Identification of a Novel Keratin 9 Missense Mutation in a Chinese Family with Epidermolytic Palmoplantar Keratoderma.

BACKGROUND/AIMS: Epidermolytic palmoplantar keratoderma (EPPK) is an autosomal dominant genodermatosis. It is characterized by diffuse yellow keratoses on the palmoplantar epidermis, with an erythematous border. The keratin 9 gene (KRT9) and less frequently the keratin 1 gene (KRT1) are responsible for EPPK. This study aims to identify and analyse genetic defects responsible for EPPK in a Han Chinese pedigree. METHODS: A four-generation Han Chinese pedigree containing five individuals affected with EPPK was recruited. Exome sequencing, Sanger sequencing, and bioinformatics tools were conducted to identify the mutation in this pedigree. HaCaT cells were transfected with either wild-type or mutated KRT9. Confocal laser immunofluorescence assay, imaging processing, and statistical analysis were performed to evaluate wild-type and mutant KRT9 groups. RESULTS: A novel heterozygous c.1369C>T transition (p.Leu457Phe) in exon 6 of the KRT9 gene was identified in four patients. It co-segregated with the disorder in the family. Functional analysis showed that withdrawal of the filament network from the cell periphery and particle formation were present in about 10% of Leu457Phe-transfected HaCaT cells, while approximately 3% of cells transfected with wild-type KRT9 showed this phenotype. The particles in mutant group were larger than that in wild-type group (P-value < 0.05). CONCLUSION: The variant may be the disease-causing missense mutation and produce dominant negative effects by interrupting keratin network formation. This study indicates the pathogenic role of the KRT9 gene mutation in this pedigree with EPPK, and may be helpful in genetic counseling, prenatal diagnosis and gene-targeted therapies of EPPK.

Identification of an ND4 Mutation in Leber Hereditary Optic Neuropathy.

SIGNIFICANCE: We identified a missense mutation, m.11778G>A (p.R340H), in the mitochondrially encoded NADH dehydrogenase 4 gene (ND4) in eight patients and three asymptomatic carriers, even though the incidence of this has been considered low in Chinese population. These results have implications for the families' genetic counseling and clinical management. PURPOSE: Leber hereditary optic neuropathy (LHON OMIM 535000) is one of the most common inherited optic neuropathies. The aim of this study was to identify the genetic cause in two Han Chinese families with LHON. METHODS: We used Sanger sequencing to identify the genetic cause of two Han Chinese families from Hunan, China, with LHON. RESULTS: The patients in these two families presented with typical LHON, with male patients experiencing more severe phenotypes. A missense mutation, m.11778G>A (p.R340H), in the ND4 gene was identified in eight patients and three asymptomatic carriers, even though the incidence of this has been considered low in Chinese population. CONCLUSIONS: Eight of 11 family members (72.7%) manifested some vision loss, which is far higher percentage than reported in other studies. The variant is predicted to be the disease-causing mutation and results in seriously abnormal function of complex I subunits of the mitochondrial respiratory chain. These results have implications for the families' genetic counseling and clinical management and help to develop new LHON target-gene therapy strategies.

Identification of a novel GJA3 mutation in congenital nuclear cataract.

PURPOSE: Congenital cataract is a visual impairment that needs correction as early as possible after birth. This study aimed to identify whether genetic defects exist in a Chinese Han pedigree with congenital nuclear cataract. METHODS: A family consisting of six members and three patients with nuclear cataract spanning three generations and 100 unrelated ethnically matched normal subjects were recruited in this study. Exome sequencing was performed in the 24-year-old proband, and Sanger sequencing was then conducted in other family members and 100 normal controls. RESULTS: A novel missense variant, c.428G>A (p.G143E), in the gap junction protein-alpha 3 gene (GJA3) was identified in three patients of the family but unidentified in three family members without lens opacity and 100 normal controls. CONCLUSIONS: A novel missense mutation, c.428G>A (p.G143E), in the GJA3 gene, localized to the cytoplasmic loop, was suggested to be the genetic cause of congenital nuclear cataract, which further expands the gene mutation spectrum. Our findings suggest that exome sequencing is a powerful and cost-effective tool to discover mutation(s) in disorders with high genetic and clinical heterogeneity. Further functional studies in the GJA3 gene mutations may help uncover pathogenic mechanisms of congenital cataract and therefore provide a possible genetic therapy for this disorder.

Identification of a GJA3 mutation in a Chinese family with congenital nuclear cataract using exome sequencing.

Congenital cataract, a clinically and genetically heterogeneous lens disorder is defined as any opacity of the lens presented from birth and is responsible for approximately 10% of worldwide childhood poor vision or blindness. To identify the genetic defect responsible for congenital nuclear cataract in a four-generation Chinese Han family, exome and direct Sanger sequencings were conducted and a missense variant c.139G>A (p.D47N) in the gap junction protein-alpha 3 gene (GJA3) was identified. The variant co-segregated with patients of the family and was not observed in unaffected family members or normal controls. The above findings indicated that the variant was a pathogenic mutation. The mutation p.D47N was found in the first extracellular loop (El) domain of GJA3 protein. Our data suggest that exome sequencing is a powerful tool to discover mutation(s) in cataract, a disorder with high genetic heterogeneity. Our findings may also provide new insights into the cause and diagnosis of congenital nuclear cataract and have implications for genetic counseling.

[A retrospective study on the progression of myopia in school-age children].

OBJECTIVE: To study the progression of myopia in school-age children over the past 12 years and factors influencing myopia progression. METHODS: A total of 4569 cases of 5 to 12-year-old children who had refractive examinations in the Third Xiangya Hospital, Central South University between January 2000 and December 2011 were enrolled in this study. The children had no family history of congenital high myopia or other eye diseases. Myopia progression was evaluated when the children were re-examined. The refractive state of each child was measured with cyclopiegic retinoscopy. RESULTS: The mean spherical equivalent (SE) myopia was-2.0±1.7 D between January 2000 and December 2011. There was no statistical difference in yearly myopia progression between different years. The average age of the myopic children decreased from 10.1 in 2000 to 8.9 years old in 2011 (P<0.05). Mean myopia progression was -0.6±0.7 D per year from 2000 to 2011. Myopia progression reduced gradually in 5 to 8-year-olds (P<0.05), however, it accelerated between ages 9 and 11 years. Myopia progression in 10- and 11-year-olds was significantly greater than in 7- and 8-year-olds (P<0.01). The multiple linear regression analysis demonstrated that age and baseline myopic refraction were positively related to myopia progression. CONCLUSIONS: There was no obvious change in the yearly myopia progression of the children over the past 12 years. The mean age of myopia occurrence became younger with time. More preventive measures are needed to ward off high myopia in children with moderate myopia, especially those aged over 10 years.

Path planning of a manipulator based on an improved P_RRT* algorithm.

Aiming to build upon the slow convergence speed and low search efficiency of the potential function-based rapidly exploring random tree star (RRT*) algorithm (P_RRT*), this paper proposes a path planning method for manipulators with an improved P_RRT* algorithm (defined as improved P_RRT*), which is used to solve the path planning problem for manipulators in three-dimensional space. This method first adopts a random sampling method based on a potential function. Second, based on a probability value, the nearest neighbour node is selected by the nearest Euclidean distance to the random sampling point and the minimum cost function, and in the expansion of new nodes, twice expansion methods are used to accelerate the search efficiency of the algorithm. The first expansion adopts the goal-biased expansion strategy, and the second expansion adopts the strategy of random sampling in a rectangular area. Then, the parent node of the new node is reselected, and the path is rerouted to obtain a clear path from the initial point to the target point. Redundant node deletion and the maximum curvature constraint are used to remove redundant nodes and minimize the curvature on the generated path to reduce the tortuosity of the path. The Bezier curve is used to fit the processed path and obtain the trajectory planning curve for the manipulator. Finally, the improved P_RRT* algorithm is verified experimentally in Python and the Robot Operating System (ROS) and compared with other algorithms. The experimental results verify the effectiveness and superiority of the improved algorithm.

[Influence of near-work and outdoor activities on myopia progression in school children].

OBJECTIVE: To study the influence of near-work and outdoor activities on myopia progression in school children. METHODS: Eighty 7-11-year-old school children with myopia were randomly assigned into an intervention group (n=41) and a control group (n=39). The children in the intervention group did near- and middle-vision activities less than 30 hrs per week and more outdoor activities than 14-15 hrs per week. Myopia progression was observed regularly over 2 years after which ophthalmologists administered questionnaires regarding near-vision work (reading, writing and using computer), middle-vision work (watching TV and extracurricular learning activities), outdoor activities, using nature light, wearing glasses, etc. RESULTS: The annual mean myopia progression (0.38 ± 0.15 D) in the intervention group was significantly lower than that in the control group (0.52 ± 0.19 D; P<0.01). The children in the two groups spent similar amounts of time in near-vision activities, but the children in the intervention group spent less time in middle-vision activities (P<0.01) and more outdoor activities (13.7 ± 2.4 vs 6.2 ± 1.6 hrs/wk; P<0.01). When considering all children in the study, there were 4 factors that significantly correlated with less myopia progression: more outdoor activities, more time spent wearing glasses, more time spent in natural light and less time using a computer. When analyzing the intervention group separately, more outdoor activity was inversely correlated with myopia progression (t=-2.510, P<0.05). Separate analysis of the control group indicated that more time wearing glasses was correlated with less myopia progression (t=-3.115, P<0.05). CONCLUSIONS: Myopia progression in school children may be slowed by more outdoor activities, more time spent in natural light and more time wearing corrective glasses.

Fabrication and characterization of ZnTiO3/Zn2Ti3O8/ZnO ternary photocatalyst for synergetic removal of aqueous organic pollutants and Cr(VI) ions.

Highly efficient photocatalysts have great development prospects in wastewater treatment, especially in the degradation of organic pollutants and reduction of inorganic heavy metal ions. Herein, a Z-scheme ZnTiO3/Zn2Ti3O8/ZnO ternary photocatalyst was prepared by the solvothermal-calcination method and the influence of the content of tetrabutyl titanate precursor and different reaction temperature on the crystal phase structures, photoelectrochemical properties and photocatalytic activities of the samples were investigated. Due to its unique Z-scheme structure and suitable band gap position, which is favorable for the efficient migration and separation of photo-generated electrons and holes and the improvement of photocatalytic redox reaction capability, the samples show excellent performance for the degradation of organic pollutants and reduction of heavy metal Cr(VI) ions. Based on a series of characterization analyses, a possible Z-scheme photocatalytic mechanism is proposed. This work provides a simple preparation method for fabrication of multivariate heterojunction photocatalyst for degradation of organic pollutants and removal of heavy metal ions.

Heterozygous RHO p.R135W missense mutation in a large Han-Chinese family with retinitis pigmentosa and different refractive errors.

Retinitis pigmentosa (RP), the most common type of inherited retinal degeneration causing blindness, initially manifests as severely impaired rod function followed by deteriorating cone function. Mutations in the rhodopsin gene (RHO) are the most common cause of autosomal dominant RP (adRP). The present study aims to identify the disease-causing mutation in a numerous, four-generation Han-Chinese family with adRP detected by whole exome sequencing and Sanger sequencing. Afflicted family members present classic adRP along with heterogeneous clinical phenotypes including differing refractive errors, cataracts, astigmatism and epiretinal membranes. A missense mutation, c.403C>T (p.R135W), in the RHO gene was identified in nine subjects and it co-segregated with family members. The mutation is predicted to be disease-causing and results in rhodopsin protein abnormalities. The present study extends the genotype-phenotype relationship between RHO gene mutations and adRP clinical findings. The results have implications for familial genetic counseling, clinical management and developing RP target gene therapy strategies.

Identification of a Novel Mutation in the ABCA4 Gene in a Chinese Family with Retinitis Pigmentosa Using Exome Sequencing.

Retinitis pigmentosa (RP) is a group of hereditary, degenerative retinal disorders characterized by progressive retinal dysfunction, outer retina cell loss, and retinal tissue atrophy. It eventually leads to tunnel vision and legal, or total blindness. Here we aimed to reveal the causal gene and mutation contributing to the development of autosomal recessive RP (arRP) in a consanguineous family. A novel homozygous mutation, c.4845delT (p.K1616Rfs*46), in the ATP-binding cassette subfamily A member 4gene ( ABCA4 ) was identified. It may reduce ABCA4 protein activity, leading to progressive degeneration of both rod and cone photoreceptors. The study extends the arRP genotypic spectrum and confirms a genotype-phenotype relationship. This study may also disclose some new clues for RP genetic causes and pathogenesis, as well as clinical and genetic diagnosis. The research findings may contribute to improvement in clinical care, therapy, genetic screening, and counseling.

Identification of a missense mutation in the tyrosinase gene in a Chinese family with oculocutaneous albinism type 1.

Oculocutaneous albinism (OCA) is a group of heterogeneous and autosomal recessive disorders characterized by a reduction or complete loss of melanin biosynthesis in melanocytes. OCA type 1 (OCA1) is the most severe and common form of OCA, and is caused by mutations in the tyrosinase gene (TYR). The present study aimed to identify the genetic cause of OCA1 in a four­generation consanguineous Chinese Han family. Complete physical examinations were performed and blood samples were collected from five members of the family and 100 unrelated healthy controls. Exome sequencing was conducted in the proband, followed by verification in other family members, using Sanger sequencing. Patients in the family presented with typical OCA1 features, including hypopigmentation of the skin and hair, and distinctive ocular changes. A homozygous missense variant, c.896G>A (p.R299H), in the TYR gene was identified in two patients, which co­segregated with disease in the family. This variant was not present in the 100 healthy controls. These results expand the number of mutations identified to be responsible for OCA1 in the Chinese Han population, and may have implications for genetic counseling and clinical management of the disease.

A homozygous MYO7A mutation associated to Usher syndrome and unilateral auditory neuropathy spectrum disorder.

Usher syndrome (USH) is an autosomal recessive disorder characterized by sensorineural hearing loss, progressive visual loss and night blindness due to retinitis pigmentosa (RP), with or without vestibular dysfunction. The purpose of this study was to detect the causative gene in a consanguineous Chinese family with USH. A c.3696_3706del (p.R1232Sfs*72) variant in the myosin VIIa gene (MYO7A) was identified in the homozygous state by exome sequencing. The co­segregation of the MYO7A c.3696_3706del variant with the phenotype of deafness and progressive visual loss in the USH family was confirmed by Sanger sequencing. The variant was absent in 200 healthy controls. Therefore, the c.3696_3706del variant may disrupt the interaction between myosin VIIa and other USH1 proteins, and impair melanosome transport in retinal pigment epithelial cells. Notably, bilateral auditory brainstem responses were absent in two patients of the USH family, while distortion product otoacoustic emissions were elicited in the right ears of the two patients, consistent with clinical diagnosis of unilateral auditory neuropathy spectrum disorder. These data suggested that the homozygous c.3696_3706del variant in the MYO7A gene may be the disease­causing mutation for the disorder in this family. These findings broaden the phenotype spectrum of the MYO7A gene, and may facilitate understanding of the molecular pathogenesis of the disease, and genetic counseling for the family.

AgI/TiO2 nanobelts monolithic catalyst with enhanced visible light photocatalytic activity.

AgI nanoparticles (NPs) have been decorated on the TiO2 nanobelts (NBs) immobilized on a metal Ti substrate by a simple impregnating-precipitation method. The as-achieved AgI/TiO2 monolithic catalyst exhibits a high and stable visible photocatalytic activity toward acid orange II (AO-II) degradation, which is attributed to the suitable energy band match of AgI NPs and TiO2 NBs, leading to the efficient transfer of photo-generated electrons. In addition, it was found that ·O2(-) radicals and h(+) are the main reactive species for the degradation of AO-II under visible light irradiation. A reasonable photocatalytic mechanism of AgI/TiO2 photocatalyst toward AO-II degradation was discussed. This monolithic catalyst provides an advantage over the drawback encountered with powder suspension.

Clinical description and genome wide linkage study of Y-sutural cataract and myopia in a Chinese family.

PURPOSE: To describe the clinical characteristics of a Y-sutural cataract associated with myopia in a large Chinese family and to identify the causative gene and mutation. METHODS: An autosomal dominant Y-sutural cataract and myopia were identified in members of a large family of Han ethnicity living in southern China. Ophthalmological examinations were performed and a medical history was taken. Blood samples were collected for DNA isolation. A genome wide scan was performed using markers spaced at about 10 cM intervals for genotyping and two point linkage analysis. Candidate genes were sequenced. RESULTS: Bilateral lens opacities, the only sign of cataract in early childhood and the most prominent sign in all affected individuals, involved the entire anterior Y and posterior inverted Y sutures, showing a feather duster like appearance. The Y-sutural cataract in this family mapped to an 11.4 cM (13.5 Mb) region between D3S3606 and D3S1309 on chromosome 3q22 with a maximum lod score of 5.7 at theta=0 for D3S1292. Sequence analysis of the beaded filament structural protein 2 (BFSP2) gene identified a previously described c.697_699delGAA (E233del) mutation which was present in all individuals with Y-sutural cataract but not in unaffected individuals and controls. Myopia, observed in 10 out of 12 cataract patients and significantly higher than that in unaffected offspring and siblings (1 out of 8), was independently mapped to a 61.2 cM (59 Mb) region between D3S3606 and D3S1262 on 3q21.3-q27.2 with maximum lod score of 3.79. CONCLUSIONS: This Y-sutural cataract is caused by an E233del mutation in BFSP2 which provides additional evidence supporting mutations in BFSP2 as a cause for cataract and demonstrates phenotypic variability in cataracts caused by BFSP2. The Y-sutural opacity in the lens might be the typical and earliest sign for cataract caused by the BFSP2 mutation. In addition, these results demonstrate a myopia susceptibility locus in this region, which might also be associated with the mutation in BFSP2.

Heterogeneous phenotype in a family with the FERM domain-containing 7 gene R335X mutation.

OBJECTIVE: Infantile nystagmus (IN) is characterized by bilateral involuntary, periodic, and predominantly ocular oscillations. In this article, we describe a mutation screen conducted on a 4-generation family in which 4 patients were affected with X-linked IN (XLIN). DESIGN: Experimental study. PARTICIPANTS: A 4-generation Chinese Han family including 4 symptomatic members with IN and 200 normal male controls. METHODS: DNA was extracted from peripheral blood, and the FERM domain-containing 7 gene (FRMD7) was amplified on DNA samples of all the available family members. The mutation screen was conducted by performing direct DNA sequencing. RESULTS: A nonsense mutation (R335X) in the FRMD7 gene was identified in 4 male patients and an asymptomatic female member. CONCLUSIONS: Although the R335X mutation in the FRMD7 gene has been previously described, the clinical features, including both disease penetrance and severity, among individuals with FRMD7 mutation in our family vary greatly. One female member with the heterozygous R335X mutation had no clinical manifestation of the disease. This incomplete penetrance suggests that random X-chromosome inactivation may play a role in the pathogenesis of IN, and that loss of functional FRMD7 may account for the development of this disorder. Our findings may be helpful in the genetic counseling of patients with nystagmus.

Novel RS1 mutations associated with X-linked juvenile retinoschisis.

To identify mutations in the retinoschisin (RS1) gene in families with X-linked retinoschisis (XLRS). Twenty families with XLRS were enrolled in this study. All six coding exons and adjacent intronic regions of RS1 were amplified by polymerase chain reaction (PCR). The nucleotide sequences of the amplicons were determined by Sanger sequencing. Ten hemizygous mutations in RS1 were detected in patients from 14 of the 20 families. Four of the ten mutations were novel, including c:176G>A (p:Cys59Tyr) in exon 3, c:531T>G (p:Tyr177X), c:607C>G (p:Pro203Ala) and c:668G>A (p:Cys223Tyr) in exon 6. These four novel mutations were not present in 176 normal individuals. The remaining six were recurrent mutations, including c:214G>A (p:Glu72Lys), c:304C>T (p:Arg102Trp), c:436G>A (p:Glu146Lys), c:544C>T (p:Arg182Cys), c:599G>A (p:Arg200His) and c:644A>T (p:Glu215Val). Our study expanded the mutation spectrum of RS1 and enriches our understanding of the molecular basis of XLRS.

Evaluation of the ELOVL4, PRPH2 and ABCA4 genes in patients with Stargardt macular degeneration.

Mutations in the ATP-binding cassette, subfamily A, member 4 (ABCA4), elongation of very long chain fatty acids 4 (ELOVL4) and peripherin-2 (PRPH2) genes have been identified in patients with Stargardt macular degeneration (STGD). The aim of this study was to investigate which of these genes is responsible for susceptibility in Chinese patients. A total of 41 probands with STGD or suspected STGD were enrolled in the study. The coding regions and adjacent intronic sequences of the ELOVL4 and PRPH2 genes and 3 coding exons of the ABCA4 gene were amplified by polymerase chain reaction (PCR). The nucleotide sequences of the amplicons were determined by Sanger sequencing. Three novel heterozygous missense mutations in the ABCA4 gene were identified: c:2633C>A (p:Ser878X), c:5646G>A (p:Met1882Ile) and c:6389T>A (p:Met2130Lys). These mutations were not present in 176 normal individuals and were predicted to be pathogenic. Two benign variations were found: a reported variation, c:5682G>C in ABCA4 and a novel variation, c:699G>A in ELOVL4. In addition, 5 single nucleotide polymorphisms (SNPs: rs3812153, rs7764439, rs390659, rs434102 and c:929G>A) were detected in ELOVL4 and PRPH2. The c:929G>A variation has not been previously reported. We conclude that no pathogenic variations in ELOVL4 and PRPH2 were detected in the Chinese STGD patients. Our results imply that ABCA4 is more likely to be significant in Chinese STGD patients.

Novel SOX2 mutation associated with ocular coloboma in a Chinese family.

OBJECTIVES: To report a novel SOX2 (OMIM 184429) mutation in a Chinese family and to describe its ocular and extraocular clinical features. METHODS: Ocular and systemic examinations were performed, and genomic DNA was prepared from peripheral leukocytes. The coding exons and the adjacent intronic sequence of SOX2 were analyzed by cycle sequencing. RESULTS: A novel heterozygous c.695C>A (p.Thr232Asn) mutation in SOX2 was identified in a Chinese family in which both the father and the son had iris and chorioretinal uveal colobomas. In addition, cataracts were noted in the father but not in the son. Other anomalies were not found in the father but were present in the son, including brain arachnoid cyst, microcornea, retrobulbar colobomatous orbital cyst, and penoscrotal hypospadias. This mutation was not detected in the unaffected mother and 103 unaffected control individuals. CONCLUSIONS: Mutation in SOX2 is associated with typical ocular coloboma and probably other anomalies in this Chinese family. Arachnoid cyst has not been reported in individuals with the SOX2 mutation. CLINICAL RELEVANCE: The results remind us that ocular coloboma may be accompanied by arachnoid cyst and may be associated with SOX2 mutation, which will be helpful for improving diagnosis and patient care.