Characterization of N6-methyladenosine long non-coding RNAs in sporadic congenital cataract and age-related cataract

AIM:To characterize the N6-methyladenosine(m6A)modification patterns in long non-coding RNAs(lncRNAs)in sporadic congenital cataract(CC)and age-related cataract(ARC).METHODS:Anterior capsule of the lens were collected from patients with CC and ARC.Methylated RNA immunoprecipitation with next-generation sequencing and RNA sequencing were performed to identify m6A-tagged lncRNAs and lncRNAs expression.Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses and Gene Ontology annotation were used to predict potential functions of the m6A-lncRNAs.RESULTS:Large amount of m6A peaks within lncRNA were identified for both CC and ARC,while the level was much higher in ARC(49870 peaks)than that in CC(18688 peaks),yet those difference between ARC in younger age group(ARC-1)and ARC in elder age group(ARC-2)was quite slight.A total of 1305 hypermethylated and 1178 hypomethylated lncRNAs,as well as 182 differential expressed lncRNAs were exhibited in ARC compared with CC.On the other hand,5893 hypermethylated and 5213 hypomethylated lncRNAs,as well as 155 significantly altered lncRNA were identified in ARC-2 compared with ARC-1.Altered lncRNAs in ARC were mainly associated with the organization and biogenesis of intracellular organelles,as well as nucleotide excision repair.CONCLUSION:Our results for the first time present an overview of the m6A methylomes of lncRNA in CC and ARC,providing a solid basis and uncovering a new insight to reveal the potential pathogenic mechanism of CC and ARC.

Aberrant expression of COL4A1 in age-related cataract and its effect on cell proliferation,apoptosis and gene expression changes

AIM:To evaluate the regulation of the aberrant expression of collagen typeⅣalpha 1 chain(COL4A1)in the development of age-related cataract(ARC).METHODS:Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blot analysis were employed to evaluate the expression of COL4A1 in ARC patients and healthy controls.The proliferation,apoptosis,cell cycle and epithelial-mesenchymal transition(EMT)of human lens epithelial cell(HLE-B3)were further analyzed under the condition of COL4A1 gene silence.Alteration of gene expression at mRNA level after knockdown COL4A1 were also evaluated by qRT-PCR on HLE-B3 cells.RESULTS:The aberrant expression of COL4A1 was identified a clinically associated with the ARC.Silencing of COL4A1 promoted the apoptosis and inhibited the proliferation of HLE-B3 by blocking the cell cycle.Moreover,COL4A1 gene silence didn’t affect the cytoskeleton of HLE-B3 but down-regulated the Collagen typeⅣAlpha 2 Chain(COL4A2),paired box 6(PAX6),procollagen-lysine 2-oxoglutarate 5-dioxygenases 1(PLOD1)and procollagenlysine 2-oxoglutarate 5-dioxygenases 2(PLOD2)expression levels in HLE-B3 cells.Silencing the COL4A1 gene induced EMT of the HLE-B3 cells by promoting the transforming growth factor beta(TGF-β)expression.CONCLUSION:Silencing of COL4A1 induces S-phase arrest,also inhibits the proliferation and enhance HLE-B3 apoptosis and EMT,and down-regulates the expression of COL4A2,PAX6,PLOD1 and PLOD2.Thus,the expression alteration of COL4A1 may play a critical role in the pathogenesis of ARC.

Upregulation of histone H3 caused by CRYAA may contribute to the development of age-related cataract

Objective:Age-relate cataract(ARC)is a disease of the eyes with no effective drugs to prevent or treat patients.The aim of the present study is to determine whether histone H3,αA-crystallin(CRYAA),β-galactosidase(GLB1),and p53 are involved in the pathogenesis of ARC.Methods:A total of 99 anterior lens capsules(ALCs)of patients with ARC of various nuclear grades,ultraviolet models of ALCs,and two human lens epithelial cell lines(FHL-124 and SRA01/04)were used,and the expression of histone H3,CRYAA,GLB1,and p53 were detected by immunoblotting and reverse transcription and real time-quantitative polymerase chain reaction.The association between CRYAA with histone H3,GLB1,and p53 was assessed in FHL-124 and SRA01/04 cells following CRYAA overexpression.Results:Histone H3 and p53 in ALCs of patients with ARC were up-regulated in a grade-dependent manner,and the expression of CRYAA showed a positive association with histone H3,p53,and GLB1.In UV models of ALCs and human lens epithelial cell lines,the expression levels of histone H3,cell apoptosis factors(Bax/Bcl-2,cleaved caspase-3),and inflammation factors(interleukin-6,tumor necrosis factor-α)were all up-regulated.Furthermore,transfection of CRYAA in FHL-124 cells induced overexpression of histone H3.Conclusion:CRYAA-mediated upregulation of histone H3 may be involved in the pathogenesis of ARC.p53 may also have a role in ARC development,but not via the CRYAA-histone H3 axis.The results of the present study may assist in improving our understanding of the pathogenesis of ARC and in identifying potential targets for treatment.

Cigarette smoking,body mass index associated with the risks of age-related cataract in male patients in northeast China

AIM: To determine the association between cigarettes smoking, body mass index (BMI) and the risk of age-related cataract (ARC) in middle-aged and elderly men in Northeast China. METHODS: A hospital-based case control study was conducted. Cases (n =362) were men who had surgically treated ARC, 45-85 years old; controls frequency-matched (n=362) were men who had been admitted to the same hospital as cases for other diseases not related with eye diseases. Cases and controls were matched with 1:1. The cases and controls were interviewed during their hospital stay, using a structured interviewer-administrated questionnaire that included information on sociodemographic characteristics, socioeconomic, lifestyle habits (tobacco smoking and alcohol consumption, etc.), anthropometric measures, personal medical history, and family history of ARC in first-degree relatives, and simultaneously BMI was calculated. The odds ratios (OR) and 95% confidence intervals (CI) of ARC were estimated using multiple logistic regression models. RESULTS: After adjusting for age and multiple potential confounders, higher BMI was associated with an increased risk of ARC. Cigarette smoking, years smoking or moderate cigarette smoking (1-29 cigarettes per day) had no relation with the risk of ARC (P>0.05), although patients smoking >= 30 cigarettes per day had an elevated risk of ARC as compared with the non-smokers (OR=1.55, 95% CI; 1.16-2.85, P=0.026). Higher BMI was associated with an increased risk of ARC. Both overweight and obesity was associated with an obviously increased risk for surgically ARC (OR=1.55, 95% CI:1.02-1.98, P=0.015 and OR=1.71, 95% CI:1.32-2.39, P=0.013 respectively) compared to normal BMI. Then participants were grouped into quartiles of BMI (Q1 to Q4), compared to controls in the lowest quartile, the ORfor cases in the highest quartile of BMI was 1.54 (OR=1.54, 95% CI: 1.08-2.46, P=0.022). The results of univariate analysis showed cigarette smoking was not associated with ARC formation for men with lower or normal BMI (P >0.05). Compared to the non-smokers, for men of overweight or obesity, cigarette smoking was associated with a significantly increased risk for surgically ARC (OR=2.00, 95% CI: 1.49-6.65, P=0.003 and OR=1.66, 95% CI: 1.63-13.21, P=0.002 respectively). Similarly, smokers in the highest quartile of BMI had approximately 1.5 times the risk of ARC as non-smokers in the lowest quartile (OR =1.46, 95% CI: 1.06-5.29, P<0.001). Followed multivariate models revealed that the association had never changed. CONCLUSION: Current cigarette smoking is positively related to ARC only among those who smoking 30 or more cigarettes per day. For men who are both overweight and obesity, cigarette smoking is associated with a significantly increased risk for ARC.

Relationship between dietary macronutrient intake and the risk of age-related cataract in middle-aged and elderly patients in northeast China

AIM:To examine the association between dietary macronutrient intake and the risk of age-related cataract(ARC)in middle-aged and elderly men.METHODS:A hospital-based case-control study was conducted from December 2009 to November 2011.Cases(n=360)were patients with cataract aged 45-85years old,and controls(n=360)were patients who had been admitted to the same hospital for diseases not related with cataract.All subjects were interviewed using a structured interviewer-administrated questionnaire that included information on socio-demographic characteristics,lifestyle habits and detailed medical history,simultaneously,the dietary intakes of nutrients were collected via a valid semi-quantitative food frequency questionnaire(FFQ).The odds ratios(OR)and corresponding 95%confidence intervals(CI)of three types of ARC were estimated using multiple logistic regression models.RESULTS:After adjusting for multiple potential confounders,total dietary intake of carbohydrate was positively associated with cortical cataract,compared to controls in the lowest quartile,and the OR for cases in the highest quartile of intake was 2.471(95%CI:1.348-6.043,P=0.027).Higher dietary intakes of protein were protective for posterior subcapsular cataract(PSC)(OR=0.528,95%CI:0.148-0.869,P=0.023).Dietary fat intake was not associated with any type of cataract,however,participants in the highest quartile of polyunsaturated fatty acids intake had 2.7 times the risk of nuclear cataract as did those in the lowest quartile(OR=2.742,95%CI:1.790-4.200,P=0.033).CONCLUSION:A high intake of carbohydrate and polyunsaturated fatty acid may increase the odds of cortical and nuclear cataract,respectively,whereas high intake of protein,especially animal protein,may protect against PSC cataract.It is possible that dietary changes of target population may reduce the risk of ARC.

miR-211 regulates the antioxidant function of lens epithelial cells affected by age-related cataracts

AIM： To investigate the effects and mechanism of miR-211 in mediating the antioxidant function of lens epithelial cells affected by age-related cataracts. METHODS： Real-time quantitative polymerase chain reaction （RT-qPCR） was used to detect miR-211 expression in the anterior lens capsules of healthy people, the anterior lens capsules of patients with age-related cataracts, and human epithelial cell line （SRA01/04） cells exposed to oxidative stress. A 2＇, 7＇-dichloro-fluorescein diacetate （DCFH-DA） probe was used to measure the levels of endogenous reactive oxygen species （ROS） in human lens epithelial cells （hLECs） exposed to 400 pmol/L H2O2 for lh. SRA01/04 cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls. After 72h, these cells were exposed to 400 IJmollL H2O2 for lh, then p53 and Bax mRNA expression were measured using RT-qPCR. p53 and Bax protein expression were also measured by Western blotting analysis. Finally, cell viability was assessed using an MTS assay. RESULTS： Compared to the control group, expression of miR-211 in the anterior lens capsules of age-related cataract patients and in SRA01/04 cells exposed to oxidative stress was significantly increased （P〈0.001）. Levels of endogenous ROS were significantly elevated in hLECs exposed to oxidative stress （P〈0.001）. Compared to the mimic control group, the hLECs in the miR-211 mimic group expressed significantly higher levels of p53 and Bax mRNA and protein while cell viability was significantly reduced （P〈0.001）. Conversely, p53 and Bax mRNA and protein expression were significantly reduced in the miR-211 inhibitor group as compared to the control group, while the cells in this group had much higher levels of call viability （P〈0.001）. CONCLUSION： miR-211 is upregulatsd in the anterior lens capsules of age-related cataract patients, miR-211 decreased the antioxidative stress capacity of lens epithelial cells by upregulating p53 and Bax, while inhibiting cell proliferation and repair. This finding suggests that miR-211 may play a key role in the development of age-related cataracts.

Screening of methylation genes in age-related cataract

AIM： To analyze and screen the methylation status of whole-genome in age-related cataract samples. METHODS： Anterior lens capsule samples were collected from age-related cortical cataract patients over 50 years of age with LOCS III score of nuclear color ≥4 along with control subjects. DNAs were extracted and subjected to methylation microarray for the identification of methylated genes employing the high-throughput sequencing approach. RESULTS： Compared with the control group, 843 sites were found methylated, including 802 hypermethylation sites with 542 corresponding genes, 41 demethylation sites with 29 corresponding sites. COL4 A1, GJA3, SIPA1 L3 were confirmed by mass spectrometry, the results were consistent with high-throughput sequencing. CONCLUSION： DNA methylation microarrays is an efficient way for screening the aberrantly methylated genes. In this study, we are able to screen a few age-related cataract genes such as COL4 A1, GJA3, and SIPA1 L3 for their aberrant methylation patterns in cataract patients however further work is warranted to understand the significance of these findings.

A pilot exome-wide association study of age-related cataract in Koreans

Age-related cataract （ARC） is the most common cause of visual impairment and blindness worldwide. A previous study reported that genetic factors could explain approximately 50% of the heritability of cataract. However, a genetic predisposition to ARC and the contributing factors have not yet been elucidated in the Korean population. In this study, we assessed the influence of genetic polymorphisms on the risk of ARC in Koreans, including 156 cataract cases and 138 healthy adults. We conducted an exome-wide association study using Illumina Human Exome-12v1.2 platform to screen 244,770 single nucleotide polymorphisms （SNPs）. No SNPs reached exome-wide significance level of association （P 〈 l x 10 6）. B3GNT4 rs7136356 showed the most significant association with ARC （P = 6.54x 10 5）. Two loci （MUC16 and P2RY2） among the top 20 ARC-associated SNPs were recognized as probably linked to cata- ractogenesis. Functions of these genes were potentially related to regulating dehydration or homeostasis of the eyes, and showed a potential association with dry eye disease. This finding suggests that mucin- and dry eye disease-related genes may play a significant role in cataractogenesis. Our study provides insight into the genetic predisposition of ARC in Koreans. Additional studies with larger sample sizes are required to confirm the results of this study.

Prevalence and risk factors on age-related cataract and surgery in adults over 50 years old in Binhu District, Wuxi, China

AIM:To investigate the prevalence and risk factors of age-related cataract(ARC),ARC surgery procedures,and postoperative vision results among adults over 50 years old in the Binhu District of Wuxi City,China.METHODS:Thirty basic sampling units were analyzed via a cluster random sampling method.Detailed medical histories were collected and eye examinations were performed.Cataract prevalence and surgical procedures were quantified.RESULTS:Among the 6150 participants,1421 cataract cases were diagnosed and prevalence was 23.1%.The prevalence of cortical,nuclear,and posterior subcapsular cataracts increased with age(P<0.001).Cataract prevalence was significantly higher among elderly,female,or illiterate individuals and people with hypertension,diabetes,and a history of smoking and drinking(all P<0.05).As participant age increased and education level decreased,the frequency of cataract blindness surgeries gradually decreased,but without statistical significance within groups(P>0.05).The odds ratio of cataract patients who had or did not have cataract surgery was 3.15(87/28)and the frequency of cataract blindness surgery was 75.7%(87/115).Poor visual outcomes was in 107 eyes(40.7%)after cataract surgery.Poor vision was mostly caused by uncorrected reflective errors(30.9%)and ocular comorbidities(41.1%).The prevalence of cataract surgery complications was 5.7%(15/263).Surgical complications and posterior capsular opacification were avoidable factors facilitating poor vision.CONCLUSION:ARC,especially in females and illiterateindividuals,presents a public health problem in this district.Poor visual outcomes after cataract surgery are frequent.High-quality cataract surgeries and treatment of ocular comorbidities are vital.

Effect of the posterior corneal surface on total corneal astigmatism in patients with age-related cataract

AIM： To explore the effect of the posterior astigmatism on total corneal astigmatism and evaluate the error caused by substituting the corneal astigmatism of the simulated keratometriy （simulated K） for the total corneal astigmatism in age-related cataract patients. METHODS： A total of 211 eyes with age-related cataract from 164 patients （mean age： 66.8±9.0y, range： 45-83y） were examined using a multi-colored spot reflection topographer, and the total corneal astigmatism was measured. The power vector components J0 and J45 were analyzed. Correlations between the magnitude difference of the simulated K and total cornea astigmatism （magnitude differenceSimK-Tca）, anterior J0, and absolute meridian difference （AMD） between the anterior and posterior astigmatisms were calculated. To compare the astigmatism of the simulated K and total cornea both in magnitude and axial orientation, we drew double-angle plots and calculated the vector difference between the two measures using vector analysis. A corrective regression formula was used to adjust the magnitude of the simulated K astigmatism to approach that of the total cornea. RESULTS： The magnitude differenceSimK-Tca was positively correlated with the anterior corneal J0 （Spearman’s rho= 0.539; P〈0.001） and negatively correlated with the AMDR （Spearman’s rho=-0.875, P〈0.001）. When the anterior J0 value was larger than 1.3 D or smaller than -0.8 D, the errors caused by determining the total corneal astigmatism with the karatometric calculation tended to be greater than 0.25 D. An underestimation by 16% was observed for against the rule （ATR） astigmatism and an overestimation by 9% was observed for with the rule （WTR） astigmatism when ignoring the posterior measurements. CONCLUSION： Posterior corneal astigmatism should be valued for more precise corneal astigmatism management, especially for higher ATR astigmatism of the anterior corneal surface. We suggest a 9% reduction in the magnitude of the simulated K in eyes with WTR astigmatism, and a 16% addition of the magnitude of the simulated K in eyes with ATR astigmatism.

The impact of GJA3 SNPs on susceptibility to age-related cataract

AIM: To determine the association of gap junction protein alpha 3(GJA3) gene tag single-nucleotide polymorphisms(SNPs) with susceptibility to age-related cataract(ARC).METHODS: In total, 486 ARC patients were matched with 500 healthy controls. All the participants underwent complete ophthalmic examinations. Haplotype-tagging SNPs of GJA3 gene were selected from the HapMap Beijing Han Chinese population. Genomic DNA was extracted from the peripheral blood leukocytes of all the subjects.Under three different genetic models: dominant, recessive,and additive, the association between SNPs and ARC was examined. After adjusting for age and sex, the genetic effects of the GJA3 SNPs were evaluated with logistic regression analysis.RESULTS: Four tag GJA3 SNPs(rs6490519, rs9506430,rs9509053, and rs9552089) were included in the present study. None of the SNPs showed a significant relationship with an altered risk of total ARC under the dominant,recessive, or additive models. In the subgroup analysis,rs9506430 had a significant effect on the formation of a posterior subcapsular cataract(P=0.002, OR: 0.227, 95%CI:0.088-0.590) under the recessive model.CONCLUSION: Our study indicates that GJA3 variants may influence the development of posterior subcapsular cataracts. Further studies need to be designed to confirm this possibility.

Association of IGF1R polymorphisms(rs1546713) with susceptibility to age-related cataract in a Han Chinese population

AIM:To explore the susceptible association between the insulin-like growth factor-1 receptor(IGF1 R)single nucleotide polymorphism(SNP)and age-related cataract(ARC),and investigate the underlying mechanisms in human lens epithelium(HLE)cells.METHODS:Totally 1190 unrelated participants,comprising 690 ARC patients and 500 healthy individuals in Han Chinese population were recruited and genotyped for target SNP.Theχ2-test was used to detect genotypic distribution between the patient and control groups and the logistic regression was performed to adjust the age and gender.Meanwhile,different biological experimental methods,such as cell counting kit 8(CCK-8)assay,flow cytometry,quantitative real time polymerase chain reaction(Q-PCR)and Western blot,were used to detect cell viability,cell cycle progression and apoptosis in HLE cells or IGF1 R knockdown HLE cells.RESULTS:The rs1546713 in IGF1 R gene was identified(P=0.046,OR:1.606,95%CI:1.245-2.071),which shown a significant relevance with ARC risk under the dominant model.The results demonstrated that IGF1 R knockdown inhibited cell proliferation by inducing cell cycle arrested at S phase and promoting apoptosis.Mechanistically,the cell cycle blocked at S phase was linked with the alterations of cyclin A,cyclin B,cyclin E and P21.The pro-apoptosis function of IGF1 R may related with stimulating the activation of Caspase-3 and altering the expression levels of apoptotic proteins,including Bcl-2,Bax and Caspase-3.CONCLUSION:This study first report that IGF1 R polymorphisms may affect susceptibility to ARCs in Han Chinese population and provide new clues to understand the pathogenic mechanism of ARCs.Notably,IGF1 R is likely a potential target for ARC prevention and treatment.

Physical activity and risk of age-related cataract

AIM: To summarize quantitatively the prospective association between physical activity and age-related cataract(ARC) risk.METHODS: Pub Med, Embase, Web of Science, and Cochrane Library were systematically searched for all relevant follow up studies until July 2019. Multivariableadjusted relative risks(RRs) and corresponding 95% confidence intervals(CIs) from individual studies were used to calculate the overall summary estimates. The doseresponse relationship was assessed using generalized least-squares trend estimation.RESULTS: Six prospective cohort studies, involving 19 173 cases in 6.2-12.1 y follow up of 171 620 participants, were included in the analysis. Increased physical activity was significantly associated with reduced risk of ARC by 10%(RR: 0.90;95%CI: 0.81, 0.99, P=0.04). Stratified analysis by assessment method for physical activity suggest that studies using metabolic equivalent(MET) per day tended to report a slightly stronger association with ARC(RR: 0.85;95%CI: 0.81, 0.90, P<0.001) than studies which assessed activity by weekly activity(RR: 0.96;95%CI: 0.89, 1.03, P=0.24). Dose-response analysis indicated that the risk of ARC decreased by 2%(RR: 0.98;95%CI: 0.98, 0.99, P<0.001) for every 6 METs per day increase in activity.CONCLUSION: The findings from this Meta-analysis provide additional evidence that increased physical activity is inversely associated with ARC risk dose-responsively.

Use of Cataract Surgery in Urban Beijing: a Post Screening Follow-up of the Elderly with Visual Impairment due to Age-related Cataract

Objective To understand the perception for the use of cataract surgical services in a population acceptors and non-acceptors of cataract surgery in urban Beijing. Methods From a community-based screening program a total of 158 patients with presenting visual acuity of less than 6/18 on either eye due to age-related cataract were informed about the possibility of surgical treatment. These patients were interviewed and re-examined 36 to 46 months after initial screening. The main reasons for not accepting surgery were obtained using a questionnaire. Vision function and vision-related quality of life scores were assessed in those who received and did not receive surgery. Results At the follow-up examination 116 of the 158 patients were available and 36 （31.0%） had undergone cataract surgery. Cases who chose surgery had higher education level than those who did not seek surgery （OR=2.64, 95% CI： 1.08-6.63, P=0.02）. There were no significant differences in vision function （P=0.11） or quality of life scores （P=0.16） between the surgery group and the non-surgery group. Main reasons for not having surgery included no perceived need （50.0%）, feeling of being ＂too old＂ （19.2%）, and worry about the quality of surgery （9.6%）. Cost was seeking surgery. cited by 1 （1.9%） subject as the main reason for not

The J-shape association of serum total IgE levels with age-related cataract

AIM： To address the association between serum total IgE levels and age-related cataract in adults. METHODS： The study participants consisted of 1052 adults aged 40y or older in the Korean National Health and Nutrition Examination Survey 2010. We performed multivariable logistic regression analyses using the quartile cut-points of total IgE levels. RESULTS： The odds ratios （ORs） for nuclear and any cataract with ≥267 kU/L of serum IgE levels were 1.75 [95% confidence intervals （CI）, 1.04-2.96] and 2.00 （95%CI, 1.22-3.27）, respectively, comparing to 35-87 kU/L. Interestingly, participants with ≤35 kU/L of IgE levels （OR, 1.67; 95%CI, 1.02-2.72） also had higher risk for any cataract than those with 35-87 kU/L. The risk for any cataract （OR, 1.48; 95%CI, 1.03-2.13） was higher in participants with high total IgE levels （〉150 kU/L）, comparing to normal participants. CONCLUSION： Our findings indicate a J-shaped relationship between serum IgE levels and age-related cataract.

miR-211 promotes lens epithelial cells apoptosis by targeting silent mating-type information regulation 2 homolog 1 in age-related cataracts

AIM： To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS： This study used real-time quantitative polymerase chain reaction（RT-q PCR） to measure the expression of miR-211 and its predicted target gene [silent matingtype information regulation 2 homolog 1（SIRT1）] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line（SRA01/04） cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72 h after transfection, miR NA and protein expression of SIRT1 were measured using RT-qP CR and Western blotting; then cells were exposed to 200 μmol/L H2O2 for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.RESULTS： Compared to the control group, expression of miR-211 was significantly increased（P〈0.001）, the miR NA and protein expression of SIRT1 were significantly decreased（P〈0.001） in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miR NA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased（P〈0.001）. In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased（P〈0.001）. A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.CONCLUSION： miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.

Lens Epithelial Cell Proliferation and Cell Density in Human Age-related Cataract

Purpose: To discuss the potential effect of the lens epithelial cell proliferation inage-related cataract.Methods: In vitro cell proliferation was assayed by MTT method to evaluate the lensepithelial cell density, index, and proliferation capacity in normal lens and all kinds ofage-related cataract. Capsulotomy specimens from all kinds of patients who underwentcataract phacoemulsification extraction surgery were compared with the lens epithelialspecimens from non-cataract lenses of Eye Bank eyes.Results:Lens epithelial cell density of central anterior capsule(LECD) in female normallens was higher than that in male, LECD in nuclear cataract (> NⅢ) was higher thanthat in normal lens, but in the mature cortical cataract, LECD was lower. Mitotic indexof three kinds of age-related cataracts in vivo had no statistical difference, neither didcell proliferation capacity of cultivated cells in vitro.Conclusion: The individual difference of lens epithelial cell density and proliferationcapacity in vivo may be

DNA hypermethylation of COL4A1 in ultraviolet-Binduced age-related cataract models in vitro and in vivo

AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB respectively.The MTT assay was utilized to evaluate cell proliferation.Flow cytometry was employed for analysis of cell apoptosis and cell cycle.COL4A1 expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction(RTPCR).The localization of COL4A1 in HLEB3 cells was determined by immunofluorescence.The methylation status of CpG islands located in COL4A1 promoter was verified using bisulfite-sequencing PCR(BSP).DNMTs and TETs mRNA levels was examined by RT-PCR.RESULTS:UVB exposure decreased HLEB3 cells proliferation,while increased the apoptosis rate and cells were arrested in G0/G1 phase.COL4A1 expression was markedly inhibited in UVB treated cells compared to the controls.Hypermethylation status was detected in the CpG islands within COL4A1 promoter in HLEB3 cells subjected to UVB exposure.Expressions of DNMTs including DNMT1/2/3 were elevated in UVB treated HLEB3 cells compared to that in the controls,while expressions of TETs including TET1/2/3 showed the opposite trend.Results from the UVB treated rat model further confirmed the decreased expression of COL4A1,hypermethylation status of the CpG islands at promoter of COL4A1 and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.CONCLUSION:DNA hypermethylation of COL4A1 promoter CpG islands is correlated with decreased COL4A1 expression in UVB induced HLEB3 cells and anterior lens capsules of rats.

Association of age at diagnosis of diabetes with subsequent risk of age-related ocular diseases and vision acuity

BACKGROUND The importance of age on the development of ocular conditions has been reported by numerous studies.Diabetes may have different associations with different stages of ocular conditions,and the duration of diabetes may affect the development of diabetic eye disease.While there is a dose-response relationship between the age at diagnosis of diabetes and the risk of cardiovascular disease and mortality,whether the age at diagnosis of diabetes is associated with incident ocular conditions remains to be explored.It is unclear which types of diabetes are more predictive of ocular conditions.AIM To examine associations between the age of diabetes diagnosis and the incidence of cataract,glaucoma,age-related macular degeneration(AMD),and vision acuity.METHODS Our analysis was using the UK Biobank.The cohort included 8709 diabetic participants and 17418 controls for ocular condition analysis,and 6689 diabetic participants and 13378 controls for vision analysis.Ocular diseases were identified using inpatient records until January 2021.Vision acuity was assessed using a chart.RESULTS During a median follow-up of 11.0 years,3874,665,and 616 new cases of cataract,glaucoma,and AMD,respectively,were identified.A stronger association between diabetes and incident ocular conditions was observed where diabetes was diagnosed at a younger age.Individuals with type 2 diabetes(T2D)diagnosed at<45 years[HR(95%CI):2.71(1.49-4.93)],45-49 years[2.57(1.17-5.65)],50-54 years[1.85(1.13-3.04)],or 50-59 years of age[1.53(1.00-2.34)]had a higher risk of AMD independent of glycated haemoglobin.T2D diagnosed<45 years[HR(95%CI):2.18(1.71-2.79)],45-49 years[1.54(1.19-2.01)],50-54 years[1.60(1.31-1.96)],or 55-59 years of age[1.21(1.02-1.43)]was associated with an increased cataract risk.T2D diagnosed<45 years of age only was associated with an increased risk of glaucoma[HR(95%CI):1.76(1.00-3.12)].HRs(95%CIs)for AMD,cataract,and glaucoma associated with type 1 diabetes(T1D)were 4.12(1.99-8.53),2.95(2.17-4.02),and 2.40(1.09-5.31),respectively.In multivariable-adjusted analysis,individuals with T2D diagnosed<45 years of age[β95%CI:0.025(0.009,0.040)]had a larger increase in LogMAR.Theβ(95%CI)for LogMAR associated with T1D was 0.044(0.014,0.073).CONCLUSION The younger age at the diagnosis of diabetes is associated with a larger relative risk of incident ocular diseases and greater vision loss.

Biomaterial engineering strategies for modeling the Bruch's membrane in age-related macular degeneration

Age-related macular degeneration,a multifactorial inflammatory degenerative retinal disease,ranks as the leading cause of blindness in the elderly.Strikingly,there is a scarcity of curative therapies,especially for the atrophic advanced form of age-related macular degeneration,likely due to the lack of models able to fully recapitulate the native structure of the outer blood retinal barrier,the prime to rget tissue of age-related macular degeneration.Standard in vitro systems rely on 2D monocultures unable to adequately reproduce the structure and function of the outer blood retinal barrier,integrated by the dynamic interaction of the retinal pigment epithelium,the Bruch's membrane,and the underlying choriocapillaris.The Bruch's membrane provides structu ral and mechanical support and regulates the molecular trafficking in the outer blood retinal barrier,and therefo re adequate Bruch's membrane-mimics are key for the development of physiologically relevant models of the outer blood retinal barrie r.In the last years,advances in the field of biomaterial engineering have provided novel approaches to mimic the Bruch's membrane from a variety of materials.This review provides a discussion of the integrated properties and function of outer blood retinal barrier components in healt hy and age-related macular degeneration status to understand the requirements to adequately fabricate Bruch's membrane biomimetic systems.Then,we discuss novel materials and techniques to fabricate Bruch's membrane-like scaffolds for age-related macular degeneration in vitro modeling,discussing their advantages and challenges with a special focus on the potential of Bruch's membrane-like mimics based on decellularized tissue.