal control group, the viabilities of the cells were insignificantly changed in the 50 μ mol/L CoCl2 group and 100 μmol/L CoCl2 group (all at P＞0.05) ,but the cell viabilities were significantly reduced in the 200 μμmol/L CoCl2 group and 300 μmol/L CoCl2 group in various time points (all at P＜0.05).Hoechst 33342 staining showed that the apoptotic cells with nuclear condensation and high green fluorescence intensity were obtained in the hypoxia groups.The relative expression levels of ANXA2 mRNA were significantly lower in the hypoxic groups than those in the normal control group (all at P ＜ 0.05).The relative expression levels of ANXA2 protein were significantly lower in the hypoxia 3-,6-, 12-and 24-hour group than those in the normal control group (all at P＜ 0.05).Apoptotic cells were seen in the hypoxic 3-hour group, hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group compared with the normal control group, showing the bright blue fluorescence in cellular nucleus for hoechst 33342.The relative expressing levels of ANXA2 mRNA in the cells were 0.80±0.14,0.67±0.33, 0.49±0.17 and 0.39±0.02 in the hypoxic 3-hour group,hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group, which were significantly declined in comparison with the normal control group, with a statistically difference among the groups (F=434.354, P =0.000).The relative expression values of ANXA2 protein were 0.552 6±0.012 3,0.425 9± 0.033 4,0.344 9 ± 0.017 8 and 0.382 7 ± 0.022 1 in the hypoxic 3-hour group, hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group,which were remarkably lower than 0.602 1 ±0.001 4 in the normal control group, showing considerably difference among the groups (F =3.057, P =0.000).ANXA2 proteins were highly expressed in the cellular nucleus and less expressed in the cell membrane and cytoplasm in the normal cells.Compared with the normal control group, the ANXA2 protein showed weak expression in the hypoxia group and primarily in the cytoplasm.Conclusions The expression of ANXA2 down-regulates in hypoxic mouse RGC-5 cells,which may participate in the apoptosis process of RGCs in high glucose environment.
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Objective: To investigate the effect of losartan on angiotensin II (Ang II) expression and myocardial remodeling in myocardial infarction (MI) rats’ model. Methods: A total of 32 SD male rats were divided into 4 groups, Sham operation group, MI group, MI with losartan 10mg/(kg·d) group and MI with losartan 20mg/(kg·d). n=8 in each group. MI model was established and the electrocardiogram changes before and after MI were recorded, hemodynamic indexes were detected at 4 weeks after MI, pathological changes of myocardial tissue were examined by HE staining. The myocardial mRNA and protein expressions of ACE2 and Ang II were detected by RT-PCR and Western Blot analysis. Results: Compared with Sham operation group, MI group showed increased LVMI and decreased LVEF P Conclusion: Losartan could increase ACE2 expression and therefore, inhibit Ang II expression and improve the ventricular remodeling in MI rats’ model.
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Background Heparanase degrade heparan sulfate side chains of heparan sulfate proteoglycans in the extracellular matrix.Heparanase induces angiogenesis and likely promotes the vascularization of tumor.ObjectiveThe present study is to investigate the expression of heparanase and perlecan in retinas with oxygen-induced retinopathy.Methods Sixty-five clean neonatal C57BL/6J mice were raised in a hyperbaric oxygen box with a volume percentage of 75%±2% for 5 days and then returned to the normal air room.Another 65 matched mice were raised in the normal environment as controls.Evans blue was infused by the superior vena cava in all the mice on postnatal days 12,13,17,21 and 30,afterwards fluorescein angiography was performed and then the mice were sacrificed.The retinas of mice were isolated and prepared and the retinal vessels were examined under a fluorescent microscope and optical microscope.Heranase and perlecan mRNA was detected using reverse transcription PCR (RT-PCR).Heranase and perlecan proteins were detected by Western blot.The analysis of variance was used to compare the mRNA and the protein levels of heranase and perlecan between the experimental and control groups.Results The expression of heparanase mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=16.303,P=0.000;F_(time)=18.614,P=0.000;F_(interaction)=11.299,P=0.000),and the expression of heparanase mRNA was significantly enhanced in mice from postnatal days 12,13,17 and 21 compared with normal control mice (P=0.001,0.000,0.000,0.001,respectively).The expression of heparanase protein in the retinas of different ages of mice and the different groups followed the same tendency(F_(group)=458.134,P=0.000;F_(time)=78.466,P=0.000;F_(interaction)=71.398,P=0.000).The expression of perlecan mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=7.351,P=0.013;F_(time)=9.098,P=0.000;F_(interaction)=3.349,P=0.000),and increase in differences also were clearly seen in mice from postnatal days 13,17 and 21 compared with normal control mice (P=0.048,0.000,0.003,respectively).Conclusion The expression of heparanase and perlecan is associated with the development and progression of retinal neovascularization,and perlecan and heparanase together produce a synergistic effect.Heparanase and perlecan may participate in the angiogenesis of oxygen-induced retinopathy.
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ObjectiveTo explore the surgical techniques, efficacy and the management of complicated ocular trauma with anterior-posterior segment complications such as cyclodialysis. Methods Fifty-five patients (55 eyes) with complicated ocular trauma were enrolled in this study. Among them, there were 35 cases with eyeball contusion and 20 cases of eyeball rupture. Preoperative visual acuity was from no light perception to 0. 15, intraocular pressure (IOP) ranged from one to 10 mm Hg (1 mm Hg=0. 133 kPa).Cyclodialysis, vitreous hemorrhage or retinal detachment were revealed by B-ultrasound and ultrasound biomicroscopy (UBM). Cyclodialysis clefts ranged from one to 12 clock-hours. All patients underwent 3-port pars plana vitrectomy with gas/silicone oil tamponade and ciliary body reattachment by cryotherapy (cyclodialysis clefts ≤ three clock-hours) or suture fixation ( cyclodialysis clefts ＞ three clock-hours).Healing after surgical trauma, visual acuity, intraocular pressure, intraocular hemorrhage, ciliary body and retinal reattachment were followed up. Results In one month after surgery, UBM showed 54 patients out of 55 patients had good ciliary body reattachment. Gonioscopy revealed cyclodialysis still existed in one patient, and this was cured by a second surgical suture fixation. In three months after surgery, ocular trauma healed in all 55 patients, the visual acuity ranged from no light perception to 0. 15, with a best corrected visual acuity of 0.8. The ciliary body and retina had good reattachment. IOP of 52 patients was normal; IOP of three patients was still lower than 10 mm Hg. Three patients had secondary glaucoma which was treated by glaucoma surgery. Conclusions Complicated ocular trauma with cyclodialysis can be treated with vitrectomy and cryotherapy or transscleral sutures. The procedure is safe and effective.
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Objective To analyze the causes and pathogens of infectious endophthalmitis in ourhospital.Methods The clinical data and laboratory findings of 282 inpatients with infectiousendophthalmitis were retrospectively reviewed.There were 206 males(73.05%)and 76 females(26.95%)with a mean age of(36.0±11.2)years(range from 2 to 79 years).Except 14 patients who lost theireyeballs as of serious conditions.pathogen examinations of vitreous specimen had been performed for allremaining 268 cases.The clinieal and laboratory data were statistically analyzed by Chi-square and Kappaconsistency tests,when P<0.05 the difference should be considered as statistically significant.ResultsThe 4 major causes of infectious endophthalmitis of those 282 patients included ocular trauma(177cases,62.77%),infectious keratitis(61 cases,21.63%),eye surgery(32 cases,11.35%),andendogenous endophthalmitis(12 cases,4.25%).19.21% of the patients in ocular trauma group was 2-13years old,55.74% of the patients in infectious keratitis group was 14-50 years old,and 60.61% of thepatients in eye surgery group was 51-79 years old.The major pathogens were fungus and staphylococcieach accounted for 30.63% of all cases.Most of the pathogenic bacteria were sensitive to ciprofloxaein(81.81%)and gentamicin(77.92%).Conclusions The major reason of infectious endophthalmitis wasocular trauma,and the major pathogens were fungus and staphylococci.Those findings may help us totreat infectious endophthalmitis patients appropriately.
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Objective To investigate the clinical features of polypoidal choroidal vasculopathy. Methods The clinical manifestations and the findings of fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA) of 7 eyes in 6 patients with polypoidal choroidal vasculopathy were analyzed retrospectively. Results The ophthalmoscopic examination of PCV in this series of patients revealed orange-red lesions mainly in macular region in early stage, and then hemorrhage, exudation, edema, serous and (or) hemorrhagic detachment of retinal pigment epithelium in affected portions in advanced stage, and retinal and choroidal atrophy in late stage. FFA discovered the vascular dilation at the border of the choroidal vascular network in 1 eye, dotted hyperfluorescence in 6 eyes, and patches of hyperfluorescence in late phase. ICGA disclosed a vascular branching network in choroid with polypoid pattern of the terminal path of the vessels of network in early phase in 5 eyes, and the typical dotted or clustered polypoidal hyperfluorescence in 7 eyes in late phase. Conclusion The characteristic findings of FFA and ICGA are very diagnostic for PCV.