Xinxing Guo

Comprehensive Mutation Analysis by Whole-Exome Sequencing in 41 Chinese Families With Leber Congenital Amaurosis

PURPOSE Leber congenital amaurosis (LCA) is a genetically heterogeneous disease with, to date, 19 identified causative genes. Our aim was to evaluate the mutations in all 19 genes in Chinese families with LCA. METHODS LCA patients from 41 unrelated Chinese families were enrolled, including 25 previously unanalyzed families and 16 families screened previously by Sanger sequencing, but with no identified mutations. Genetic variations were screened by whole-exome sequencing and then validated using Sanger sequencing. RESULTS A total of 41 variants predicted to affect protein coding or splicing was detected by whole-exome sequencing, and 40 were confirmed by Sanger sequencing. Bioinformatic and segregation analyses revealed 22 potentially pathogenic variants (17 novel) in 15 probands, comprised of 3 of 16 previously analyzed families and 12 of 25 (48%) previously unanalyzed families. In the latter 12 families, mutations were found in CEP290 (three probands); GUCY2D (two probands); and CRB1, CRX, RPE65, IQCB1, LCA5, TULP1, and IMPDH1 (one proband each). Based on the results from 87 previously analyzed probands and 25 new cases, GUCY2D, CRB1, RPGRIP1, CEP290, and CRX were the five most frequently mutated genes, which was similar to the results from studies in Caucasian subjects. CONCLUSIONS Whole-exome sequencing detected mutations in the 19 known LCA genes in approximately half of Chinese families with LCA. These results, together with our previous results, demonstrate the spectrum and frequency of mutations of the 19 genes responsible for LCA in Han Chinese individuals. Whole-exome sequencing is an efficient method for detecting mutations in highly heterogeneous hereditary diseases. Chinese Abstract.

Effect of Ginkgo Biloba on Visual Field and Contrast Sensitivity in Chinese Patients With Normal Tension Glaucoma: A Randomized, Crossover Clinical Trial

PURPOSE We evaluated the effect of ginkgo biloba extract on visual field defect and contrast sensitivity in a Chinese cohort with normal tension glaucoma. METHODS In this prospective, randomized, placebo-controlled crossover study, patients newly diagnosed with normal tension glaucoma, either in a tertiary glaucoma clinic (n = 5) or in a cohort undergoing routine general physical examinations in a primary care clinic (n = 30), underwent two 4-week phases of treatment, separated by a washout period of 8 weeks. Randomization determined whether ginkgo biloba extract (40 mg, 3 times per day) or placebo (identical-appearing tablets) was received first. Primary outcomes were change in contrast sensitivity and mean deviation on 24-2 SITA standard visual field testing, while secondary outcomes included IOP and self-reported adverse events. RESULTS A total of 35 patients with mean age 63.7 (6.5) years were randomized to the ginkgo biloba extract-placebo (n = 18) or the placebo-ginkgo biloba extract (n = 17) sequence. A total of 28 patients (80.0%, 14 in each group) who completed testing did not differ at baseline in age, sex, visual field mean deviation, contrast sensitivity, IOP, or blood pressure. Changes in visual field and contrast sensitivity did not differ by treatment received or sequence (P > 0.2 for all). Power to have detected a difference in mean defect as large as previously reported was 80%. CONCLUSIONS In contrast to some previous reports, ginkgo biloba extract treatment had no effect on mean defect or contrast sensitivity in this group of normal tension glaucoma patients. (http://www.chictr.org number, ChiCTR-TRC-08000724).

Childhood gene-environment interactions and age-dependent effects of genetic variants associated with refractive error and myopia: The CREAM Consortium

Myopia, currently at epidemic levels in East Asia, is a leading cause of untreatable visual impairment. Genome-wide association studies (GWAS) in adults have identified 39 loci associated with refractive error and myopia. Here, the age-of-onset of association between genetic variants at these 39 loci and refractive error was investigated in 5200 children assessed longitudinally across ages 7–15 years, along with gene-environment interactions involving the major environmental risk-factors, nearwork and time outdoors. Specific variants could be categorized as showing evidence of: (a) early-onset effects remaining stable through childhood, (b) early-onset effects that progressed further with increasing age, or (c) onset later in childhood (N = 10, 5 and 11 variants, respectively). A genetic risk score (GRS) for all 39 variants explained 0.6% (P = 6.6E–08) and 2.3% (P = 6.9E–21) of the variance in refractive error at ages 7 and 15, respectively, supporting increased effects from these genetic variants at older ages. Replication in multi-ancestry samples (combined N = 5599) yielded evidence of childhood onset for 6 of 12 variants present in both Asians and Europeans. There was no indication that variant or GRS effects altered depending on time outdoors, however 5 variants showed nominal evidence of interactions with nearwork (top variant, rs7829127 in ZMAT4; P = 6.3E–04).

The epidemics of myopia: Aetiology and prevention

&NA; There is an epidemic of myopia in East and Southeast Asia, with the prevalence of myopia in young adults around 80–90%, and an accompanying high prevalence of high myopia in young adults (10–20%). This may foreshadow an increase in low vision and blindness due to pathological myopia. These two epidemics are linked, since the increasingly early onset of myopia, combined with high progression rates, naturally generates an epidemic of high myopia, with high prevalences of “acquired” high myopia appearing around the age of 11–13. The major risk factors identified are intensive education, and limited time outdoors. The localization of the epidemic appears to be due to the high educational pressures and limited time outdoors in the region, rather than to genetically elevated sensitivity to these factors. Causality has been demonstrated in the case of time outdoors through randomized clinical trials in which increased time outdoors in schools has prevented the onset of myopia. In the case of educational pressures, evidence of causality comes from the high prevalence of myopia and high myopia in Jewish boys attending Orthodox schools in Israel compared to their sisters attending religious schools, and boys and girls attending secular schools. Combining increased time outdoors in schools, to slow the onset of myopia, with clinical methods for slowing myopic progression, should lead to the control of this epidemic, which would otherwise pose a major health challenge. Reforms to the organization of school systems to reduce intense early competition for accelerated learning pathways may also be important. HighlightsThere is an epidemic of myopia in the developed countries of East and Southeast Asia.A related epidemic of high myopia is due to early onset myopia and rapid myopic progression.There is a new and highly prevalent form of high myopia, which is acquired rather than genetic.Intense education and limited time outdoors play major causal roles in both epidemics.These modifiable risk factors are already being used in schools to contain the epidemics.

Normative Distribution of Visual Acuity in 3- to 6-Year-Old Chinese Preschoolers: The Shenzhen Kindergarten Eye Study

PURPOSE To document the distribution of uncorrected visual acuity (UCVA) in a defined population of Chinese preschoolers and to discuss its implications for vision referral. METHODS Preschoolers aged 3 to 6 years old were recruited from kindergartens in Shenzen. Uncorrected visual acuity was estimated by using Early Treatment Diabetic Retinopathy Study Tumbling E charts, followed by cycloplegic refraction and ocular examination. The reference population was defined as children without clinically significant ocular abnormalities, with spherical equivalent refraction greater than -0.50 to less than +2.00 diopters (D), astigmatism less than 0.75 D, and anisometropia less than 2.00 D. The age-specific UCVA cutoffs were defined by the line where the single-sided 95th percentile of the reference population fell. RESULTS A total of 483 of the 1255 children enrolled were considered the reference population. The monocular UCVA cutoff fell on the line of 20/63 at age 3, 20/50 at age 4, and 20/40 at ages 5 and 6. Using no better than these lines as criteria generated referral rates of 9.4% to 27.8% in the general population at different ages, and detected 83.3% and more than 90.0% of those with myopia and amblyopia, respectively. Using uncorrected interocular difference of two or more lines referred 3.6% to 4.3% of the population but identified only approximately 20.0% of those with amblyopia. CONCLUSIONS Visual acuity is still developing in preschoolers even at age 6. Most children with myopia and amblyopia can be identified with age-specific, monocular UCVA cutoffs in vision screening using Tumbling E charts, with tolerable false-positive rates. Further studies are needed to define the age at which children without significant refractive errors reach 20/20 UCVA.

ALMS1 null mutations: a common cause of Leber congenital amaurosis and early-onset severe cone-rod dystrophy.

In our previous studies, mutations in known candidate genes were detected in approximately 50% of Chinese patients with various forms of retinal degeneration. The next stage, identifying additional causative mutations in patients with various forms of genetic eye diseases based on whole exome sequencing of 1220 samples, revealed frequent homozygous or compound heterozygous null mutations in ALMS1, which are known to associate with Alström syndrome as well as individuals diagnosed with Leber congenital amaurosis (LCA) or early‐onset severe cone–rod dystrophy (CORD) without signs of systemic phenotypes except that one had a congenital heart abnormity. Sanger sequencing, co‐segregation analysis and analysis of normal individuals identified a total of 13 null mutations in ALMS1 in 11 probands, including 4 probands with homozygous mutations and 7 with compound heterozygous mutations. Follow‐up examinations revealed absent or mild systemic manifestations of Alström syndrome in those available: 9 of 15 patients in 11 families. These findings not only expand the spectrum of phenotypes associated with ALMS1 mutations but also suggest that ALMS1 should be regarded as a candidate causative gene in patients diagnosed with isolated LCA and early‐onset severe CORD.

Age-Related Changes of Intraocular Pressure in Elderly People in Southern China: Lingtou Eye Cohort Study

Purpose To study age-related changes of intraocular pressure (IOP) and assess the cohort effect in both cross-sectional and longitudinal settings among elderly Chinese adults. Methods Participants were enrolled from the Lingtou Eye Cohort Study with Chinese government officials aged 40 years and older at baseline and received physical check-up and ocular examinations from 2010 to 2012. IOP was measured using a non-contact tonometer according to standardized protocols, as well as systolic blood pressure (SBP), diastolic blood pressure (DBP) and body mass index (BMI). Participants who had attended IOP measurements in both 2010 and 2012 were included in this study. Cross-sectional association of IOP with age was assessed using multivariate liner regression analyses and based on the data of 2010. Longitudinal changes in IOP were assessed by paired t-test. Results A total of 3372 subjects were enrolled in the current analysis (2010 mean [SD] age, 61.9 [7.1] years; 60.2% men). The mean IOP in 2010 was 15.4±2.3 mmHg for women and 15.2±2.3 mmHg for men with an intersex difference (P = 0.029). Cross-sectional analysis showed that IOP was negatively associated with age (P = 0.003, β = -0.033 for women and P<0.001, β = -0.061 for men) adjusted for baseline SBP, DBP and BMI. Paired t-test suggested that IOP was higher in the year 2012 than 2010 in women (P = 0.006) but did not change significantly in men within 2 years (P = 0.345). In addition, the 2-year changes of IOP were not associated with age adjusted for baseline IOP in 2010 (P = 0.249). Conclusion Cross-sectional data suggests that IOP is lower in people with older age. Longitudinal data does not support such findings and thus the identified decreasing pattern with age in cross-sectional analysis is likely caused by cohort effects.

Six-year changes in refraction and related ocular biometric factors in an adult Chinese population

Purpose To investigate longitudinal changes in refraction and biometry in Chinese adults. Design Population-based prospective cohort study. Methods 1817 subjects aged ≥ 35 years were randomly recruited from Yuexiu district, Guangzhou, China in 2008. Of which 1595 (87.8%) were reexamined in 2010 and 1427 (78.5%) were reexamined in 2014. Non-cycloplegic automated refraction and visual acuity test were performed at baseline and the 6-year follow-up examination for all participants. In addition, 50% of the participants were randomly selected for axial length (AL), anterior chamber depth (ACD) and lens thickness (LT) measurements using non-contact partial coherence laser interferometry. Lens power (LP) was calculated with the Bennett’s equation. Results A total of 1300 participants were included in current analysis (2008 mean [SD] age, 51.4 [10.6] years; 54.5% women). Mean change in spherical equivalence (SE) was +0.24 (95% confidence interval [CI], +0.19 to +0.30), +0.51 (95% CI, +0.46 to +0.57), +0.26 (95% CI, +0.15 to +0.38) and -0.05 (95% CI, -0.21 to +0.10) diopters (D) for individuals in the age groups of 35 to 44, 45 to 54, 55 to 64 and 65+ years at baseline, respectively. Corneal power, AL and LT increased while ACD and LP decreased during the follow-up. Baseline SE and changes in biometric factors could explain 97.2% of the variance in longitudinal SE change while LP solely could explain 65.2%. Six-year mean change in cylinder power was -0.16 (95% CI, -0.19 to -0.13) D, the axis of astigmatism changed from “with-the-rule” to “against-the-rule” in 16.4% of the participants and to “oblique” in 0.9%. Conclusions This study confirms a hyperopic shift in the elderly before 65 years old and a myopic shift thereafter. Longitudinal refraction change could be well explained by corresponding biometry changes, especially LP. There is also a shift to “against-the-rule” astigmatism for the adult population.

Progression of Near Vision Loss and Incidence of Near Vision Impairment in an Adult Chinese Population

PURPOSE To investigate the progression of near vision loss and the cumulative incidence of near vision impairment (NVI) 6 years after initial examination of an urban Chinese cohort. DESIGN Population-based, prospective cohort study. PARTICIPANTS People aged ≥35 years examined at baseline in the Yuexiu District of Guangzhou, China. METHODS Participants examined at baseline were invited for 2-year and 6-year follow-up examinations in 2010 and 2014, respectively. Examinations included noncycloplegic autorefraction and binocular near visual acuity (NVA) with and without current near correction measured at 40 cm using a LogMAR ETDRS near vision tumbling E chart. Those with uncorrected binocular NVA (UCNVA) ≤20/40 underwent subjective refraction to obtain best-corrected binocular NVA (BCNVA). MAIN OUTCOME MEASURES Change in UCNVA between baseline and 2014 follow-up examinations and the 6-year cumulative incidence of vision impairment based on 3 definitions: NVA ≤20/40, ≤20/50, and ≤20/63. RESULTS Among the 1817 baseline participants, 1595 (87.8%) were reexamined in 2010 and 1427 (78.5%) in 2014. Mean vision loss between baseline and the 2014 follow-up was 1.54 (±1.74) lines of UCNVA. Vision loss was associated with age 80 years or older, less education, and better baseline UCNVA. The 6-year cumulative incidence of uncorrected binocular NVI (UCNVI) across the 3 vision impairment definitions was 55.2% (95% confidence interval [CI], 46.1%-64.3%), 51.3% (95% CI, 44.0%-58.7%), and 42.4% (95% CI, 35.5%-49.3%), respectively. With best-corrected binocular NVI (BCNVI), incidence was 6.89% (95% CI, 4.28%-9.50%), 5.17% (95% CI, 2.89%-7.44%), and 2.62% (95% CI, 1.11%-4.12%), respectively. A higher incidence of UCNVI was associated with worse baseline UCNVA for all 3 impairment definitions. Similarly, incidence of BCNVI was associated with worse baseline BCNVA, but also with older age and education at the primary level or less. Gender was not significant for either UCNVI or BCNVI. CONCLUSIONS Approximately half of those aged 35 years or older develop UCNVI in 6 years, the overwhelming majority of whom can be corrected with spectacles. Cost-effective strategies to provide spectacles to this at-risk population remains an issue requiring further study.

Significant Axial Elongation with Minimal Change in Refraction in 3- to 6-Year-Old Chinese Preschoolers: The Shenzhen Kindergarten Eye Study

PURPOSE To document the distribution of ocular biometry and to evaluate its associations with refraction in a group of Chinese preschoolers. DESIGN Population-based cross-sectional study. PARTICIPANTS A total of 1133 preschoolers 3 to 6 years of age from 8 representative kindergartens. METHODS Biometric measurements including axial length (AL), anterior chamber depth (ACD), and corneal radius of curvature (CR) were obtained from partial-coherence laser interferometry (IOL Master; Carl Zeiss Meditec, Oberkochen, Germany) before cycloplegia. Lens power (LP) and AL-to-CR ratio were calculated. Cycloplegic refraction (3 drops of 1% cyclopentolate) was measured using an autorefractor (KR8800; Topcon Corp., Tokyo, Japan), and spherical equivalent refraction (SER) was calculated. Biometric and refractive parameters were assessed as a function of age and gender. Multiple regression analysis was performed to explore the associations between refraction and ocular biometry. MAIN OUTCOME MEASURES Ocular biometric distributions and their relationships to refraction. RESULTS Among the 1127 children (99.5%) with successful cycloplegic refraction, mean SER was 1.37±0.63 diopters (D). Prevalence of myopia increased from 0% at 3 years of age to 3.7% (95% confidence interval, 1.0%-6.5%) at 6 years of age. Biometric parameters followed Gaussian distributions with means of 22.39±0.68 mm for AL, 7.79±0.25 mm for CR, and 24.61±1.42 D for calculated LP; and non-Gaussian distributions with means of 3.34±0.24 mm for ACD and 2.88±0.06 for AL-to-CR ratio. Axial length, ACD, and AL-to-CR ratio increased from 3 to 6 years of age, CR remained stable, whereas LP declined. Overall, SER declined slightly. For the SER variance, AL explained 18.6% and AL-to-CR ratio explained 39.8%, whereas AL, CR, and LP accounted for 80.0% after adjusting for age and gender. CONCLUSIONS Young Chinese children are predominantly mildly hyperopic, with a low prevalence of myopia by the age of 6 years. An increase of 1 mm in AL was associated with only 0.45 D of myopic change. Decreases in LP reduce the myopic shifts that normally would be associated with increases in AL, and thus play a key role in refractive development in this age group.