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  • 1
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    Points of View on the Tools for Genome/Gene Editing
    MDPI AG ; 2021
    In:  International Journal of Molecular Sciences Vol. 22, No. 18 ( 2021-09-13), p. 9872-
    Details
    In: International Journal of Molecular Sciences, MDPI AG, Vol. 22, No. 18 ( 2021-09-13), p. 9872-
    Abstract: Theoretically, a DNA sequence-specific recognition protein that can distinguish a DNA sequence equal to or more than 16 bp could be unique to mammalian genomes. Long-sequence-specific nucleases, such as naturally occurring Homing Endonucleases and artificially engineered ZFN, TALEN, and Cas9-sgRNA, have been developed and widely applied in genome editing. In contrast to other counterparts, which recognize DNA target sites by the protein moieties themselves, Cas9 uses a single-guide RNA (sgRNA) as a template for DNA target recognition. Due to the simplicity in designing and synthesizing a sgRNA for a target site, Cas9-sgRNA has become the most current tool for genome editing. Moreover, the RNA-guided DNA recognition activity of Cas9-sgRNA is independent of both of the nuclease activities of it on the complementary strand by the HNH domain and the non-complementary strand by the RuvC domain, and HNH nuclease activity null mutant (H840A) and RuvC nuclease activity null mutant (D10A) were identified. In accompaniment with the sgRNA, Cas9, Cas9(D10A), Cas9(H840A), and Cas9(D10A, H840A) can be used to achieve double strand breakage, complementary strand breakage, non-complementary strand breakage, and no breakage on-target site, respectively. Based on such unique characteristics, many engineered enzyme activities, such as DNA methylation, histone methylation, histone acetylation, cytidine deamination, adenine deamination, and primer-directed mutation, could be introduced within or around the target site. In order to prevent off-targeting by the lasting expression of Cas9 derivatives, a lot of transient expression methods, including the direct delivery of Cas9-sgRNA riboprotein, were developed. The issue of biosafety is indispensable in in vivo applications; Cas9-sgRNA packaged into virus-like particles or extracellular vesicles have been designed and some in vivo therapeutic trials have been reported.
    Type of Medium: Online Resource
    ISSN: 1422-0067
    URL: Article
    DOI: 10.3390/ijms22189872
    Language: English
    Publisher: MDPI AG
    Publication Date: 2021
    detail.hit.zdb_id: 2019364-6
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  • 2
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    The Role of SCL Isoforms in Embryonic Hematopoiesis
    MDPI AG ; 2023
    In:  International Journal of Molecular Sciences Vol. 24, No. 7 ( 2023-03-29), p. 6427-
    Details
    In: International Journal of Molecular Sciences, MDPI AG, Vol. 24, No. 7 ( 2023-03-29), p. 6427-
    Abstract: Three waves of hematopoiesis occur in the mouse embryo. The primitive hematopoiesis appears as blood islands in the extra embryonic yolk sac at E7.5. The extra embryonic pro-definitive hematopoiesis launches in late E8 and the embryonic definitive one turns on at E10.5 indicated by the emergence of hemogenic endothelial cells on the inner wall of the extra embryonic arteries and the embryonic aorta. To study the roles of SCL protein isoforms in murine hematopoiesis, the SCL-large (SCL-L) isoform was selectively destroyed with the remaining SCL-small (SCL-S) isoform intact. It was demonstrated that SCL-S was specifically expressed in the hemogenic endothelial cells (HECs) and SCL-L was only detected in the dispersed cells after budding from HECs. The SCLΔ/Δ homozygous mutant embryos only survived to E10.5 with normal extra embryonic vessels and red blood cells. In wild-type mouse embryos, a layer of neatly aligned CD34+ and CD43+ cells appeared on the endothelial wall of the aorta of the E10.5 fetus. However, the cells at the same site expressed CD31 rather than CD34 and/or CD43 in the E10.5 SCLΔ/Δ embryo, indicating that only the endothelial lineage was developed. These results reveal that the SCL-S is sufficient to sustain the primitive hematopoiesis and SCL-L is necessary to launch the definitive hematopoiesis.
    Type of Medium: Online Resource
    ISSN: 1422-0067
    URL: Article
    DOI: 10.3390/ijms24076427
    Language: English
    Publisher: MDPI AG
    Publication Date: 2023
    detail.hit.zdb_id: 2019364-6
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  • 3
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    Effects of Personal Exposures to Micro- and Nano-Particulate Matter, Black Carbon, Particle-Bound Polycyclic Aromatic Hydrocarbons, and Carbon Monoxide on Heart Rate Variability in a Panel of Healthy Older Subjects
    MDPI AG ; 2019
    In:  International Journal of Environmental Research and Public Health Vol. 16, No. 23 ( 2019-11-23), p. 4672-
    Details
    In: International Journal of Environmental Research and Public Health, MDPI AG, Vol. 16, No. 23 ( 2019-11-23), p. 4672-
    Abstract: As a non-invasive method, heart rate variability (HRV) has been widely used to study cardiovascular autonomous control. Environmental epidemiological studies indicated that the increase in an average concentration of particulate matter (PM) would result in a decrease in HRV, which was related to the increase of cardiovascular mortality in patients with myocardial infarction and the general population. With rapid economic and social development in Asia, how air pollutants, such as PM of different sizes and their components, affect the cardiovascular health of older people, still need to be further explored. The current study includes a 72 h personal exposure monitoring of seven healthy older people who lived in the Taipei metropolitan area. Mobile equipment, a portable electrocardiogram recorder, and the generalized additive mixed model (GAMM) were adopted to evaluate how HRV indices were affected by size-fractionated PM, particle-bound polycyclic aromatic hydrocarbons (p-PAHs), black carbon (BC), and carbon monoxide (CO). Other related confounding factors, such as age, sex, body mass index (BMI), temperature, relative humidity (RH), time, and monitoring week were controlled by fixed effects of the GAMM. Statistical analyses of multi-pollutant models showed that PM2.5–10, PM1, and nanoparticle (NP) could cause heart rate (HR), time-domain indices, and frequency-domain indices to rise; PM1–2.5 and BC would cause the frequency-domain index to rise; p-PAHs would cause HR to rise, and CO would cause time-domain index and frequency-domain index to decline. In addition, the moving average time all fell after one hour and might appear at 8 h in HRVs’ largest percentage change caused by each pollutant, results of which suggested that size-fractionated PM, p-PAHs, BC, and CO exposures have delayed effects on HRVs. In conclusion, the results of the study showed that the increase in personal pollutant exposure would affect cardiac autonomic control function of healthy older residents in metropolitan areas, and the susceptibility of cardiovascular effects was higher than that of healthy young people. Since the small sample size would limit the generalizability of this study, more studies with larger scale are warranted to better understand the HRV effects of simultaneous PM and other pollution exposures for subpopulation groups.
    Type of Medium: Online Resource
    ISSN: 1660-4601
    URL: Article
    DOI: 10.3390/ijerph16234672
    Language: English
    Publisher: MDPI AG
    Publication Date: 2019
    detail.hit.zdb_id: 2175195-X
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  • 4
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    Impacts of In-Cabin Exposure to Size-Fractionated Particulate Matters and Carbon Monoxide on Changes in Heart Rate Variability for Healthy Public Transit Commuters
    MDPI AG ; 2019
    In:  Atmosphere Vol. 10, No. 7 ( 2019-07-17), p. 409-
    Details
    In: Atmosphere, MDPI AG, Vol. 10, No. 7 ( 2019-07-17), p. 409-
    Abstract: To evaluate the cardiovascular impact of traffic-related pollutant exposure on healthy young adults, the research team has collected the primary data of in-cabin exposure to air pollutants and heart rate variability (HRV). Twenty young healthy college students were recruited in Taipei metropolitan area. In addition to electrocardiogram, personal exposure to air pollutants, i.e., particulate matter (PM) and carbon monoxide (CO), and weather conditions, including temperature and relative humidity (RH), on campus, bus, and mass rapid transit were monitored continuously. The following HRV parameters were evaluated using generalized additive mixed model to adjust for personal and meteorological variables: heart rate (HR), the square root of the mean of the sum of the squares of differences between adjacent normal-to-normal (NN) intervals (r-MSSD), the standard deviation of all NN intervals (SDNN), the percentage of successive NN interval differences greater than 50 ms (pNN50), low-frequency power (LF), high-frequency power (HF), total power (TP), and LF/HF. They were assessed to find out the association between in-cabin exposure and HRV parameters. Compared with the HRV parameters measured on campus, the percent changes in r-MSSD, SDNN, pNN50+1, LF, HF, and TP decreased when the participants were in public transits. After adjusting for all locations, 5 min moving averages of PM2.5–10 and PM1 were significantly associated with the increase in the percent changes in HR and SDNN. Additionally, 5 min moving averages of PM2.5–10 exposure were significantly associated with the decrease in the percent change in HF, while it was significantly associated with the increase of the percent change in LF/HF. The reduction of the percent change in HR was also found to be significantly associated with 5 min CO moving averages. To conclude, current analyses have shown that size-fractionated PMs and CO exposure in public transits might lead to significant changes of HRV parameters for healthy young adults.
    Type of Medium: Online Resource
    ISSN: 2073-4433
    URL: Article
    DOI: 10.3390/atmos10070409
    Language: English
    Publisher: MDPI AG
    Publication Date: 2019
    detail.hit.zdb_id: 2605928-9
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  • 5
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    Methylmercury Induces Mitochondria- and Endoplasmic Reticulum Stress-Dependent Pancreatic β-Cell Apoptosis via an Oxidative Stress-Mediated JNK Signaling Pathway
    MDPI AG ; 2022
    In:  International Journal of Molecular Sciences Vol. 23, No. 5 ( 2022-03-05), p. 2858-
    Details
    In: International Journal of Molecular Sciences, MDPI AG, Vol. 23, No. 5 ( 2022-03-05), p. 2858-
    Abstract: Methylmercury (MeHg), a long-lasting organic pollutant, is known to induce cytotoxic effects in mammalian cells. Epidemiological studies have suggested that environmental exposure to MeHg is linked to the development of diabetes mellitus (DM). The exact molecular mechanism of MeHg-induced pancreatic β-cell cytotoxicity is still unclear. Here, we found that MeHg (1-4 μM) significantly decreased insulin secretion and cell viability in pancreatic β-cell-derived RIN-m5F cells. A concomitant elevation of mitochondrial-dependent apoptotic events was observed, including decreased mitochondrial membrane potential and increased proapoptotic (Bax, Bak, p53)/antiapoptotic (Bcl-2) mRNA ratio, cytochrome c release, annexin V-Cy3 binding, caspase-3 activity, and caspase-3/-7/-9 activation. Exposure of RIN-m5F cells to MeHg (2 μM) also induced protein expression of endoplasmic reticulum (ER) stress-related signaling molecules, including C/EBP homologous protein (CHOP), X-box binding protein (XBP-1), and caspase-12. Pretreatment with 4-phenylbutyric acid (4-PBA; an ER stress inhibitor) and specific siRNAs for CHOP and XBP-1 significantly inhibited their expression and caspase-3/-12 activation in MeHg-exposed RIN-mF cells. MeHg could also evoke c-Jun N-terminal kinase (JNK) activation and reactive oxygen species (ROS) generation. Antioxidant N-acetylcysteine (NAC; 1mM) or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox; 100 μM) markedly prevented MeH-induced ROS generation and decreased cell viability in RIN-m5F cells. Furthermore, pretreatment of cells with SP600125 (JNK inhibitor; 10 μM) or NAC (1 mM) or transfection with JNK-specific siRNA obviously attenuated the MeHg-induced JNK phosphorylation, CHOP and XBP-1 protein expression, apoptotic events, and insulin secretion dysfunction. NAC significantly inhibited MeHg-activated JNK signaling, but SP600125 could not effectively reduce MeHg-induced ROS generation. Collectively, these findings demonstrate that the induction of ROS-activated JNK signaling is a crucial mechanism underlying MeHg-induced mitochondria- and ER stress-dependent apoptosis, ultimately leading to β-cell death.
    Type of Medium: Online Resource
    ISSN: 1422-0067
    URL: Article
    DOI: 10.3390/ijms23052858
    Language: English
    Publisher: MDPI AG
    Publication Date: 2022
    detail.hit.zdb_id: 2019364-6
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  • 6
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    Electronic Payment Behaviors of Consumers under Digital Transformation in Finance—A Case Study of Third-Party Payments
    MDPI AG ; 2023
    In:  Journal of Risk and Financial Management Vol. 16, No. 8 ( 2023-07-25), p. 346-
    Details
    In: Journal of Risk and Financial Management, MDPI AG, Vol. 16, No. 8 ( 2023-07-25), p. 346-
    Abstract: In the digital era, new financial technologies and big data are accelerating the development of financial transactions. With the rise of e-commerce transactions, the financial industry has come to recognize that banking as a service can be seamlessly integrated into any scenario, thanks to disruptive innovation driven by electronic and third-party payments. This study aims to examine the consumer acceptance of third-party payment systems offered by electronic payment platforms for e-commerce, as well as their continued usage in the context of digital transformation in finance. This study employed the questionnaire survey method, and it distributed questionnaires to consumers who have used third-party payment systems. A total of 332 valid questionnaires were collected. The results indicate that user acceptance of innovative technologies and various external variables (e.g., the user’s external environment, internal characteristics, and information system quality) were significantly positively correlated with perceived usefulness, perceived ease of use, and behavioral intention regarding the electronic payment behaviors of consumers. Based on the empirical results, this study proposes important managerial implications for the financial industry and e-commerce platforms in promoting electronic payment innovation.
    Type of Medium: Online Resource
    ISSN: 1911-8074
    URL: Article
    DOI: 10.3390/jrfm16080346
    Language: English
    Publisher: MDPI AG
    Publication Date: 2023
    detail.hit.zdb_id: 2739117-6
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  • 7
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    Efficacy of Myopia Control and Distribution of Corneal Epithelial Thickness in Children Treated with Orthokeratology Assessed Using Optical Coherence Tomography
    MDPI AG ; 2022
    In:  Journal of Personalized Medicine Vol. 12, No. 2 ( 2022-02-14), p. 278-
    Details
    In: Journal of Personalized Medicine, MDPI AG, Vol. 12, No. 2 ( 2022-02-14), p. 278-
    Abstract: The association between myopia control efficacy in children treated with orthokeratology and corneal epithelial thickness is still unknown. The aim of this study was to explore the corneal epithelial thickness and its association with axial length changes in children treated with orthokeratology. This retrospective cohort study enrolled children aged from 9 to 15 years who had received orthokeratology for myopia control and had been followed up for at least 1 year. Anterior segment optical coherence tomography was performed to generate wide epithelial thickness maps of the patients. Annual axial length changes were calculated from the axial length at 6 months after the initiation of orthokeratology lens wear and at final measurements. Corneal epithelial thickness data were obtained from 24 sectors and a central 2 mm zone of the wide epithelial thickness map. Associations between annual axial length changes and corneal epithelial thickness for each sector/zone of the wide epithelial thickness map, and orthokeratology treatment data were determined by generalized estimating equations. Finally, a total of 83 eyes of 43 patients (mean age 11.2 years) were included in the analysis. The mean annual axial length change was 0.169 mm; when regressing demographic and ortho-k parameters to mean annual axial length changes, age and target power were both negatively associated with them (β = −14.43, p = 0.008; β = −0.26, p = 0.008, respectively). After adjusting for age and target power, the annual axial length changes were positively associated with the corneal epithelium thickness of IT1, I1, SN2, and S2 sectors of the wide epithelial thickness map, and negatively with that of the I3 sector. In conclusion, we identified associations between annual axial length changes and the corneal epithelium thickness of certain sectors in children treated with orthokeratology. This may facilitate the design of orthokeratology lenses with enhanced efficacy for myopia control.
    Type of Medium: Online Resource
    ISSN: 2075-4426
    URL: Article
    DOI: 10.3390/jpm12020278
    Language: English
    Publisher: MDPI AG
    Publication Date: 2022
    detail.hit.zdb_id: 2662248-8
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  • 8
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    Magnolol, A Novel Antagonist of Thrombin and PAR-1, Inhibits Thrombin-Induced Connective Tissue Growth Factor (CTGF) Expression in Vascular Smooth Muscle Cells and Ameliorate Pathogenesis of Restenosis in Rats
    MDPI AG ; 2020
    In:  Applied Sciences Vol. 10, No. 23 ( 2020-12-05), p. 8729-
    Details
    In: Applied Sciences, MDPI AG, Vol. 10, No. 23 ( 2020-12-05), p. 8729-
    Abstract: Restenosis and destructive vascular remodeling are the main reasons for treatment failure in patients undergoing percutaneous coronary intervention (PCI). In this study, we explored the efficacy of magnolol (a traditional Chinese medicine) in the treatment of restenosis. The results of this study showed that the activities of thrombin and PAR-1 (protease-activated receptor 1) were significantly decreased by the treatment of magnolol. Based on protein docking analysis, magnolol exhibits its potential to bind to the PAR-1 active site. In addition, thrombin-induced connective tissue growth factor (CTGF) expression and the upstream of CTGF such as JNK-1 (but not JNK-2), c-Jun, and AP-1 were distinctly inhibited by magnolol (50 μM) in vascular smooth muscle cells (VSMC). For the functional assay, magnolol (50 μM) significantly inhibited the migration of VSMC, and rats treated with magnolol (13 mg/kg/day) after balloon angioplasty has observed a significant reduction in the formation of common arterial neointima. In conclusion, we identified a novel mechanism by which magnolol acts as the thrombin activity inhibitor and may be the PAR-1 antagonist. In accordance with these functions, magnolol could decrease thrombin-induced CTGF expression in VSMCs via PAR-1/JNK-1/AP-1 signaling.
    Type of Medium: Online Resource
    ISSN: 2076-3417
    URL: Article
    DOI: 10.3390/app10238729
    Language: English
    Publisher: MDPI AG
    Publication Date: 2020
    detail.hit.zdb_id: 2704225-X
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  • 9
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    Antiviral Activity of Compound L3 against Dengue and Zika Viruses In Vitro and In Vivo
    MDPI AG ; 2020
    In:  International Journal of Molecular Sciences Vol. 21, No. 11 ( 2020-06-05), p. 4050-
    Details
    In: International Journal of Molecular Sciences, MDPI AG, Vol. 21, No. 11 ( 2020-06-05), p. 4050-
    Abstract: Dengue virus (DENV) and Zika virus (ZIKV) are mosquito-borne flaviviruses that cause severe illness after infection. Currently, there are no specific or effective treatments against DENV and ZIKV. Previous studies have shown that tyrosine kinase activities and signal transduction are involved in flavivirus replication, suggesting a potential therapeutic strategy for DENV and ZIKV. In this study, we found that compound L3 can significantly reduce viral protein expression and viral titers in HEK-293, MCF-7, HepG2, and Huh-7 cells and exhibits superior therapeutic efficacy against flaviviral infection compared to other tyrosine kinase inhibitors. In addition, compound L3 can decrease endogenous HER2 activation and inhibit the phosphorylation of the HER2 downstream signaling molecules Src and ERK1/2, the levels of which have been associated with viral protein expression in MCF-7 cells. Moreover, silencing HER2 diminished DENV-2 and ZIKV expression in MCF-7 cells, which suggests that HER2 activity is involved in flavivirus replication. Furthermore, in DENV-2-infected AG129 mice, treatment with compound L3 increased the survival rates and reduced the viremia levels. Overall, compound L3 demonstrates therapeutic efficacy both in vitro and in vivo and could be developed as a promising antiviral drug against emerging flaviviruses or for concurrent DENV and ZIKV outbreaks.
    Type of Medium: Online Resource
    ISSN: 1422-0067
    URL: Article
    DOI: 10.3390/ijms21114050
    Language: English
    Publisher: MDPI AG
    Publication Date: 2020
    detail.hit.zdb_id: 2019364-6
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  • 10
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    Blood Aluminum Levels in Patients with Hemodialysis and Peritoneal Dialysis
    MDPI AG ; 2022
    In:  International Journal of Environmental Research and Public Health Vol. 19, No. 7 ( 2022-03-24), p. 3885-
    Details
    In: International Journal of Environmental Research and Public Health, MDPI AG, Vol. 19, No. 7 ( 2022-03-24), p. 3885-
    Abstract: Background. This retrospective observational study attempted to examine the prevalence of abnormal blood aluminum levels in dialysis patients, and to explore the association of pathogenic factors, such as demographic, clinical, laboratory as well as the use of phosphate binding drugs, drugs for secondary hyperparathyroidism and erythropoiesis-stimulating drugs with the blood aluminum levels. Methods. The study included 1175 patients (874 hemodialysis and 301 peritoneal dialysis), recruited from Chang Gung Memorial Hospital in November 2020. Patients were stratified into two groups by their blood aluminum levels, as normal ( 〈 2 µg/dL, n = 1150) or abnormal (≥2 µg/dL, n = 25). Results. The patients aged 60.4 ± 13.2 years and were dialyzed for 8.6 ± 8.1 years. The average blood aluminum level was 1.0 ± 0.4 µg/dL. Patients with abnormal blood aluminum levels received more sevelamer than patients with normal blood aluminum level (p = 0.014). Patients with abnormal blood aluminum levels had higher platelet count (p = 0.001), triglyceride (p 〈 0.001) and total iron binding capacity (p = 0.003) than patients with normal blood aluminum levels. Moreover, the cardiothoracic ratio was higher in patients with abnormal blood aluminum levels than patients with normal blood aluminum levels (p = 0.003). Conclusions. The prevalence of abnormal blood aluminum levels was low at 2.2%. Nevertheless, the linking of cardiothoracic ratio of more than 0.5 as well as elevated blood platelet count and triglyceride level with blood aluminum levels are interesting, and warranted more researches in this area.
    Type of Medium: Online Resource
    ISSN: 1660-4601
    URL: Article
    DOI: 10.3390/ijerph19073885
    Language: English
    Publisher: MDPI AG
    Publication Date: 2022
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