Browsing by Author "Yuan, Chiun-Jye"

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A general strategy to achieve ultra-high gene transfection efficiency using lipid-nanoparticle composites
(2014-09-01)
Vankayala, Raviraj
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Chiang, Chi-Shiun
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Chao, Jui-I
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Yuan, Chiun-Jye
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Lin, Shyr-Yeu
Gene therapy provides a new hope for previously "incurable" diseases. Low gene transfection efficiency, however, is the bottle-neck to the success of gene therapy. It is very challenging to develop non-viral nanocarriers to achieve ultra-high gene transfection efficiencies. Herein, we report a novel design of "tight binding-but-detachable" lipid-nanoparticle composite to achieve ultrahigh gene transfection efficiencies of 60 similar to 82%, approaching the best value (similar to 90%) obtained using viral vectors. We show that Fe@CNPs nanoparticles coated with LP-2000 lipid molecules can be used as gene carriers to achieve ultra-high (60-80%) gene transfection efficiencies in HeLa, U-87MG, and TRAMP-C1 cells. In contrast, Fe@CNPs having surface-covalently bound N,N,N-trimethyl-N-2-methacryloxyethyl ammonium chloride (TMAEA) oligomers can only achieve low (23-28%) gene transfection efficiencies. Similarly ultrahigh gene transfection/expression was also observed in zebrafish model using lipid-coated Fe@CNPs as gene carriers. Evidences for tight binding and detachability of DNA from lipid-nanoparticle nanocarriers will be presented. (C) 2014 Elsevier Ltd. All rights reserved.
A rapid and selective method for monitoring the growth of coliforms in milk using the combination of amperometric sensor and reducing of methylene blue
(2009-09-07)
Lee, Yuan-Geuy
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Wu, Hung-Yi
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Hsu, Chuan-Liang
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Liang, Hong-Jen
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Yuan, Chiun-Jye
A novel method for monitoring the growth of coliforms in milk was developed based on measuring the current change in an amperometric sensor. The sensor consists of a circuit with a homemade potentiostat and a pair of electrodes. The electrode was immersed in milk samples containing methylene blue with various concentrations of bacterial inoculums. The microbial metabolism led to the reduction of methylene blue resulting in a change of current. The time required to identify readily detectable change (detection time, DT) provided an approximate measurement of the amount of microorganisms in the initial inoculums. The sensor system used in this study has the selectivity towards coliform bacteria such as Escherichia coli and Enterobacter aerogenes. The calibration curve of DT against concentration of coliform showed a linear correlation coefficient (R(2) = 0.9192) over the range of 10(2)-10(8) CFU/mL. The sensor was able to detect the coliform bacteria at initial concentrations of 10(5) CFU/mL within 6 h, making it suitable for use in real-time monitoring of bacterial growth. This system has potential application in the detection of coliform concentration in milk at dairy farms when a proper selective media is designed. (C) 2009 Elsevier B.V. All rights reserved.
Activation of rabbit TLR9 by different CpG-ODN optimized for mouse and human TLR9
(2012-09-01)
Liu, Jin
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Xu, Congfeng
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Liu, Yi-Ling
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Matsuo, Hanako
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Hsieh, Rebecca Pe-Feng
"Synthetic CpG-oligodeoxynucleotides (CpG-ODN) are potent adjuvants that accelerate and boost antigen-specific immune responses. Toll-like receptor 9 (TLR9) is the cellular receptor for these CpG-ODN. Previous studies have shown species-specific activation of mouse TLR9 (mTLR9) and human TLR9 (hTLR9) by their optimized CpG-ODN. The interaction between rabbit TLR9 (rabTLR9) and CpG-ODN, however, has not been previously investigated. Here, we cloned and characterized rabTLR9 and comparatively investigated the activation of the rabbit, mouse, and human TLR9 by CpG-ODN. The complete open reading frame of rabTLR9 encodes 1028 amino acid residues, which share 70.6% and 75.5% of the identities of mTLR9 and hTLR9, respectively. Rabbit TLR9 is preferentially expressed in immune cells rich tissues, and is localized in intracellular vesicles. While mTLR9 and hTLR9 displayed species-specific recognition of their optimized CpG-ODN, rabbit TLR9 was activated by these CpG-ODN without any preference. This result suggests that rabTLR9 has a broader ligand-recognition profile than mouse and human TLR9. (C) 2012 Elsevier Ltd. All rights reserved."
Application of SiO2-poly(dimethylsiloxane) hybrid material in the fabrication of amperometric biosensor
(2006-07-21)
Ho, Wei-Jen
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Yuan, Chiun-Jye
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Reiko, Ohara
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生物科技學系
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Department of Biological Science and Technology
Silica xerogel was widely used in the development of biosensors by coupling the desired biological components to various transducers. Unfortunately, the application of xerogels is limited due to their poor mechanical properties and poor structural maintenance of entrapped biomaterials. In this study, the availability of poly(dimethylsiloxane) (PDMS)-modified silica sol-gel (TEOS/PDMS Ormosil) glass in the immobilization of enzymes and its application in the fabrication of amperometric biosensor was investigated. We demonstrated that most of activity of encapsulated horseradish peroxidase can be preserved in PDMS-modified SiO2 glass compared with conventional silica xerogel. The synthesized monoliths exhibited high transparency and crack-free. The enzyme electrode based on glucose oxidase encapsulated TEOS/PDMS Ormosil glass on Pd electrode was fabricated and characterized electrochemically. The characteristics of the biosensor were studied by cyclic voltammetry and chronoamperometry. The biosensor exhibited a series of good properties: high sensitivity (1.63 mu A mM(-1) analyte), short response time and high reproducibility. The results indicate that TEOS/PDMS Ormosil glass has great potential to the immobilization of biomaterials as well as the fabrication of biosensors. (c) 2006 Elsevier B.V. All rights reserved.
Blosignaling of mammalian Ste20-related kinases
(2008-07-01)
Ling, Pin
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Lu, Te-Jung
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Yuan, Chiun-Jye
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Lai, Ming-Derg
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生物科技學系
Sterile 20 (ste20) protein is an upstream ser/thr kinase in yeast, and several mammalian Ste20-like (MST) kinases have been identified. This review focuses on the signal transduction, interacting proteins, and potential biological function of MST1, 2, 3, and 4 kinases, since several novel signal pathways of these kinases have been characterized recently. MST1 and MST2 kinases play an important role in cell growth and apoptosis, and the signal pathways involves many important molecules including RAS, AKT, and FOXO3. MST3 and MST4 have similar kinase domain, but have opposite effects on apoptosis and transformation. The downstream signaling molecules of these two kinases are beginning to be elucidated. Based on the expression pattern and signal pathways, we will discuss the perspective biological functions of four MST family kinases in cancer, immune, cardiovascular, and brain function. (c) 2007 Elsevier Inc. All rights reserved.
Comparison of amperometric biosensors fabricated by palladium sputtering, palladium electrodeposition and Nafion/carbon nanotube casting on screen-printed carbon electrodes
(2007-01-15)
Lee, Chung-Hun
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Wang, Shih-Chang
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Yuan, Chiun-Jye
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Wen, Meng-Fang
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Chang, Ku-Shang
Different strategies, including palladium electrodeposition (Pd-CV), I'd sputtering (Pd-S) and Nafion-solubilized carbon nanotube casting (Nafion/CNT), were used to modify screen-printed carbon electrodes (SPCEs) for the fabrication of amperometric enzyme biosensors. The electrochemical properties of the bare and modified SPCEs and the optimal conditions for surface modification were determined. The electrochemical response of the bare SPCE to H2O2 under the potential of 0.3 V could be improved about 100-fold by Pd modification by electrodeposition or sputtering. By contrast, the electrochemical response of the bare SPCE was enhanced by only about 11-fold by Nafion/CNT casting. Moreover, the Pd-CV-SPCEs exhibited better reproducibility of electrochemical response (a relative standard deviation (R.S.D.)< 6.0%) than freshly prepared Pds-SPCEs (R.S.D. > 10%). The glucose biosensor fabricated from Pd-modified electrodes could be stored for up to 108 days without loosing significant activity. The Pd-CV-SPCE also showed very reliable signal characteristics upon 50 consecutively repeated measurements of ascorbic acid. The electrocatalytic detection of the Pd-SPCE was combined with additional advantages of resistance to surface fouling and hence good stability. In conclusion, this study demonstrated that deposition of Pd thin film on SPCEs by electrodeposition or sputtering provided superior enhancement of electrochemical properties compared to Nafion/CNT-SPCEs. Despite their high electrochemical response, Pds-SPCEs required an activation process to improve stability and Pd-CV-SPCEs suffered from poor between electrode reproducibility. (c) 2006 Elsevier B.V. All rights reserved.
Development of biosensor for protein tyrosine kinases
(2010-04-01)
Wei, Lan-Yi
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Yuan, Chiun-Jye
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生物科技學系
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Department of Biological Science and Technology
Development of biosensor for protein tyrosine kinases
(2012-04-01)
Wei, Lan-Yi
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Yuan, Chiun-Jye
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生物科技學系
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Department of Biological Science and Technology
Development of biosensor for protein tyrosine kinases
(2012-04-01)
Wei, Lan-Yi
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Yuan, Chiun-Jye
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生物科技學系
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Department of Biological Science and Technology
Development of biosensor for serine/threonine kinases
(2012-04-01)
Lee, Jhao-hong
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Yuan, Chiun-Jye
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生物科技學系
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分子醫學與生物工程研究所
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Department of Biological Science and Technology
Development of biosensor for serine/threonine kinases
(2012-04-01)
Lee, Jhao-hong
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Yuan, Chiun-Jye
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生物科技學系
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分子醫學與生物工程研究所
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Department of Biological Science and Technology
Development of biosensor for serine/threonine kinases
(2010-04-01)
Lee, Jhao-hong
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Yuan, Chiun-Jye
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生物科技學系
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分子醫學與生物工程研究所
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Department of Biological Science and Technology
Development of CpG-Oligodeoxynucleotides for Effective Activation of Rabbit TLR9 Mediated Immune Responses
(2014-09-30)
Chuang, Tsung-Hsien
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Lai, Chao-Yang
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Tseng, Ping-Hui
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Yuan, Chiun-Jye
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Hsu, Li-Chung
CpG-oligodeoxynucleotides (CpG-ODN) are potent immune stimuli being developed for use as adjuvants in different species. Toll-like receptor 9 (TLR9) is the cellular receptor for CpG-ODN in mammalian cells. The CpG-ODN with 18-24 deoxynucleotides that are in current use for human and mouse cells, however, have low activity with rabbit TLR9. Using a cell-based activation assay, we developed a type of CpG-ODN containing a GACGTT or AACGTT motif in 12 phosphorothioate-modified deoxynucleotides with potent stimulatory activity for rabbit TLR9. The developed CpG-ODN have higher activities than other developed CpG-ODN in eliciting antigen-nonspecific immune responses in rabbit splenocytes. When mixed with an NJ85 peptide derived from rabbit hemorrhagic disease virus, they had potent activities to boost an antigen-specific T cell activation and antibody production in rabbits. Compared to Freund\'s adjuvant, the developed CpG-ODN are capable of boosting a potent and less toxic antibody response. The results of this study suggest that both the choice of CpG-motif and its length are important factors for CpG-ODN to effectively activate rabbit TLR9 mediated immune responses.
Development of Single-Vector Tet-on Inducible Systems with High Sensitivity to Doxycycline
(2012-07-01)
Chao, Jiun-Shuan
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Chao, Chun-Chieh
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Chang, Chu-Li
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Chiu, Yi-Rong
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Yuan, Chiun-Jye
Single-vector Tet-on systems were developed to enable the tight regulation of transgenes in mammalian cells with a low dosage of doxycycline. Both the regulatory and the responsive units were integrated in a single vector and separated by a short DNA segment (214 bp). In the developed single-vector Tet-on systems, a high level of expression of the transgene can be induced by doxycycline at a concentration of as low as 1 ng/ml, which is 500-1,000 times lower than that usually utilized in other Tet-on systems. The single-vector Tet-on system developed here exhibited 3.5-10.8 times greater inducibility of the transgene in response to doxycycline than did a dual-vector system from a commercial source. Further studies indicate that the basal activity of Tet-on systems depends greatly on the strength of the promoter that controls the transactivator. The basal activity of Tet-on systems was high when the transactivator that was directed by the human cytomegalovirus promoter, and it was almost undetectable when the transactivator was placed under the control of a moderate strength mouse mammary tumor virus promoter. Moreover, the introduction of selectable markers allows the developed single-vector Tet-on systems to facilitate the generation of conditional transgenic cells and animals with high inducibility, low basal activity and detrimental effects of the long-term administration of doxycycline.
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] .
Effect of anions on the oxidation and reduction of hydrogen peroxide on the gold nanoparticle-deposited carbon fiber paper electrode
(2015-10-20)
Lin, Chen-Han
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Wei, Lan-Yi
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Lee, Jhao-Hong
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Lien, Chun-Lung
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Lu, Chung-Hsiang
The nanostructured gold has been shown to catalyze the oxidation of hydrogen peroxide (H2O2), an important analyte for clinical diagnosis. However, little is known about the effect of anions on its catalytic activity. In this study the gold nanoparticles (AuNP) of average size of 30-80 nm were electrodeposited on the carbon fiber paper (CFP) electrode to explore the effect of anions, such as phosphate, sulfate, nitrate, acetate, chloride and hydroxide ions, on the AuNP-catalyzed H2O2-oxidation. In phosphate solution the AuNP-deposited CFP (AuNP/CFP) exhibited high catalytic activity to H2O2. However, the catalytic activity of AuNP was severed blocked in solutions containing chloride, sulfate, nitrate, and acetate ions. Although hydroxide was shown to facilitate the catalysis of AuNP, the oxidation of H2O2 was greatly interfered by the gold oxide formation in 0.05 M NaOH. A strong dipole moment between adsorbed anions, such sulfate, nitrate and acetate, and AuNP is suggested to block the access of H2O2 molecules to the surface of AuNPs. This postulation is demonstrated by the finding that the electrochemical active surface area of AuNP/CFP is 0.90 +/- 0.02 cm(2), 0.79 +/- 0.08 cm(2), 0.66 +/- 0.09 cm(2), 0.45 +/- 0.04 cm(2), 0.42 +/- 0.08 cm(2) and 0.15 +/- 0.08 cm(2), in 0.05 M hydroxide, phosphate, bicarbonate, nitrate, acetate and sulfate solutions, respectively. These results indicate that the microenvironment near the surface of nanostructured Au is important to determine the catalytic activity of Au to H2O2. (C) 2015 Elsevier Ltd. All rights reserved.
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).
Enzyme-encapsulated silica nanoparticle for cancer chemotherapy
(2012-04-01)
Chiu, Yi-Rong
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Ho, Wei-Jen
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Chao, Jiun-Shuan
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Yuan, Chiun-Jye
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生物科技學系
A novel horseradish peroxidase-encapsulated silica nanoparticle (SNP) was generated in this study under relatively mild conditions. The generated enzyme-encapsulated SNP were relatively uniform in size (average 70 +/- 14.3 nm), monodispersed, and spherical, as characterized by transmission electron microscopy and scanning electron microscopy. The horseradish peroxidase encapsulated in silica nanoparticle exhibits biological properties, such as a pH-dependent activity profile and k(m) value, similar to that of free enzymes. Furthermore, enzyme-encapsulated SNP exhibited good operational stability for the repetitive usage with a relative standard deviation of 5.1 % ( n = 10) and a high stability for long term storage (>60 days) at 4 degrees C. The feasibility of using enzyme-encapsulated SNP in prodrug cancer therapy was also demonstrated by its capability to convert the prodrug indole-3-acetic acid into cytotoxic peroxyl radicals and trigger the death of tumor cells. These results indicate that the developed enzyme-encapsulated SNP has potential in the applications of prodrug cancer therapy.
Exploration and characterization of the memcapacitor and memristor properties of Ni-DNA nanowire devices
(2017-09-01)
Chu, Hsueh-Liang
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Lai, Jian-Jhong
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Wu, Li-Ying
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Chang, Shen-Lin
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Liu, Chia-Ming
A 2-mu m-long Ni ion-chelated DNA molecule (Ni-DNA) was found for the first time to possess both memcapacitor and memristor properties; this Ni-DNA molecule is known as a dual memory circuit element (memelement). As a memelement, the state of impedance on Ni-DNA is proportional to the unit number of Ni ions containing a base pair complex (Ni-bp), which is determined by the previously applied external voltage. Interestingly, the impedances of Ni-DNA change in response to a change in the sweeping frequencies of the external bias. Our simulation results also indicate that changes in the effective resistance and capacitance of Ni-bp may be attributed to changes in the Ni ion redox species in the Ni-bp of a Ni-DNA nanowire. Therefore, the working mechanism of a nanowire-type memcapacitor and memristor is revealed. In summary, the Ni-DNA nanowire is shown to be a multi-dimensional memory device, whose memory state depends on the length of DNA and applied external voltages/frequencies.
Fabrication of a carbon fiber paper as the electrode and its application toward developing a sensitive unmediated amperometric biosensor
(2011-02-15)
Yuan, Chiun-Jye
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Wang, Chung-Liang
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Wu, Teng Yang
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Hwang, Kuo-Chu
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Chao, Wei-Chi
Carbon fiber paper (CFP), a material frequently used as the diffusion layer in fuel cells, was found recently to exhibit a potential as an electrode for the development of sensitive, unmediated biosensors. After nitrogen plasma treatment, the CFP exhibited a quasi-reversible behavior to the redox couple (e.g., ferricyanide) with an electron transfer rate constant of 7.2 x 10(-3) cm s(-1). This rate constant is approximately double that of a Pt-electrode and is much higher than that of many carbon-based electrodes. The unmediated CFP-based tyrosinase biosensor fabricated for this study exhibited an optimal working potential and operating pH value of -0.2 V and 6.5, respectively. Compared to other unmediated tyrosinase biosensors, the CFP-based tyrosinase biosensor offers a high sensitivity for the monitoring of phenolic compounds (17.8, 7.1, 5.2 and 3.7 mu A mu M(-1) cm(-2) for catechol, phenol, bisphenol and 3-aminophenol, respectively). The lowest detection limit for catechol, phenol, bisphenol and 3-aminophenol was 2, 5, 5 and 12 nM, respectively. Furthermore, this biosensor exhibited a good repeatability, a fast response time (around 105), and a wide linear dynamic range of detection for phenolic compounds. (C) 2010 Elsevier B.V. All rights reserved.