Up-regulation of P-gp via NF-κB activation confers protection against oxidative damage in the retinal pigment epithelium cells.

Dysfunction of retinal pigment epithelial (RPE) cells has been associated with the pathogenesis of age-related macular degeneration in relation to increased oxidative stress, subsequent mitochondrial dysfunction and cell death. Permeability-glycoprotein (P-gp), encoded by the multidrug resistance 1 gene (MDR1), is an active efflux pump involved in cell homeostasis and nuclear factor κB (NF-κB) shows potential involvement in P-gp regulation due to its binding to the promoter domains of MDR1 gene. This study sought to determine the role of P-gp expression regulated by NF-κB in RPE cells during oxidative stress. The human RPE D407 cells were exposed to increasing concentrations of hydrogen peroxide (H2O2) for 24 h. The small-interfering RNA (siRNA) transfection was used to down-regulate P-gp and NF-κB, and the expressions of P-gp and NF-κB p65 were determined by quantitative real-time PCR, western blot and immunofluorescence. The activity of NF-κB was detected by luciferase reporter assay. Mitochondrial membrane potential and cell death rate were detected by flow cytometry. We found that H2O2 exposure caused increasing rate of cell death and induced an elevated expression of P-gp as well as NF-κB activation and nucleus translocation in D407 cells. Inhibiting or silencing NF-κB led to a decrease in the oxidative-induced expression of P-gp. Down-regulation of P-gp by siRNA transfection further impaired the mitochondrial membrane potential and cell death rate in oxidative cells. Moreover, inhibition/knockdown of NF-κB decreased the high rate of cell death caused by H2O2. In conclusion, P-gp can provide moderate cytoprotection for the human RPE cells by ameliorating the mitochondrial dysfunction and NF-κB activation may be a potential regulator of P-gp expression response to oxidative stress.

The treatment and risk factors of retinopathy of prematurity in neonatal intensive care units.

BACKGROUND: Retinopathy of prematurity (ROP) is a vascular proliferative disorder of the developing retina and a significant cause of childhood blindness around the world. The incidence of ROP is affected by many factors, and the incidence rate varies from country to country. The purpose of this study is to report the incidence and risk factors of ROP in neonatal intensive care unit (NICU) of Guangzhou First People's Hospital in China. METHODS: A retrospective review was performed on 436 premature infants who were consecutive ROP screened in the NICU of Guangzhou First People's Hospital from March 2013 to October 2017. The single-factor analysis and the logistic multivariate regression analysis were used to detect risk factors of ROP. RESULTS: Total 436 premature infants were consecutive ROP screened, 138 (31.65%) were found ROP, and 61(13.99%) were treated. The single-factor analysis revealed that the incidence of ROP was associated with multiple births, gestational age, birth weight, mechanical ventilation, intravascular hemolysis, the number of operations and blood culture results. The logistic multivariate regression analysis revealed that gestational age; birth weight, mechanical ventilation, minimum SaO2 and daily weight gain were independent risk factors for ROP onset. Forty-nine patients underwent retinal laser photocoagulation with recurrence 20 patients. Twelve patients underwent anti-VEGF drug (Ranibizumab) via intraocular injection with 5 patients of recurrence. CONCLUSIONS: The incidence of ROP in NICU of Guangzhou China will match those in middle-income countries, but higher than high-income countries. Anti-VEGF drugs could be preferred as a good treatment method for zone 1 ROP and aggressive posterior ROP.

Pre-Transplant Platelet Refractoriness and Alternative Donors Are Associated With Cytomegalovirus Retinitis in Hematopoietic Stem Cell Transplantation for Severe Aplastic Anemia.

Background: Cytomegalovirus retinitis is a severe, vision-threatening opportunistic infection in an immunodeficient population. Reports on cytomegalovirus retinitis in hematopoietic stem cell transplant recipients due to severe aplastic anemia have been scant. This study assessed the risk of cytomegalovirus retinitis in relation to the pre-transplant status of severe aplastic anemia patients. Methods: We conducted a retrospective nested case-control study of cytomegalovirus retinitis among severe aplastic anemia patients receiving allogeneic hematopoietic stem cell transplants in a tertiary care institution that attends severe aplastic anemia patients from southern China from January 1, 2013 to December 31, 2018. Each cytomegalovirus retinitis case was matched with four controls without cytomegalovirus retinitis by age and gender. Thirteen pre-transplant parameters were chosen to compare the risk factor levels between the cases and controls. Multivariable logistic regressions were used to estimate the odds ratios (ORs) and 95% confidence intervals (CIs). Results: A total of 361 severe aplastic anemia patients received hematopoietic stem cell transplants in the study period 2013-2018 in our medical institution, and 31 (8.58%) developed cytomegalovirus retinitis. Cytomegalovirus retinitis was diagnosed in the median of 148 days after transplantation. We confirmed platelet refractoriness more frequently in cases than in controls (p = 0.0005). Compared with human leukocyte antigen-matched sibling donors, alternative donors were significantly more prone to cytomegalovirus retinitis (p = 0.0009). After stepwise selection in multivariate logistic regression, platelet refractoriness (OR 5.41, 95% CI 1.98-15.39), haploidentical donor (OR 7.46, 95% CI 2.19-34.87), and unrelated donor (OR 8.38, 95% CI 2.30-41.34) were associated with an increased risk of cytomegalovirus retinitis. Conclusions: Pre-transplant platelet refractoriness and alternative donors were significant predictors of cytomegalovirus retinitis in severe aplastic anemia recipients. These results highlight the importance of accounting for existing risks while developing prevention strategies and preemptive treatment for severe aplastic anemia recipients. We recommend that the platelet count be closely monitored and thrombopoietin be properly applied during the period when cytomegalovirus retinitis is prone to occur.

Low-Reflectivity Drusen With Overlying RPE Damage Revealed by Spectral-Domain OCT: Hint for the Development of Age-Related Macular Degeneration.

Purpose: To observe the relationship between the characteristic changes in the drusen morphology revealed by the spectral-domain optical coherence tomography (SD-OCT) and the progression of age-related macular degeneration (AMD). Methods: A total of 380 drusen in 45 eyes in 35 patients with the intermediate drusen were longitudinally followed up every 6 months by SD-OCT for a period of 24 months. The drusen were divided into the dynamic group and stable group according to the following parameters: number, volume, concurrent retinal pigment epithelium (RPE)/ellipsoid zone (EZ) damage, and the development of advanced AMD. The morphological characteristics of the progressive or stable drusen were further analyzed. Odds ratios (ORs) and the risk for the drusen progression were calculated. Results: The level of interobserver and intraobserver agreement for each drusen tomographic morphological parameters ranged from 82.7 to 90%. At the end of an average follow-up of 15.92 ± 6.99 months, six patients developed choroidal neovascularization and no patients developed geographic atrophy. Finally, 139 drusen changed and 241 drusen remained stable. The drusen with low reflectivity (p < 0.001; OR: 5.26; 95% CI: 2.24-12.36), non-homogeneity without a core (p < 0.001; OR: 4.31; 95% CI: 2.08-8.92), RPE damage (p < 0.001; OR: 28.12; 95% CI: 9.43-83.85), and the EZ damage (p < 0.001; OR: 14.01; 95% CI: 5.28-37.18) were significantly associated with active change; the drusen with low reflectivity (p = 0.01; OR: 2.95; 95% CI: 1.29-6.75) and decreased overlying RPE reflectivity (p < 0.001; OR: 21.67; 95% CI: 9.20-51.02) were the independent predictors for progression. The drusen with high reflectivity were significantly associated with stabilization (p = 0.03; OR: 0.17; 95% CI: 0.04-0.84). Conclusion: Spectral-domain optical coherence tomography is an optimized, accurate, and efficient method to follow-up the drusen. The intermediate non-exudative AMD prognosis of the patient was most strongly correlated with the drusen reflectivity and disruption of the overlying RPE layer. The drusen with low reflectivity and overlying RPE damage were more likely to progress and required frequent follow-up.

Factors for Visual Acuity Improvement After Anti-VEGF Treatment of Wet Age-Related Macular Degeneration in China: 12 Months Follow up.

Purpose: To evaluate the treatment solutions and effectiveness of intravitreal ranibizumab (RBZ) or conbercept in patients with wet age-related macular degeneration (wAMD) in a real-life setting in China. Methods: The medical records of 368 patients with wAMD who started RBZ or conbercept treatment between 1 May 2014 and 30 April 2018 were evaluated. All patients were defined on fundus angiography at baseline to determine the subtype of AMD (PCV or CNV). We report visual acuity (VA) and central retinal thickness (CRT) measurements at baseline and 12 months. Results: The average number of anti-VEGF injections was 2.1 ± 1.2. The BCVA improvement of these two groups was similar with a difference of 1.00 letter (95% CI: -1.4~3.4, p = 0.8505). At the end of the study, a BCVA increase of at least 5 letters was determined to be a satisfactory efficacy endpoint. Several factors were related to the possible improvement in the satisfactory efficacy endpoint, including female sex (OR 2.07, 95% CI 1.22~3.51), number of injections (OR 1.40, 95% CI 1.12~1.75) and VA change at the first month (OR 13.75, 95% CI 7.41~25.51). Additionally, some factors were related to the possible reduction in the satisfactory efficacy endpoint, including diabetes (OR 0.27, 95% CI 0.10~0.73) and disease history (OR 0.75, 95% CI 0.57~0.98). Conclusion: Our study demonstrates that anti-VEGF drugs can effectively improve BCVA and reduce CRT in AMD patients. Sex, number of injections, VA change at the first month, diabetes and disease history are the most important factors affecting visual acuity.

[Clinical value of preoperative comprehensive evaluation of cataract surgery in age-related cataract].

OBJECTIVE: To evaluate the clinical results, the selection of time and procedures of surgery and preoperative evaluation of the phacoemulsification or small incision cataract surgery with intraocular lens implantation (PSI) in senile cataract patients with cardiovascular disease. METHODS: The authors performed a retrospective study in 218 senior patients (255 eyes) with cardiovascular disease, which were treated by PSI in the past 5 years. RESULTS: Thirteen cases (14 eyes) did not have PSI due to surgical contraindication or impossible to tolerate the operation or impossible to obtain an improvement of visual acuity after operation. Two hundred and five cases (241 eyes) with cardiovascular disease were treated with PSI. After follow up for 1 - 18 months (mean 6.3 months), 165 cases (193 eyes) had a best corrected visual acuity (BCVA) equal or more than 20/60 (80.5%); 32 cases (39 eyes) had BCVA of 20/400 or less than 20/60 (15.6%); and 8 cases (9 eyes) had BCVA less than 20/400 (3.9%). There were no severe complications during the operation. One case developed inhalation pneumonia postoperatively and was transferred to internal medicine for rescue. CONCLUSIONS: Senile cataract patients with cardiovascular disease must be evaluated before performing PSI. The advantages of general hospital should be utilized to treat patients with cardiovascular disease before, during and after the operation. The use of sedatives, adrenalin and steroids must be individualized; high-risk patients should be monitored by EKG and supplied by constant inhalation of low dose oxygen to minimize the surgical risk. In senile cataract patients with cardiovascular disease and IV grade or more lens nucleus, small incision cataract surgery with intraocular lens implantation is better than phacoemulsification with intraocular lens implantation.

[Urea soluble fraction of bovine melanin associated antigen and experimental study on its uveitogenic activity].

OBJECTIVE: The purpose of this study was to investigate the uveitogenic activity of autoantigen in the bovine iris-ciliary body. METHODS: Urea soluble fraction of boveine melanin associated antigen (USF-BMAA) was isolated from the bovine iris-ciliary body biochemically and determined by SDS-PAGE as well as amino acid analysis. Lewis rats and F344 rats were immunized with USF-BMAA emulsified with equal volume complete Freud's adjuvant and Bordetella pertussis. RESULTS: A strongly stained protein band was observed in the USF-BMAA by SDS-PAGE whose molecular weight is approximate 64 000. Amino acid analysis of USF-BMAA showed that it contains 17 kinds of amino acids with high content of Glu, Leu and Asp. The experimental melanin associated antigen-induced uveitis (EMIU) was successfully incited in both eyes of the Lewis and F344 rats. The inflammation was mainly located in the anterior uvea, and spontaneously recovered. Mild focal choroiditis was present in the rats with severe lesion. However, the inflammation was not observed in the retina and pineal gland. CONCLUSIONS: USF-BMAA may be the major part of autoantigen of the uveal tract with uveitogenic activity. Unlike experimental autoimmune uveoretinitis (EAU) which is incited by the retinal soluble antigen (S-Ag) in the rats, no involvement of the retina and pineal gland is found in EMIU.

Atomic force microscopy analysis of progenitor corneal epithelial cells fractionated by a rapid centrifugation isolation technique.

PURPOSE: To investigate the use of atomic force microscopy (AFM) to image the three groups of corneal epithelial cells fractionated by a novel rapid centrifugation isolation technique. METHODS: Epithelial cells harvested from primary cultures of rabbit limbal rings were centrifuged onto uncoated dishes, first at 1400 rpm and then at 1800 rpm. The adherent cells after centrifugation at 1400 rpm (ATC1), the adherent cells at 1800 rpm (ATC2) and the non-adherent cells at 1800 rpm (NAC) were investigated for BrdU retention and were subjected to contact mode AFM and Transmission Electron Microscopy (TEM). RESULTS: Compared with unfractionated cells, the ATC1 group, accounting for about 10% of the whole population, was enriched in BrdU label-retaining cells. There were dramatic overall shape, surface membrane and intra-cellular ultrastructure differences noted among ATC1, ATC2 and NAC populations. The whole cell roughness measurements were 21.1±1.5 nm, 79.5±3.4 nm and 103±4.6 nm for the ATC1, ATC2 and NAC groups, respectively. The mero-nucleus roughness measurements were 34.2±1.7 nm, 13.0±0.8 nm and 8.5±0.5 nm in the ATC1, ATC2 and NAC populations, respectively. CONCLUSIONS: AFM was found to be a good tool for distinguishing among the three groups of cells. BrdU label retention, the AFM parameters and TEM together suggest that the ATC1, ATC2 and NAC populations may be progenitor corneal epithelial cells, transit amplifying cells and terminal differentiation cells, respectively.