ObjectiveTo observe the correlation between posterior myopic retinoschisis(MRS) and posterior scleral staphyma (PS) in pathological myopia (PM), and to preliminarily explore the influencing factors of MRS.MethodsA retrospective case series study. From November 2016 to November 2019, 38 patients with PM with MRS diagnosed in Henan Eye Hospital & Henan Eye Institute from were included in the study. There were 10 males and 28 females; 13 patients were binocular and 25 patients were monocular. The average age was (49±13) years old. BCVA, retinoscopy optometry, frequency domain OCT, three-dimensional magnetic resonance imaging (3D-MRI) examination and axial length (AL) measurement were performed. According to the frequency domain OCT inspection results, MRS was divided into inner splitting, outer splitting and mixed splitting; based on the 3D-MRI scan results, PS was divided into broad macula, narrow macula,discoid, nasal, subdisc and other types. The correlation between MRS and PS was tested by χ2 test or Fisher exact test.ResultsAmong 60 eyes, 58 eyes (96.77%) of MRS combined with PS. Among them, the wide macula, narrow macula, discoid, nasal, subdisc, and other types were 30 (51.72%), 19 (32.75%), 1 (1.72%), 2 (3.48%), 2 (3.48%) and 4 (6.85%) eyes; inner split, outer split, and mixed split were 10 (17.24%), 24 (41.38%), 24 (41.38%) eyes. Of the 19 eyes with narrow macular PS, MRS involved the fovea in 16 eyes; of the 39 eyes with PS of other forms, MRS involved the fovea in 22 eyes. There was a statistically significant difference between the narrow macular type and other types involving foveal eyes (P=0.044). The correlation between MRS involving the fovea and narrow macular PS was moderate (Cramer's V=0.275). The ages of patients with inner split, outer split, and mixed split were 44±12, 56±10, and 44±13 years, respectively. Patients with inner splitting were younger than those with outer splitting, and those with outer splitting were older than those with inner splitting and mixed splitting. The differences were statistically significant (P=0.010, 0.010, 0.060).ConclusionPM with MRS mostly occur in PS-affected eyes, and mainly macular PS (wide macula, narrow macula).
ObjectiveTo investigate the feasibility of immoribund skin fibroblast cell line derived from Leber′s hereditary optic neuropathy (LHON) patients as a cell model. MethodsA basic research. Two LHON patients and 2 healthy volunteers were recruited from Department of Ophthalmology of Genetic Clinic of Henan Provincial Eye Hospital. The skin tissue of participants was obtained, and the 4 immortalized skin fibroblasts were constructed by SV40 virus infection, including 2 LHON patient cells (LHON-1 and LHON-2 cells) and 2 healthy volunteers cells (NC-1 and NC-2 cells). Mitochondrial morphology in cells was observed by electron microscope. The levels of reactive oxygen species (ROS), nicotinamide adenine dinucleotide-oxidation state (NAD+), nicotinamide adenine dinucleotide-reduction state (NADH) and adenosine triphosphate (ATP) in fibroblasts were detected. Cellular oxygen consumption was measured by seahorse mitochondrial pressure assay. Cell viability was detected using cell counting kit-8 (CCK8). One-way ANOVA was performed to compare the levels of ROS, NAD+, NADH and ATP in LHON and NC cells, as well as basal oxygen consumption, maximal oxygen consumption, ATP-coupled oxygen consumption, and cell viability. ResultsCompared with NC-1 and NC-2, the number of mitochondrial crest in LHON fibroblasts was significantly reduced, indicating abnormal mitochondrial morphology. Biochemical analysis showed that ROS levels in LHON cells increased, but NAD+/NADH and ATP levels decreased, and the oxygen consumption was significantly inhibited, indicating the presence of mitochondrial damage and respiratory dysfunction. The results of CCK-8 detection showed that the survival ability of LHON-1 and LHON-2 cells was worse under stress conditions. ConclusionImmortalized skin fibroblast cell lines from LHON patients presented mitochondrial dysfunction.
ObjectiveTo identify the pathogenic genes and mutations in a family with Usher syndrome type 2.MethodsA three-generation family including 7 individuals was enrolled in this study. There were 2 male patients and 5 unaffected individuals. All participants was underwent related ophthalmologic examination, including best corrected visual acuity, slit-lamp, indirect ophthalmoscopy, electroretinogram (ERG), optical coherence tomography and visual field test. DNA was extracted from 3 ml peripheral venous blood of all participants. A total of 136 hereditary retinal disease target genes were screened and the DNA sequence was performed by Next-generation sequence analysis. Then the suspected mutations compared with databases to identify the suspected mutations, which should be verified with non-affected family members and 100 normal subjects by PCR and Sanger sequence.ResultsThe sequence result showed that 2 patients, the proband and his brother, carried complex heterozygous mutations in the USH2A gene: c.5459T＞C (p.M1820T) in exon 27, c.802G＞A (p.G268R) in exon 5 and c.1190T＞A (p.I397K) in exon 7. The c.5459T＞C and c.1190T＞A mutations in USH2A have not been reported in the literature and database. Although their mother carried c.5459T＞C (p.M1820T) and c.802G＞A (p.G268R), and their father carried c.1190T＞A (p.I397K) heterozygous mutations, the parents did not present phenotype. These mutations were not detected in other normal family members. The result was supported by co-segregation analysis.ConclusionThe heterozygous mutations c.5459T＞C (p.M1820T), c.1190T＞A (p.I397K) and c.802G＞A (p.G268R) in USH2A gene cause Usher syndrome in this family.
ObjectiveTo observe the gene mutations and clinical phenotypes in patients with Usher syndrome type 2 (USH2) and retinitis pigmentosa (RP).Methods From August 2018 to January 2019, 4 patients and 11 normal family members from 3 families of USH2 and RP who visited Henan Eye Hospital were enrolled in the study. Detailed medical history was obtained and visual acuity, fundus color photography, OCT, visual field, full field ERG examination were performed. Among the three families, pedigree 1 was diagnosed with USH2, pedigree 2 and pedigree 3 were diagnosed with RP. The peripheral venous blood of patients and their family members were collected, and the whole genomic DNA was extracted. Targeted capture next generation sequencing analysis was performed on these members, and Sanger sequencing and family co-segregation were verified.ResultsIn the family F1, the proband had symptoms of RP and sensorineural deafness. Sequencing revealed two heterozygous frameshift variants: c.13877-13880 del AGAC (p. Q4626P) in exon 64 and c.798 del T (p. F266L) in exon 5 of USH2A. Both patients of family 2 and 3 showed RP signs without deafness. Two heterozygous variants c.15178T＞C (p. S5060 P) in exon 70 and c.6986C＞A (p. P2329H) in exon 37, and a pathogenic heterozygous variant c.5836C＞T (p. R1946X) in exon 29 of USH2A were identified in family F2. A heterozygous missense variant c.14951C＞T (p. P4984L) in exon 68 and a variant c.11156G＞A (p. R3719H) in exon 57 of USH2A were found in family F3. The results of conservation analysis showed that the corresponding amino acid sites of USH2A p.Q4626P, p.F266L, p.S5060P, p.P2329H and p.P4984L were highly conserved in many species. Among these 7 pathogenic variants detected, M1-M4 and M6 were novel.ConclusionsMutation USH2A gene are the main cause of USH2 and non-syndromic RP. Different variants affect protein translation and synthesis, consequently causing different clinical phenotypes.
ObjectiveTo identify the causative gene in a family affected with Usher syndrome (USH) with retinitis pigmentosa sine pigmento (RPSP) and to analyze the genotype-phenotype correlation.MethodsA retrospective clinical study. A 9-year-old girl with RPSP type 1F USH diagnosed in the ophthalmology clinic of Henan Provincial People's Hospital in November 2019 and her parents were included in the study. The patient had bilateral night blindness for more than 4 years, she suffered from hearing loss 7 years, and is currently binaural sensorineural deafness. The best corrected visual acuity in both eyes was 0.5+. There was showed no obvious pigmentation on the fundus. The visual acuity of the peripheral field of vision decreased. Optical coherence tomography showed that the outer layer of the peripheral retina became thinner and the ellipsoid band disappeared. On electroretinogram examination, the rod and cone system response was severely decreased. The clinical phenotype of the parents of the child were normal. The peripheral venous blood of the child and his parents were extracted, the whole genome DNA was extracted, the custom developed targeted capture kit (PS400) was used, and the next-generation sequencing technology was used to detect genetic mutations. The suspected pathogenic mutation sites were verified by Sanger; co-segregation was performed among family members. The pathogenicity of variants were evaluated according to the interpretation standards and guidelines of sequence variants. Bioinformatics techniques were used to assess the impact of variants on encoded proteins.ResultsThe results of genetic testing showed that the proband detected the PCDH15 gene c.4109dupA (p.K1370fs) (M1), c.17dupA (p.Y6_L7delinsX) (M2) compound heterozygous mutation sites, verified by Sanger sequencing, the mutations were in the family in a state of co-segregation. According to the evaluation of sequence variation interpretation standards and guidelines, M1 and M2 were pathogenic variants of the PCDH15 gene. M1 led to a complete change in the transmembrane structure of the encoded protein, and M2 caused the gene to only translate 6 amino acids, which predicted that the PCDH15 protein cannot be synthesized. According to the clinical phenotype, gene mutation pathogenicity and protein structure prediction, the final clinical diagnosis was PCDH15-related type 1F.ConclusionsPCDH15 genes c.4109dupA and c.17dupA are the pathogenic mutation sites of USH in this family. These compound heterozygous new mutations lead to the failure of normal synthesis of PCDH15 protein, which leads to ocular and ear manifestations.
ObjectiveTo study the research hot spots of ophthalmology-related coronavirus disease 2019 (COVID-19). MethodsPubMed database as the data source, the literatures of ophthalmology-related COVID-19 published on January 1, 2020 to February 22, 2022 were collected, limited to Medline included, the language type was limited to English and Chinese, and 1 592 literatures were included. By reading the titles and abstracts, the literatures of meeting notice, editor's note, etc. and the literature that was not quite relevant with ophthalmology-related COVID-19 were removed, and finally 1 547 literatures were included. Bibliographic Items Co-occurrence Matrix Builder (BICOMB 2.02 software) was used to collect the frequency of major Mesh terms/subheadings and the frequency of major Mesh terms after removing the subheadings, and the number of included articles published in the top 10 journals by the number of ophthalmology-related COVID-19 articles was recorded. VosViewer 1.6.18 software was used for cluster analysis of collaborator network and major Mesh terms, and the publication status and country or region distribution of active authors of ophthalmology-related COVID-19 were recorded. ResultsOf the 1 547 literatures, the active authors were mainly from India, Italy, Singapore, Spain, and Hong Kong, China, and so on; the top 10 journals published 617 articles in total (39.88%, 617/1 547). The high frequency major Mesh terms/subheadings included COVID-19, viral pneumonia, coronavirus infection, eye diseases/epidemiology, complications, prevention & control, diagnosis, virology, and Severe acute respiratory syndrome coronavirus 2, betacoronavirus/isolation & purification, ophthalmology/education, organization & administration, telemedicine, delivery of health care/organization & administration, and mucormycosis/diagnosis, etc. After taking out the subheadings, the high frequency of major Mesh terms also included conjunctivitis, orbital disease, retinal diseases, neuromyelitis optica, retinal vein occlusion, myopia and other eye diseases, eye diseases-related systemic diseases, such as multiple sclerosis and Miller Fisher syndrome, therapy and prevention-related drugs, such as hydroxyl chloroquine, angiogenesis inhibitors, and vaccination. ConclusionsOphthalmology-related COVID-19 researches have received extensive attention worldwide, COVID-19 is associated with multiple ocular diseases of anterior and posterior segments. COVID-19-related mucormycosis, hydroxychloroquine and possible retinal toxicity, and possible ocular adverse effects associated with vaccination are also noteworthy.
ObjectiveTo compare the quantitative measurements of the retinal capillary nonperfusion areas in a cohort of proliferative diabetic retinopathy (PDR) patients with fluorescein fundus angiography (FFA) and swept source optical coherence tomography angiography (SS-OCTA), and to determine the intrapersonal variability between examiners.MethodsA cross-sectional study. Eighteen eyes of eleven PDR patients diagnosed in Department of ophthalmology of Henan Provincial People's Hospital from September 2019 to January 2020 were included in this study. FFA was performed using Spectralis HRA+OCT (Germany Heidelberg Company) from and SS-OCTA was performed using VG200D (China Vision Micro Image Corporation). SS-OCTA was used to collect images of retinal layer, superficial capillary plexus (SCP) and deep capillary plexus (DCP). The same observation area was 80°×60° for SS-OCTA and 55° for FFA with both setting centered on the fovea. The forty-nine retinal capillary nonperfusion areas were observed. The area measurement was completed independently by three examiners. Paired sample t test or paired sample Wilcoxon test were used to compare the measured values of retinal capillary nonperfusion areas between the two examination methods and among the three examiners.ResultsThere was no significant difference in the retinal layer, SCP and DCP nonperfusion area measured by FFA and SS-OCTA among the three examiners (P＞0.05), and the consistency is good (consistency correlation coefficient＞0.9, P＜0.05). The nonperfusion area measured by FFA was 0.786 mm2. The median nonperfusion area of retinal layer and SCP measured by SS-OCTA were 0.787 mm2 and 0.791 mm2, respectively, and the average nonperfusion area of DCP was 0.878±0.366 mm2. The nonperfusion area of retinal layer and SCP measured by FFA and SS-OCTA showed no statistically significant difference (P=0.054, 0.198). The nonperfusion area of DCP measured by SS-OCTA was significantly larger than that of FFA, and the difference was statistically significant (P＜0.001). The results of repeatability analysis showed that 93.88% (46/49) of the DCP nonperfusion area data measured by SS-OCTA were greater than those measured by FFA.ConclusionThe retinal nonperfusion area of DCP in PDR patients measured by SS-OCTA is larger than that of FFA.
ObjectiveTo study the efficiency and difference of the artificial intelligence (AI) system based on fundus-reading in community and hospital scenarios in screening/diagnosing diabetic retinopathy (DR) among aged population, and further evaluate its application value. MethodsA combination of retrospective and prospective study. The clinical data of 1 608 elderly patients with diabetes were continuously treated in Henan Eye Hospital & Henan Eye Institute from July 2018 to March 2021, were collected. Among them, there were 659 males and 949 females; median age was 64 years old. From December 2018 to April 2019, 496 elderly diabetes patients were prospectively recruited in the community. Among them, there were 202 males and 294 female; median age was 62 years old. An ophthalmologist or a trained endocrinologist performed a non-mydriatic fundus color photographic examination in both eyes, and a 45° frontal radiograph was taken with the central fovea as the central posterior pole. The AI system was developed based on the deep learning YOLO source code, AI system based on the deep learning algorithm was applied in final diagnosis reporting by the "AI+manual-check" method. The diagnosis of DR were classified into 0-4 stage. The 2-4 stage patients were classified into referral DR group. ResultsA total of 1 989 cases (94.5%, 1 989/2 104) were read by AI, of which 437 (88.1%, 437/496) and 1 552 (96.5%, 1 552/1 608) from the community and hospital, respectively. The reading rate of AI films from community sources was lower than that from hospital sources, and the difference was statistically significant (χ2=51.612, P<0.001). The main reasons for poor image quality in the community were small pupil (47.1%, 24/51), cataract (19.6%, 10/51), and cataract combined with small pupil (21.6%, 11/51). The total negative rate of DR was 62.4% (1 241/1 989); among them, the community and hospital sources were 84.2% and 56.3%, respectively, and the AI diagnosis negative rate of community source was higher than that of hospital, and the difference was statistically significant (χ2=113.108, P<0.001). AI diagnosis required referral to DR 20.2% (401/1 989). Among them, community and hospital sources were 6.4% and 24.0%, respectively. The rate of referral for DR for AI diagnosis from community sources was lower than that of hospitals, and the difference was statistically significant (χ2=65.655, P<0.001). There was a statistically significant difference in the composition ratio of patients with different stages of DR diagnosed by AI from different sources (χ2=13.435, P=0.001). Among them, community-derived patients were mainly DR without referral (52.2%, 36/69); hospital-derived patients were mainly DR requiring referral (54.9%, 373/679), and the detection rate of treated DR was higher (14.3%). The first rank of the order of the fundus lesions number automatically identified by AI was drusen (68.4%) and intraretinal hemorrhage (48.5%) in the communities and hospitals respectively. Conclusions It is more suitable for early and negative DR screening for its high non-referral DR detection rate in the community. Whilst referral DR were mainly found in hospital scenario.
ObjectiveThe clinical phenotypes and pathogenicity of isolated cone-rod dystrophy (CORD) caused by two novel complex heterozygous variants of the CEP290 gene were analyzed using high-resolution multi-mode imaging and gene detection techniques. MethodsA retrospective study. Two patients and two family members from a CORD family who were diagnosed by genetic testing at Henan Provincial People's Hospital in December 2021 were included in the study. All subjects underwent best-corrected visual acuity (BCVA), color fundus photography, autofluorescence, swept-source optical coherence tomography (SS-OCT), adaptive optics fundus imaging, static threshold field, full field and multiple electroretinogram (ERG) examination, as well as other systemic examinations throughout the body. The peripheral venous blood of the subjects was collected, and the whole genome DNA was extracted. DNA sequencing was performed using the Inherited Retinal Disease Kit PS400, and Sanger verification and pedigree co-segregation analysis were performed on the suspected pathogenic mutation sites. Validation was performed by Sanger sequencing, pathogenicity analysis was performed in accordance with the American College of Medical Genetics and Genomics (ACMG) guidelines. Conservation of variation among different species was analyzed by GERP++, Clustal Omega and Weblogo. ResultsBoth patients were male, and their ages were 21 and 29 years old, respectively. The right eye and left eye about BCVAs were 0.7, 0.4 and 0.3, 0.4, respectively. The full field and multiple electroretinogram ERG showed a decreased function of cones and rods, especially cones. SS-OCT showed thinning of the outer nuclear layer of macular, and attenuation of ellipsoid zone reflectivity in B-scan. Adaptive optics fundus imaging examination showed that the arrangement of cone cells in the fovea of the fovea was disordered and the density decreased, and the retinal pigment epithelial cells were seen through the atrophy of cone cells in some areas at 10°visual angle. No obvious abnormality was found in other systemic examinations of the whole body. Genetic testing showed that 2 novel compound heterozygous variants c.950T >A (p.Leu317*) (M1) and c.4144_4149del (p.Tyr1382_Glu1383del) (M2) in CEP290 were found in two patients. The first variant was predicted to be harmful in MutationTaster and CADD. GERP++ showed highly conserved among different species. The pathogenicity of the variant was suspected to be likely pathogenic according to ACMG guidelines. The pathogenicity of the second variant was uncertain significance. The parents of the proband had no similar ocular abnormalities. Verified by Sanger sequencing, it was consistent with co-separation in the family. ConclusionsPatients with pure CORD caused by CEP290 gene mutation still retain better vision when the cone structure is abnormal, the density is decreased, and the function of cone and rod cells is decreased. CEP290 M1 and M2 are newly discovered nonsense mutations and newly discovered deletion mutations, which expanded the causative gene spectrum of pure CORD.
ObjectiveTo observe the correlation between retinal capillary density and retinal thickness in the macula and spherical equivalent (SE) in school-age children. MethodsA cross-sectional study. From May to December 2022, 182 school-age children who visited the ophthalmology department of the First Affiliated Hospital of Zhengzhou University were included . There were 95 males and 87 females. The age ranged from 6 to 12 years, and the spherical equivalent (SE) was +0.50 to －6.00 D. They were divided into three groups based on the SE of the right eyes: 54 eyes in emmetropia group (+0.50≤SE＜－0.50 D), 71 eyes in low myopia group (－0.50≤SE＜－3.00 D), and 57 eyes in moderate myopia group (－3.00≤SE≤－6.00 D). The macular area of 6 mm×6 mm was scanned using swept-source optical coherence tomography angiography and was divided into three concentric rings centered on the fovea, including the macular central fovea (0-1 mm diameter), inner ring (1-3 mm diameter) and outer ring (3-6 mm diameter). The retinal thickness and blood flow density of superficial vascular plexus (SVP) and deep vascular plexus (DVP) in different zones within 6 mm of the macular area were measured. The relationships between SE and SVP, DVP and retinal thickness in each ring region were investigated by univariate and multivariate linear regression analyses, smooth curve fitting, and threshold effects. ResultsThere were significant differences in the SVP (F=6.64, 26.06, 22.69) and DVP (F=7.97, 25.01, 5.09) of macular central fovea, inner ring and outer ring among the emmetropia, low myopia and moderate myopia groups (P＜0.05). Univariate linear regression analysis showed that the SVP (β=－0.56, －1.17, －0.79) and DVP (β=－1.03, －0.93, －0.45) of the three regions were positively correlated with SE (P＜0.05). After smooth curve fitting, threshold effect analysis and multivariate linear regression analysis, the SVP and DVP in the macular central fovea were linearly positively correlated with SE (β=－0.91, －1.40; P＜0.05), and SVP and DVP in the inner ring and outer ring showed an inverted U-shaped curve relationship with SE with the inflection (＜3.00 D). When the SE was less than ＜3.00 D, the SVP and DVP in the inner ring and outer ring were positively correlated with SE (P＜0.05). When the SE was higher than －3.00 D, except for the DVP in the inner ring region, the other parameters were negatively correlated with SE (P＜0.05). There were significant differences in retinal thickness of the inner ring and outer ring (F=5.47, 16.36; P＜0.05), and no significant difference in the macular central fovea among the emmetropia, low and moderate myopia groups (F=2.16, P＞0.05). By using univariate and multivariate linear regression analyses, the retinal thickness in the inner ring and outer ring were negatively correlated with SE (β =1.99, 3.05; P＜0.05). However, no correlation was found between retinal thickness and SE in the macular central fovea (β=－1.65, P＞0.05). ConclusionsIn school-age children with SE between +0.50 D and －6.00 D, the retinal capillaries density of the macular central fovea gradually increase, and increase first and then decrease in the inner ring and outer ring with increasing SE. The retinal thickness of inner ring and outer ring gradually decrease and not change significantly in the macular central fovea.