Application of pediatric ocular trauma score in pediatric open globe injuries.

AIM: To evaluate the predictive value of pediatric penetrating ocular trauma score (POTS) on the visual outcome in children with open globe injury. METHODS: A retrospective study in 90 children (60 males and 30 females) aged 1-15y (average, 7.48±2.86y) with penetrating ocular trauma was performed. Each patient's POTS was calculated. The effects of POTS on final visual acuity (FVA) were examined. Correlation between factors affecting POTS and the FVA was established. RESULTS: All patients presented with single-eye trauma. The follow-up time was 3-21mo (average, 10.23±3.54mo). Among the 90 cases of penetrating wounds, 71 eyes (78.89%) were injured in Zone I (wound involvement limited to the cornea, including the corneoscleral limbus), 17 eyes (18.89%) were injured in Zone II (wound involving the sclera and no more posterior than 5 mm from the corneoscleral limbus), and 2 eyes (2.22%) were injured in Zone III (wound involvement posterior to the anterior 5 mm of the sclera). Analysis of POTS and FVA showed important correlation between them (r=0.414, P=0.000). Initial visual acuity (P=0.00), age (P=0.02), injury location (P=0.002), traumatic cataract (P=0.00), vitreous hemorrhage (P=0.027), retinal detachment (P=0.003), and endophthalmitis (P=0.03) were found to be statistically significant factors for the FVA outcome. CONCLUSION: Ocular trauma presents serious consequences and poor prognosis in children. The POTS may be a reliable prognostic tool in children with open globe injury.

microRNA-218 Inhibits Oxygen-induced Retinal Neovascularization via Reducing the Expression of Roundabout 1.

BACKGROUND: The mechanisms of pathological retinal neovascularization (RNV) remain unknown. Several microRNAs were reported to be involved in the process of RNV. Oxygen-induced retinopathy (OIR) is a useful model to investigate RNV. Our present work explored the expression and the role of microRNA-128 (miR-218) in oxygen-induced RNV. METHODS: OIR was used to establish RNV model. The expression level of miR-218 in the retina from OIR mice was assessed by quantitative real-time reverse transcriptase polymerase chain reaction. Fluorescein angiography was performed in retinae of OIR mice, and RNV was quantified by hematoxylin and eosin staining to evaluate the effect of pCDH-CMV-miR-218 intravitreal injection on RNV in OIR mice. Roundabout 1 (Robo1) expression was detected by Western blotting in mouse retinal vascular endothelial cells expressing a high or low level of miR-218 and retinal tissues from OIR mice. Cell migration was evaluated by scratch wound assay. RESULTS: In OIR mice, the expression level of miR-218 was significantly down-regulated (P = 0.006). Retinal Robo1 expression was significantly increased at both mRNA and protein levels (P = 0.001, 0.008; respectively). miR-218 intravitreal injection inhibited retinal angiogenesis in OIR mice, and the restoration of miR-218 in retina led to down-regulation of Robo1. CONCLUSIONS: Our experiments showed that restoration of miR-218 inhibited retinal angiogenesis via targeting Robo1. MiR-218 contributed to the inhibition of retinal angiogenesis and miR-218 might be a new therapeutic target for preventing RNV.

Small interference RNA targeting vascular endothelial growth factor gene effectively attenuates retinal neovascularization in mice model.

BACKGROUND: The mechanism of retinal neovascularization is not understood completely. Many growth factors are involved in the process of retinal neovascularization, such as vascular endothelial growth factor (VEGF) and pigment epithelium-deprived factor (PEDF), which are the representatives of angiogenic and antiangiogenic molecules respectively. Oxygen induced retinopathy (OIR) is a useful model to investigate retinal neovascularization. The present study was conducted to investigate the feasibility of small interference RNA (siRNA) targeting VEGF gene in attenuating oxygen induced retinopathy (OIR) by regulating VEGF to PEDF ratio (VEGF/PEDF). METHODS: In vitro, cultured EOMA cells were transfected with VEGF-siRNA (psi-HI(TM)/EGFP/VEGF siRNA) and Lipofectamine(TM) 2000 for 24, 48, and 72 hours, respectively. Expression of VEGF mRNA was evaluated by real time polymerase chain reaction (PCR) and the level of VEGF protein was analyzed by Western blotting. In vivo, OIR model mice were established, the mice (C57BL/6J) received an intra-vitreal injection of 1 µl of mixture of psi-HI(TM)/EGFP/VEGF siRNA and Lipofectamine 2000. Expressions of retinal VEGF and PEDF protein were measured by Western blotting, retinal neovascularization was observed by fluorescein angiography, and quantified. RESULTS: In vitro psi-HI(TM)/EGFP/VEGF siRNA treatment significantly reduced VEGF mRNA and protein expression. In vivo, with decreased VEGF and VEGF-PEDF ratio, significant attenuation of neovascular tufts, avascular regions, tortuous, and dilated blood vessels were observed in the interfered animals. CONCLUSIONS: VEGF plays an important role in OIR, and the transfection of VEGF-siRNA can effectively downregulate VEGF expression in vivo, accompanied by the downregulation of VEGF-PEDF ratio, and simultaneous attenuation of retinal neovascularization was also observed. These findings suggest that VEGF/PEDF may serve as a potential target in the treatment of retinal neovascularization and RNA interference targeting VEGF expression, which represents a possible therapeutic strategy.

[Expression of vascular endothelial growth factor and pigment epithelium derived factor in mouse oxygen-induced retinopathy and its significance].

OBJECTIVE: To investigate the expression and significance of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in oxygen-induced mouse retinopathy. METHODS: This experiment was a control experiment study. Newborn C57BL-6 mice were exposed to hyperoxia, and then returned to normoxia to induce retinal neovascularization. Mice were sacrificed at postnatal days 12, 14 and 17 and the retina were processed for RT-PCR, Western Blot and fluorescein angiography. Analysis of variance and Dunnett's t3 was used to compare the mRNA and protein level of VEGF and PEDF between experiment and control groups. Statistical difference was considered significant at a P value less than 0.05. RESULTS: At any of the time points tested, there was significant difference at VEGF protein level (A value) between the experiment (0.47 +/- 0.12, 2.15 +/- 0. 46, 5.49 +/- 0.97) and control (l.81 +/- 0.50, 0.90 +/- 0.05, 0.88 +/- 0.91) groups (P = 0.009, 0.010, 0.000, respectively); the same situation occurred at PEDF protein level (P = 0.002, 0.046, 0.000, respectively). At postnatal day 12, 14 and 17, a significant difference at VEGF protein level was observed among the different experiment groups (P = 0.002, 0.001, 0.000, respectively); the same situation occurred at PEDF protein level (P = 0.009, 0.010, 0.000, respectively). There was significant difference at VEGF mRNA level between the experiment and control groups (P = 0.001, 0.000, 0.001); at postnatal day 12 and 14, the same situation occurred at PEDF protein level (P = 0.001, 0.000, respectively), but there was no significant difference at postnatal day 17 (P = 0.612). At postnatal day 12, 14 and 17, a significant difference at VEGF mRNA level was observed among different experiment groups (P = 0.000, 0.001, 0.000, respectively); the same situation occurred at PEDF mRNA level (P = 0.000, 0.001, 0.000, respectively). The time course of the decrease of PEDF was consistent with the increase of VEGF expression. VEGF/PEDF ratio change was correlated with the development and progression of retinal neovascularization. The time course of PEDF mRNA down-regulation was consistent with the VEGF mRNA up-regulation, similar with the changes of the protein. The change of VEGF and PEDF mRNA was prior to that of the protein. CONCLUSIONS: One of the mechanisms for development of retinal neovascularization is the changes of VEGF\PEDF level in the retina.

[Fluorescein angiography of retinal neovascularization model in mice].

OBJECTIVE: To investigate a high molecular weight fluorescein angiography in the application of retinal neovascularization model in mouse. METHODS: retinal neovascularization model was induced by exposure mouse to an environment containing high concentration of oxygen. High molecular weight fluorescein isothiocyanate dextran were perfused through the left ventricle directly, then the mouse eyes were enucleated and fixed with 4% paraformaldehyde. The retina was separated from the eyecup and flat mounting was performed on a gelatin coated slide. The vasculature was examined under fluorescent microscope. RESULT: The whole retinal vasculature was clearly visualized under fluorescent microscope. By focusing on different layer of the tissue, superficial, deep vascular plexus and connecting vessels also could be distinguished. The neovascular response occurred at the junction between the vascular and the avascular retina. CONCLUSION: High molecular weight fluorescein angiography can be applied for retinal neovascularization evaluation.