Objective To investigate the protective effects of riluzole, a sustained activator of K2P subfamily member TRAAK potassium channel, in human retinal pigment epithelium (hRPE) cells with oxidative induce by tert-butyl hydroperoxide (t-BHP) in vitro, and to evaluate the possible involvement of K2P in the cytoprotective function of retina degeneration diseases. Methods The third to fifth passage of the primary cultured hRPE cells were used in the following experiments．hRPE cells were divided into seven groups: normal control group.t-BHP (300 mu;mol/L) group.t-BHP with riluzole (2, 5, 10, 20 mu;mol/L) group and riluzole (10 mu;mol/L) group. The apoptosis was measured by the 3(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, annexinV/PI double staining flow cytometry. Changes of cells and nuclei morphology were observed under a phase contrast microscope and a fluorescence microscope after 4prime;, 6-diamidino-2-phenylindole (DAPI) staining. Immunofluorescence 1abelling was carried out to analysis the expression of TRAAK. Results After 24 hours incubation with 300 mu;mol/L t-BHP, the cells viability decreased to (58.7plusmn;12.2)% as compared to the normal control groups. The cell viability of t-BHP with riluzole group at different concentrations was higher than the t-BHP group, while 10 mu;mol/L riluzole showed maximally protective effect on hRPE death induced by t-BHP（t=4.84.P＜0.05）. Riluzole remarkably decreased pyknotic nucleus and cell swelling when compared with t-BHP group. Morphology of cells was fusiform with the uniform elliptic nuclei in normal and riluzole group. The Results of annexinV/PI double staining flow cytometry showed that ratio of normal cells were （97.6plusmn;1.3）%, （70.3plusmn;7.0）%, （86.9plusmn;5.2）%, （93.9plusmn;1.5）% in normal group.t-BHP group.t-BHP with riluzole group and riluzole group respectively. The ratio significant decreased in t-BHP group when it was compared with the other groups (t=7.53, 4.59, 6.49, respectively.P＜0.05). By contrast with normal group and riluzole group, the ratio of normal cells in t-BHP with riluzole group had no statistical significance（t=2.94, 1.91, respectively.P＞0.05）. Riluzole (10 mu;mol/L) also significantly decreased the ratio of early stage apoptotic cells from (25.50plusmn;8.02)% to (1.20plusmn;0.72)% in t-BHP injured groups （t=7.13，P＜0.05）. The ratio of early stage apoptotic cells significant decreased in t-BHP group when it was compared with the normal group and riluzole group （t=7.07, 5.94, respectively.P＜0.05）. By comparison with normal group and riluzole group, there are no statistical significance in t-BHP with riluzole group（t=0.06, 1.18, respectively.P＞0.05）. The mean gray values of TRAAK expression were 0.040plusmn;0.003, 0.041plusmn;0.001, 0.049plusmn;0.001, 0.055plusmn;0.001 in normal group.t-BHP group.t-BHP with riluzole group and riluzole group respectively. TRAAK density was significantly higher in t-BHP with riluzole group and riluzole group（t=7.40, 12.70, respectively.P＜0.05）. Conclusions Riluzole can protect hRPE cells against oxidative injury-induced cell death at early apoptosis stage. The mechanism may relate to that riluzole can promote the expression of K2P TRAAK potassium channel.
Objective To evaluate the efficacy and safety of intravitreal anti-vascular endothelial growth factor (VEGF) combined with photodynamic therapy (PDT) vs. photodynamic therapy for polypoidal choroidal vasculopathy (PCV).Methods A computerized search was conducted in Pubmed, OVID, Chinese Biological Medicine Database（CBM）,China National Knowledge Infrastructure （CNKI） by using key words ldquo;polypoidal choroidal vasculopathy, photodynamic therapy, intravitreal anti-VEGFrdquo; in Chinese and/or English combined with manually searching of bibliographies of pertinent articles, journals and literature reference proceedings. Randomized controlled trials (RCT) and non-RCT were collected. The search time was ranged from establishment of each database to September, 2011. The search was no 1imitation in language. The best corrected visual acuity (BCVA),resolution and recurring of lesions, decrease or complete resolution of pigment epithelial detachment （PED）,visual extinction or blindness rate,the rate of subretinal hemorrhage were analyzed by RevMan 5.0 software. Results In total, one RCT and four non-RCTs (273 patients) were included in the meta-analysis involving 148 patients in single treatment group and 125 patients in combined treatment group. The results of metaanalyses showed that there was no significant difference between two groups in the mean logarithm of minimal angle of resolution BCVA at six months ［standard mean difference=0.01, 95% confidence interval (CI): -0.12- 0.14,P=0.84］and 12 months ［standard mean difference = 0.04, 95%CI: -0.16-0.25,P=0.69 after treatment. There was no significant difference between two groups in the resolution of lesions ［odds ratio (OR)=1.38,95%CI:0.74-2.55,P=0.31］ at the months after treatment and decrease or complete resolution of PED (OR=0.67,95%CI:0.12-3.69,P=0.65) at 12 months after treatment. There was no significant difference between two groups in the recurring of lesions (OR=1.14, 95% CI:0.58-2.24,P=0.71) and lost of ge; three lines vision or blindness rate (OR=1.20, 95%CI:0.34-4.18,P=0.78) at 12 months after treatment. The rate of subretinal hemorrhage in combine treatment group was significant lower than single treatment group (OR=0.41, 95%CI:0.18 -0.94,P=0.04). Conclusions The incidence of subretinal hemorrhage occurred in patients with PCV after intravitreal anti-VEGF combined with PDT is much lower than that after single PDT.But the visual improvement, resolution of lesions and recurring of lesions of combined treatment need further studied to see if it is better than single PDT.
Objective To evaluate the application value of intraocular biopsy in the diagnosis of atypical intraocular lesions. Methods The clinical data of 31 patients (31 eyes) with atypical intraocular lesions were retrospectively analyzed. All patients received intraocular biopsy including anterior chamber puncture, vitreous puncture and vitreous biopsy followed by pathological cell examination. Cytological examination was immediately performed for all biopsy fluids or tissues; biopsy times, the positive detecting rate and independent pathological diagnosis rate were analyzed. Intraoperative and postoperative complications were observed. Eyeballs with biopsy-suggested malignancy lesions were enucleated and underwent histopathological analysis. The biopsy results and histopathological results were compared and analyzed.Result Thirty-one eyes received 35 times of biopsy operation in total. The available samples harvested from 29 patients through 31 operations were valid for pathological cell examination，the positive detecting rate was 88.6%. Among the 31 eyes, 12 eyes had malignant lesions; 15 eyes had benign lesions; two eyes were diagnosed with benign lesions initially, but corrected to malignant through the second biopsy;the lesions in two eyes were not determined by biopsy. Among the 29 eyes with valid biopsy, 23 eyes were diagnosed independently by pathological examination; the diagnosis of the other six eyes was made based on pathological examination and clinical features. The independent pathological diagnosis rate was 71.4%. The complications included intraocular bleeding in five eyes, retinal detachment in three eyes and more serous inflammation in one eye. The sensitivity for diagnosis of malignant lesions was 85.7% and the specificity was 100.0%. The predictive value of positive test was 100.0% and the negative one was 86.7%.Conclusion Intraocular biopsy has important values in the diagnosis of atypical intraocular lesions.
Objective To evaluated the efficacy of vitreoretinal surgery for X-linked retinoschisis (XLRS) and its complications. Methods Twentyone XLRS patients (27 eyes) with retinal detachment or vitreous hemorrhage who were treated by vitreoretinal surgery were enrolled in this study. There were microcystislike splitting in all the eyes. The mean visual acuity was 0.11±0.09 and the mean area of macular splitting was (1.09±0.56) mm2. Among the eyes, there were 12 eyes with rhegmatogenous retinal detachment, five eye with traction retinal detachment, six eyes with vitreous hemorrhage and four eyes with retinal detachment and vitreous hemorrhage. All the patients underwent a standard threeport pars plana vitrectomy. Internal limiting membrane peeling, laser photocoagulation, and C3F8 gas or silicone oil tamponade were carried out in different condition. The follow-up was 9-122 months (average 51 months). The preoperative visual acuity and anatomic structure of retina were observed. Results The mean visual acuity at last visit was increased to 0.26±0.15, the difference was significant (t=-6.320, P=0.000). It improved in 20 eyes (74.1%), remained unchanged in seven eyes (25.9%). The retina remained attached in 27 eyes. The mean area of macular splitting was decreased to (0.29±0.21) mm2, the difference was significant (t=10.358, P=0.000). The complications were found in four eyes (14.8%) which including two eyes with proliferative vitroretinopathy and traction retinal detachment six and eight months after surgery, one eye with cataract four months after surgery, and one eye with vitreous hemorrhage 15 months after surgery. The retina remained attached in these four eyes after reoperation. Conclusion Vitreoretinal surgery can significantly improve visual acuity, resume the anatomic structure of retina.
Objective To investigate the correlation between mutation genotypes and phenotypes of X-linked retinoschisis (XLRS) patients. Methods 33 male XLRS patients, 26 female carriers and 100 normal subjects were enrolled in this study. All 33 XLRS patients were bilateral, which included 18 patients from 8 families and 15 sporadic patients. Among 66 XLRS eyes, there are microcystis-like foveal splitting in 49 eyes (74.2%), lamellar macular splitting in 43 eyes (65.2%), peripheral splitting in 32 eyes (48.5%), retinal detachment in 17 eyes (25.8%), and vitreous hemorrhage in 8 eyes (13.6%). Electroretinogram was performed on 42 eyes which showed decreased amplitude of b-wave. The 6 exons of RS1 gene were amplified by polymerase chain reaction and then directly sequenced.The correlation analysis was performed between mutation genotypes and phenotypes. Results There were 19 RS1 gene mutations including 6 novel mutations (p.Gly70Cys, p.Trp112Arg, p.Arg156Trp, p.His207ProfsX56, p.Arg209AlafsX28, p.Cys223Tyr). There was no correlation between mutation genotypes and phenotypes (chi;2=0.731, 3.438, 0.820, 3.208, 1.992; P＞0.05 ).Conclusions RS1 gene mutation is a major cause of XLRS. The RS1 mutation genotype is not correlated with phenotype, so that the prognosis cannot be predicted by the genotypes.