Objective To observe the protective effect of polypyrimidine bundle-binding proteinrelated splicing factor (PSF) over-expression on RPE cell injury induced by advanced glycation end products (AGEs).Methods The human RPE cells cultured in vitro were divided into three groups:normal control group (N group),blank control group (N + AGEs group),empty vector control group (Vec + AGEs group),and PSF high expression group (PSF + AGEs).group).RPE cells in N group were routinely cultured;RPE cells in N + AGEs group were only transfected but did not introduce any exogenous genes combined with AGEs induction;Vec +AGEs group and PSF + AGEs group were transfected with pcDNA The empty vector or pcDNA-PSF eukaryotic expression plasmid was introduced into RPE cells and induced by AGEs.Except the N group,the other 3 groups of cells were transfected accordingly,and were stimulated with 150 μg/ml AGEs for 72 h after 24 h.HE staining and Hoechst 33258 staining were used to observe the effect of high PSF expression on the morphological changes of RPE cells;ROS level detection was used to analyze the effect of PSF high expression on the ROS expression of RPE cells induced by AGEs;MTT colorimetric method was used to detect the high PSF expression Effects on the viability of RPE cells;Western blot was used to detect the effects of different time and dose of PSF on the expression of heme oxygenase 1 (HO-1).Results HE staining and Hoechst 33258 staining observation showed that the cells in group N were full in shape,the nucleus was round,the cytoplasm was rich,and the staining was uniform;the cells in N + AGEs group and Vec + AGEs group were reduced in size,the eosinophilic staining was enhanced,and the nucleus was densely densely stained.Pyrolysis and even fragmentation;the morphology of cells in the PSF + AGEs group was still full,the cytoplasm staining was more uniform,and the nucleus staining was uniform.The results of MTT colorimetry showed that high expression of PSF can effectively improve the viability of RPE cells,but this effect can be effectively antagonized by ZnPP,and the difference is statistically significant (F=33.26,P＜0.05).DCFH-DA test results showed that compared with the N + AGEs group and Vec + AGEs group,the ROS production in PSF + AGEs group decreased,the difference was statistically significant (F=1 1.94,P＜0.05).Western blot analysis showed that PSF protein upregulated HO-1 expression in a time-and dose-dependent manner.The relative expression level of HO-1 at 24,48,and 72 h after PSF protein was significantly higher than that at 0 h,and the difference was statistically significant (F=164.91,P＜0.05).The relative expression level of HO-1 under the action of 0.1,0.5,1.0,1.5,and 2.0 μg PSF protein was significantly higher than 0.0 μg,and the difference was statistically significant (F=104.82,P＜0.05).Conclusion PSF may inhibit the production of ROS by up-regulating the expression of HO-1,thus protecting the RPE cells induced by AGEs.

Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by t test. Results:Compared with the control group, cell viability ( t=12.73, 16.26, P=0.000 2, <0.000 1), cell migration rate ( t=28.17, 37.48, P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant ( t=16.36, 69.35, P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4). Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.

Objective:To evaluate and compare the postoperative visual quality after phacoemulsification combined with toric (ART) and non-toric (ReSTOR) multifocal intraocular lens (IOL) implantation.Methods:A cohort study was conducted.Thirty-nine cataract patients (50 eyes) who underwent phacoemulsification combined with ART IOL implantation were enrolled as ART group, and 32 patients (41 eyes) who received ReSTOR IOL implantation were enrolled as ReSTOR group in Tianjin Medical University Eye Hospital from January 2017 to January 2018.Three months after surgery, the uncorrected distance visual acuity (UDVA), uncorrected intermediate visual acuity (UIVA) and uncorrected near visual acuity (UNVA), manifest refraction, defocus curve, contrast sensitivity (CS), modulation transfer function cutoff, Strehl2D ratio (SR), objective scattering index, OQAS values under 100%, 20%, 9% contrasts (OV 100%, OV 20%, OV 9%), total aberrations, total lower-order aberrations, total higher-order aberrations, spherical aberrations, coma and trefoil aberrations of the two groups were tested and compared.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Tianjin Medical University Eye Hospital (No.2019KY[L]-04). Written informed consent was obtained from each subject prior to entering the cohort.Results:UDVA at 3 months after surgery was (0.07±0.09)LogMAR and (0.09±0.12)LogMAR in ART group and ReSTOR group, and CDVA was (-0.01±0.07)LogMAR and (-0.01±0.07)LogMAR, and UIVA was (0.23±0.11)LogMAR and (0.22±0.13)LogMAR, and UNVA was (0.11±0.15)LogMAR and (0.06±0.11)LogMAR.UNVA was slightly better in ReSTOR group than ART group, and the difference was statistically significant ( t=2.085, P=0.040). The mean depth of focus was (4.12±0.79)D in ART group and (4.24±0.95)D in ReSTOR group.The postoperative residual astigmatism (0.32±0.31)D was significantly lower than preoperative corneal astigmatism (1.27±0.40)D in ART group ( t=13.209, P<0.001). CS values at 6, 12, and 18 c/d under photopic without glare, 12 and 18 c/d under photopic with glare, 3, 6, and 12 c/d under scotopic without glare in ART group were slightly lower than those in ReSTOR group, showing statistically significant differences (all at P<0.05) without clinical significance.SR and OV 20% values in ART group were 0.14±0.05 and 0.55±0.24, which were slightly lower than 0.17±0.06 and 0.66±0.29 in ReSTOR group, with statistically significant differences ( t=-2.012, P=0.048; t=-2.557, P=0.043). Total aberrations and coma aberration under 5 mm pupil diameter in ART group were 0.88(0.59, 1.13)μm and 0.21(0.13, 0.30)μm, which were higher than 0.58(0.47, 0.74)μm and 0.10(0.08, 0.21)μm in ReSTOR group, showing statistically significant differences ( Z=-2.073, P=0.038; Z=-2.101, P=0.036). Conclusions:Cataract phacoemulsification combined with ART IOL implantation can provide good vision and visual quality while correcting preoperative corneal astigmatism, and the resolution in dim light or low to medium spatial frequencies is slightly weaker than eyes implanted with ReSTOR IOL.

Objective To investigate the inhibitory effect oflentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygeninduced retinopathy (OIR).Methods One hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group,simple OIR model group,OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group),with 16,32,32 and 32 mice,respectively.When the mice were 7 days old,the mice in the normal control group were fed in a routine environment,and the mice in the OIR model group,Vec group and PSF group were established OIR model.The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1 × 10~ TU/ml) at the age of 12 days.No injection was performed in the normal control group and simple OIR group.RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina.Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase1 (HO-1).Western blot analysis was applied to detect the protein expression ofNrf2,HO-1 and PSF.Results Of the normal control group,simple OIR model group,Vec group and PSF group,the number of pre-retinal neovascular cell nuclei were 0.00,14.36 ± 5.50,15.67 ± 4.96,8.13 ± 2.09,the non-perfusion area were 0.00％,(35.71 ± 2.81)％,(36.57 ± 4.53)％,(15.33 ± 4.75)％,respectively.The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87,165.70;P＜0.05).Compared with the normal control group,there were more pre-retinal neovascular cell nucleis and larger nonperfusion area in the simple OIR model group and Vec group (P＜0.05).Compared with the simple OIR model group and Vec group,there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P＜0.05).Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2,HO-1 (F=53.66,83.54) and protein expression ofNrf2,HO-1 and PSF (F=58.38,52.69,24.79) among 4 groups were significant (P＜0.05).The rnRNA expression ofNrf2,HO-1 and protein expression of Nrf2,HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P＜0.05).The mRNA expression ofNrf2,HO-1 and protein expression ofNrf2,HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P＜0.05).model group and Vec group (P＜0.05).Conclusion Intravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.