Variant in EZR leads to defects in lens development.

BACKGROUND: Congenital cataract is a common cause of blindness. Genetic factors always play important role. MATERIAL AND METHODS: This study identified a novel missense variant (c.1412C>T (p.P471L)) in the EZR gene in a four-generation Chinese family with nuclear cataract by linkage analysis and whole-exome sequencing. A knockout study in zebrafish using transcription activator-like effector nucleases was carried out to gain insight into candidate gene function. RESULTS: Conservative and functional prediction suggests that the P-to-L substitution may impair the function of the human ezrin protein. Histology showed developmental delays in the ezrin-mutated zebrafish, manifesting as multilayered lens epithelial cells. Immunohistochemistry revealed abnormal proliferation patterns in mutant fish. CONCLUSIONS: The study suggests that ezrin may be involved in the enucleation and differentiation of lens epithelial cells.

Glucose transporter 3 (GLUT3) promotes lactylation modifications by regulating lactate dehydrogenase A (LDHA) in gastric cancer.

OBJECTIVES: Glucose transporter 3 (GLUT3) plays a major role in glycolysis and glucose metabolism in cancer cells. We aimed to investigate the correlation between GLUT3 and histone lactylation modification in the occurrence and progression of gastric cancer. MATERIALS AND METHODS: We initially used single-cell sequencing data to determine the expression levels of GLUT3 and lactate dehydrogenase A (LDHA) in primary tumor, tumor-adjacent normal, and metastasis tumor tissues. Immunohistochemistry analysis was conducted to measure GLUT3, LDHA, and L-lactyl levels in gastric normal and cancer tissues. Transwell and scratch assays were performed to evaluate the metastatic and invasive capacity of gastric cancer cell lines. Western blotting was used to measure L-lactyl and histone lactylation levels in gastric cancer cell lines. RESULTS: Single-cell sequencing data showed that GLUT3 expression was significantly increased in primary tumor and metastasis tumor tissues. In addition, GLUT3 expression was positively correlated with that of LDHA expression and lactylation-related pathways. Western blotting and immunohistochemistry analyses revealed that GLUT3 was highly expressed in gastric cancer tissues and cell lines. GLUT3 knockdown in gastric cancer cell lines inhibited their metastatic and invasive capacity to various degrees. Additionally, the levels of LDHA, L-lactyl, H3K9, H3K18, and H3K56 significantly decreased after GLUT3 knockdown, indicating that GLUT3 affects lactylation in gastric cancer cells. Moreover, LDHA overexpression in a GLUT3 knockdown cell line reversed the levels of lactylation and EMT-related markers, and the EMT functional phenotype induced by GLUT3 knockdown. The in vivo results were consistent with the in vitro results. CONCLUSIONS: This study suggests the important role of histone lactylation in the occurrence and progression of gastric cancer, and GLUT3 may be a new diagnostic marker and therapeutic target for gastric cancer.

The role of mip in the development of lens in zebrafish.

Major intrinsic protein (MIP) functions as a water channel and a cell-junction molecule in the vertebrate eye lens. The pathogenic mechanism behind the loss of MIP function in the lens, which leads to degraded optical quality and cataract formation, is still unclear. In this study, a zebrafish model with the mipb mutant was produced. The expression of mipb mRNA and protein was dramatically reduced in the mutant. Immunological analysis reveals that loss function of mip leads to the diffuse distribution of ZL-1 in the mutant lens. Furthermore, in situ hybridization reveals that mip knockout results in a decrease in the transcripts of beaded filament structural protein 2 (Bfsp2) in the lens. Histology study shows that lens fibers in the mutants are less uniform in shape and the fiber arrangement is disrupted. The presented data provides evidence for the essential role of mipb in the development of lens fibers. The absence of mipb during lens formation is likely to result in aberrant lens fiber formation and impaired lens function.

Correction: A liquid-crystal-based immunosensor for the detection of cardiac troponin I.

Correction for 'A liquid-crystal-based immunosensor for the detection of cardiac troponin I' by Chunli Xia et al., Analyst, 2020, 145, 4569-4575, DOI: 10.1039/D0AN00425A.

A liquid-crystal-based immunosensor for the detection of cardiac troponin I.

Cardiac troponin I (cTnI) is one of the most sensitive and specific markers of myocardial cell injury, which can detect even minor myocardial damages. It is recognized as the main biochemical marker of the rapid diagnosis of acute myocardial infarction (AMI) and acute coronary syndrome (ACS). In this study, a label-free biosensor that utilizes the birefringence property of a nematic liquid crystal (LC) for the detection of cTnI is demonstrated. A chemically sensitive film with specific molecular recognition ability was decorated on the surface of a substrate, and the LC molecules were arranged in a vertically oriented order under the influence of the sensitive film, and a dark background signal was obtained using a polarizing optical microscope. When the antigen-antibody specifically binds to form a stronger acting force, the orientation of the LC molecules changes, resulting in a bright optical appearance. This LC-based immunosensor not only has the advantages of a facile structure, low cost and excellent specificity but also high sensitivity (a low detection limit of 1 pg ml-1), and has a promising future in biomedical related fields.

Long noncoding RNA LINC-PINT regulates laryngeal carcinoma cell stemness and chemoresistance through miR-425-5p/PTCH1/SHH axis.

Functional, noncoding RNA of about 200 nucleotides in length are known as long noncoding RNA (lncRNA). Advances in -omics have revolutionized the information with respect to the coding and noncoding regions of the genome. Several studies have illustrated the role of lncRNA in cell growth and cancer. Profiling and bioinformatic studies of laryngeal cancer has identified LINC-PINT as one of the lncRNA. However, the functional aspects of the deregulation have not been studied in laryngeal tumors. In this study, LINC-PINT expression in normal and tumor tissues were studied. Using a bioinformatic approach, microRNA (miRNA) targets of LINC-PINT and gene targets of the miRNA were determined. The impact of LINC-PINT on cell proliferation and chemoresistance was determined. Further through a set of silencing and re-expression studies phenotype rescue was studied. LINC-PINT expression was downregulated in laryngeal tumors. LINC-PINT targeted miR-425-5p by three sites. miR-425-5p also targeted PTCH1 a protein of the Hedgehog pathway. Downregulation of LINC-PINT was associated with increased cancer stemness and chemoresistance to cisplatin. Our results indicate a probable role of LINC-PINT in the pathology of laryngeal tumors. LINC-PINT re-expression in laryngeal tumors may be explored for reversion of cancer cell stemness and also for rescue of drug resistance phenotype.

Refractive Errors in Northern China Between the Residents with Drinking Water Containing Excessive Fluorine and Normal Drinking Water.

The purpose of this study was to evaluate the refractive errors and the demographic associations between drinking water with excessive fluoride and normal drinking water among residents in Northern China. Of the 1843 residents, 1415 (aged ≥40 years) were divided into drinking-water-excessive fluoride (DWEF) group (>1.20 mg/L) and control group (≤1.20 mg/L) on the basis of the fluoride concentrations in drinking water. Of the 221 subjects in the DWEF group, with 1.47 ± 0.25 mg/L (fluoride concentrations in drinking water), the prevalence rates of myopia, hyperopia, and astigmatism were 38.5 % (95 % confidence interval [CI] = 32.1-45.3), 19.9 % (95 % CI = 15-26), and 41.6 % (95 % CI = 35.1-48.4), respectively. Of the 1194 subjects in the control group with 0.20 ± 0.18 mg/L, the prevalence of myopia, hyperopia, and astigmatism were 31.5 % (95 % CI = 28.9-34.2), 27.6 % (95 % CI = 25.1-30.3), and 45.6 % (95 % CI = 42.8-48.5), respectively. A statistically significant difference was not observed in the association of spherical equivalent and fluoride concentrations in drinking water (P = 0.84 > 0.05). This report provides the data of the refractive state of the residents consuming drinking water with excess amounts of fluoride in northern China. The refractive errors did not result from ingestion of mild excess amounts of fluoride in the drinking water.

A novel MIP gene mutation analysis in a Chinese family affected with congenital progressive punctate cataract.

Congenital cataracts are one of the leading causes of visual impairment and blindness in children, and genetic factors play an important role in their development. This study aimed to identify the genetic defects associated with autosomal dominant congenital progressive punctate cataracts in a Chinese family and to explore the potential pathogenesis. Detailed family history and clinical data were recorded, and all the family members' blood samples were collected for DNA extraction. Linkage analysis was performed by microsatellite markers that are associated with punctate cataracts, and logarithm (base 10) of odds (LOD) scores were calculated using the LINKAGE program. Positive two-point LOD scores were obtained at markers D12S1622 (Zmax = 2.71 at θ = 0.0), D12S1724 (Zmax = 2.71 at θ = 0.0), and D12S90 (Zmax = 2.71 at θ = 0.0), which flank the major intrinsic protein of lens fiber (MIP) gene on chromosomal region 12q13. Direct sequencing of the encoding region of the MIP gene revealed a novel mutation (G>D) in exon 4 at nucleotide 644, which caused a substitution of glycine to aspartic acid at codon 215 (p.G215D) for the MIP protein. The mutation cosegregated with all patients with congenital progressive punctate cataracts, but it was absent in the healthy members. Bioinformatics analysis predicted that the mutation affects the function of the MIP protein. The wild type (WT) and G215D mutant of MIP were transfected with green fluorescent protein (GFP) into Hela cells separately, and it was found that the G215D mutant was aberrantly located in the cytoplasm instead of in the plasma membrane. In summary, our study presented genetic and functional evidence linking the new MIP mutation of G215D to autosomal dominant congenital cataracts, which adds to the list of MIP mutations linked to congenital progressive punctate cataracts.

Major intrinsic protein (MIP) polymorphism is associated with age-related cataract in Chinese.

PURPOSE: Age-related cataract (ARC) is a complex multi-factorial disorder involving several genetic and environmental factors. The major intrinsic protein of lens fiber gene (MIP) encodes the most abundant junctional membrane protein in the mature lens and plays a critical role in maintainace of lens normal structure and internal circulation. To determine the relationship between single nucleotide polymorphisms (SNPs) in MIP and the susceptibility to ARC in a Chinese population, we conducted this case-control study. METHODS: A total of 164 unrelated ARC patients and 132 normal controls were involved in the study. All participants completed full physical and ophthalmic examinations and provided a blood sample for DNA extraction. Seven SNPs (rs2269348, rs61759527, c.-4T>C, rs77163805, rs74641138, rs35033450, and rs36032520) in MIP were amplified by polymerase chain reaction (PCR) and then sequenced. Statistical analysis was performed using SNPstats. RESULTS: Polymorphisms rs61759527, rs77163805, rs35033450, and rs36032520 were not detected in all 296 subjects. There were no statistical differences in genotype or allele frequency of rs2269348 and rs74641138 between ARC cases and controls. But in c.-4C>T, cataract patients had a higher TC genotype and C allele frequencies (p=0.0018 and p=0.017, respectively) compared to healthy controls. The haplotype CCG of rs2269348, c.-4T>C and rs74641138 also exhibited a significantly higher distribution in cases than controls (OR=8.83, p=0.0024). CONCLUSIONS: Our findings indicate that the genotype TC in polymorphism c.-4T>C and haplotype CCG of rs2269348, c.-4T>C, and rs74641138 in MIP may attach an additional genetic risk factor for ARC in Chinese. This is the first association study about SNPs in MIP and susceptibility to ARC in Chinese population.

A novel mutation in the connexin 46 (GJA3) gene associated with congenital cataract in a Chinese pedigree.

PURPOSE: To identify the potential pathogenic mutation in a three-generation Chinese family with congenital nuclear pulverulent cataracts. METHODS: A three-generation pedigree was recruited for our study. Three patients and four healthy members of the family underwent a comprehensive clinical examination. Genomic DNA extracted from peripheral blood was amplified using the polymerase chain reaction (PCR) method and the exons of all candidate genes were sequenced. RESULTS: When sequencing the encoding regions of the candidate genes, a novel mutation (c.559C>T) was identified in the gap junction protein alpha 3 (GJA3) gene, which resulted in the substitution of highly conserved proline by serine at codon 187 (P187S). There was no noticeable nucleotide polymorphism in other candidate genes. The mutation co-segregated with all patients, but was absent in the healthy members and 100 normal individuals. CONCLUSIONS: The present study identified a novel mutation (c.559C>T) in the GJA3 gene associated with autosomal dominant pulverulent cataracts in a Chinese family. As the first report to relate p.P187S mutation in GJA3, it expands the mutation spectrum of GJA3 in association with congenital cataracts.

Polymorphisms of the DNA repair genes XPD and XRCC1 and the risk of age-related cataract development in Han Chinese.

PURPOSE: Epidemiological evidence suggests that ultraviolet (UV) irradiation and oxidative stress play an important role in age-related cataract pathogenesis. UV irradiation and oxidative stress can produce a wide range of DNA damage. Polymorphisms of DNA repair enzymes may affect repair efficiency and the role of DNA repair mechanisms has received attention recently in age-related cataract pathogenesis. In this case-control study, The aim was to determine the frequency of polymorphisms in two DNA repair enzyme genes, xeroderma pigmentosum complementation group D (XPD) codon 751 and x-ray cross-complementing group 1 (XRCC1) codon 399, in patients with age-related cataract and in healthy controls. METHODS: Polymerase chain reaction and restriction fragment length polymorphisms were used to analyze XPD T751G and XRCC1 G399A in 180 patients with age-related cataract and 174 healthy individuals as controls. RESULTS: There was a significant difference between the case and control groups in the XRCC1399 genotype. The odds ratio of the XRCC1 G/A polymorphism compared with the G/G wild-type genotype was 1.92 (95% CI = 1.17-3.15, p = 0.01). Moreover, individuals who carried at least one A-allele (G/A or A/A) had a 1.68-fold increased risk of developing age-related cataract compared with those who carried the G/G wild type genotype (OR = 1.68; 95% CI: 1.05-2.68, p = 0.030). No statistically significant difference was found in the genotypic and allelic distributions of the polymorphisms in the XPD gene between the groups. CONCLUSIONS: These results suggest that polymorphisms in XRCC1 codon 399 may be associated with the development of age-related cataract in Han Chinese.

Genetic analysis of the FOXL2 gene using quantitative real-time PCR in Chinese patients with blepharophimosis-ptosis-epicanthus inversus syndrome.

PURPOSE: The purpose of this study was to identify the mutation(s) or deletion(s) of the forkhead box protein L2 (FOXL2) gene in Chinese patients with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES). METHODS: Genomic DNA extracted from peripheral blood was collected from two Chinese families and from one sporadic case. PCR direct sequencing and quantitative real-time PCR-based copy number screening for the whole exon of FOXL2 were performed. RESULTS: Direct sequencing revealed an indel mutation c.50C→TA in the sporadic case which resulted in a frameshift generating 78 novel amino acids and terminating prematurely at codon 95. Deletions in the FOXL2 gene were confirmed by quantitative real-time PCR (q-real-time PCR) in two families in which intragenic mutations were excluded by direct sequencing. These changes containing deletions and a de novo mutation were not detected either in the non-carrier relatives or in 100 normal controls. CONCLUSIONS: This study identified two deletions and a de novo mutation in the FOXL2 gene in Chinese BPES patients. This is the first study to report FOXL2 gene deletions detected by q-real-time PCR in this ethnic group. This technique enriches the diagnostic methods of molecular genetics in BPES patients. The de novo mutation expands the mutation spectrum of FOXL2.

A novel mutation in GJA8 causing congenital cataract-microcornea syndrome in a Chinese pedigree.

PURPOSE: To identify the underlying genetic defect in a four-generation family of Chinese origin with autosomal dominant congenital cataract-microcornea syndrome (CCMC). METHODS: All individuals in the study underwent a full clinical examination and the details of history were collected . Genomic DNA extracted from peripheral blood was amplified by polymerase chain reaction (PCR) method and the exons of all candidate genes were sequenced. RESULTS: Direct sequencing of the encoding regions of the candidate genes revealed a heterozygous mutation c.592C-->T in exon 2 of the gap junction protein, alpha 8 (GJA8) gene. This mutation was responsible for the familial disorder through the substitution of a highly conserved arginine to tryptophan at codon 198 (p.R198W). This change co-segregated with all affected members of the family, but was not detected either in the non-carrier relatives or in the 100 normal controls. CONCLUSIONS: This report is the first to relate p.R198W mutation in GJA8 with CCMC. The result expands the mutation spectrum of GJA8 in associated with congenital cataract and microcornea, and implies that this gene has direct involvement with the development of the lens as well as the other anterior segment of the eye.

A missense mutation in CRYBA4 associated with congenital cataract and microcornea.

PURPOSE: To identify mutations in a Chinese family with congenital cataract and microcornea. METHODS: Detailed family history and clinical data were recorded. Genomic DNA was extracted from leukocytes of venous blood of the patients and noncarriers in this family along with 100 normal individuals. All six exons of crystallin, beta A4 gene (CRYBA4) were amplified by PCR methods and direct sequencing. RESULTS: We identified a c.225G>T sequence change that led to an amino acid substitution G64W in the CRYBA4-induced protein in two patients of this family; this nucleotide substitution was not detected in the other individuals. CONCLUSIONS: A novel missense mutation in CRYBA4 was identified in our study. It expands the mutation spectrum of CRYBA4 and provides useful information to the study of molecular pathogenesis of cataract and microcornea.