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Lee Y.-B.,Yongkang Veterans Hospital | Ko K.-C.,Yongkang Veterans Hospital | Shi M.-D.,Yongkang Veterans Hospital | Shi M.-D.,Kaohsiung Medical University | And 5 more authors.
Journal of Food Science | Year: 2010

This study first investigates the anti-metastatic effect of α-mangostin on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) expressions in human breast adenocarcinoma cells, MCF-7. First, the result demonstrated α-mangostin could inhibit TPA-induced abilities of the adhesion, invasion, and migration by cell-matrix adhesion assay and Boyden chamber assay. Data also showed α-mangostin could inhibit the activation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) involved in the downregulation the enzyme activities, protein, and messenger RNA levels of MMP-2 and MMP-9 induced by TPA. Next, α-mangostin also strongly inhibited TPA-induced degradation of inhibitor of kappaBα (IκBα) and the nuclear levels of nuclear factor kappa B (NF-κB), c-Fos, and c-Jun. Also, a dose-dependent inhibition on the binding abilities of NF-κB and activator protein-1 (AP-1) by α-mangostin treatment was further observed. Further, the treatment of specific inhibitor for ERK (U0126) to MCF-7 cells could inhibit TPA-induced MMP-2 and MMP-9 expressions along with an inhibition on cell invasion and migration. Presented data reveal that α-mangostin is a novel, effective, antimetastatic agent that functions by downregulating MMP-2 and MMP-9 gene expressions. © 2009 Institute of Food Technologists ®.


Huang G.-J.,China Medical University at Taichung | Hsieh W.-T.,China Medical University at Taichung | Chang H.-Y.,China Medical University at Taichung | Chang H.-Y.,Yongkang Veterans Hospital | And 4 more authors.
Journal of Agricultural and Food Chemistry | Year: 2011

The inhibitory activity from the isolated component of the fruiting body Phellinus merrillii (PM) was evaluated against α-glucosidase and lens aldose reductase from Sprague-Dawley male rats and compared to the quercetin as an aldose reductase inhibitor and acarbose as an α-glucosidase inhibitor. The ethanol extracts of PM (EPM) showed the strong α-glucosidase and aldose reductase activities. α-Glucosidase and aldose reductase inhibitors were identified as hispidin (A), hispolon (B), and inotilone (C), which were isolated from EtOAc-soluble fractions of EPM. The above structures were elucidated by their spectra and comparison with the literatures. Among them, hispidin, hispolon, and inotilone exhibited potent against α-glucosidase inhibitor activity with IC50 values of 297.06 ± 2.06, 12.38 ± 0.13, and 18.62 ± 0.23 μg/mL, respectively, and aldose reductase inhibitor activity with IC50 values of 48.26 ± 2.48, 9.47 ± 0.52, and 15.37 ± 0.32μg/mL, respectively. These findings demonstrated that PM maybe a good source for lead compounds as alternatives for antidiabetic agents currently used. The importance of finding effective antidiabetic therapeutics led us to further investigate natural compounds. © 2011 American Chemical Society.


Chen C.-C.,Chia Nan University of Pharmacy and Science | Lien H.-Y.,Yongkang Veterans Hospital | Lien H.-Y.,Tungnan University | Hsu Y.-J.,Chi Mei Medical Center | And 3 more authors.
Journal of Applied Physiology | Year: 2010

Epidemiologic studies showed that men treated with statins appear to have a lower incidence of sudden death than men without statins. However, the specific factor for this remained disappointingly elusive. We assessed whether pravastatin enhanced connexin43 expression after myocardial infarction through attenuation of endothelin-1. Twenty-four hours after ligation of the anterior descending artery, male Wistar rats were randomized to vehicle, pravastatin, mevalonate, bosentan, or a combination of pravastatin and mevalonate or pravastatin and bosentan for 4 wk. Myocardial endothelin- 1 levels were significantly elevated in vehicle-treated rats at the border zone compared with sham-operated rats. Myocardial connexin43 expression at the border zone was significantly decreased in vehicle-treated infarcted rats compared with sham-operated rats. Attenuated connexin43 expression was blunted after administration of pravastatin, as assessed by immunofluorescence analysis, Western blotting, and real-time quantitative RT-PCR of connexin43. Bosentan enhanced connexin43 amount in infarcted rats and did not have additional beneficial effects on pravastatintreated rats. Arrhythmic scores during programmed stimulation in vehicle- treated rats were significantly higher than scores in those treated with pravastatin. In contrast, the beneficial effects of pravastatin-induced connexin43 were abolished by the addition of mevalonate and a protein kinase C inducer. In addition, the amount of connexin43 showed significant increase after addition of bisindolylmaleimide, implicating that protein kinase C is a relevant target in endothelin-1-mediated connexin43 expression. Thus chronic use of pravastatin after infarction, resulting in enhanced connexin43 amount by attenuation of mevalonate-dependent endothelin-1 through a protein kinase C-dependent pathway, may attenuate the arrhythmogenic response to programmed electrical stimulation. © 2010 the American Physiological Society.


Meir Y.-J.,Chang Gung University | Weirauch M.T.,University of Toronto | Yang H.-S.,YongKang Veterans Hospital | Chung P.-C.,Chang Gung University | And 2 more authors.
BMC Biotechnology | Year: 2011

Background: DNA transposons have emerged as indispensible tools for manipulating vertebrate genomes with applications ranging from insertional mutagenesis and transgenesis to gene therapy. To fully explore the potential of two highly active DNA transposons, piggyBac and Tol2, as mammalian genetic tools, we have conducted a side-by-side comparison of the two transposon systems in the same setting to evaluate their advantages and disadvantages for use in gene therapy and gene discovery.Results: We have observed that (1) the Tol2 transposase (but not piggyBac) is highly sensitive to molecular engineering; (2) the piggyBac donor with only the 40 bp 3'-and 67 bp 5'-terminal repeat domain is sufficient for effective transposition; and (3) a small amount of piggyBac transposases results in robust transposition suggesting the piggyBac transpospase is highly active. Performing genome-wide target profiling on data sets obtained by retrieving chromosomal targeting sequences from individual clones, we have identified several piggyBac and Tol2 hotspots and observed that (4) piggyBac and Tol2 display a clear difference in targeting preferences in the human genome. Finally, we have observed that (5) only sites with a particular sequence context can be targeted by either piggyBac or Tol2.Conclusions: The non-overlapping targeting preference of piggyBac and Tol2 makes them complementary research tools for manipulating mammalian genomes. PiggyBac is the most promising transposon-based vector system for achieving site-specific targeting of therapeutic genes due to the flexibility of its transposase for being molecularly engineered. Insights from this study will provide a basis for engineering piggyBac transposases to achieve site-specific therapeutic gene targeting. © 2011 Meir et al; licensee BioMed Central Ltd.


PubMed | Chang Bing Show Chwan Memorial Hospital, Chang Gung University, Yongkang Veterans Hospital, Chang Gung Institute of Technology and Kaohsiung Veterans General Hospital
Type: Journal Article | Journal: Human & experimental toxicology | Year: 2014

Celecoxib has been shown to have antitumor effect in previous studies but the mechanisms are unclear. The effect of celecoxib on cytosolic Ca(2+) concentrations ([Ca(2+)]i) and viability in HA59T human hepatoma cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)]i. Celecoxib at concentrations of 10-50 M induced a [Ca(2+)]i rise in a concentration-dependent manner. The response was reduced by 80% by removing Ca(2+). Celecoxib induced Mn(2+) influx, leading to quenching of fura-2 fluorescence. Celecoxib-evoked Ca(2+) entry was suppressed by nifedipine, econazole, SK&F96365, and protein kinase C modulators. In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin nearly abolished celecoxib-induced [Ca(2+)]i rise. Incubation with celecoxib abolished thapsigargin-induced [Ca(2+)]i rise. Inhibition of phospholipase C with U73122 abolished celecoxib-induced [Ca(2+)]i rise. At 1-50 M, celecoxib inhibited cell viability by less than 20%, which was not reversed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N, N, N, N-tetraacetic acid/acetoxy methyl (BAPTA/AM). Celecoxib (10-50 M) also induced apoptosis. In sum, in HA59T hepatoma cells, celecoxib induced a [Ca(2+)]i rise by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via protein kinase C-sensitive store-operated Ca(2+) channels. Celecoxib also caused cell death via apoptosis.


Wu C.-L.,National Cheng Kung University | Su S.-B.,Southern Taiwan University of Science and Technology | Su S.-B.,Chi Mei Medical Center | Lien H.-Y.,Yongkang Veterans Hospital | And 2 more authors.
Burns | Year: 2012

Objective: To evaluate the role of the chemical burns caused by hydroxide ion in the fatal effects of tetramethylammonium ion (TMA) in dermal exposure to tetramethylammonium hydroxide (TMAH), we conducted a rat study consisting of two-step treatments with dermal exposure to NaOH and tetramethylammonium chloride (TMACl). Methods: In the first step, NaOH or saline was administered in the gauze on the shaved skin for 5 min, and in the second step, TMAH, TMACl, or saline was administered in the same way. The mean blood pressure (MBP), heart rate (HR), and survival in rats were compared among seven groups. Results: Dermal exposure to saline and then 2.75 M TMACl introduced limited and temporary non-fatal effects. Exposure to 2.75 M NaOH and then saline had almost no effects and caused no deaths. Treatments with more concentrated NaOH or TMACl resulted in suppressions of MBP and HR, and deaths were observed after the dosing of TMACl. Conclusion: The toxicity of dermal exposure to TMA alone is limited, but fatal effects can be introduced by pre-treatment with hydroxide ion. Therefore, the chemical burn caused by hydroxide ion plays an essential role in the toxicity, implicating that effective neutralizing may help decreasing the fatality rate. © 2012 Elsevier Ltd and ISBI. All rights reserved.


Chien S.-T.,Kaohsiung Armed Forces General Hospital | Lin S.-S.,Kaohsiung Armed Forces General Hospital | Wang C.-K.,E Chyun Dermatology Clinic | Wang C.-K.,Chung Hwa University of Medical Technology | And 3 more authors.
Molecular and Cellular Biochemistry | Year: 2011

Lung cancer is one of the most common malignancies in the world and its metastasis is the major cause of death in cancer patients. Acacetin (5,7-dihydroxy-4′-methoxyflavone), a flavonoid compound, has anti-peroxidative and anti-inflammatory effects. The effect of acacetin on invasion and migration in human NSCLC A549 cells was investigated. First, the result demonstrated acacetin could exhibit an inhibitory effect on the abilities of the adhesion, morphology/actin cytoskeleton arrangement, invasion, and migration by cell-matrix adhesion assay, immunofluorescence assay, Boyden chamber assay, and wound-healing assay. Molecular data showed that the effect of acacetin in A549 cells might be mediated via sustained inactivation of the phosphorylation of mixed-lineage protein kinase 3 (MLK3), mitogen-activated protein kinase kinases 3/6 (MKK3/6), and p38α MAPK signal involved in the downregulation of the expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and urokinase-type plasminogen activator (u-PA). Next, acacetin significantly decreased in the phosphorylation and degradation of inhibitor of kappaBα (IκBα), and the nuclear levels of nuclear factor kappa B (NF-κB), c-Fos, and c-Jun. Also, the treatment with acacetin to A549 cells also leads to a concentration-dependent inhibition on the binding abilities of NF-κB and activator protein-1 (AP-1). Furthermore, the treatment of specific inhibitor for p38 MAPK (SB203580) to A549 cells could cause reduced activities of MMP-2/9 and u-PA. In addition, acacetin significantly decreased the levels of phospho-p38α MAPK, MMP-2/9, and u-PA in p38α-cDNA-transfected cells concomitantly with a marked reduction on cell invasion and migration. Our results revealed the anti-migration and anti-invasion effects of acacetin, which may act as a promising therapeutic agent for the treatment of lung cancer. © 2011 Springer Science+Business Media, LLC.


Lee T.-M.,Taipei Medical University | Lin C.-C.,Chi Mei Medical Center | Lien H.-Y.,Yongkang Veterans Hospital | Lien H.-Y.,Tungnan University | And 2 more authors.
Journal of Cellular and Molecular Medicine | Year: 2012

Downward remodelling of gap junctional proteins between myocytes may trigger ventricular arrhythmia after myocardial infarction. We have demonstrated that ATP-sensitive potassium (KATP) channel agonists attenuated post-infarction arrhythmias. However, the involved mechanisms remain unclear. The purpose of this study was to determine whether KATP channel agonists can attenuate arrhythmias through preserving protein kinase C (PKC)-ε-dependent connexin43 level after myocardial infarction. Male Wistar rats after ligating coronary artery were randomized to either vehicle, nicorandil, pinacidil, glibenclamide or a combination of nicorandil and glibenclamide or pinacidil and glibenclamide for 4 weeks. To elucidate the role of PKCε in the modulation of connexin43 level, carbachol and myristoylated PKCε V1-2 peptide were also assessed. Myocardial connexin43 level was significantly decreased in vehicle-treated infarcted rats compared with sham. Attenuated connexin43 level was blunted after administering KATP channel agonists, assessed by immunofluorescent analysis, Western blotting, and real-time quantitative reverse transcription-PCR of connexin43. Arrhythmic scores during programmed stimulation in the KATP channel agonists-treated rats were significantly lower than those treated with vehicle. The beneficial effects of KATP channel agonists were blocked by either glibenclamide or 5-hydroxydecanoate. Addition of the PKC activator, phorbol 12-myristate 13-acetate and the specific PKCε agonist, carbachol, blocked the effects of nicorandil on connexin43 phosphorylation and dye permeability. The specific PKCε antagonist, myristoylated PKCε V1-2 peptide, did not have additional beneficial effects on connexin43 phosphorylation compared with rats treated with nicorandil alone. Chronic use of KATP channel agonists after infarction, resulting in enhanced connexin43 level through a PKCε-dependent pathway, may attenuate the arrhythmogenic response to programmed electrical stimulation. © 2011 The Authors Journal of Cellular and Molecular Medicine. © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.


Lee Y.-Y.,I - Shou University | Lee Y.-Y.,YongKang Veterans Hospital | Lin J.L.,I - Shou University
Patient Education and Counseling | Year: 2011

Objective: To examine the impact of trust on patient outcomes (satisfaction, HbA 1C, physical and mental health-related quality of life (HRQoL)) and to investigate the role of decision-making preferences in the trust-outcome relationship. Methods: We conducted a one-year longitudinal analysis of 614 type 2 diabetic patients (mean age: 59.3 years; mean disease duration: 6.7 years). Patients' self-administered questionnaires and medical record were used for the research. Multiple regression analyses were conducted to investigate the relationship among variables during a 12-month follow-up. Further, we used latent growth modeling (LGM) to assess changes in health outcomes and to examine how these changes were related to trust. Results: Regression analyses revealed that trust was positively related to glycemic control, physical HRQoL, and satisfaction at 12 months. Patients with higher decision-making preferences experienced a greater increase in subsequent satisfaction. The results of LGM showed that higher levels of trust were associated with greater increases in physical HRQoL. Conclusion: Trust contributes to improvements in health outcomes. The relationship between trust and satisfaction may be stronger among patients with higher decision-making preferences. Practice implications: For healthcare providers, efforts should be made to cultivate patients' trust and enhance their decision-making preferences to maximize satisfaction and improve outcomes. © 2010 Elsevier Ireland Ltd.


Chen P.-Y.,Chung Hwa University of Medical Technology | Yin L.-T.,Chung Hwa University of Medical Technology | Shi M.-D.,Yongkang Veterans Hospital | Lee Y.-C.,Chung Hwa University of Medical Technology
Life Science Journal | Year: 2013

In this research, the drift and temperture characteristics of extended gate field effect transistor (EGFET) based on tin oxide/indium tin oxide (SnO2/ITO) sensing gate were investigated. This separate structure EGFET is formed by dividing an ion sensitive membrane from the field effect transistor. This separative ion sensitive membrane has the advantages of simple structure, fabrication and encapsulation. Accordingly, the field effect transistor does not need to put into solution, so we can realize the pure characteristics of optical and temperature of SnO2 sensing film. The device exhibited a drift amount in pH2,4,6,8 and 10 are 0.884, 1.58, 1.71, 1.8 and 2.51 mV/hr, respectively. The temperature coefficient of sensitivity of SnO2/ITO glass EGFET is 0.31 (mV/pH)/°CThe pH value with a zero temperature coefficient, which can be defined as pHZTC, is found as pH2.5. © 2013. Marsland Press, Zhengzhou University.

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