Reduction of ETV1 is Identified as a Prominent Feature of Age-Related Cataract.

PURPOSE: To identify the inactive genes in cataract lenses and explore their function in lens epithelial cells (LECs). METHODS: Lens epithelium samples obtained from both age-related cataract (ARC) patients and normal donors were subjected to two forms of histone H3 immunoprecipitation: H3K9ac and H3K27me3 chromatin immunoprecipitation (ChIP), followed by ChIP-seq. The intersection set of "active genes in normal controls" and "repressed genes in cataract lenses" was identified. To validate the role of a specific gene, ETV1, within this set, quantitative polymerase chain reaction (qPCR), western blot, and immunofluorescence were performed using clinical lens epithelium samples. Small interference RNA (siRNA) was utilized to reduce the mRNA level of ETV1 in cultured LECs. Following this, transwell assay and western blot was performed to examine the migration ability of the cells. Furthermore, RNA-seq analysis was conducted on both cell samples with ETV1 knockdown and control cells. Additionally, the expression level of ETV1 in LECs was examined using qPCR under H2O2 treatment. RESULTS: Six genes were identified in the intersection set of "active genes in normal controls" and "repressed genes in ARC lenses". Among these genes, ETV1 showed the most significant fold-change decrease in the cataract samples compared to the control samples. After ETV1 knockdown by siRNA in cultured LECs, reduced cell migration was observed, along with a decrease in the expression of ß-Catenin and Vimentin, two specific genes associated with cell migration. In addition, under the oxidative stress induced by H2O2 treatment, the expression level of ETV1 in LECs significantly decreased. CONCLUSIONS: Based on the findings of this study, it can be concluded that ETV1 is significantly reduced in human ARC lenses. The repression of ETV1 in ARC lenses appears to contribute to the disrupted differentiation of lens epithelium, which is likely caused by the inhibition of both cell differentiation and migration processes.

Comparative study of FLACS vs conventional phacoemulsification for complex cataracts in vitrectomized eyes.

PURPOSE: To compare the visual outcomes and complications of femtosecond laser-assisted cataract surgery (FLACS) and conventional phacoemulsification (CPS) in patients with prior pars plana vitrectomy (PPV). SETTING: Department of Ophthalmology, Eye and Ears, Nose, and Throat Hospital of Fudan University, Shanghai, China. DESIGN: Prospective randomized controlled cohort study. METHOD: According to the Lens Opacity Classification System III, cataract patients with nucleus grade 3 to 6 after prior PPV were randomized to 2 groups (FLACS or CPS). Endothelial cell density (ECD), central corneal thickness (CCT), and corrected distance visual acuity (CDVA) were evaluated preoperatively and at 1 day, 3 days, 1 week, 2 weeks, 1 month, and 3 months postoperatively. Intraoperative parameters and intraoperative and postoperative complications were recorded. RESULTS: 86 eyes (45 and 41 in the FLACS and CPS groups, respectively) were included and analyzed, among which 43 had hard nuclei (grade 5 or 6). Cumulative dissipated energy was lower and ultrasound time was shorter among FLACS patients with hard nuclei ( P < .001). CDVA was similar in both groups at the final visit ( P > .05). ECD decreased in both groups but was lower among CPS patients with hard nuclei throughout the follow-up period ( P < .05). CCT had returned to preoperative levels at the final visit in both groups; however, eyes with hard cataract in the FLACS group recovered sooner. No serious complications occurred in either group. CONCLUSIONS: FLACS is beneficial in improving surgical safety and providing quicker visual rehabilitation in vitrectomized eyes with hard nuclear cataracts.

An Optical Coherence Tomography-Based Deep Learning Algorithm for Visual Acuity Prediction of Highly Myopic Eyes After Cataract Surgery.

BACKGROUND: Due to complicated and variable fundus status of highly myopic eyes, their visual benefit from cataract surgery remains hard to be determined preoperatively. We therefore aimed to develop an optical coherence tomography (OCT)-based deep learning algorithms to predict the postoperative visual acuity of highly myopic eyes after cataract surgery. MATERIALS AND METHODS: The internal dataset consisted of 1,415 highly myopic eyes having cataract surgeries in our hospital. Another external dataset consisted of 161 highly myopic eyes from Heping Eye Hospital. Preoperative macular OCT images were set as the only feature. The best corrected visual acuity (BCVA) at 4 weeks after surgery was set as the ground truth. Five different deep learning algorithms, namely ResNet-18, ResNet-34, ResNet-50, ResNet-101, and Inception-v3, were used to develop the model aiming at predicting the postoperative BCVA, and an ensemble learning was further developed. The model was further evaluated in the internal and external test datasets. RESULTS: The ensemble learning showed the lowest mean absolute error (MAE) of 0.1566 logMAR and the lowest root mean square error (RMSE) of 0.2433 logMAR in the validation dataset. Promising outcomes in the internal and external test datasets were revealed with MAEs of 0.1524 and 0.1602 logMAR and RMSEs of 0.2612 and 0.2020 logMAR, respectively. Considerable sensitivity and precision were achieved in the BCVA < 0.30 logMAR group, with 90.32 and 75.34% in the internal test dataset and 81.75 and 89.60% in the external test dataset, respectively. The percentages of the prediction errors within ± 0.30 logMAR were 89.01% in the internal and 88.82% in the external test dataset. CONCLUSION: Promising prediction outcomes of postoperative BCVA were achieved by the novel OCT-trained deep learning model, which will be helpful for the surgical planning of highly myopic cataract patients.

Risk factors of intraocular lens dislocation following routine cataract surgery: a case-control study.

Clinical relevance: Intraocular lens dislocation usually causes severe visual symptoms and even blindness in patients. It is important for eye care practitioners to be able to identify patients susceptible to intraocular lens dislocation.Background: To investigate the risk factors of intraocular lens dislocation following routine cataract surgery.Methods: This retrospective case-control study included patients diagnosed with intraocular lens dislocation between January 2007 and March 2018. Controls matched to the cases by age and time of routine cataract surgery in a ratio of 3:1 were randomly selected from patients without intraocular lens dislocation. The medical records of the patients were reviewed and analysed. Independent t-test, chi-square test, and logistic regression analysis were employed to identify risk factors of intraocular lens dislocation.Results: In total, 127 cases (with intraocular lens dislocation) and 381 controls (without intraocular lens dislocation) were evaluated. Among cases, the proportion of men was significantly higher than that of women (p = 0.012). High myopia was significantly associated with all types of intraocular lens dislocation; it was also a risk factor for in-the-bag intraocular lens dislocation. Lens subluxation was significantly associated with the in-the-bag and late intraocular lens dislocations. Intraoperative and post-operative vitrectomy, rather than pre-operative vitrectomy, were associated with intraocular lens dislocation. Post-operative trauma was significantly associated with in-the-bag intraocular lens dislocation; pre-operative and post-operative trauma were significantly associated with out-of-the-bag intraocular lens dislocation. Male sex (odds ratio (OR) = 2.996, p = 0.006), pre-operative trauma (OR = 7.861, p = 0.014), high myopia (OR = 11.268, p < 0.001), and lens subluxation (OR = 13.431, p = 0.018) were the risk factors of intraocular lens dislocation.Conclusion: Male sex, pre-operative trauma, high myopia, and lens subluxation were the main predisposing factors for intraocular lens dislocation in Chinese Han population.

Toric Intraocular Lens Implantation in the Correction of Moderate-To-High Corneal Astigmatism in Cataract Patients: Clinical Efficacy and Safety.

METHODS: A total of 57 cataract patients (57 eyes) with regular corneal astigmatism (≥2.57 D) were enrolled in this retrospective cohort study. Phacoemulsification with toric IOL implantation was performed for all patients. The uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA) were recorded before and one year after surgery, and statistical analysis of preoperative corneal astigmatism, postoperative residual astigmatism, aberrations, IOL rotation, and related factors was performed to evaluate the efficacy, safety, and stability of toric IOLs in correcting moderate-to-high corneal astigmatism. RESULTS: One year after surgery, visual acuity was significantly improved compared with that before surgery (preoperative log MAR 0.87 ± 0.34 vs. postoperative log MAR 0.31 ± 0.26, p < 0.001), and the self-reported spectacle independence rate was 68.42%. The total residual astigmatism was 1.18 ± 0.85 D, which was significantly less than the preoperative value (3.41 ± 0.99 D) (p < 0.001). The degree of toric IOL rotation was 4.93 ± 3.02°, and 54.39% of patients had a lens rotation of less than 5°. The IOLs of 5.26% (3 eyes) of patients rotated more than 10°, and these patients received glasses instead of undergoing IOL repositioning. CONCLUSIONS: Toric IOL implantation provided optimal vision outcomes and low spectacle dependence during a one-year follow-up period. The results from our study show that toric IOL implantation is a safe and effective option for cataract patients with moderate-to-high corneal astigmatism.

Analysis of corneal astigmatism and aberration in chinese congenital cataract and developmental cataract patients before cataract surgery.

BACKGROUND: To investigate the distribution of corneal astigmatism, aberration, and biometric parameters in Chinese congenital cataract (CC) /developmental cataract patients before cataract surgery. METHODS: We evaluated eyes of CC /developmental cataract patients scheduled for cataract surgery from January 2016 to September 2019. Astigmatism, aberrations, and biometric parameters were measured with the Pentacam Scheimpflug imaging device (Pentacam HR, Oculus). Cataract was diagnosed and classified by slit-lamp examination after full mydriasis. RESULTS: We evaluated 538 eyes in 356 patients. The mean values of anterior corneal astigmatism (ACA), posterior corneal astigmatism (PCA), and total corneal astigmatism (TCA) were determined as 1.98 ± 1.06 D (range 0.0‒4.8 D), 0.49 ± 0.26 D (range 0.0‒1.9 D), and 2.09 ± 1.19 D (range 0.2‒8.8 D), respectively. ACA and TCA ≥ 1.25 D was present in 379 eyes (70.3%) and 392 eyes (72.8%), respectively. PCA between 0.25 D and 0.75 D was found in 380 eyes (70.6%). There was a statistically significant positive linear correlation between lower-order aberrations root mean square values (LOA RMS) and corneal astigmatism (CA). Furthermore, in terms of distribution of central cornea thickness, anterior chamber depth, ACA, PCA, and TCA in different types of cataracts, ACA was highest in patients with zonular cataracts. Finally, we found anterior corneal measurements may overestimate WTR astigmatism, underestimate ATR astigmatism, and underestimate oblique astigmatism, respectively. CONCLUSIONS: Most CC /developmental cataract patients had moderate to high astigmatism and ACA accounted for the largest proportion in the zonular group. This can provide a basis for planning of CC/developmental cataract surgery by ophthalmologists in clinical practice.

Three-Year Outcomes of Cionni-Modified Capsular Tension Ring Implantation in Children Under 8 Years Old With Ectopia Lentis.

PURPOSE: This study evaluated visual outcomes and complications at 3 years post-implantation of a Cionni-modified capsular tension ring (MCTR) with an intraocular lens (IOL) in ectopia lentis patients ≤8 years old. DESIGN: Prospective clinical cohort study. METHODS: Included were 101 eyes from 57 patients <8 years of age, who underwent surgery for nontraumatic ectopia lentis between November 2015 and December 2016. Exclusion criteria were planned IOL fixation in the ciliary sulcus, severe intraoperative complications, and incomplete follow-up. All eyes received in-the-bag implantation of a posterior IOL and Cionni-MCTR. Posterior capsulectomy and anterior vitrectomy were performed through the pars plana in 23 eyes of children <5 years of age. Patients were examined at 1 day, and 1, 6, and 12 months, and at 2 and 3 years postoperatively. Outcome; measurements included best-corrected visual acuity (BCVA), intraocular pressure (IOP), IOL centration, and posterior capsule opacification (PCO). RESULTS: In all eyes, BCVA improved significantly after surgery, especially during the first 12 months (P < .05). Three years post-operatively, 44 eyes had BCVA 0.9 or better. Prophylactic Nd:YAG laser capsulotomy was performed 3 months post-surgery in 24 eyes; 34 eyes underwent this; procedure 6 months post-surgery because of PCO. A second surgery was; warranted in 4 eyes because of severe IOL decentration and combined anterior capsule contraction. No severe postoperative complications, such as retinal detachment or endophthalmitis, occurred. CONCLUSIONS: Implantation of in-the-bag IOL with Cionni MCTR is effective for visual rehabilitation in young children with ectopia lentis. A close follow-up of these patients is necessary to monitor IOL centration and stability.

Clinical Characteristics of Intraocular Lens Dislocation in Chinese Han Populations.

PURPOSE: To investigate the clinical characteristics of patients with intraocular lens (IOL) dislocation after IOL implantation in Chinese Han populations. METHODS: The medical records of patients with IOL dislocation were retrospectively reviewed from January 2007 to December 2017, and a total of 312 patients (male: 231, female: 97) (328 eyes) were included in this study. The axial length (AL), IOL power, and the time interval between cataract surgery and IOL dislocation as well as the ocular conditions associated with IOL dislocation were recorded. The IOL dislocation was classified and graded based on its relationship with the capsule and the position of the dislocated IOL. RESULTS: The mean time between original cataract surgery and IOL dislocation was 5.63 ± 5.13 years; IOL dislocation occurred in up to 56.1% (184 eyes) of the eyes within 5 years. Trauma was found in 136 eyes (41.5%); pars plana vitrectomies were performed in 61 eyes (18.6%), and high myopia was detected in 108 eyes (32.9%). A total of 243 eyes (74.1%) had out-of-the-bag IOL dislocations, while 85 eyes (25.9%) had in-the-bag IOL dislocations. There was a statistically significant difference in the constituent ratio of trauma between in-the-bag dislocation and out-of-the-bag dislocation (Pearson's chi2 = 33.3992, P < 0.001); ocular blunt traumas were significantly higher in in-the-bag dislocations, while open-globe injuries were significantly higher in out-of-the-bag dislocations. A statistically significant difference was found for the ratio of patients with AL longer than 30 mm between in-the-bag dislocation and out-of-the-bag dislocation (Pearson's chi2 = 9.7355, P < 0.002). CONCLUSIONS: In Chinese Han populations, the most common IOL dislocation is out-of-the-bag dislocation; the most common risk factors were trauma, long axial length, and eyes undergoing pars plana vitrectomy; a minimum follow-up of 5 years is suggested for IOL dislocation-predisposed eyes undergoing cataract surgery.

Influences of the three-dimensional parameters of pterygium on corneal astigmatism and the intraocular lens power calculation.

Pterygium morphology had great effect on corneal astigmatism and intraocular lens (IOL) power calculation in cataract patients. However, previous studies all focused on the pterygium surface parameters, the invasion degree or cross-sectional area of the pterygia into the corneal stroma were neglected. We studied the effect of three-dimensional parameters of pterygium on corneal astigmatism and IOL power prediction. We enrolled 81 eyes of 81 patients with primary nasal pterygium, measured the corneal astigmatism (Pentacam HR) and predicted IOL power change (IOLmaster500) before and after pterygium surgery. The three-dimensional parameters of pterygium (length, width, area, height and invasion cross-sectional area) were measured by slit lamp photography and Scheimpflug images. After pterygium surgery, corneal astigmatism decreased from 4.35 ± 4.24 to 1.07 ± 0.95 D and total corneal refractive power increased from 43.02 ± 1.96 to 43.95 ± 0.95 D (both P < 0.001). The predicted IOL power decreased from 22.87 ± 2.82 to 21.71 ± 2.85 D (P < 0.001) after surgery. Notably, 34 eyes (41.98%) had ≥3.0 D of pterygium induced astigmatism (PIA), and 33 eyes (40.74%) had ≥1.0 D of predicted IOL power change. PIA was independently influenced by the pterygium surface area (r = 0.43, P < 0.001) and cross-sectional area (r = 1.25, P = 0.018), while the predicted IOL power change was independently affected by the pterygium width (r = 0.70, P < 0.001). Cataract surgeons could evaluate the effects of a pterygium according to its three-dimensional parameters and prepare an optimal surgical strategy for cataract combined pterygium patients.

Evaluation of the antioxidant effects of different histone deacetylase inhibitors (HDACis) on human lens epithelial cells (HLECs) after UVB exposure.

BACKGROUND: To compare the protective effects of the histone deacetylase inhibitors (HDACis) ß-hydroxybutyrate (ßOHB), trichostatin A (TSA), suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA) on human lens epithelial cells(HLECs) following ultraviolet-B (UVB) exposure. METHODS: HLECs were divided into subgroups: four HDACi groups, a control group, a UVB-treated group and a DMSO group (cells treated with DMSO and UVB irradiation). In the HDACi groups, HLECs were cultured with different concentrations of HDACis 12 h prior to UVB irradiation. The protective effects of the HDACis were evaluated by assessing apoptosis rates, cell activity and expression levels of genes associated with apotosis (caspase-3, Bcl-2, BAX, SOD1, FOXO3A and MT2). The levels of superoxide dismutase (SOD), reactive oxygen species (ROS), malondialdehyde (MDA) and total antioxidant capacity (T-AOC) were detected in order to evaluate oxidative stress. RESULTS: The results showed that SAHA (1 µmol/L, 2 µmol/L) and TSA (0.2 µmol/L) had mild protective effects on cell viability. ßOHB (4 mmol/L) and TSA (0.2 mol/L) demonstrated protective effects on BCL-2 expression. TSA (0.2 mol/L) showed protective effects on SOD1 expression. TSA (0.2 mol/L) and SAHA (1 µmol/L) suppressed BAX and caspase-3 expression. TSA (0.2 mol/L, 0.8 mol/L) and SAHA (1 µmol/L, 2 µmol/L) suppressed the expression of FOXO3A and MT2. SOD levels were increased after treatment with ßOHB (4 mmol/L), SAHA (8 µmol/L) and TSA (0.1 mol/L, 0.2 mol/L). T-AOC levels were increased in UVB-treated HLECs after treatment with SAHA (2 µmol/L). MDA levels decreased in UVB-treated HLECs following treatment with TSA (0.2 mol/L, 0.8 mol/L). ROS levels decreased in UVB-treated HLECs following treatment with ßOHB (4 mmol/L), SAHA (1 µmol/L, 2 µmol/L) and TSA (0.2 mol/L). Western blotting results demonstrated that SOD1 levels significantly increased in the ßOHB (4 mmol/L), SAHA (1 µmol/L, 2 µmol/L), TSA (0.1 mol/L, 0.2 mol/L) and VPA (5 mmol/L) groups. Only SAHA (1 µmol/L) had an anti-apoptotic effect on UVB-treated HLECs. CONCLUSIONS: Our findings indicate that low concentrations of HDACis (1 µmol/L of SAHA) mildly inhibit oxidative stress, thus protecting HLECs from oxidation. These results may suggest that there is a possibility to explore the clinical applications of HDACis for treatment and prevention of cataracts.

Uveitis-glaucoma-hyphema syndrome associated with an in-the-bag square-edge intraocular lens.

A 54-year-old woman presented with recurrent redness and blurred vision of the left eye with elevated intraocular pressure (IOP) for one year. She was treated as "iridocyclitis" and ``Posner-Schlossman syndrome'' at the local hospitals. However, the patient developed intermittent ocular inflammation and hyphema. Patient had a cataract surgery and intraocular lens (IOL) implantation in the left eye one year before at the local hospital. A diagnostic procedure was performed and the possible pathogenesis was discussed.

Comparison of long-term decentration and tilt in two types of multifocal intraocular lenses with OPD-Scan III aberrometer.

PURPOSE: To compare the long-term decentration and tilt in two multifocal intraocular lenses (MfIOLs) with an OPD-Scan III aberrometer. METHODS: Eighty cataract patients who underwent uneventful MfIOL implantation (42 with AcrySof® IQ ReSTOR® SN6AD1 and 38 with AMO Tecnis® ZMB00) were enrolled. At 1 year after surgery, a postoperative visual acuity evaluation included the measurement of uncorrected distance visual acuity, corrected distance visual acuity, distance-corrected near visual acuity, and distance-corrected intermediate visual acuity. OPD-Scan III aberrometer was used to collect the decentration, tilt, and high-order aberration (HOA) data. Significance was tested with Student's t test, Mann-Whitney U test, paired t test, and χ2 test RESULTS: The average intraocular tilt was lower in the SN6AD1 group than in the ZMB00 group, whereas the average decentration of the two groups did not differ significantly. The mean total ocular HOAs, ocular trefoil, total internal HOAs, and spherical aberrations were significantly lower in the SN6AD1 group than in the ZMB00 group. In both the SN6AD1 and ZMB00 groups, the intraocular tilt was directly proportional to the total ocular HOAs, coma, and spherical aberration. However, there was no significant correlation between decentration and any type of HOA in SN6AD1 group, whereas decentration correlated positively with total ocular HOAs, coma, and spherical aberration in ZMB00 group. CONCLUSION: Both MfIOLs significantly improve the visual acuity of the patients. However, tilt was lower in the SN6AD1 group than in the ZMB00 group 1 year after surgery, which provide fewer HOAs and better visual quality.

Structural basis for neutralization of Japanese encephalitis virus by two potent therapeutic antibodies.

Japanese encephalitis virus (JEV), closely related to dengue, Zika, yellow fever and West Nile viruses, remains neglected and not well characterized 1 . JEV is the leading causative agent of encephalitis, and is responsible for thousands of deaths each year in Asia. Humoral immunity is essential for protecting against flavivirus infections and passive immunization has been demonstrated to be effective in curing disease2,3. Here, we demonstrate that JEV-specific monoclonal antibodies, 2F2 and 2H4, block attachment of the virus to its receptor and also prevent fusion of the virus. Neutralization of JEV by these antibodies is exceptionally potent and confers clear therapeutic benefit in mouse models. A single 20 µg dose of these antibodies resulted in 100% survival and complete clearance of JEV from the brains of mice. The 4.7 Å and 4.6 Å resolution cryo-electron microscopy structures of JEV-2F2-Fab and JEV-2H4-Fab complexes, together with the crystal structure of 2H4 Fab and our recent near-atomic structure of JEV 4 , unveil the nature and location of epitopes targeted by the antibodies. Both 2F2 and 2H4 Fabs bind quaternary epitopes that span across three adjacent envelope proteins. Our results provide a structural and molecular basis for the application of 2F2 and 2H4 to treat JEV infection.

Potential of CeCl3@mSiO2 nanoparticles in alleviating diabetic cataract development and progression.

Cataract is a major cause of visual impairment for diabetic patients. It is imperative to develop efficient therapeutic agents against diabetic cataract (DC) because diabetes confers higher risk for complications after cataract surgery. We have previously reported the role of CeCl3 loaded mesoporous silica (CeCl3@mSiO2) nanoparticles in reducing the oxidative stress of lens epithelial cells. However, the potential of CeCl3@mSiO2 in preventing diabetic cataract development remains unclear. In this study, we applied CeCl3@mSiO2 nanoparticles with a size of 87.6±8.9nm to streptozotocin-induced diabetic cataract rat model by intraperitoneal injection. Our results showed that CeCl3@mSiO2 efficiently ameliorated the progression of DC. Consistent with antioxidant effect of CeCl3@mSiO2in vitro, administration of CeCl3@mSiO2 significantly abrogated hyperglycemia-mediated upregulation of advanced glycation end products, lipid peroxidation and protein carbonylation in animal lens. Taken together, our study provides a potential nanodrug to manage the development of DC.

miR-1307 promotes the proliferation of prostate cancer by targeting FOXO3A.

microRNAs have emerged as important regulators in various cancers, including prostate cancer. In this study, we investigated the role of miR-1307 in cell proliferation of prostate cancer. We found miR-1307 was overexpressed in prostate cancer cells and tissues, overexpression of miR-1307 significantly promoted cell proliferation and tumorigenesis in vitro investigated by MTT assay, colony formation assay and soft agar growth assay, meanwhile overexpression of miR-1307 inhibited cell cycle inhibitors p21 and p27 both in mRNA and protein levels. Knockdown of miR-1307 reduced these effects, confirming miR-1307 promotes prostate cancer cell proliferation. FOXO3A (Forkhead box protein O3a) was the target of miR-1307, miR-1307 directly bound to the 3'UTR of FOXO3A. Simultaneous knockdown of miR-1307 and FOXO3A promoted cell proliferation of prostate cancer. In summary, our results suggested miR-1307 contributed to prostate cancer proliferation by targeting FOXO3A.

Distribution of gene mutations in sporadic congenital cataract in a Han Chinese population.

PURPOSE: This study aimed to investigate the genetic effects underlying non-familial sporadic congenital cataract (SCC). METHODS: We collected DNA samples from 74 patients with SCC and 20 patients with traumatic cataract (TC) in an age-matched group and performed genomic sequencing of 61 lens-related genes with target region capture and next-generation sequencing (NGS). The suspected SCC variants were validated with MassARRAY and Sanger sequencing. DNA samples from 103 healthy subjects were used as additional controls in the confirmation examination. RESULTS: By filtering against common variants in public databases and those associated with TC cases, we identified 23 SCC-specific variants in 17 genes from 19 patients, which were predicted to be functional. These mutations were further confirmed by examination of the 103 healthy controls. Among the mutated genes, CRYBB3 had the highest mutation frequency with mutations detected four times in four patients, followed by EPHA2, NHS, and WDR36, the mutation of which were detected two times in two patients. We observed that the four patients with CRYBB3 mutations had three different cataract phenotypes. CONCLUSIONS: From this study, we concluded the clinical and genetic heterogeneity of SCC. This is the first study to report broad spectrum genotyping for patients with SCC.

Generation of Human Lens Epithelial-Like Cells From Patient-Specific Induced Pluripotent Stem Cells.

Cataractogenesis begins from the dynamic lens epithelial cells (LECs) and adjacent fiber cells. LECs derived from cell lines cannot maintain the crystalline expression as the primary LECs. The current study aimed to efficiently generate large numbers of human LECs from patient-specific induced pluripotent stem cells (iPSCs). Anterior lens capsules were collected from cataract surgery and were used to culture primary hLECs. iPSCs were induced from these primary hLECs by lentiviral transduction of Oct4, Sox2, Klf4, and c-Myc. Then, the generated iPSCs were re-differentiated into hLECs by the 3-step addition of defined factor combinations (Noggin, BMP4/7, bFGF, and EGF) modified from an established method. During the re-differentiation process, colonies of interest were isolated using a glass picking tool and cloning cylinders based on the colony morphology. After two steps of isolation, populations of LEC-like cells (LLCs) were generated and identified by the expression of lens marker genes by qPCR, western blot and immunofluorescence staining. The study introduced a modified protocol to isolate LLCs from iPSCs by defined factors in a short time frame. This technique could be useful for mechanistic studies of lens-related diseases. J. Cell. Physiol. 231: 2555-2562, 2016. © 2016 Wiley Periodicals, Inc.

The efficacy and complications of black diaphragm intra-ocular lens implantation in patients with congenital aniridia.

PURPOSE: To evaluate the efficacy and complications of black diaphragm intra-ocular (BDI) lens implantation in patients with congenital aniridia. METHODS: Twenty patients underwent BDI lens implantation for the treatment of congenital aniridia from January 1999 to December 2012. Fifteen patients (23 eyes) were enrolled in our study, and the mean follow-up period was 26 months. Patient demographics, clinical evaluations [visual acuity (VA), best-corrected visual acuity (BCVA), intra-ocular pressure measurement (IOP), corneal endothelial cell density (ECD) and ultrasound biomicroscopy (UBM)], complications and treatments were analysed for each patient. RESULTS: Sixteen eyes (70.1%) had a BCVA better than 20/200 after BDI lens implantation, and photophobia obviously decreased in all patients. Six eyes (26.09%) developed secondary glaucoma after BDI lens implantation, and one eye underwent glaucoma surgery. Corneal decompensation occurred in two eyes (8.70%), one of which was complicated by glaucoma. Two eyes (8.70%) developed visual axis opacity (VAO) after surgery. One patient (4.35%) had limbal stem cell failure, and another patient (4.35%) had an eccentric BDI lens. Intra-ocular pressure measurement elevation and ECD reduction were found after BDI lens implantation. CONCLUSIONS: Black diaphragm intra-ocular lens implantation can effectively improve VA, decrease photophobia and resolve cosmetic issues in most congenital aniridia eyes. Glaucoma, corneal decompensation and VAO were the major long-term complications of BDI lens implantation in patients with congenital aniridia. All patients should be managed attentively because of high risk of complications and followed long term to achieve favourable outcomes.

Long-term efficacy and complications of black diaphragm intraocular lens implantation in patients with traumatic aniridia.

AIMS: To determine the efficacy and complications of long-term black diaphragm intraocular (BDI) lens implantation in patients with traumatic aniridia. METHODS: This study analysed the demographics, clinical evaluations and treatments of 95 patients with BDI lens implantation and 75 patients who wore rigid gas-permeable (RGP) contact lenses (CLs). The clinical examinations included evaluations of visual acuity (VA), best-corrected visual acuity (BCVA), intraocular pressure measurements (IOP), and corneal endothelial cell density (ECD). Ultrasound biomicroscopy (UBM) was used to observe the haptic lens positions. RESULTS: In the BDI group, light sensitivity dramatically decreased postoperatively in all patients, and 78 patients (82.1%) had a BCVA score >20/200. A total of 36 eyes (37.89%) developed elevated IOP after the BDI lens implantation, and 12 eyes (12.63%) underwent glaucoma laser surgery. Corneal decompensation occurred in nine eyes (9.47%), three of which developed glaucoma. Twelve patients (12.63%) experienced limbal stem cell failure, and 16 patients (16.84%) had eccentric BDI lenses. All patients in the RGP group had a BCVA score ≥20/200. Only seven eyes (9.3%) had cornea complications. There were no statistically significant differences in either the IOP or ECD after wearing the RGP CLs. CONCLUSIONS: Glaucoma, corneal decompensation, limbal stem cell failure, and eccentric BDI lenses appeared to be the major long-term complications in BDI lens implantation. BDI lens implantation could effectively improve postoperative outcomes by reducing glare disability, improving visual performance, and addressing cosmetic issues. Because the implantation of the BDI lens in traumatic aniridia had a high incidence of complications, this treatment should be used with caution, and the long-term complications should be seriously considered.

Efficient generation of lens progenitor cells from cataract patient-specific induced pluripotent stem cells.

The development of a technique to induce the transformation of somatic cells to a pluripotent state via the ectopic expression of defined transcription factors was a transformational event in the field of regenerative medicine. The development of this technique also impacted ophthalmology, as patient-specific induced pluripotent stemcells (iPSCs) may be useful resources for some ophthalmological diseases. The lens is a key refractive element in the eye that focuses images of the visual world onto the retina. To establish a new model for drug screening to treat lens diseases and investigating lens aging and development, we examined whether human lens epithelial cells (HLECs) could be induced into iPSCs and if lens-specific differentiation of these cells could be achieved under defined chemical conditions. We first efficiently reprogrammed HLECs from age-related cataract patients to iPSCs with OCT-4, SOX-2, and KLF-4. The resulting HLEC-derived iPS (HLE-iPS) colonies were indistinguishable from human ES cells with respect to morphology, gene expression, pluripotent marker expression and their ability to generate all embryonic germ-cell layers. Next, we performed a 3-step induction procedure: HLE-iPS cells were differentiated into large numbers of lens progenitor-like cells with defined factors (Noggin, BMP and FGF2), and we determined that these cells expressed lens-specific markers (PAX6, SOX2, SIX3, CRYAB, CRYAA, BFSP1, and MIP). In addition, HLE-iPS-derived lens cells exhibited reduced expression of epithelial mesenchymal transition (EMT) markers compared with human embryonic stem cells (hESCs) and fibroblast-derived iPSCs. Our study describes a highly efficient procedure for generating lens progenitor cells from cataract patient HLEC-derived iPSCs. These patient-derived pluripotent cells provide a valuable model for studying the developmental and molecular biological mechanisms that underlie cell determination in lens development and cataract pathophysiology.