Browsing by Author "Lin, Ching-Min"

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A Lipo-PEG-PEI complex for encapsulating curcumin that enhances its antitumor effects on curcumin-sensitive and curcumin-resistance cells
(2012-04-01)
Lin, Yu-Ling
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Liu, Yen-Ku
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Tsai, Nu-Man
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Hsieh, Jui-Hung
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Chen, Chia-Hung
A cationic liposome-PEG-PEI complex (LPPC) was used as a carrier for the encapsulation of hydrophobic curcumin to give curcumin/LPPC. Curcumin/LPPC had an average size less than 270 nm and a zeta potential of approximately 40 mV. The LPPC encapsulation efficiency for curcumin was about 45%. The authors found it surprising that the cytotoxic activity of the curcumin/LPPC was fivefold higher than curcumin when tested on curcumin-sensitive cells and 20-fold more active against curcumin-resistant cells. Curcumin/LPPC treatment caused a cell cycle arrest at G2/M phase, which rapidly resulted in apoptosis. The increased cytotoxic activity of curcumin/LPPC is likely attributable to its rapid accumulation in the cell. In vivo, administration of curcumin/LPPC inhibited about 60 - 90% of tumor growth in mice bearing CT-26 or B16F10 cells. These results demonstrate LPPC encapsulation technology is able to enhance the effects of antitumor drugs. Use of this technology may provide a new tool for cancer therapy, especially for drug-resistant cancer.
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
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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.
Antibodies against Helicobacter pylori heat shock protein 60 aggravate HSP60-mediated proinflammatory responses
(2011-08-01)
Liao, Kuang-Wen
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Lin, Chen-Si
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Chen, Wen-Liang
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Yang, Chu-Ting
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Lin, Ching-Min
Anti-Helicobacter pylori heat shock protein 60 (HpHSP60) antibodies are Usually found in H. pylori-infected patients and are known to be associated with the progression of gastric diseases. However, the effects of these antibodies on the functions of HpHSP60 have not been identified. This study aims to investigate the effects of the interaction between anti-HSP60 antibodies and HpHSP60 on inflammatory responses. Anti-HpHSP60 polyclonal sera and monoclonal antibodies (mAbs) were produced to evaluate their effects on HpHSP60-induced IL-8 and TNF-alpha activity. The results indicated that anti-HpHSP60 polyclonal sera collected from patients infected with H. pylori or from rabbit and mice immunized with HpHSP60 could significantly enhance HpHSP60-mediated IL-8 and TNF-alpha secretion from monocytic THP-1 cells. Similar effects were also found with anti-HpHSP60 mAbs. Further analysis revealed that this phenomenon was only carried out by anti-HpHSP60 antibody but not by other non-specific mAbs. Moreover, the non-specific mAbs decreased the synergism of HpHSP60 and anti-HpHSP60 mAbs in proinflammatory cytokine induction. Herein, we have examined the role of anti-HpHSP60 antibody in host immune responses for the first time. This study demonstrated that H. pylori HSP60/mAbs could modulate helico-bacterial pathogenesis by increasing IL-8 and TNF-alpha production. The pathogen-specific antibodies may execute potential immune functions rather than recognize or neutralize microbes. (C) 2011 Elsevier Ltd. All rights reserved.
Inhibitory Effects of Chloroform Extracts Derived from Corbicula fluminea on the Release of Pro-inflammatory Cytokines
(2012-04-25)
Lin, Ching-Min
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Lin, Yu-Ling
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Tsai, Nu-Man
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Wu, Hsin-Yi
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Ho, Shu-Yi
Corbicula fluminea, the primary freshwater bivalve cultivated in Taiwan, was formerly used as a remedy for hepatitis. Recent reports indicate that C. fluminea has many bioactivities, but it remains unknown whether C. fluminea affects inflammation. This study explored the anti-inflammatory activity of C.fluminea. C. fluminea was first treated with chloroform to obtain clam chloroform extracts (CCEs). On the basis of the assay for the release of pro-inflammatory cytokines in vitro and in vivo, the results show that the CCEs significantly lowered the release of lipopolysaccharide (LPS)-induced pro-inflammatory cytokines. Additionally, the CCEs reduced LPS-induced organ damage. Real-time polymerase chain reaction analysis suggested that CCEs inhibit the LPS-induced mRNA expression of interleukin-1 beta and tumor necrosis factor-alpha. Western blot analysis indicated that the CCEs increased expression of I kappa B and attenuated the phosphorylation of I kappa B. Gas chromatography mass spectrometry suggests that phytosterols and fatty adds are responsible for the anti-inflammatory properties of CCEs. Taken together, CCEs have the potential to be developed as an anti-inflammatory functional food.
Label-free protein quantitation by dielectric spectroscopy of dual-frequency liquid crystal
(2019-03-01)
Lin, Ching-Min
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Wu, Po-Chang
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Lee, Mon-Juan
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Lee, Wei
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光電學院
A dual-frequency-liquid-crystal (DFLC)-based biosensor was developed and its frequency-dependent dielectric properties were exploited to detect and quantitate a protein standard, bovine serum albumin (BSA). By analyzing the dielectric spectra of DFLC in the presence of BSA at various concentrations, we found that the difference in dielectric permittivity between the high- and low-frequency regimes is correlated to BSA concentration, thereby permitting a DFLC-based protein quantitative method. The dielectric properties of DFLCs are fundamental in the design of liquid crystal displays and fast-switching devices. Results from this study demonstrate the extended potential of the frequency-revertible dielectric anisotropy of DFLC in biosensing and protein quantitation.
Liposome-based polymer complex as a novel adjuvant: enhancement of specific antibody production and isotype switch
(2012)
Chen, Chia-Hung
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Lin, Yu-Ling
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Liu, Yen-Ku
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He, Pei-Juin
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Lin, Ching-Min
The aim of vaccination is to induce appropriate immunity against pathogens. Antibody-mediated immunity is critical for protection against many virus diseases, although it is becoming more evident that coordinated, multifunctional immune responses lead to the most effective defense. Specific antibody (Ab) isotypes are more efficient at protecting against pathogen invasion in different locations in the body. For example, compared to other Ab isotypes, immunoglobulin (Ig) A provides more protection at mucosal areas. In this study, we developed a cationic lipopolymer (liposome-polyethylene glycol-polyethyleneimine complex [LPPC])-adjuvant that strongly adsorbs antigens or immunomodulators onto its surface to enhance or switch immune responses. The results demonstrate that LPPC enhances uptake ability, surface marker expression, proinflammatory cytokine release, and antigen presentation in mouse-phagocytes. In contrast to Freund's adjuvant, LPPC preferentially activates-Th1-immunity against antigens in vivo. With lipopolysaccharides or CpG oligodeoxynucleotides, LPPC dramatically enhances the IgA or IgG2A proportion of total Ig, even in hosts that have developed Th2 immunities and high IgG1 serum titers. Taken together, the results demonstrate that the LPPC adjuvant not only increases the immunogenicity of antigens but also modulates host immunity to produce an appropriate Ab isotype by combining with immunomodulators.
miRTarBase update 2014: an information resource for experimentally validated miRNA-target interactions
(2014-01-01)
Hsu, Sheng-Da
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Tseng, Yu-Ting
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Shrestha, Sirjana
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Lin, Yu-Ling
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Khaleel, Anas
MicroRNAs (miRNAs) are small non-coding RNA molecules capable of negatively regulating gene expression to control many cellular mechanisms. The miRTarBase database (http://mirtarbase.mbc.nctu.edu.tw/) provides the most current and comprehensive information of experimentally validated miRNA-target interactions. The database was launched in 2010 with data sources for >100 published studies in the identification of miRNA targets, molecular networks of miRNA targets and systems biology, and the current release (2013, version 4) includes significant expansions and enhancements over the initial release (2010, version 1). This article reports the current status of and recent improvements to the database, including (i) a 14-fold increase to miRNA-target interaction entries, (ii) a miRNA-target network, (iii) expression profile of miRNA and its target gene, (iv) miRNA target-associated diseases and (v) additional utilities including an upgrade reminder and an error reporting/user feedback system.
The inhibitory effect of 7, 7 ''-dimethoxyagastisflavone on the metastasis of melanoma cells via the suppression of F-actin polymerization
(2017-09-01)
Lin, Ching-Min
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Lin, Yu-Ling
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Ho, Shu-Yi
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Chen, Pin-Rong
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Tsai, Yi-Hsuan
7,7"-Dimethoxyagastisflavone (DMGF), a biflavonoid isolated from Taxus x media cv. Hicksii, induces apoptotic and autophagic cell death. However, whether DMGF suppresses tumor metastasis is unclear. The aim of this study was to investigate the anti-metastatic activities of DMGF on the metastatic processes of melanoma cells in vivo and in vitro. A transwell assay showed that DMGF could effectively attenuate the motility of B16F10 cells, and the results of real-time PCR revealed that DMGF also suppressed the expressions of matrix metalloproteinase-2 (MMP-2). Moreover, DMGF did not influence tube formation but inhibited the migration of endothelial cells. Furthermore, animal models were used to monitor the effects of DMGF on tumor metastasis, and all models showed that DMGF significantly suppressed the metastatic behaviors of B16F10 cells, including intravasation, colonization, and invasion of the lymphatic duct. In addition, DMGF could also reduce the densities of the blood vessels in the tumor area in vivo. Further investigation of the molecular mechanisms of anti-metastatic activity revealed that DMGF can down-regulate the levels of key modulators of the Cdc42/Rac1 pathway to interfere in F-actin polymerization and suppress the formation of lamellipodia by reducing the phosphorylation of CREB. These data suggested that DMGF presents anti-metastatic activities in B16F10 melanoma cells. Here, we demonstrated that DMGF can inhibit the metastasis of highly invasive melanoma cancer cells through the down-regulation of F-actin polymerization. Considering these findings, DMGF may be further developed to serve as a chemoprevention drug for patients with metastatic melanoma.
Utilization of I kappa B-EGFP Chimeric Gene as an Indicator to Identify Microbial Metabolites with NF-kappa B Inhibitor Activity
(2010-12-01)
Lin, Yu-Ling
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Chen, Yen-Shun
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Hsieh, Jui-Hung
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Lin, Ching-Min
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Wu, Hsin-Yi
NF-kappa B regulates several important expressions, such as cytokine release, anti-apoptosis, adhesion molecule expression, and cell cycle processing. Several NF-kappa B inhibitors have been discovered as an anti-tumor or anti-inflammatory drug. The activity of NF-kappa B transcription factor is negatively regulated by I kappa B binding. In this study, I kappa B assay system was established and I kappa B-EGFP fusion protein was used as an indicator to monitor the effects of substances on the I kappa B degradation. The results indicated that the chosen hydroquinone could inhibit the I kappa B degradation and cause the cell de-attachment from the bottom of culture plate. In addition, this system could also monitor the I kappa B degradation of microbial metabolite of natural mixtures of propolis. Thus, the I kappa B assay system may be a good system for drug discovery related to microbial metabolite.