Browsing by Author "Chang, Chia-Ching"

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A Unique and Potent Protein Binding Nature of Liposome Containing Polyethylenimine and Polyethylene Glycol: A Nondisplaceable Property
(2011-06-01)
Liu, Yen-Ku
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Lin, Yu-Ling
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Chen, Chia-Hung
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Lin, Ching-Min
;
Ma, Kuan-Lun
Most of the currently available targeting vectors are produced via the linkage of targeting molecules. However, the coupling process is complicated, and the covalent linkage may attenuate the activity of certain targeting molecules. In this study, we have developed a cationic liposome complexed with polyethylenimine and polyethylene glycol polymers (LPPC) that can capture various proteins without covalent conjugation. Characterizations of prepared LPPC revealed that the maximal-binding capacity was about 170 mu g of bovine serum albumin to 40 mu g of sphere-shaped LPPC (180 nm). The proteins were essentially located at or near the surface when analyzed by atomic force or transmission electron microscopy. We demonstrate that polyethylenimine was an essential component to bind the proteins. Upon the saturation of captured proteins, a given protein could not be displaced by other additional proteins and still retained its biological activity. Using a variety of functional proteins, we show some typical examples of the utility of incorporated beta-glucuronidase and antibodies onto the LPPC. The beta-glucuronidase can be used for the study of antigen-antibody interactions, whereas in studies with the antibody complex, we used anti-CD3 as an agonist to stimulate the proliferation of peripheral blood mononuclear cells via a receptor-mediated mechanism and anti-VEGFR for cell staining. In conclusion, the prepared LPPC can provide a platform to capture biologically and biochemically functional proteins on its surface for various applications, such as cell signaling, cell profiling, noncovalent enzyme-linked immunoassays, and others not mentioned. Biotechnol. Bioeng. Biotechnol. Bioeng. 2011; 108: 1318-1327. (C) 2010 Wiley Periodicals, Inc.
Activation of p38 mitogen-activated protein kinase by celecoxib oppositely regulates survivin and gamma-H2AX in human colorectal cancer cells
(2007-07-01)
Hslao, Po-Wen
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Chang, Chia-Ching
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Liu, Huel-Fang
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Tsai, Chuan-Mei
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Chiu, Ted H.
Cancer cells express survivin that facilitates tumorigenesis. Celecoxib has been shown to reduce human colorectal cancers. However, the role and regulation of survivin by celecoxib in colorectal carcinoma cells remain unclear. Treatment with 40-80 mu M celecoxib for 24 h induced cytotoxicity and proliferation inhibition via a concentration-dependent manner in RKO colorectal carcinoma cells. Celecoxib blocked the survivin protein expression and increased the phosphorylation of H2AX at serine- 193 (gamma-H2AX). The survivin gene knockdown by transfection with a survivin siRNA revealed that the loss of survivin correlated with the expression of gamma-H2AX. Meanwhile, celecoxib increased caspase-3 activation and apoptosis. Celecoxib activated the phosphorylation of p38 mitogen-activated protein (MAP) kinase. The phosphorylated proteins of p38 MAP kinase and gamma-H2AX were observed in the apoptotic cells. SB203580, a specific p38 MAP kinase inhibitor, protected the survivin protein expression and decreased the levels of gamma-H2AX and apoptosis in the celecoxib-exposed cells. The blockade of survivin expression increased the celecoxib-induced cytotoxicity; conversely, overexpression of survivin by transfection with a survivin -expressing vector raised the cancer cell proliferation and resisted the celecoxib-induced cell death. Our results provide for the first time that p38 MAP kinase participates in the down-regulation of survivin and subsequently induces the activation of gamma-H2AX for mediating apoptosis following treatment with celecoxib in human colorectal cancer cells. (c) 2007 Elsevier Inc. All rights reserved.
Alpha-bungarotoxin binding to target cell in a developing visual system by carboxylated nanodiamond
(2008-05-21)
Liu, Kuang-Kai
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Mei-Fang Chen
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Chen, Po-Yi
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Lee, Tony J. F.
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Cheng, Chia-Liang
Biological molecules conjugating with nanoparticles are valuable for applications including bio-imaging, bio-detection, and bio-sensing. Nanometer-sized diamond particles have excellent electronic and chemical properties for bio-conjugation. In this study, we manipulated the carboxyl group produced on the surface of nanodiamond (carboxylated nanodiamond, cND) for conjugating with alpha-bungarotoxin (alpha-BTX), a neurotoxin derived from Bungarus multicinctus with specific blockade of alpha7-nicotinic acetylcholine receptor (alpha 7-nAChR). The electrostatic binding of cND-alpha-BTX was mediated by the negative charge of the cND and the positive charge of the alpha-BTX in physiological pH conditions. Sodium dodecyl sulfate-polyacrylamide gel analysis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF-MS) spectra displayed that alpha-BTX proteins were conjugated with cND particles via non-covalent bindings. The green fluorescence of the cND particles combining with the red fluorescence of tetramethylrhodamine-labeled a-BTX presented a yellow color at the same location, which indicated that alpha-BTX proteins were conjugated with cND particles. Xenopus laevis's oocytes expressed the human alpha 7-nAChR proteins by microinjection with alpha 7-nAChR mRNA. The cND-alpha-BTX complexes were bound to alpha 7-nAChR locating on the cell membrane of oocytes and human lung A549 cancer cells analyzed by laser scanning confocal microscopy. The choline-evoked alpha 7-nAChR-mediated inward currents of the oocytes were blocked by cND-alpha-BTX complexes in a concentration-dependent manner using two-electrode voltage-clamp recording. Furthermore, the fluorescence intensity of cND-alpha-BTX binding on A549 cells could be quantified by flow cytometry. These results indicate that cND-conjugated alpha-BTX still preserves its biological activity in blocking the function of alpha 7-nAChR, and provide a visual system showing the binding of alpha-BTX to alpha 7-nAChR.
An unopened knot protein: YbeA
(2010-04-01)
Chang, Yu-Juan
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Yeh, Yi-Chen
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Ho, Yen-Peng
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Chang, Chia-Ching
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生物科技學系
Bio-imaging of alpha-bungarotoxin binding to target cell by carboxylated nanodiamond
(2008-04-01)
Chao, Jui-I
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Liu, Kuang-Kai
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Chen, Mei-Fang
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Chen, Po-Yi
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Lee, Tony J. F.
Biocompatible and detectable carboxylated nanodiamond on human cell
(2007-08-15)
Liu, Kuang-Kai
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Cheng, Chia-Liang
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Chang, Chia-Ching
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Chao, Jui-I
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生物科技學系
Surface-modified carboxylated nanometre-sized diamond (cND) has been applied for the conjugation of biological molecules such as DNA and protein. In this study, we evaluated the biocompatibility and detection of cNDs and carbon nanotubes on human lung A549 epithelial cells and HFL-1 normal fibroblasts. Treatment with 5 or 100 nm cND particles, 0.1 - 100 mu g ml(-1), did not reduce the cell viability and alter the protein expression profile in lung cells; however, carbon nanotubes induced cytotoxicity in these cells. The cNDs particles were accumulated in A549 cells, which were observed by atomic force microscopy and laser scanning confocal microscopy. Both 5 and 100 nm cNDs particles exhibited the green fluorescence and were ingested into cells. Moreover, the fluorescence intensities were increased in cells via a concentration-dependent manner after treatment with 5 and 100 nm cNDs, which can be detected by flow cytometer analysis. The fluorescence intensities of 5 nm cNDs were relative higher than 100 nm cNDs in cells at equal concentration treatment. The observation demonstrated that cND-interacting with cell is detectable by a confocal microscope, flow cytometer and atomic force microscope. These nanoparticles may be useful for further biomedical applications based on the properties of uptake ability, detectability and little cytotoxicity in human cells.
Calcium dependent p21 (cip1/waf1) stabilizer cyclin I (Ccni)
(2008-04-01)
Chu, Hsueh-Liang
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Chen, Tzu-Hsuan
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Yeh, Yi-Chen
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Tsai, Chuan-Mei
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Wu, Chang-You
Carboxylated nanodiamond-mediated CRISPR-Cas9 delivery of human retinoschisis mutation into human iPSCs and mouse retina
(2020-01-01)
Yang, Tien-Chun
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Chang, Chia-Yu
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Yarmishyn, Aliaksandr A.
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Mao, Yen-Shiang
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Yang, Yi-Ping
Nanodiamonds (NDs) are considered to be relatively safe carbon nanomaterials used for the transmission of DNA, proteins and drugs. The feasibility of utilizing the NDs to deliver CRISPR-Cas9 system for gene editing has not been clearly studied. Therefore, in this study, we aimed to use NDs as the carriers of CRISPR-Cas9 components designed to introduce the mutation in RS1 gene associated with Xlinked retinoschisis (XLRS). ND particles with a diameter of 3 nm were functionalized by carboxylation of the surface and covalently conjugated with fluorescent mCherry protein. Two linear DNA constructs were attached to the conjugated mCherry: one encoded Cas9 endonuclease and GFP reporter, another encoded sgRNA and contained insert of HDR template designed to introduce RS1 c.625C>T mutation. Such nanoparticles were successfully delivered and internalized by human iPSCs and mouse retinas, the efficiency of internalization was significantly improved by mixing with BSA. The delivery of ND particles led to introduction of RS1 c.625C>T mutation in both human iPSCs and mouse retinas. Rs1 gene editing in mouse retinas resulted in several pathological features typical for XLRS, such as aberrant photoreceptor structure. To conclude, our ND-based CRISPR-Cas9 delivery system can be utilized as a tool for creating in vitro and in vivo disease models of XLRS. Statement of significance X-linked retinoschisis (XLRS) is a prevalent hereditary retinal disease, which is caused by mutations in RS1 gene, whose product is important for structural organization of the retina. The recent development of genome editing techniques such as CRISPR-Cas9 significantly improved the prospects for better understanding the pathology and development of treatment for this disease. Firstly, gene editing can allow development of appropriate in vitro and in vivo disease models; secondly, CRISPR-Cas9 can be applied for gene therapy by removing the disease-causative mutation in vivo. The major prerequisite for these approaches is to develop safe and efficient CRISPR-Cas9 delivery system. In this study, we tested specifically modified nanodiamonds for such a delivery system. We were able to introduce Rs1 mutation into the mouse retina and, importantly, observed several XLRS-specific effects. (C) 2019 Acta Materialia Inc. Published by Elsevier Ltd.
Characterizing the polymeric status of Helicobacter pylori heat shock protein 60
(2009-10-16)
Lin, Ching-Yi
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Huang, Yu-Shan
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Li, Chi-Han
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Hsieh, Yuan-Ting
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Tsai, Nu-Man
Helicobacter pylori heat shock protein 60 (HpHsp60) was first identified as an adhesion molecule associated with H. pylori infection. Here we have analyzed the structure of HpHsp60 via amino acid BLAST, circular dichroism, and electrophoresis and the results indicate that most recombinant HpHsp60 molecules exist as dimers or tetramers, which is quite different from Escherichia coli Hsp60. Treatment of human monocytic cells THP-1 with HpHsp60 was found to up-regulate a panel of cytokines including IL-1 alpha, IL-8, IL-10, IFN-gamma, TNF-alpha, TGF-beta, GRO, and RANTES. Carboxymethylated HpHsp60 molecules with a switched oligomeric status were able to further enhance NF-kappa B-mediated IL-8 and TNF-alpha secretion in THP-1 cells compared to unmodified HpHsp60 molecules. These results indicated that the oligomeric status of HpHsp60s might have an important role in regulating host inflammation and thus help facilitate H. pylori persistent infection. (C) 2009 Elsevier Inc. All rights reserved.
Design and synthesis of pyridine-pyrazole-sulfonate derivatives as potential anti-HBV agents
(2016)
Chuang, Hong
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Huang, Lin-Chiang Sherlock
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Kapoor, Mohit
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Liao, Yi-Jen
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Yang, Cheng-Lin
Hepatitis B virus (HBV) is an infectious disease, which can cause acute and chronic infections. Every year, over 7.5 million persons die due to HBV. No effective drug exists for the treatment of HBV. Thus, we designed and synthesized 16 new pyridine-pyrazole-sulfonate compounds containing pyridine-SCH2-pyrazole and pyridine-pyrazole derivatives. Their structures were characterized by H-1-NMR, C-13-NMR, IR, mass spectroscopy, and high performance liquid chromatography. All the compounds were evaluated for their anti-HBV activities and a structure-activity relationship (SAR) in HepG2 2.2.15 cells was established. We found that the pyridine-pyrazole derivatives could inhibit the HBV gene expression and viral DNA replication. Among these compounds, 2-[3-(2-nitrophenylsulfonyl)oxy-5-pyrazol-yl]pyridine 19d has shown the most potent inhibitory activity with an IC50 value of 9.19 mu M and high selectivity index, SI (TC50/IC50) of 35.46. Hence, we believe our compounds could serve as a potential lead compounds for anti-HBV drug development.
Development of a Graphene Oxide-Incorporated Polydimethylsiloxane Membrane with Hexagonal Micropillars
(2018-09-01)
Lin, Yi-Ying
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Chien, Yueh
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Chuang, Jen-Hua
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Chang, Chia-Ching
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Yang, Yi-Ping
Several efforts have been made on the development of bioscaffolds including the polydimethylsiloxane (PDMS) elastomer for supporting cell growth into stable sheets. However, PDMS has several disadvantages, such as intrinsic surface hydrophobicity and mechanical strength. Herein, we generated a novel PDMS-based biomimetic membrane by sequential modifications of the PMDS elastomer with graphene oxide (GO) and addition of a hexagonal micropillar structure at the bottom of the biomembrane. GO was initially homogenously mixed with pure PDMS and then was further coated onto the upper surface of the resultant PDMS. The elastic modulus and hydrophilicity were significantly improved by such modifications. In addition, the development of hexagonal micropillars with smaller diameters largely improved the ion permeability and increased the motion resistance. We further cultured retinal pigment epithelial (RPE) cells on the surface of this modified PDMS biomembrane and assayed its biocompatibility. Remarkably, the GO incorporation and coating exhibited beneficial effect on the cell growth and the new formation of tight junctions in RPE cells. Taken together, this GO-modified PDMS scaffold with polyhexagonal micropillars may be utilized as an ideal cell sheet and adaptor for cell cultivation and can be used in vivo for the transplantation of cells such as RPE cells.
Development of a Growth- Hormone- Conjugated Nanodiamond Complex for Cancer Therapy
(2014-05-01)
Chu, Hsueh-Liang
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Chen, Hung-Wei
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Tseng, Shin-Hua
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Hsu, Ming-Hua
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Ho, Li-Ping
It is highly desirable to develop a therapeutic, observable nanoparticle complex for specific targeting in cancer therapy. Growth hormone (GH) and its antagonists have been explored as cancer cell-targeting molecules for both imaging and therapeutic applications. In this study, a low toxicity, biocompatible, therapeutic, and observable GH-nanoparticle complex for specifically targeting growth hormone receptor (GHR) in cancer cells was synthesized by conjugating GH with green fluorescence protein and carboxylated nanodiamond. Moreover, we have shown that this complex can be triggered by laser irradiation to create a nanoblast and induce cell death in the A549 non-small-cell lung cancer cell line via the apoptotic pathway. This laser-mediated, cancer-targeting platform can be widely used in cancer therapy.
Development of nordihydroguaiaretic acid derivatives as potential multidrug-resistant selective agents for cancer treatment
(2015-01-01)
Huang, Lin-Chiang Sherlock
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Chuang, Hong
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Kapoor, Mohit
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Hsieh, Cheng-Ying
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Chou, Shih-Ching
In this research, we designed and synthesized a new series of nordihydroguaiaretic acid (NDGA) derivatives for multidrug resistance (MDR) research. A methylsulfonyl NDGA derivative, ((2R,3S)-2,3-dimethylbutane-1,4-diyl)bis(benzene-4,1,2-triyl) tetramethanesulfonate (5d), was found to inhibit MDR1 gene expression and suppress drug resistantMES-SA/Dx5 cells. Moreover, the combination of 5d and doxorubicin/terameprocol (M4N) showed a profound synergistic effect on inhibition of drug resistant cancer cells, suggesting that 5d is a potential adjuvant applied with doxorubicin or terameprocol in cancer treatment.
Direct Observation of Single Molecule Conformational Change of Tight-Turn Paperclip DNA Triplex in Solution
(2009-10-01)
Liu, Ching-Ping
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Wey, Ming-Tsai
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Chang, Chia-Ching
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Kan, Lou-Sing
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生物科技學系
DNA triplex modulates gene expression by forming stable conformation in physiological condition. However, it is not feasible to observe this unique molecular structure of large molecule with 54 oligodeoxynucleotides directly by conventional nuclear magnetic approach. In this study, we observed directly single molecular images of paperclip DNA triplexes formation in a buffer solution of pH 6.0 by atomic force microscopy (AFM). Meanwhile, a diffuse "tail" of unwound DNA was observed in pH 8.0 solution. This designable approach in visualizing the overall structures and shapes of oligo-DNAs at the single molecular level, by AFM, is applicable to other biopolymers as well.
Direct visualization of triplex DNA molecular dynamics by fluorescence resonance energy transfer and atomic force microscopy measurements
(2008-04-01)
Chang, Chia-Ching
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Lin, Po-Yen
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Chang, Chia-Seng
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Kan, Lou-Sing
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交大名義發表
Direct visualization of triplex DNA molecular dynamics by fluorescence resonance energy transfer and atomic force microscopy measurements
(2007-11-12)
Chang, Chia-Ching
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Lin, Po-Yen
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Chen, Yen-Fu
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Chang, Chia-Seng
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Kan, Lou-Sing
We have detected the dynamics of 17-mer DNA triplex dissociation mechanism at the molecular level. Fluorescence resonance energy transfer (FRET) was used as an indicator of intermolecular interaction in nanometer range, whereas atomic force microscopy (AFM) was employed to address single molecule with sub-angstrom precision. The maximum rupture force of DNA triplex was found at pH 4.65, consistent with macroscopic observations. These results indicated that the FRET together with an AFM detection system could be used to reveal the DNA triplex interaction in nanometer scale unambiguously. (c) 2007 American Institute of Physics.
Directly Thiolated Modification onto the Surface of Detonation Nanodiamonds
(2014-05-28)
Hsu, Ming-Hua
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Chuang, Hong
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Cheng, Fong-Yu
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Huang, Ying-Pei
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Han, Chien-Chung
An efficient method for modifying the surface of detonation nanodiamonds (5 and 100 nm) with thiol groups (-SH) by using an organic chemistry strategy is presented herein. Thiolated nanodiamonds were characterized by spectroscopic techniques, and the atomic percentage of sulfur was analyzed by elemental analysis and X-ray photoelectron spectroscopy. The conjugation between thiolated nanodiamonds and gold nanoparticles was elucidated by transmission electron microscopy and UV-vis spectrometry. Moreover, the material did not show significant cytotoxicity to the human lung carcinoma cell line and may prospectively be applied in bioconjugated technology. The new method that we elucidated may significantly improve the approach to surface modification of detonation nanodiamonds and build up a new platform for the application of nanodiamonds.
DNA Mismatch Detection by Metal Ion Enhanced Impedance Analysis
(2009-10-01)
Jangjian, Peng-Chung
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Liu, Tzeng-Feng
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Tsai, Chuan-Mei
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Li, Mei-Yi
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Tsai, Ming-Shih
Conventional diagnoses of genetic mutation and disease depend on the analysis of DNA sequences. However, mismatches in the DNA sequences are difficult to detect using traditional sequencing. Doping metal ions, such as nickel, into the short DNA (28 mer similar to 30 mer), markedly reduces its electrical resistance, which is measured by electrical impedance analysis. The change in resistance that is caused by a mismatched base-pair in short DNA can also be monitored by this approach. In this study, the resistance increased exponentially with the number of mismatched base-pairs. Accordingly, an intuitive and direct method for evaluating the number of DNA mismatches can possibly be developed.
DNA-based nanowires and nanodevices
(2017-01-01)
Pandian, Sureshbabu Ram Kumar
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Yuan, Chiun-Jye
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Lin, Chung-Ching
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Wang, Wen-Hung
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Chang, Chia-Ching
DNA (deoxyribonucleic acid) is a highly versatile biopolymer that has been a recent focus in the field of nanomachines and nanoelectronics. DNA exhibits many properties, such as high stability, adjustable conductance, vast information storage, self-organising capability and programmability, making it an ideal material in the applications of nanodevices, nanoelectronics and molecular computing. Even though native DNA has low conductance, it can easily be converted into a potential conductor by doping metal ions into the base pairs. Nickel ions have been employed to tune DNA into conducting polymers. Doping of nickel ions within DNA (Ni-DNA) increases the conductivity of DNA by at least 20 folds compared with that of native DNA. Further studies showed that Ni-DNA nanowires exhibit characteristics of memristors, making them a potential mass information storage system. In summary, DNA molecules have promising applications in a variety of fields, including nanodevices, nanomachines, nanoelectronics, organic solar cells, organic light emitting diodes and biosensors. [GRAPHICS] .
Enhanced bioconjugation on sputtered palladium nano-thin-film electrode
(2019-03-04)
Chang, Chia-Yu
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Chen, Wei
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Su, Chien-Hao
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Chang, Pang-Chia
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Huang, Yun-Tzu
A highly reactive surface with an enhanced ability for chemical bonding relies on the presence of specifically coordinated atoms and step edges at the surface. In this study, an electrode with a unique Stranski-Krastanov-like thin film, with an epitaxial sputtering of a palladium (Pd) nanoparticle double layer on the polyethylene terephthalate substrate, was developed. On the surface of this flexible Pd-nano-thin-film (NTF) electrode with a (1 1 1) containing surface, DNA probes can be quickly immobilized in as short a period as 20 min, which is 24 times faster than that on the gold electrode. A DNA-based anticancer compound (ACC) sensing and screening process that would use the DNA functionalized Pd-NTF electrode as the biosensor was then proposed. Interestingly, the developed biosensor could detect DNA and ACCs, such as doxorubicin, tetra-O-methyl nordihydroguaiaretic acid, and Taxol via interactions with solutions containing 1 mu l ACCs within 11 min, and the sensitivity of the ACC solution is similar to 0.1 mu M (similar to 36 pg per-test), as detected by electrochemical impedance spectroscopy. Moreover, this highly reactive surface can be used in regular sensors and other interfaces, in scientific applications. (C) 2019 Author(s).