Highly efficient capture approach for the identification of diverse inherited retinal disorders.

Our study presents a 319-gene panel targeting inherited retinal dystrophy (IRD) genes. Through a multi-center retrospective cohort study, we validated the assay's effectiveness and clinical utility and characterized the mutation spectrum of Taiwanese IRD patients. Between January 2018 and May 2022, 493 patients in 425 unrelated families, all initially suspected of having IRD without prior genetic diagnoses, underwent detailed ophthalmic and physical examinations (with extra-ocular features recorded) and genetic testing with our customized panel. Disease-causing variants were identified by segregation analysis and clinical interpretation, with validation via Sanger sequencing. We achieved a read depth of >200× for 94.2% of the targeted 1.2 Mb region. 68.5% (291/425) of the probands received molecular diagnoses, with 53.9% (229/425) resolved cases. Retinitis pigmentosa (RP) is the most prevalent initial clinical impression (64.2%), and 90.8% of the cohort have the five most prevalent phenotypes (RP, cone-rod syndrome, Usher's syndrome, Leber's congenital amaurosis, Bietti crystalline dystrophy). The most commonly mutated genes of probands that received molecular diagnosis are USH2A (13.7% of the cohort), EYS (11.3%), CYP4V2 (4.8%), ABCA4 (4.5%), RPGR (3.4%), and RP1 (3.1%), collectively accounted for 40.8% of diagnoses. We identify 87 unique unreported variants previously not associated with IRD and refine clinical diagnoses for 21 patients (7.22% of positive cases). We developed a customized gene panel and tested it on the largest Taiwanese cohort, showing that it provides excellent coverage for diverse IRD phenotypes.

Confined Self-Assembly of Block Copolymers within Emulsion Droplets: A Perspective.

When the self-assembly of block copolymers (BCPs) occurs within organic emulsion droplets in the aqueous phase, the strong structural frustration of BCP chains causes the formation of a series of well-regulated BCP particles that cannot be obtained from the self-assembly of BCPs in the bulk state or solution. In this Perspective, we review the recent progress of the self-assembly of BCPs confined in emulsion droplets. The governing factors of the structure and morphology of the as-prepared BCP particles are summarized. In addition, the applications of the as-prepared BCP particles in photonic crystals and drug release are discussed. Finally, we also give a forward-looking perspective on future challenges in this field.

Knockdown of POLQ interferes the development and progression of hepatocellular carcinoma through regulating cell proliferation, apoptosis and migration.

BACKGROUND: DNA Polymerase Theta (POLQ) is a DNA polymerase involved in error-prone translesion DNA synthesis (TLS) and error-prone repair of DNA double-strand breaks (DSBs), whose function in hepatocellular carcinoma has not been investigated. METHODS: In the present study, both the data collected from the Cancer Genome Atlas (TCGA) and our group's results showed higher POLQ expression in HCC tissues than the para-cancerous tissues, which was associated with higher malignancy and poor prognosis. POLQ knockdown HCC cell model (shPOLQ) was constructed along with the corresponding negative control (shCtrl) through lentivirus infection for loss-of-function study. RESULTS: We found that, upon knockdown of POLQ, the proliferation and migration of HCC cells decreased and apoptosis percentage increased. Moreover, the percentage of cells in G2 phase significantly increased in shPOLQ group compared with shCtrl group. Xenografts in mice grafted with shPOLQ cells grew much slower than that transplanted with shCtrl cells, and expressed lower Ki67 level. Furthermore, an apoptosis-related signaling array was used to explore the involvement of downstream signaling pathways, suggesting the enhanced phosphorylation of HSP27 and JNK, and the de-activation of mTOR, PRAS40, ERK1/2 and STAT3 pathways. CONCLUSIONS: Collectively, our study revealed that POLQ may participate in the development of HCC, depletion of which may be a promising treatment strategy for HCC.

Extracellular cyclophilin A induces cardiac hypertrophy via the ERK/p47phox pathway.

Excessive reactive oxygen species (ROS) are a critical driver of cardiac hypertrophy developing into heart failure. Cyclophilin A (CyPA), a member of the cyclophilin family, has been highlighted as a main secreted ROS-induced factor. The mechanism by which extracellular CyPA interacts with cardiomyocytes is unclear. We showed that extracellular CyPA is upregulated in cardiac hypertrophy rats and expressed around hypertrophic cardiomyocytes. Cell experiments further confirmed that extracellular CyPA induces H9c2 cardiomyocytes hypertrophy via ROS generation. Extracellular CyPA-induced ROS is derived from nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase, and extracellular CyPA activates p47phox membrane translocation through ERK1/2 pathway. When blocking extracellular matrix metalloproteinase inducer (EMMPRIN), most of the extracellular CyPA effects were significantly inhibited. The current study shows that extracellular CyPA is one of the key factors linking oxidative stress and cardiac hypertrophy, and may be a potential target for cardiac hypertrophy therapy.

Role of CyPA in cardiac hypertrophy and remodeling.

Pathological cardiac hypertrophy is a complex process and eventually develops into heart failure, in which the heart responds to various intrinsic or external stress, involving increased interstitial fibrosis, cell death and cardiac dysfunction. Studies have shown that oxidative stress is an important mechanism for this maladaptation. Cyclophilin A (CyPA) is a member of the cyclophilin (CyPs) family. Many cells secrete CyPA to the outside of the cells in response to oxidative stress. CyPA from blood vessels and the heart itself participate in a variety of signaling pathways to regulate the production of reactive oxygen species (ROS) and mediate inflammation, promote cardiomyocyte hypertrophy and proliferation of cardiac fibroblasts, stimulate endothelial injury and vascular smooth muscle hyperplasia, and promote the dissolution of extracellular matrix (ECM) by activating matrix metalloproteinases (MMPs). The events triggered by CyPA cause a decline of diastolic and systolic function and finally lead to the occurrence of heart failure. This article aims to introduce the role and mechanism of CyPA in cardiac hypertrophy and remodeling, and highlights its potential role as a disease biomarker and therapeutic target.

Inactivation of p53 in pterygium influence miR-200a expression resulting in ZEB1/ZEB2 up-regulation and EMT processing.

Loss of p53 function has been linked to progression of pterygium. MiR-200a is known to be controlled by p53. Here, we hypothesize that expression of miR-200a and downstream ZEB1/ZEB2 genes are regulated epithelial-mesenchymal transition (EMT) involved in the pathogenesis and recurrence of pterygium. For this study, 120 primary pterygial samples were collected. Immunohistochemistry and real-time RT-PCR were performed to determine the expression of p53, p53 down-stream EMT associated protein and miR-200a. The molecular correlation of p53, miR-200a and downstream genes were confirmed using primary pterygium cells (PECs). Expression of miR-200a in pterygium tissues was significantly lower than in conjunctiva controls (p = 0.015). Up-regulated miR-200a levels were positively correlated with and p53 protein expression (p < 0.001). The miR-200a downstream ZEB1/ZEB1 protein expression were negative correlated with miR-200a expression. Cell model studies demonstrated that miR-200a controlled the EMT of PECs through up-regulated ZEB1, ZEB2 and Snail gene expression. Our study demonstrated that inactivation of p53 in pterygium may influence miR-200a, resulting in ZEB1/ZEB2 up-regulation and EMT processing of pterygium. Therefore, we suggest that expression of miR-200a play an important role in EMT processing and recurrence of pterygium.

Endotoxin-induced acute lung injury in mice is protected by 5,7-dihydroxy-8-methoxyflavone via inhibition of oxidative stress and HIF-1α.

Up to date, the morbidity and mortality rates of acute lung injury (ALI) still rank high among clinical illnesses. Endotoxin, also called lipopolysaccharide (LPS), induced sepsis is the major cause for ALI. Beneficial biological effects, such as antioxidation, anti-inflammation, and neuroprotection was found to express by 5,7-dihydroxy-8-methoxyflavone (DHMF). The purpose of present study was to investigate the potential protective effects of DHMF and the possibile mechanisms involved in LPS-induced ALI. In our experimental model, ALI was induced in mice by intratracheal injection of LPS, and DHMF at various concentrations was injected intraperitoneally for 30 min prior to LPS administration. Pretreatment with DHMF inhibited not only the histolopatholgical changes occurred in lungs but also leukocytes infiltration in LPS-induced ALI. Decreased activity of antioxidative enzymes (AOE) such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) caused by LPS was reversed by DHMF. LPS-induced lipid peroxidation HIF-1α accumulation, NF-κB phosphorylation, and IκBα degradation were all inhibited by DHMF. In addition, LPS-induced expression of proinflammatory mediators such as TNF-α and IL-1ß were also inhibited by 5,7-dihydroxy-8-methoxyflavone. These results suggested that the protective mechanisms of DHMF on endotoxin-induced ALI might be via up-regulation of antioxidative enzymes, inhibition of NFκB phosphorylation, and HIF-1α accumulation. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1700-1709, 2016.

Maternal coffee consumption during pregnancy and risk of childhood acute leukemia: a metaanalysis.

OBJECTIVE: This study was undertaken to explore the association between maternal coffee consumption during pregnancy and childhood acute leukemia (AL). STUDY DESIGN: The PubMed database was used to search studies up to May 5, 2013, and the lists of references of retrieved articles were also screened to identify additional relevant studies. Studies were included if they reported the odds ratio and corresponding 95% confidence interval (CI) of childhood AL, including childhood acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML), with respect to maternal coffee consumption during pregnancy. RESULTS: Compared with non/lowest drinkers, the combined odds ratio regarding the relationship of maternal coffee consumption during pregnancy and childhood AL was 1.22 (95% CI, 1.04-1.43) for ever drinkers, 1.16 (95% CI, 1.00-1.34) for low to moderate-level drinkers, and 1.72 (95% CI, 1.37-2.16) for high-level drinkers. When analysis was conducted by subtypes of childhood AL, maternal coffee consumption (high-level drinkers vs non/lowest drinkers) was statistically significantly associated with childhood ALL (1.65; 95% CI, 1.28-2.12) and childhood AML (1.58; 95% CI, 1.20-2.08). We observed the linear dose-response relationship of coffee consumption and childhood AL (P for nonlinearity = .68), including childhood ALL and childhood AML; with increased coffee consumption, the risk of childhood AL increased. CONCLUSION: The findings of the metaanalysis suggest that maternal coffee consumption during pregnancy may increase the risk of childhood AL. Because of limited studies, further prospective studies are urgently needed to explore the adverse effect of coffee consumption on childhood AL.

Vascular endothelial growth factor gene polymorphism and protein expression in the pathogenesis of pterygium.

BACKGROUND AND AIMS: Vascular endothelial growth factor (VEGF) gene expression has been linked to cancer progression. Here we hypothesise that the polymorphism and protein expression of VEGF are correlated with the pathogenesis and therapy response of pterygium. METHODS: 60 pterygial and 121 normal conjunctival samples were collected to determine the genotypes and protein expression of VEGF. Primary pterygium cells (PECs) were used to confirm the effect of the VEGF polymorphism on the angiogenesis of pterygium. RESULTS: 48 (83.3%) pterygial specimens tested positive for VEGF protein expression, which was significantly higher than in the control groups (16.7%, p<0.0001). The frequency of the 936 C>T variant, but not the -2578C>A variant, was significantly higher in the pterygium group compared with the control group. VEGF protein expression was significantly higher in the 936 C/C group than in the 936 C/T and T/T groups (p=0.001). The results of our cell model showed that PECs with the C/C genotype had a higher angiogenesis ability and higher response to the antiangiogenesis drug bevacizumab than cells with the C/T and T/T genotypes. CONCLUSIONS: We suggest that VEGF could be used as a target for pterygium therapy in patients with the 936C>T genotype.

Myopization factors affecting urban elementary school students in Taiwan.

PURPOSE: To investigate factors that may contribute to the myopization of urban elementary school students in Taiwan. METHODS: Grades 1 to 6 students of the same racial background (n = 1894; mean age, 6.3-11.3 years) in three schools, located in Tamsui, Taichung, and Tainan, were refracted to obtain the best corrected visual acuity. The refractive power needed for best corrected visual acuity was used for subsequent statistical analysis. On behalf of their children, parents also completed a questionnaire on six categories of potential myopization variables. Correlation between these variables and the increase or decrease in the refractive error was assessed. The predictive value of each variable was also calculated based on linear regression analysis. RESULTS: The overall mean refractive error in grades 1 to 6 was -0.37, -0.68, -1.33, -1.60, -1.90, and -2.51 D, respectively. The prevalence of myopia (-1.00 D or more minus) showed a significant difference between grades 2 and 3 and, again, between grades 5 and 6. In addition, 20 potential modulating factors were evaluated; 65.9% of the change in the refractive error could be explained by four: (1) lag in optimal correction, defined as a -1.00-D deficit between new refractive error and current optical correction; (2) outdoor spectacle wear; (3) spectacles for different working distances; and (4) hours spent on reading and writing on weekdays. In contrast, outdoor time and the intake frequency of 36 food items both held very low predictive values of 0.2% and 2.5%, respectively. CONCLUSIONS: Each variable associated with the refractive error has a different predictive value, either positive or negative. Ultimately, the interplay of these variables decides the outcome of the pattern and the degree of school myopia.

CYP1A1 protein activity is associated with allelic variation in pterygium tissues and cells.

BACKGROUND: A thymine/cytosine point mutation in the MSP I restriction site of cytochrome P450 1A1 (CYP1A1) has been linked to susceptibility to smoking-related cancers and is reported to result in increased enzyme activity. Therefore, we sought to determine whether allelic variation of CYP1A1 is associated with protein expression and protein activity in pterygium. METHODS: We collected 150 pterygium samples and 50 normal conjunctiva samples, which served as controls. DNA samples were extracted from blood cells and then subjected to real-time ploymerase chain reaction (PCR) to determine CYP1A1 genotype. CYP1A1 protein expression was determined by immunohistochemical staining with a monoclonal antibody for CYP1A1. Pterygium epithelial cells (PECs), cultured in a serum-free culture medium, real-time PCR, western blot and enzyme-linked immunosorbent assay (ELISA) were used to understand the effect of CYP1A1 allelic variation in protein expression and activity. RESULTS: Forty-eight (33.3%) pterygium specimens tested positive for CYP1A1 protein expression. CYP1A1 protein expression was significantly greater in the pterygium group than in the control group (p<0.0001). In addition, CYP1A1 protein expression was associated with allelic variation. CYP1A1 protein expression was significantly greater in the m2/m2 group than in the m1/m1and m1/m2 groups (p=0.006). In the cell model, CYP1A1 protein expression and b[a]P 7,8-diol 9,10-epoxide (BPDE)-like DNA adducts increased in CYP1A1 m2/m2 (genotype T/T) PEC cells as compared with m1/m2 (genotype C/T) and m1/m1 (genotype C/C) cells. CONCLUSIONS: CYP1A1 expression in pterygium correlates with allelic variation and can be used as an independent risk marker.

Characterising visual deficits in children of an urban elementary school in Taiwan.

PURPOSE: The aim was to screen children from Grades 1 to 6 in an urban elementary school in Central Taiwan for visual deficits and associated parameters and, as an extension, to examine the acceptance of cycloplegic therapy as well as the lag in optimal vision correction. METHODS: Of 900 students in one school, 731 participated in the study, with parental consent. Data from 694 students, who had also completed a vision correction history were analysed. In addition to body height and weight, the screening included vision, non-cycloplegic autorefraction and distance retinoscopy, axial length and functional testing. RESULTS: There was a decrease in students with vision of 1.0 or better from 55.8 per cent in Grade 1 to 20.0 per cent in Grade 6. The decreases between Grades 2 and 3 and Grades 5 and 6 were significant. These trends were in general agreement with those based on refractive error and axial length. The students had abnormal functional findings including: stereoscopic vision, 9.2 per cent; cover tests, 14.1 per cent; pupillary responses, 13.8 per cent; and less commonly in extraocular muscular functions (3.0 per cent) and colour vision (5.2 per cent). A full 40 per cent of students received cycloplegic therapy with 25 per cent dropping out for various reasons. These cases were generally associated with lower vision and higher myopia. A lag between subnormal vision and optical correction was also observed with 55.1 per cent apparently not optimally corrected. Other parameters, including body height, weight and body mass index were not correlated with vision or refractive error. CONCLUSIONS: Age-dependent increase in the prevalence of myopia appears to continue despite the common practice of topical cycloplegic therapy in Taiwan. Timely correction of the refractive error is also lacking. While maintaining a visual acuity of 1.0 or better for all students at all times is not possible, this lag might be shortened by more frequent screening and/or direct provision of optical aids.