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      • KCI등재

        Preconcentration-enhanced Immunosensing for Whole Human Cancer Cell Lysate based on a Nanofluidic Preconcentrator

        Hsuan Franziska Wu,Tamara G. Amstislavskaya,Pin-Hsuan Chen,Ting-Feng Wu,Yu-Hung Chen,Chun-Ping Jen 한국바이오칩학회 2016 BioChip Journal Vol.10 No.3

        Sample preconcentration is an important step that increases the accuracy of subsequent detection, especially for samples with extremely low concentrations. Due to the overlap of electrical double layers in a nanofluidic channel, the concentration polarization effect can be generated by applying an electric field. A nonlinear electrokinetic flow is induced, which results in the fast accumulation of proteins in front of the induced ionic depletion zone, the so-called exclusion- enrichment effect. In this way, a protein sample can be driven by electroosmotic flow and accumulated at a specific location. In the present study, a nanofluidic preconcentrator fabricated with the help of junction gap electric breakdown was integrated with microelectrodes for immunoassay. The preconcentration chip for proteins was fabricated using simple standard soft lithography with a polydimethylsiloxane replica. Human galectin-1 proteins from the cell lysate of T24 cells were concentrated and immunoassayed in the proposed microchip. The capability of the proposed microchip for concentrating multiple proteins from cell lysates and immunoassays after preconcentration was demonstrated. Immunosensing was evaluated by measurements of both fluorescence intensities and impedance, which proved the enhancement of preconcentration for immunoassay.

      • KCI등재

        Systematically Studying Dissolution Process of 3D Printed Acrylonitrile Butadiene Styrene (ABS) Mold for Creation of Complex and Fully Transparent Polydimethylsiloxane (PDMS) Fluidic Devices

        Chen Pin-Chuan,Chou Ching Chan,Chiang Chung Hsuan 한국바이오칩학회 2021 BioChip Journal Vol.15 No.2

        Our major objective in this study was to create complex, three-dimensional, and fully transparent polydimethylsiloxane(PDMS) fluidic device by revising the previously reported fabrication process and to systematically study the influence of each fabrication step to the final PDMS fluidic device. The current fabrication process adopted fused deposition modeling (FDM) 3D printers to create molds of acrylonitrile butadiene styrene (ABS) for use in PDMS casting, then solvent solution was used to dissolve the ABS mold embedded inside the PDMS device and a transparent PDMS device was created for experiments. However, it is quite challenging to ensure the complete removal of ABS molds inside the long, curly, and narrow channels. Ultrasonication was added into our fabrication process to improve the efficacy of dissolving ABS molds inside the channels and conclusions can be derived from these experiments: (1) ultrasonication-assisted dissolution is an effective approach to the complete removal of ABS molds embedded inside these long, curly, and narrow channels (for example, the mixer demonstrated herein had a diameter of 2 mm and length of 162 mm); (2) the application of solvent vapor polishing to 3D-printed molds is highly effective in reducing the surface roughness of the molds (8 ~ 10 μm before polishing to 038 ~ 0.5 μm after polishing) and important to preserve the transparency of the resulting PDMS devices; (3) ensuring the circulation of fresh solvent solution is critical to shorten the dissolution process.

      • SCOPUSKCI등재

        Melatonin acts synergistically with pazopanib against renal cell carcinoma cells through p38 mitogen-activated protein kinase-mediated mitochondrial and autophagic apoptosis

        ( Chien-pin Lai ),( Yong-syuan Chen ),( Tsung-ho Ying ),( Cheng-yen Kao ),( Hui-ling Chiou ),( Shao-hsuan Kao ),( Yi-hsien Hsieh ) 대한신장학회 2023 Kidney Research and Clinical Practice Vol.42 No.4

        Background: Mounting evidence indicates that melatonin has possible activity against different tumors. Pazopanib is an anticancer drug used to treat renal cell carcinoma (RCC). This study tested the anticancer activity of melatonin combined with pazopanib on RCC cells and explored the underlying mechanistic pathways of its action. Methods: The 786-O and A-498 human RCC cell lines were used as cell models. Cell viability and tumorigenesis were detected with the MTT and colony formation assays, respectively. Apoptosis and autophagy were assessed using TUNEL, annexin V/propidium iodide, and acridine orange staining with flow cytometry. The expression of cellular signaling proteins was investigated with western blotting. The in vivo growth of tumors derived from RCC cells was evaluated using a xenograft mouse model. Results: Together, melatonin and pazopanib reduced cell viability and colony formation and promoted the apoptosis of RCC cells. Furthermore, the combination of melatonin and pazopanib triggered more mitochondrial, caspase-mediated, and LC3-II-mediated autophagic apoptosis than melatonin or pazopanib alone. The combination also induced higher activation of the p38 mitogen-activated protein kinase (p38MAPK) in the promotion of autophagy and apoptosis by RCC cells than melatonin or pazopanib alone. Finally, tumor xenograft experiments confirmed that melatonin and pazopanib cooperatively inhibited RCC growth in vivo and predicted a possible interaction between melatonin/pazopanib and LC3-II. Conclusion: The combination of melatonin and pazopanib inhibits the growth of RCC cells by inducing p38MAPK-mediated mitochondrial and autophagic apoptosis. Therefore, melatonin might be a potential adjuvant that could act synergistically with pazopanib for RCC treatment.

      • KCI등재

        Ziwuliuzhu Acupuncture Modulates Clock mRNA, Bmal1 mRNA and Melatonin in Insomnia Rats

        Huang Ao,Xiao Gefang,Chen Yiliu,Hu Zuying,Lee Pin-Hsuan,Huang Yusen,Zhuang Zifeng,Zhang Yuling,Qing Peng,Zhao Canghuan 사단법인약침학회 2023 Journal of Acupuncture & Meridian Studies Vol.16 No.3

        Background: In clinics, Ziwuliuzhu acupuncture is widely considered an effective method of treating insomnia; however, there is currently limited information available regarding its possible mechanisms. Although the method of Ziwuliuzhu acupuncture possesses a unique rhythmic pattern. Objectives: In this study, we have creatively combined the traditional Chinese medicine of Ziwuliuzhu with a modern biological rhythm to investigate the internal mechanism of insomnia. Methods: Pathological tissue from the hypothalamus was analyzed using hematoxylin– eosin staining. The level of TNF (tumor necrosis factor)-α in the SCN (suprachiasmatic nucleus) area of the hypothalamus was detected in situ using the TUNEL fluorescence staining assay. The concentration of hypothalamic melatonin was detected using the enzyme-linked immunosorbent assay (ELISA). The mRNA expression of Clock and Bmal1 was measured using RT-qPCR. Results: In the Ziwuliuzhu acupuncture groups, the structural damage in the hypothalamic neurons was alleviated compared to the model group and the expression of inflammatory factors was reduced. The mRNA expression levels of Clock and Bmal1 were significantly increased (p < 0.05). The concentration of melatonin was significantly increased (p < 0.001). Although there were no significant differences between the treatment groups (diazepam group, Nazi group, Najia group, and routine group) (p > 0.05). Conclusion: Ziwuliuzhu acupuncture alleviated neuronal damage and modulated the inflammatory reaction in the hypothalamus of rats with insomnia. Moreover, Ziwuliuzhu acupuncture increased the expression levels of Clock and Bmal1 mRNA, and MT content. This study has potentially highlighted one of

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