Objective To study the influence of bilateral optic nerve transections on the expression of Nogo A/B and NgR in hypothalamic supraoptic nucleus neurocytes of rats in order to elucidate the possible regulatory mechanisms of supraoptic nucleus regeneration.Methods Twelve adult SD rats were randomly assigned into 2 groups(6 each): control group and optic nerve division group(transection of bilateral optic nerves).Sham operation was done in,rats of the control group without transection of sptic nerves.The rats' brains were removed from both groups 7 days after surgery,and then frozen sections of the brains were made.Different antibodies and fluorescent probe dyes were used to label Nogo-A/B and NgR proteins in the hypothalamic supraoptic nucleus neurocytes.And then laser-confocal microscopy was used to observe these proteins.Results No Nogo-A/B and NgR proteins were expressed in neurocytes of hypothalamic supraoptic nucleus in control group.However,Nogo-A/B and NgR proteins were positively expressed around the nuclei of neurocytes in rats of the optic nerve division group 7 days after bilateral optic nerve transection in the said area Conclusions Nogo-A/B and NgR may participate in the regeneration regulation of supraoptic nucleus neurocytes after injury.However,there must be other factors also involved in the regeneration regulation mechanisms.The existence of these factors might explain the strong activity of regeneration of supraoptic nucleus neurocytes after injury even with the existence of the major axon regeneration inhibitor Nogo-A/B and its receptor NgR in the microenvironment.

Objective　To evaluate the therapeutic effect of phacoemulsification for complicated cataract secondary to uveitis. Metho ds　Twenty-one eyes of 19 patients with complicated cataract secondary to uveitis were emulsified in situ frature technique after blunt separation of iris posterior synechiae, excision of organized membrane with scissors and st retching pupil. Results　The visual acuities with correction on e week after operation was≥ 0.5 in 16 eyes (76.2%), and ≥0.2 in 18 eyes (85.7% ). The major complications were iris damage and corneal edema. No rupture of pos terior capsule and corneal decompensation were found. After operation, a round p upil was obtained in 19 eyes (90.5%) and an irregular pupil in 2 eyes (9.5%).Conclusion　This operation is suitable for complicated cataract se condary to uveitis, and worth to popularize due to the minimization of iris dam age and postoperative inflammation in the anterior chamber, good visual acuity r ehabilitation earlier.

Objective To observe the effect of ciliary neurotrophic factor (CNTF) with different concentrations on the growth and survival of rats' retinal ganglion cells (RGC) in vitro. Methods The retinae of 15 Wistar rats which were 2 or 3 days after birth were dissociated into cell suspension with 0 05% trypsin digestion. After 3 days, cultured RGC were identified with immunohistochemistry method using anti-rat Thy-1.1 monoclonal antibody. Cultured RGC were divided into the 10, 20, 40 ng/ml CNTF group (Ⅰ,Ⅱ, and Ⅲgroup) and the control group respectively. The duration of living RGC was recorded. After 3, 5 and 7 days, the A value of living cells was tested by methylthio-tetrazole colorimetric microassay. Results The result of immunohistochemical examination showed that 90% of living cells cultured for 3 days were RGC. No protuberance or volume increase of RGC were observed in CNTF groups and the control group. The duration of the living RGC was prolonged 3 to 4 days in CNTF groups compared with the control group. The A values of living RGC at the 5th and 7th days in the CNTF groups and the control group were: 0.075 8?0.013 9 and 0.069 3?0.011 3 in I group, 0.0902?0.011 4 and 0.082 5?0.0125 in Ⅱ group, 0.079 2?0.013 3 and 0.065 3?0.008 6 in Ⅲ group, and 0.062 0?0.007 1 and 0.051 3?0.006 8 in the control group, respectively. The differences between the simultaneous CNTF and control group were significant (between Ⅱ group and the control group: P

Objective To evaluate the functional outcomes of the phacoemulsification(Phaco)with posterior continuous curvilinear capsulorhexis(PCCC),or with PCCC and anterior vitrectomy(AV)on after cataract.Design Meta analysis based on the reference searching.Participants A comprehensive electric search in MEDLINE,CNKI,CBMdisc,VIP information,CMCC,CCPD,SSreader,21dmedia and a manual search in related textbooks,journals,congress articles and their references in English and Chinese.Methods Strictly include and exclude the relevant articles from the resource with statistical criterion and get the needing data.Statistically analysed the data with software of Review Manager4.2.2,SPSS10.0.Heterogeneity of the included articles was tested that used to select appropriate effect model to Meta-analysis.Treatment effect was measured as odds ratio between Phaco with PCCC group and Phaco with PCCC+AV group.Change the selected effect model and calculate the fail-safe number(Nfs)to assess the sensitivity and specificity of the research.Main Outcome Measures Sensitivity,heterogeneity,the according confidence intervals(CI)and Nfs.Results Six studies were included,with a sum of 305 eyes.With the outcomes of heterogeneity test,the fixed effect model was used in both groups to calculate OR.The pooled OR of after cataract was 0.13(95%CI is 0.06-0.25).Sensitivity analysis demonstrated that no disproportionate influences of individual study.And Nfs=76.8,the result was confirmed.Conclusion Phacoemulsification with posterior continuous curvilinear capsulorhexis and anterior vitrectomy is superior to phacoemulsification with posterior continuous curvilinear capsulorhexis on preventing after cataract.

Objective To isolate and cultivate the retinal stem cells(RSCs) with improved method,identify the RSCs from embryonic rats,explore the RSCs' features in growth and differentiation,and research for an effective method to cultivate retinal stem cell in vitro from rat ocular tissues.Methods RSCs were isolated from rodents and humans and characterized in vitro.RSCs were isolated from SD rat embryonic retina(gestational age 18d) by shearing,blowing,digesting,centrifuging and filtering,and amplified by using serum-free DMEM/F12 medium containing basic fibroblast growth factor(bFGF),and then the cells were cultured and sub-cultivated by floating growth in vitro.After sub-cultivation,the RSCs were gathered by centrifuging,differentiation was induced with 5% fetal bovine serum(FBS),and the differentiated cells grew and adhered to the culture medium.Immunocytochemistry was employed with specific antibodies to identify RSCs and their potentiality of differentiation.The potentiality of RSCs to replace retinal cells in retinal degenerative diseases was confirmed by transplantation test.Results The primarily cultured retinal cells could grow in a floating state to,form compact cellular spheres,and they had the potential to proliferate in vitro in subsequent cultures to form new cellular spheres.After incubation in the medium with bromodeoxyuridine(BrDU),most of both primarily cultured and sub-cultivated cells expressed the neuroectodermal marker Nestin and BrDU.Differentiated RSCs expressed CD90.1(Thy1.1) after being induced.Conclusion The isolated retinal stem cells from SD embryonic rats,which are undifferentiated,multipotent and may self-renew,have the ability of proliferation and differentiation in vitro,and the differentiated RSCs possess the characteristics of retinal neural cells,including differentiation to retinal ganglion cells.

Objective To observe the corneal nerve fibres damage in different stage of diabetic retinopathy (DR) with type 2 diabetes.Methods A cross-sectional study.One hundred and twenty eyes of 120 patients with type 2 diabetes served as diabetes group.According to International Clinical Diabetic Retinopathy Disease Severity Scales (2002),diabetes patients were classified into 4 subgroups:patients without diabetic retinopathy (NDR),patients with mild or moderate non-proliferative diabetic retinopathy (mNPDR),patients with severe non-proliferative diabetic retinopathy (sNPDR) and patients with proliferative diabetic retinopathy (PDR),each subgroup has 30 eyes of 30 patients.Another 30 eyes of 30 healthy participants served as control group.All eyes were scanned with HRT3 in vivo corneal confocal microscopy.Images of sub-basal nerve plexus were quantified including nerve fiber length (NFL),nerve fiber density (NFD),nerve fiber branch density (NFB),and nerve tortuosity (NT).The correlations of corneal nerve fiber with age,duration of diabetes and glycated hemoglobin (HbA1 c) were analyzed using Spearman correlation analysis.Results NFL,NFD and NFB were found to be significantly lower in diabetic patients (F=147.315,142.586,65.898;P=0.000,0.000,0.000),NT was significantly greater in diabetic patients (F=39.431,P=0.000),when compared to control group.In diabetic patients,NFL,NFD and NFB were gradually reduced with DR severity,NT was gradually increased with DR severity.While the difference ofNFL,NFD,NFB,NT was not statistically significant between sNPDR and PDR subgroups (P＞.0.05),but was statistically significant between other subgroups (P＜0.05).Spearman correlation analysis results showed that age (r=-0.071,-0.080,0.001,0.100;P=0.391,0.328,0.991,0.224) and HbAlc (r=-0.109,-0.115,-0.126,0.025;P=0.238,0.211,0.169,0.781) had no correlation with NFL,NFD,NFB,NT.Duration of diabetes was negatively correlated with the NFL,NFD (r=-0.212,-0.264;P=0.020,0.004),positive correlated with NT (r=0.261,P=0.004),and had no correlation with NFB (r=-0.119,P=0.194).Conclusions Corneal nerve fiber loss and nerve tortuosity increased were found in patients with type 2 diabetes,and even without diabetic retinopathy.The progress of corneal neuropathy was correlated with the severity of DR,but it was not change significantly between sNPDR and PDR.

ObjectiveTo investigate the expression of paired immunoglobulin-like receptor B (PirB) in optic nerve, visual cortex, cerebella, spinal cord and sciatic nerve of normal adult mice.MethodsTwelve healthy adult BALB/c mice were randomly and equally divided into two groups.The immunohistochemistry and Western blot were used to detect the expression of PirB in the tissues described above respectively.ResultsBoth the immunohistochemistry and Western blot test revealed that the expression of PirB was positive in the optic nerve, visual cortex, cerebella and spinal cord, but negative in the sciatic nerve.The positive signals in the sections were located in the cell bodies and the neurites were observed in some of them.Western blot showed the apparent positive band of PirB in the optic nerve, visual cortex, cerebella and spinal cord rather than in the sciatic nerve.The protein expression level of PirB was relatively high in the visual cortex (P ＜0.05) but relatively low in the optic nerves (P ＜0.01).ConclusionThe PirB expresses positively in the optic nerve, indicating that PirB protein may closely correlate with the poor regeneration of the optic nerve.

Objective To explore the clinical therapeutic methods for secondary glaucoma induced by the migration of silicone oil into the anterior chamber.Methods Totally 26 cases of such secondary glaucoma,including 22 aphakic eyes and 4 phakic eyes were subjected.Silicone oil was directly taken out for the patients whose retina was reposited well after silicone oil injection for over 3 months.The aphakic patients underwent 6 o'clock position peripheral iridectomy with laser or operation on prone position,then silicon oil was reinjected,and trabeculectomy was carried out through inferior cornea.The phakic patients received silicone oil aspiration with Healon from the anterior chamber.When silicone oil remigrated into the anterior chamber,the crystal was resected,silicone oil was reinjected,and 6 o'clock position peripheral iridectomy were performed.The patients were followed up for 3 months after the intraocular pressure(IOP) reaching normal range.Results Except one patient gave up treatment,all the patients had stable and normal IOP after 3 months of follow-up.IOP was controlled through different methods in 21 aphakic eyes:5 with prone position,5 with 6'clock position peripheral iridectomy through laser or operation,4 with silicon oil reinjecting,2 with direct aspiration of silicone oil,and 3 with trabeculectomy under cornea.IOP was controled in all of phakic eyes:3 with Healon aspiration of silicone oil,1 with crystal resection and reinjection of silicone oil.Conclusion Secondary glaucoma induced by migration of silicone oil into the anterior chamber can be satisfactorily treated by many ways.

Background Collapsin response mediator protein-2 (CRMP-2) can promote the growth of axons,but CRMP-2 occurs hyperphosphorylation under the induction of cyclin-dependent kinase-5 (CDK5) after central nervous system injury,which leads to the collapse of the growth cone and hinders the repair of nervous system.Being a central nervous system tissue,whether the expressions of CRMP-2 and its phosphorylated protein (p-CRMP-2) change after optic nerve injury are rarely studied.Objective This study was to investigate the dynamic changes of CRMP-2 and p-CRMP-2 expressions in injured optic nerve tissue.Methods Forty-eight 8-or 9-week-old BALB/c mice were randomly divided into the sham operation group and postoperative 3-,7-and 14-day group.Optic nerves were exposed and clamped at retrobulbar 2 mm for 10 seconds in the right eyes during the surgery in the postoperative 3-,7-and 14-day groups,and the same operation was performed except the clamp of optic nerve in the sham operation group.The optic nerve tissue was obtained from the eyes 3,7 and 14 days after surgery.The relative expression levels of CRMP-2 mRNA and CRMP-2,p-CRMP-2 and CDK5 proteins in the tissue were detected by real-time fluorescence quantitative PCR and Western blot,respectively.The use and care of the experimental animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals of Third Military Medical University.Results No significant differences were found in the expression levels of CRMP-2 mRNA and CRMP-2 protein among the sham operation group and postoperative 3-,7-and 14-day groups (CRMP-2 mRNA:F =2.971,P =0.097;C RMP-2 protein:F=1.202,P =0.370).The relative expression levels of p-CRMP-2 protein in the optical nerve were 0.001±0.000,0.064±0.003,0.136±0.005 and 0.346±0.012,and those of CDK5 protein were 0.440±0.009,0.723±0.011,0.874±0.015 and 0.952±0.019 in the sham operation group and postoperative 3-,7-and 14-day groups respectively,showing statistically significant differences among them (p-CRMP-2:F=445.600,P ＜ 0.001;CDK5:F=186.600,P＜0.001),and the relative expression levels of p-CRMP-2 and CDK5 protein were evidently higher in the optical nerve tissue in the postoperative 3-,7-and 14-day groups than those in the sham operation group (all at P＜0.01).Conclusions There are not significant changes in the expression level of CRMP-2 in the BALB/c mice after optic nerve injury.However,the expression levels of p-CRMP-2 and CDK5 proteins are gradually upregulated as the extending of injured time.

Objective To determine the effect of paired immunoglobulin-like receptor B (PirB) on growth of retinal ganglion cells (RGCs) in vitro.Methods Expression of PirB in RGCs isolated from wild-type C57 BJ/6 mice was measured by immunofluorescence.Real-time PCR and Western blot were performed to detect PirB expressions in RGCs cultured for 1,3,5,7 and 9 days.Primary cultured RGCs were left untreated as controls,transfected with lentiviral-delivered PirB RNAi as Group A,lentiviral-delivered NgR RNAi as Group B,lentiviral-delivered PirB RNAi plus ciliary neurotrophic factor (CNTF) as Group C,lentiviral-delivered NgR plus CNTF as Group D,and only CNTF as Group E.Growth and development of RGCs were evaluated by MTT assay and morphological analysis.Results Immunofluorescence confirmed expression of PirB in RGCs,with its mRNA and protein levels changing from increase at first to decrease later with time,up to peak at days 5 and 7.PirB and NgR were respectively down-regulated in Groups A and B.Viability of RGCs was improved in Group A compared with control group [(40.2 ± 5.3) μm vs (29.1 ± 3.8) μm at day 3 ; (72.2 ± 4.2) μm vs (52.3 ± 8.2) μm at day 5,both P ＜ 0.05].By contrast,no significant difference was found in viability of RGCs among Groups B,D and E when compared to control group (P ＞ 0.05).Growth of RGCs between Groups A and C revealed insignificant difference (P ＞ 0.05).Conclusion Knockdown of PirB is beneficial for the growth of RGCs,suggesting a novel method to treat optic nerve injury.