ObjectiveTo review retinitis pigmentosa (RP)-related gene researches in China. Methods RP genes and RP gene therapy as key words were searched from those online databases including China Knowledge Resource Integrated Database of China National Knowledge Infrastructure (CNKI), National Center for Biotechnology Information (NCBI), Human Gene Mutation Database (HGMD), and Human Genome Variation Society (HGVS). Related papers written by authors from China or other countries,published from 1991 to 2011 were collected for analysis. ResultsOverall 60 RP-related genes have been identified now. Chinese researchers from 29 hospitals and research institutes confirmed 17 RP-related gene changes (239 mutations, 131 of them are pathogenic) in 300 RP families and 1572 sporadic RP patients from 1991 to 2011, as reported in 66 papers. Mutations in rhodopsin (RHO) gene, NR2E3 gene and RP1 gene were found in 2.0％, 2.9％ and 1.0％ of Chinese RP patients respectively.The relationship between genotypes and clinical phenotypes were investigated in 15 papers, including RHO gene, RDS gene, RP1 and RPGR or RP3 gene in ADRP families. Gene therapy for RP has been investigated in 4 papers. Seven hundred and eighteen papers about RP gene-mutation screening have been published from other countries,and more than 2000 variants were identified which including 352 disease-causing mutations. Mutations of RHO gene, RDS and RP1 were found in 25％-50％, 8％ and 5％-10％ of ADRP patients in Caucasian populations. RPGR mutations were found in 70％-80％ of XLRP patients in Caucasian populations, and 50％-60％ of mutations are identified in open reading frame (ORF) 15. Totally 391 papers about gene therapy for RP have been published. ConclusionsSeventeen RP-related genes have been studied and 131pathogenic mutations were found in China. Considering Chinese population, our RP research is still behind other countries.

ObjectiveTo observe the hereditary types and clinical characteristics of 137 patients with retinitis pigmentosa (RP) in Ningxia.MethodsOne hundred and thirty-seven patients with RP who diagnosed by the examinations of visual acuity, optometry, direct or indirect ophthalmoscope, visual field,optical coherence tomography (OCT) and electroretinogram were enrolled. The hereditary types and clinical characteristics were analyzed according to the family history and the results of ophthalmologic examinations.ResultsOne hundred and thirty-seven patients included 29 autosomal dominant RP (ADRP) patients from 8 families (7.4％), 16 autosomal recessive RP (ARRP) patients from 15 families (13. 9％), 10 X-linked RP (XLRP) from 3 families (2.8％), and 82 simplex RP (SRP) patients (75.9％).There were 15consanguineous marriage families out of 26 families with RP history (57.7 ％). The patients were classified as typical RP (102 patients, 74.5％) and atypical RP (35 patients, 25.5％). All the ADRP and XLRP patients showed typical clinical features of RP. Ten (62.5％) of ARRP patients and 53 (64.6％) of SRP patients had typical features of RP. Six (37.5％) of ARRP patients and 29 (35.4％) of SRP patients had atypical features of RP. Among atypical RP patients, 17 (48. 6％) patients were non-pigmented RP which including 3 patients were misdiagnosed as amblyopia during childhood. The logarithm of minimal angle of resolution (logMAR) best corrected visual acuity (BCVA) of ADRP patients was 1.04±0.51 at the age older than 51 years, while the BCVA of ARRP and XLRP patients were 0. 92+0. 61 and 1. 70±0. 02 respectively at 21 to 30 years of age. One hundred and twenty-three (89. 8%) patients suffered from varying degrees of myopia. OCT showed that the average thickness of macular fovea in ADRP patients was ( 185. 73 + 1. 23) μm at the age older than 51 years, while in ARRP and XLRP patients were (173. 21 ± 0. 98) and (170. 49+1. 15) μm respectively at 21 to 30 years of age. Conclusions ADRP and XLRP are typical RP. All atypical RP are ARRP and SRP. Non-pigmented RP are mainly seen in atypical RP which often misdiagnosed as amblyopia during childhood. The photoreceptors in macula are damaged in the early stage and the decline of visual acuity occurred at 21 to 30 years of age in patients with ARRP and XLRP. The ADRP patients has late slower decline of visual acuity and retain some visual acuity at the age older than 51 years.

Background Congenital cataract is an important cause of blindness and amblyopia in children,and about 50％ of congenital cataract is hereditary.Objective The aim of this study was to determine the diseasecausing gene of one Hui congenital cataract pedigree by using exon combined target region capture sequencing chip of eye diseases.Methods This study was approved by Ethic Committee of Ningxia People's Hospital and followed Declaration of Helsinki.One Hui congenital cataract pedigree was recruited in Ningxia Eye Hospital in 2011.All the disease history of the members in this family were collected and recorded,and the eye examinations were performed.The peripheral blood specimens were collected from family members and 300 healthy individuals for the extraction of DNA.Exon combined target region capture sequencing chip of eye diseases was used to screen the candidate diseasecausing mutations,then PCR and direct sequencing were used to confirm the disease-causing mutations.Results This H ui family included 61 members of 6 generations,and 18 patients were diagnosed in serial 5 passages,conforming to autosomal dominant inheritance pattern.Among 18 cataract patients,7 individuals were associated with nystagmus and strabismus,and 4 patients had high myopia.Eight candidate pathogenetic mutations were detected by exon combined target region capture sequencing chip of eye diseases and bioinformatics method,with 5 mutations in noncoding regions and 3 in coding regions.The mutation P24T of CYRGD gene was confirmed as pathogenic mutation of this pedigree by using PCR and direct sequencing methods.These mutations co-segregated with affected members of the family,and the mutations were not found in the unaffected family members and 300 unrelated controls.Conclusions P24T of CYRGD gene mutation is confirmed as pathogenic mutation of this pedigree.Exon combined target region capture sequencing chip provides a new approach to detect disease-causing mutations of congenital cataract with diversity clinical phenotypes.

Background Age-related macular degeneration (AMD) is the main cause of irreversible loss of central vision in old population.The incidence of AMD is increasing year by year,but the mechanism is not clearly understood.Objective This study was to investigate the association between genetic variants and the risk of AMD in Ningxia population.Methods This study was approved by Ethic Committee of Ningxia People's Hospital and complied with the Helsinki Declaration.Written informed consent was obtained from each subject.One hundred and fifty patients with AMD and 145 ethnicity-and gender-matched controls were recruited in Ningxia Eye Hospital from January 2012 to March 2013.All individuals underwent comprehensive eye examinations and genomic DNA was prepared from peripheral blood.The single nucleotide polymorphisms (SNPs) of 8 susceptibility loci in four candidate genes,including complement factor H (CFH),complement factor B (CFB),age-related maculopathy susceptibility 2 (ARMS2) and high temperature required factor A1 (HTRA1),were genotyped with Mass Array and MALDI-TOF technique by Sequenom platform.The distribution of genotype was tested for Hardy-Weinberge equilibrium (HWE).The differences of genotype distribution of allele and haplotype frequencies were compared between patients and controls using chi-squared test and the P value was significant at ＜ 0.006 level after correction of age,and the relationship of genotype distribution with AMD was evaluated by Logistic regression analysis.Measures of linkage disequilibrium (LD) was carried out by Haploview.Results All the genetypes met HWE.Seven SNPs were found to be different in the genotypic distributions and allele frequencies between patients and normal controls (all at P＜ 0.05),however,after Bonferroni correction,the differences of only four SNPs were significant between the patients and controls in the genotype and allele distributions,including the SNPs of rs10737680 and rs1410996 in CFH gene,the SNP of rs10490924 in ARMS2 gene and SNP of rs11200638 in HTRA1 gene.The allele distributions of rs800292 (Pallele =0.006,OR =1.643,95 ％ CI:1.155-2.336) in CFH and rs641153 (Pallele =0.002,OR =0.273,95 ％ CI:0.120-0.620) in CFB were significantly associated with AMD.In addition,five SNPs in CFH gene were consisted of two blocks after analysis by Haploview.In addition,five SNPs in CFH were consisted of two blocks after analysis by Haploview.The first one SNPs (including rs551397 and rs800292) and another one SNPs (including rs12124794,rs10737680) and rs1410996 were in strong linkage disequilibrium (D'=1.00).After 50 000 permutations,the GC and AT haplotypes of the first block and the AAC,TCT and ACT haplotypes in the second block were significantly different between AMD patients and controls (P =0.010,0.010,0.001,0.041 and 0.033,respectively).The allel T of rs641153 was a protective factor of AMD (P=0.002,OR =0.273,95％ CI:0.120-0.620).Conclusions The SNPs rs10737680 and rs1410996 in CFH,rs10490924 of ARMS2 gene and rs11200638 of HTRA1 gene are associated with AMD in Ningxia population.

Background Study determined that retinitis pigmentosa has a similar pathogenesis mechanism to Alzheimer disease,and activity of cyclin-dependent kinase 5 (Cdk5) and its activators participates in the degeneration of central nervous system.Roscovitine,an inhibitor of Cdk5,can suppress activity of Cdk5/p25 pathway and therefore inhibit cell apoptosis.However,the influence of roscovitine on retinitis pigmentosa(RP) is unclear.Objective This study was to investigate the expressions of p35,p25 and tau in the retinas of RCS rats.Methods Roscovitine of 4 μl was intravitreally injected in the right eyes of 12 SPF 17-day-old RCS rats,and the fellow eyes were not intervened as the control eyes.The rats were sacrificed on eighth day (postnatal 25 days) and eighteenth day (postnatal 35 days),and whole retinas were isolated to evaluate the relative expressions of Cdk5,p35,p25 and tau phosphorylation by Western blot,and the activity of Cdk5/p25 was analyzed by quantitative colorimetric assay.The results were compared between the right eyes and fellow eyes by paired t test.The use and care of the rats complied with Ethic Statement of Experimental Animal of Ningxia Medical University.Results In the eighth and eighteenth day after injection,the relative expression values (A values) of p35 in rat retinas were 1.186±0.019 and 1.069± 0.019 in the injected eyes,showing significant decreases in comparison with 1.364±0.016 and 1.214±0.008 of the fellow eyes (t =-6.294,-6.477,both at P＜0.05);the relative expression values (A values) of p25 in rat retinas were 0.312±0.009 and 0.269±0.018 in the injected eyes,which was significantly lower than 0.595±0.013 and 0.473±0.011 of the fellow eyes (t=-36.508,-11.879,both at P＜0.05).No significant difference was found in the relative expression of Cdk5 protein between the injected eyes and the fellow eyes in various time points after injection (both at P＞0.05).The activities of Cdk5/p25 were (0.003 83 ±0.000 14) mol/(s · mg) and (0.002 01 ± 0.000 11) mol/(s · mg) in the injected eyes,with significant decreases in comparison with the (0.005 47±0.000 27)mol/(s · mg)and (0.003 35±0.000 15) mol/(s · mg) of the fellow eyes (t=-9.152,P=0.000;t=-9.248,P=0.000),and the tau phosphorylation levels followed the same pattern in the eighth and eighteenth day after injection (t =-9.854,-6.744,both at P＜0.05).Conclusions Intravitreal injection of roscovitine can inhibit the activity of Cdk5/p25 and tau phosphorylation level in retinas of RCS rats to certain extend.

Background Age-related macular degeneration (AMD) is a heritable,progressive degenerative disorder that triggers central visual impairment.Research demonstrated that the single nucleotide polymorphisms (SNPs) of vascular endothelial growth factor 1 (VEGFR1) gene is associated with AMD in different population.However,the results varied among diversified ethnic origin composition and distinct regions.Objective This study was to investigate the associations between the SNPs of VEGFR1 genetic variants along with smoking exposure and the risk of AMD in Hui and Han ethnics in the Ningxia population in China.Methods A case-control study was conducted.Four hundreds and thirty-two AMD patients including 325 Han ethnic patients and 107 Hui ethnic patients were recruited from March 2011 to June 2015,and 906 ethnicity-and gender-matched age-related cataract patients were contemporaneously recruited as control group,including 698 Han ethnic patients and 208 Hui ethnic patients.Periphery blood sample of 5 ml was collected from the subjects and genomic DNA was prepared.Eight tagging SNPs loci were acquired to cover rs2281827,rs3936415,rs7337610,rs7981680,rs9554320,rs9554322,rs9582036 and rs9943922,and the genotypes of SNPs were detected by using MassARRAYTM time-of-flight mass spectrometry system.Chi-square test and multi-factor Logistic regression analysis were utilized to estimate the discrepancy of allele frequency and genotype distribution in Hui and Han AMD patients.Moreover,the correlation of AMD with smoking and age statue were further analyzed.This study protocol complied with Helsinki Declaration and was approved by Ethic Committee of Ningxia Eye Hospital.Written informed consent was obtained before any relevant medical examination.Results There were significant differences in the age between AMD group and control group in both Han and Hui ethnicity (Han:P =0.000;Hui:P =0.009).The smoking exposure was significantly different between AMD group and control group in Han ethnicity (P =0.000),and smoking was the independent risk factor of AMD disease in Han ethnicity of N ingxia region (odds ratio [OR] =2.622,95％ confidence interval [CI]:1.899-3.619).The allele frequencies of SNPs were not significantly different in the AMD patients between Han and Hui ethnicity (all at P＞0.05).However,the allele frequencies and genotype distribution of rs7337610 and rs9554322 SNPs were significantly different between the AMD group and control group in both Han and Hui ethnicity (all at P=0.00).The genotype distribution of rs9582036 and rs9943922 SNPs was significantly different between the AMD group and control group in Han ethnicity (P=0.02,0.00).Allelic G of rs7337610 was the protective factor of AMD disease in Han and Hui ethnieity (OR=0.354,95％ CI:0.288-0.435;OR=0.446,95％ CI:0.315-0.632),while allelic C of rs9554322 was the risk factor of AMD disease in Han and Hui ethnicity (OR=1.671,95％ C1:1.234-2.262;OR=3.661,95％ CI:2.156-6.218).Allelic A of rs9582036 was the risk factor of AMD disease in Han ethnicity (OR =1.477,95％ CI:1.124-1.940).Conclusions Smoking is the independent risk component for Han population with AMD.Of the eight SNPs tagged,the genotypes and alleles of rs9554322 and rs7337610 seems to confer susceptibility to AMD in both Han and Hui ethnicity,the genotypes and alleles of rs9582036 and rs9943922 confer susceptibility to AMD in only Han ethnicity.

According to the onset age, high myopia can be divided into late-onset high myopia (loHM) and early-onset high myopia (eoHM). Numerous genetic studies have shown that eoHM is different from loHM.EoHM, a special type of high myopia before school age (<7 years old), is more likely influenced by genetic factors with less contribution from environment.Therefore, individuals with eoHM are an available group for the research of pathogenesis of high myopia as a monogenic disease, and eoHM should be a unique resource in searching for genes responsible for high myopia.EoHM can be divided into nonsyndromic type which only presents with simple high myopia without any ocular or systemic abnormalities, and syndromic type that has other ocular or systemic disorders.More often, eoHM is the earliest sign of some inherited ocular diseases and the first reason for children seeking medical attention, which is an important clue for clinicians to detect underlying eye diseases.Therefore, in addition to further specific clinical examination of ocular structure and function, high attention should be paid to genetic screening of eoHM in order to promote early diagnosis and effective intervention, and long-term follow-up assessment.

Sequencing technology has evolved rapidly with the advent of high-throughput next-generation sequencing (NGS). By adopting NGS, more and more ophthalmologists and medical institutions are now performing genetic testing for molecular diagnosis and genetic research in genetic ocular disorders.Genetic testing has gradually become an indispensable test item in the diagnosis and treatment of patients with monogenetic ocular diseases and has been accompanied by new challenges in sequence interpretation due to increased complexity.As we know, NGS can detect a large number of the genetic variation data.Without strict standards to distinguish pathogenic variation from many potential functional variations in the human genome, false positive judgments of causality may be accelerated, which may hind the application and promotion of genetic diagnosis in clinical diagnosis as well as the biological understanding of diseases.The American College of Medical Genetics and Genomics (ACMG) has developed guidances for the interpretation of sequence variants and the Chinese Branch of Genetic Counseling has organized some experts in the field of genetic compiled the Chinese version of the ACMG Standards and Guidelines, which is one of the reference bases to help us interpret genetic variation.This article interpreted the ACMG Standards and Guidelines in order to provide reference for Chinese ophthalmologists in the classification of genetic variation, determination of pathogenic variation, application in clinical diagnosis and related genetic research.

Objective:To analyze the genotypes and clinical phenotypes of two families with Hermansky-Pudlak syndrome (HPS).Methods:The method of pedigree investigation was adopted.A Han Chinese HPS family and a Hui Chinese HPS family were enrolled in People's Hospital of Ningxia Hui Autonomous Region from June 2020 to May 2021.Clinical data of two probands and their phenotypically normal parents were collected.Relevant ocular and systemic examinations were carried out.Platelet dense granules in the two probands were observed with an electron microscope.DNA was extracted from peripheral venous blood collected from the subjects.The pathogenic genes were screened by whole exome sequencing.The potential disease-causing variations were analyzed by bioinformatics analysis.Validation and family cosegregation analysis of the pathogenic variations were performed by Sanger sequencing.The relationship between HPS-related gene variations and clinical characteristics was explored.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Ningxia Eye Hospital, People's Hospital of Ningxia Hui Autonomous Region (No.2016018).Written informed consent was obtained from each subject or custodian before any medical examination.Results:The two families were consistent with autosomal recessive inheritance pattern.In family 1 with a family history of consanguineous marriage, the proband had no obvious hypopigmentation on his facial skin, hair, eyebrows and eyelashes.Horizontal nystagmus, exotropia, mild visual impairment, iris atrophy, positive light transmission, orange fundus, pigment loss, macular hypoplasia, prolonged prothrombin time in laboratory examination, and a significant reduction of platelet dense granules by electron microscopy were observed.The proband in family 2 had pale brown hair and eyebrows, severe visual impairment, normal iris pigment, longer thrombin time in laboratory tests, and characteristics similar to those of the proband in family 1.A novel homozygous variant c. 2887G>T (p.E963X) was detected in the HPS3 gene of the proband in family 1.The parents of the proband from family 1 both carried a heterozygous variant c.2887G>T (p.E963X).Compound heterozygous variants were detected in HPS5 gene of the proband in family 2, c.2952-2A>C splicing variation and heterozygous deletion (a 3 144-bp deletion, located in chr11: 18302108-18305251, exon 22).The parents of the proband from family 2 carried a heterozygous variation.The three novel variations were labeled as pathogenic according to the ACMG standards and guidelines. Conclusions:Family 1 is with HPS-3 and family 2 is with HPS-5.There is a certain genotype-phenotype correspondence in the two types of HPS.

Bestrophinopathies are a group of inherited macular dystrophies caused by BEST1 gene mutations including Best vitelliform macular dystrophy, adult-onset vitelliform macular dystrophy, autosomal dominant vitreoretinochoroidopathy and autosomal recessive bestrophinopathy.The main pathological mechanism of bestrophinopathies is that the primary lesion located in the retinal pigment epithelium will affect the photoreceptor cells.The mutations in BEST1 gene not only result in macular lesions, but also potentially affect the development of eyeball, and even cause serious complications such as angle-closure glaucoma and choroidal neovascularization.It is highly heritable and clinically heterogeneous.The same pathogenic mutation site in BEST1 can lead to different clinical phenotypes, which are complex and varied and can bring great confusion to clinicians, causing misdiagnosis and missed diagnosis the disease.This review aimed to summarize and analyze the clinical manifestations of bestrophinopathies and the research progress in BEST1 gene mutations, so as to improve the understanding of clinicians toward this kind of disease and provide reference for clinical practice and future research.