Publications by authors named "Chien-Jung Chen"

9 Publications

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Bone biomarkers in koalas: validation of assays and preliminary analyses.

J Vet Diagn Invest 2020 Nov 11;32(6):856-863. Epub 2020 Sep 11.

School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia (Chen, Henning, Allavena).

Traumatic injury, including bone fracture, is, to date, one of the leading causes of koala mortality in the South East Queensland region of Australia. Further, the specialist diet of koalas, which is restricted to certain spp., may impact their normal bone physiology. Considering the dramatic koala population decline and high incidence of trauma, a greater understanding of koala bone physiology may support conservation. We retrieved from GenBank the protein sequences of parathyroid hormone (PTH), osteocalcin (OCN), and tissue-nonspecific alkaline phosphatase (TNALP) in human, dog, cattle, horse, koala, and gray short-tailed opossum. After homology was determined, plasma samples from 13 koalas were analyzed with human PTH, OCN, and bone-specific ALP (BALP) assay kits. Although koala PTH exhibited relatively low sequence homology with placental mammals, high sequence homology between humans and koalas was observed for both OCN and TNALP, and successful cross-reactivity was achieved using human enzyme immunoassay kits for detection of OCN and BALP biomarkers in koala plasma. However, we identified no correlation between OCN and BALP concentrations of healthy and trauma-affected koalas ( = 0.66 and = 0.79, respectively). Further analysis of OCN and BALP in healthy and diseased koalas will allow a better understanding of bone physiology in this unique marsupial.
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http://dx.doi.org/10.1177/1040638720957031DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649558PMC
November 2020

Electric Field-Induced Release and Measurement (EFIRM): Characterization and Technical Validation of a Novel Liquid Biopsy Platform in Plasma and Saliva.

J Mol Diagn 2020 08 1;22(8):1050-1062. Epub 2020 Jun 1.

UCLA School of Dentistry, University of California, Los Angeles, Los Angeles, California; Liquid Diagnostics LLC, San Clemente, California. Electronic address:

Electric field-induced release and measurement (EFIRM) is a novel, plate-based, liquid biopsy platform capable of detecting circulating tumor DNA containing EGFR mutations directly from saliva and plasma in both early- and late-stage patients with non-small-cell lung cancer. We investigated the properties of the target molecule for EFIRM and determined that the platform preferentially detects single-stranded DNA molecules. We then investigated the properties of the EFIRM assay and determined the linearity, linear range, precision, and limit of detection for six different EGFR variants (the four most common g.Exon19del variants), p.T790M, and p.L858R). The limit of detection was in single-digit copy number for the latter two mutations, and the limit of detection for Exon19del was 5000 copies. Following these investigations, technical validations were performed for four separate EFIRM liquid biopsy assays, qualitative and quantitative assays for both saliva and plasma. We conclude that EFIRM liquid biopsy is an assay platform that interrogates a biomarker not targeted by any other extant platform (namely, circulating single-stranded DNA molecules). The assay has acceptable performance characteristics in both quantitative and qualitative assays on both saliva and plasma.
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http://dx.doi.org/10.1016/j.jmoldx.2020.05.005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7416077PMC
August 2020

Association of injury pattern and entrapment location inside damaged buildings in the 2016 Taiwan earthquake.

J Formos Med Assoc 2019 Jan 30;118(1 Pt 2):311-323. Epub 2018 May 30.

Department of Emergency Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan. Electronic address:

Background/purpose: To explore the association of patient injury patterns and entrapped locations inside damaged buildings in the 2016 Taiwan earthquake.

Methods: A retrospective analysis was conducted using the Tainan incident registry system. Residents inside nine conjunctive, 16-story (49.3 m in height) reinforced concrete buildings were categorized as non-injured, injured, and dead. Residents were classified into different groups according to their entrapped locations in height and the severity of building damage. The field triage acuity and trauma severity among groups were compared. Statistical significance was set at the level of 0.05.

Results: There were 309 enrollees with 76 (24.6%) non-injured, 118 (38.2%) injured, and 115 (37.2%) dead. Residents either in the high floors (odds ratio [OR] = 2.9, 95% CI: 1.5-5.8, p = 0.003) or in the collapsed buildings (OR = 18.2, 95% CI: 7.6-43.6, p < 0.001) were more likely to be dead. Injured patients who were located in the high floors were more likely to have severe field triage acuities (adjusted OR = 14.7, 95% CI: 1.8-118.0, p = 0.012); intracranial hemorrhage (12.5%), intrathoracic injury (18.8%), or intra-abdominal damage (12.5%) (All p < 0.05); the need for emergency surgical intervention (31.3%, p = 0.035); and major trauma (18.8%, p = 0.001). Residents in the collapsed buildings were more likely to have a crush injury (80.0%, p < 0.001) or crush syndrome (80.0%, p < 0.001).

Conclusion: People entrapped at different heights of floors or in differently damaged buildings could have a distinct pattern of injury. Our findings may facilitate strategic approaches of patients entrapped in damaged buildings and may contribute to future training for field searches and rescues after earthquakes.
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http://dx.doi.org/10.1016/j.jfma.2018.05.012DOI Listing
January 2019

Heparin-Promoted Cellular Uptake of the Cell-Penetrating Glycosaminoglycan Binding Peptide, GBP, Depends on a Single Tryptophan.

ACS Chem Biol 2017 02 21;12(2):398-406. Epub 2016 Dec 21.

Institute of Bioinformatics and Structural Biology, National Tsing Hua University , Hsinchu 30013, Taiwan.

A 10-residue, glycosaminoglycan-binding peptide, GBP, derived from human eosinophil cationic protein has been recently designated as a potent cell-penetrating peptide. A model system containing peptide, glycan, and lipid was monitored by nuclear magnetic resonance (NMR) spectroscopy to determine the cell-penetrating mechanism. Heparin octasaccharide with dodecylphosphocholine (DPC) lipid micelle was titrated into the GBP solution. Our data revealed substantial roles for the charged residues Arg5 and Lys7 in recognizing heparin, whereas Arg3 had less effect. The aromatic residue Trp4 acted as an irreplaceable moiety for membrane insertion, as the replacement of Trp4 with Arg4 abolished cell penetration, although it significantly improved the heparin-binding ability. GBP bound either heparin or lipid in the presence or absence of the other ligand indicating that the peptide has two alternative binding sites: Trp4 is responsible for lipid insertion, and Arg5 and Lys7 are for GAG binding. We developed a molecular model showing that the two effects synergistically promote the penetration. The loss of either effect would abolish the penetration. GBP has been proven to enter cells through macropinocytosis. The GBP treatment inhibited A549 lung cancer cell migration and invasion, implying that the cellular microenvironment would be modulated by GBP internalization. The intracellular penetration of GBP leading to inhibition of epithelial cell migration and invasion depends on the presence of the tryptophan residue in its sequence compared with similar derivative peptides. Therefore, GBP shows substantial potential as a novel delivery therapeutic through rapid and effective internalization and interference with cell mobility.
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http://dx.doi.org/10.1021/acschembio.6b00864DOI Listing
February 2017

A Heparan Sulfate-Binding Cell Penetrating Peptide for Tumor Targeting and Migration Inhibition.

Biomed Res Int 2015 3;2015:237969. Epub 2015 May 3.

Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 30013, Taiwan ; Department of Medical Science, National Tsing Hua University, Hsinchu 30013, Taiwan.

As heparan sulfate proteoglycans (HSPGs) are known as co-receptors to interact with numerous growth factors and then modulate downstream biological activities, overexpression of HS/HSPG on cell surface acts as an increasingly reliable prognostic factor in tumor progression. Cell penetrating peptides (CPPs) are short-chain peptides developed as functionalized vectors for delivery approaches of impermeable agents. On cell surface negatively charged HS provides the initial attachment of basic CPPs by electrostatic interaction, leading to multiple cellular effects. Here a functional peptide (CPPecp) has been identified from critical HS binding region in hRNase3, a unique RNase family member with in vitro antitumor activity. In this study we analyze a set of HS-binding CPPs derived from natural proteins including CPPecp. In addition to cellular binding and internalization, CPPecp demonstrated multiple functions including strong binding activity to tumor cell surface with higher HS expression, significant inhibitory effects on cancer cell migration, and suppression of angiogenesis in vitro and in vivo. Moreover, different from conventional highly basic CPPs, CPPecp facilitated magnetic nanoparticle to selectively target tumor site in vivo. Therefore, CPPecp could engage its capacity to be developed as biomaterials for diagnostic imaging agent, therapeutic supplement, or functionalized vector for drug delivery.
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http://dx.doi.org/10.1155/2015/237969DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4433633PMC
March 2016

Efforts to reduce the door-to-needle time of thrombolysis in acute ischemic stroke: video-assisted therapeutic risk communication.

J Formos Med Assoc 2014 Dec 10;113(12):929-33. Epub 2014 Jan 10.

Stroke Center and Department of Neurology, Tainan Sin Lau Hospital, Tainan, Taiwan. Electronic address:

Background/purpose: Explaining the risks and benefits of recombinant tissue-plasminogen activator (rtPA) to eligible patients with acute ischemic stroke (AIS) within a few minutes is important but difficult. We examined whether a new thrombolysis program can decrease the door-to-needle (DTN) time when treating patients with AIS.

Methods: A new rtPA thrombolysis program with video assistance was adapted for patients with AIS and their families. We retrospectively compared outcome quality before (2009-2011) and after (2012) the program began. Outcomes included DTN time, the percentage of rtPA thrombolysis within 3 hours of onset in all hospitalized patients with AIS who presented within 2 hours of onset (2 hr%) and the percentage of rtPA thrombolysis in all hospitalized patients with AIS (AIS%).

Results: We recruited patients with AIS who had undergone thrombolytic therapy before (n = 18) and after (n = 14) the initiation of the new program. DTN time decreased (93 ± 24 minutes to 57 ± 14 minutes, p < 0.001) and the AIS% increased (2% to 5%, p = 0.010) after the program. The 2 hr% marginally significantly increased (18% to 33%, p = 0.080).

Conclusion: A thrombolysis program with video-assisted therapeutic risk communication decreased DTN time and increased the treatment rate of patients with AIS.
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http://dx.doi.org/10.1016/j.jfma.2013.11.012DOI Listing
December 2014

Basic amino acid residues of human eosinophil derived neurotoxin essential for glycosaminoglycan binding.

Int J Mol Sci 2013 Sep 16;14(9):19067-85. Epub 2013 Sep 16.

Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.

Human eosinophil derived neurotoxin (EDN), a granule protein secreted by activated eosinophils, is a biomarker for asthma in children. EDN belongs to the human RNase A superfamily possessing both ribonucleolytic and antiviral activities. EDN interacts with heparin oligosaccharides and heparin sulfate proteoglycans on bronchial epithelial Beas-2B cells. In this study, we demonstrate that the binding of EDN to cells requires cell surface glycosaminoglycans (GAGs), and the binding strength between EDN and GAGs depends on the sulfation levels of GAGs. Furthermore, in silico computer modeling and in vitro binding assays suggest critical roles for the following basic amino acids located within heparin binding regions (HBRs) of EDN 34QRRCKN39 (HBR1), 65NKTRKN70 (HBR2), and 113NRDQRRD119 (HBR3) and in particular Arg35, Arg36, and Arg38 within HBR1, and Arg114 and Arg117 within HBR3. Our data suggest that sulfated GAGs play a major role in EDN binding, which in turn may be related to the cellular effects of EDN.
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http://dx.doi.org/10.3390/ijms140919067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3794821PMC
September 2013

In silico prediction and in vitro characterization of multifunctional human RNase3.

Biomed Res Int 2013 17;2013:170398. Epub 2013 Jan 17.

Institute of Molecular and Cellular Biology, National Tsing Hua University, No. 101, Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan.

Human ribonucleases A (hRNaseA) superfamily consists of thirteen members with high-structure similarities but exhibits divergent physiological functions other than RNase activity. Evolution of hRNaseA superfamily has gained novel functions which may be preserved in a unique region or domain to account for additional molecular interactions. hRNase3 has multiple functions including ribonucleolytic, heparan sulfate (HS) binding, cellular binding, endocytic, lipid destabilization, cytotoxic, and antimicrobial activities. In this study, three putative multifunctional regions, (34)RWRCK(38) (HBR1), (75)RSRFR(79) (HBR2), and (101)RPGRR(105) (HBR3), of hRNase3 have been identified employing in silico sequence analysis and validated employing in vitro activity assays. A heparin binding peptide containing HBR1 is characterized to act as a key element associated with HS binding, cellular binding, and lipid binding activities. In this study, we provide novel insights to identify functional regions of hRNase3 that may have implications for all hRNaseA superfamily members.
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http://dx.doi.org/10.1155/2013/170398DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3581242PMC
September 2013

A novel cell-penetrating peptide derived from human eosinophil cationic protein.

PLoS One 2013 4;8(3):e57318. Epub 2013 Mar 4.

Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.

Cell-penetrating peptides (CPPs) are short peptides which can carry various types of molecules into cells; however, although most CPPs rapidly penetrate cells in vitro, their in vivo tissue-targeting specificities are low. Herein, we describe cell-binding, internalization, and targeting characteristics of a newly identified 10-residue CPP, denoted ECP(32-41), derived from the core heparin-binding motif of human eosinophil cationic protein (ECP). Besides traditional emphasis on positively charged residues, the presence of cysteine and tryptophan residues was demonstrated to be essential for internalization. ECP(32-41) entered Beas-2B and wild-type CHO-K1 cells, but not CHO cells lacking of cell-surface glycosaminoglycans (GAGs), indicating that binding of ECP(32-41) to cell-surface GAGs was required for internalization. When cells were cultured with GAGs or pre-treated with GAG-digesting enzymes, significant decreases in ECP(32-41) internalization were observed, suggesting that cell-surface GAGs, especially heparan sulfate proteoglycans were necessary for ECP(32-41) attachment and penetration. Furthermore, treatment with pharmacological agents identified two forms of energy-dependent endocytosis, lipid-raft endocytosis and macropinocytosis, as the major ECP(32-41) internalization routes. ECP(32-41) was demonstrated to transport various cargoes including fluorescent chemical, fluorescent protein, and peptidomimetic drug into cultured Beas-2B cells in vitro, and targeted broncho-epithelial and intestinal villi tissues in vivo. Hence this CPP has the potential to serve as a novel vehicle for intracellular delivery of biomolecules or medicines, especially for the treatment of pulmonary or gastrointestinal diseases.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0057318PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3587609PMC
August 2013