Publications by authors named "Wen-Hao Wu"

38 Publications

A rapid screening model for early predicting novel coronavirus pneumonia in Zhejiang Province of China: a multicenter study.

Sci Rep 2021 02 16;11(1):3863. Epub 2021 Feb 16.

Department of Infectious Diseases, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, No. 158 Shangtang Road, Hangzhou, 310014, Zhejiang, China.

Novel coronavirus pneumonia (NCP) has been widely spread in China and several other countries. Early finding of this pneumonia from huge numbers of suspects gives clinicians a big challenge. The aim of the study was to develop a rapid screening model for early predicting NCP in a Zhejiang population, as well as its utility in other areas. A total of 880 participants who were initially suspected of NCP from January 17 to February 19 were included. Potential predictors were selected via stepwise logistic regression analysis. The model was established based on epidemiological features, clinical manifestations, white blood cell count, and pulmonary imaging changes, with the area under receiver operating characteristic (AUROC) curve of 0.920. At a cut-off value of 1.0, the model could determine NCP with a sensitivity of 85% and a specificity of 82.3%. We further developed a simplified model by combining the geographical regions and rounding the coefficients, with the AUROC of 0.909, as well as a model without epidemiological factors with the AUROC of 0.859. The study demonstrated that the screening model was a helpful and cost-effective tool for early predicting NCP and had great clinical significance given the high activity of NCP.
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http://dx.doi.org/10.1038/s41598-021-83054-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887205PMC
February 2021

Lasso Proteins: Modular Design, Cellular Synthesis, and Topological Transformation.

Angew Chem Int Ed Engl 2020 10 24;59(43):19153-19161. Epub 2020 Aug 24.

Beijing National Laboratory for Molecular Sciences, Key Laboratory of Polymer Chemistry & Physics of Ministry of Education, Center for Soft Matter Science and Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, P. R. China.

Entangled proteins have attracted significant research interest. Herein, we report the first rationally designed lasso proteins, or protein [1]rotaxanes, by using a p53dim-entwined dimer for intramolecular entanglement and a SpyTag-SpyCatcher reaction for side-chain ring closure. The lasso structures were confirmed by proteolytic digestion, mutation, NMR spectrometry, and controlled ligation. Their dynamic properties were probed by experiments such as end-capping, proteolytic digestion, and heating/cooling. As a versatile topological intermediate, a lasso protein could be converted to a rotaxane, a heterocatenane, and a "slide-ring" network. Being entirely genetically encoded, this robust and modular lasso-protein motif is a valuable addition to the topological protein repertoire and a promising candidate for protein-based biomaterials.
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http://dx.doi.org/10.1002/anie.202006727DOI Listing
October 2020

Ischemia/hypoxia inhibits cardiomyocyte autophagy and promotes apoptosis via the Egr-1/Bim/Beclin-1 pathway.

J Geriatr Cardiol 2020 May;17(5):284-293

Department of Cardiology, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.

Background: Myocardial injury caused by microvascular obstruction (MVO) is characterized by persistent ischemia/hypoxia (IH) of cardiomyocytes after microembolization. Autophagy and Egr-1 were closely associated with various cardiovascular diseases, including MVO. Bim and Beclin-1 are the important genes for autophagy and apoptosis. We aimed to explore whether the Egr-1/Bim/Beclin-1 pathway is involved in regulating autophagy and apoptosis in IH-exposed cardiomyocytes.

Methods: Neonatal rat cardiomyocytes exposed to the IH environment were transfected with lentivirus expressing Egr-1 or Egr-1 shRNA, or further treated with 3-methyladenine (3-MA). The expressions of autophagy and apoptosis-associated genes were evaluated using RT-qPCR and Western blots assays. Autophagic vacuoles and autophagic flux were detected by transmission electron microscopy (TEM) and confocal microscope, respectively. Cell injury was assessed by lactate dehydrogenase (LDH) leakage, and apoptosis was determined by flow cytometry.

Results: IH exposure elevated Egr-1 and Bim expressions, and decreased Beclin-1 expression in rat cardiomyocytes. Egr-1 overexpression in IH-exposed cardiomyocytes significantly up-regulated the levels of Egr-1 and Bim, and down-regulated the level of Beclin-1. Egr-1 knockdown resulted in down-regulated expressions of Egr-1 and Bim, as well as up-regulated expression of Beclin-1. In addition, Egr-1 knockdown induced autophagy was suppressed by 3-MA treatments. TEM and autophagic flux experiments also confirmed that Egr-1 inhibited autophagy progression in IH-exposed cardiomyocytes. Egr-1 suppression protected cardiomyocytes from IH-induced injury, as evidenced by the positive correlations between Egr-1 expression and LDH leakage or apoptosis index in IH-exposed cardiomyocytes.

Conclusions: IH-induced cardiomyocyte autophagy and apoptosis are regulated by the Egr-1/Bim/Beclin-1 pathway, which is a potential target for treating cardiomyocyte injury caused by MVO in the IH environment.
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http://dx.doi.org/10.11909/j.issn.1671-5411.2020.05.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7276312PMC
May 2020

NMR Spectroscopic Studies Reveal the Critical Role of the Isopeptide Bond in Forming the Otherwise Unstable SpyTag-SpyCatcher Mutant Complexes.

Biochemistry 2020 06 12;59(24):2226-2236. Epub 2020 Jun 12.

Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China.

The interplay between protein folding and chemical reaction has been an intriguing subject. In this contribution, we report the study of SpyTag and SpyCatcher reactive mutants using a combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, liquid chromatography and mass spectrometry, circular dichroism, and NMR spectroscopy. It was found that the wild-type SpyCatcher is well-folded in solution and docks with SpyTag to form an intermediate that promotes isopeptide bond formation. By contrast, the double mutant SpyCatcher is disordered in solution yet remains reactive toward SpyTag, forming a well-folded covalent complex. Control experiments using the catalytically inactive mutants further reveal the critical role of the isopeptide bond in stabilizing the otherwise loose SpyTag-SpyCatcher complex, amplifying the effect of the minute sequence disparity. We believe that the synergy between protein folding and isopeptide bonding is an effective way to enhance protein stability and engineer protein-protein interactions.
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http://dx.doi.org/10.1021/acs.biochem.0c00287DOI Listing
June 2020

Transient Receptor Potential Canonical 4 and 5 Channel Antagonist ML204 Depolarized Pacemaker Potentials of Interstitial Cells of Cajal.

J Neurogastroenterol Motil 2020 09;26(4):521-528

Department of Physiology, College of Medicine, Chosun University, Gwangju, Korea.

Background/aims: To investigate an effect of ML204 (an inhibitor of transient receptor potential canonical 4 and 5 [TRPC4/5] channels) on interstitial cells of Cajal (ICCs) and therefore determine whether TRPC4/5 channels act on ICC-generated pacemaker activity.

Methods: We enforced whole cell patch clamp analysis, measurements of the intracellular Ca concentration, and reverse transcription polymerase chain reaction to determine the effect of ML204 (10 μM) or englerin A (a selective activator of TRPC4/5 channeles, 10 μM) and the existence of TRPC4/5 in mouse small intestinal ICC.

Results: Treatment of ICCs with ML204 or englerin A caused the membrane potentials to depolarize. This depolarization effect of membrane potentials by ML204 in ICCs was observed to be concentration-dependent. After treating Ca- and Na+-free solutions or flufenamic acid (a non-selective cation channel blocker), the pacemaker potentials in the ICCs were abolished. A specific anoctamin 1 channel blocker did not have any effect on the pacemaker activity in ML204-untreated control cells; however, they blocked ML204-induced pacemaker activity in ICCs. Specific primers designed against TRPC4 and TRPC5 detected the presence of TRPC4/5 in small intestinal ICCs, and the application of ML204 increased raise the frequency of Ca oscillations in ICCs, as assessed using Fluo-4 AM.

Conclusion: The results implied that ML204 could not inhibit the pacemaker activity but depolarized the membrane potential of ICCs by regulating intracellular Ca oscillations and anoctamin 1 channels.
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http://dx.doi.org/10.5056/jnm20064DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7547197PMC
September 2020

Bioinformatics analysis and verification of gene targets for benign tracheal stenosis.

Mol Genet Genomic Med 2020 06 20;8(6):e1245. Epub 2020 Apr 20.

Department of Anesthesiology, The Second Hospital of Hebei Medical University, Shijiazhuang, China.

Background: Tracheal injury could cause intratracheal scar hyperplasia which in turn causes benign tracheal stenosis (TS). With the increasing use of mechanical ventilation and ventilator, the incidence of TS is increasing. However, the molecular mechanisms of TS have not been elucidated. It is significant to further explore the molecular mechanisms of TS.

Methods: The repeatability of public data was verified. Differently expressed genes (DEGs) and most significant genes were identified between TS and normal samples. Enrichment analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were analyzed. The comparative toxicogenomics database were analyzed. TS patients were recruited and RT-qPCR were performed to verify the most significant genes.

Results: There exist strong correlations among samples of TS and normal group. There was a total of 194 DEGs, including 61 downregulated DEGs and 133 upregulated DEGs. GO were significantly enriched in mitotic nuclear division, cell cycle, and cell division. Analysis of KEGG indicated that the top pathways were cell cycle, and p53 pathway. MKI67(OMIM:176741), CCNB1(OMIM:123836), and CCNB2(OMIM:602755) were identified as the most significant genes of TS, and validated by the clinical samples.

Conclusion: Bioinformatics methods might be useful method to explore the mechanisms of TS. In addition, MKI67, CCNB1, and CCNB2 might be the most significant genes of TS.
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http://dx.doi.org/10.1002/mgg3.1245DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7284051PMC
June 2020

The effect of goal-directed fluid therapy combines closed-loop anesthesia management on postoperative rehabilitation of patients undergoing laparoscopic pancreaticoduodenectomy.

J Clin Anesth 2020 03 24;60:115-117. Epub 2019 Oct 24.

Department of Anesthesiology, The Second Hospital of Hebei Medical University, No. 215, Heping West Road, Xinhua District, Shijiazhuang 050000, PR China. Electronic address:

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http://dx.doi.org/10.1016/j.jclinane.2019.09.005DOI Listing
March 2020

Spin switching in tris(8-aminoquinoline)iron(ii)(BPh): quantitative guest-losing dependent spin crossover properties and single-crystal-to-single-crystal transformation.

Dalton Trans 2018 Dec;48(1):231-241

School of Chemistry and Material Science and Jiangsu Key Laboratory of Green Synthetic Chemistry for Functional Materials, Jiangsu Normal University, Xuzhou, Jiangsu 221116, P. R. China.

As a derivative of 2-picolylamine, which contains rich protons favouring hydrogen bond formation to assemble a variety of valuable spin crossover (SCO) compounds, 8-aminoquinoline (aqin) will be a good candidate for constructing new mononuclear bistable state compounds. With the guidance of this view, two solvated compounds [Fe(aqin)3](BPh4)2·2(CH3CN) (1·2CH3CN) and [Fe(aqin)3](BPh4)2·1.5(CH3COCH3) (2·1.5CH3COCH3) were synthesized. The structural characterization and magnetic studies demonstrate that this strategy has been successful. Single-crystal diffraction reveals that both the mononuclear compounds have facial (fac-)-configuration cations, which form hydrogen bonds using -NH2 groups with solvent molecules (acetonitrile or acetone). Subsequent magnetic measurement shows the highly sensitive solvent-dependent occurrence of a spin transition above room temperature for both compounds. Interestingly, for compound 1·2CH3CN, in the successively repeated heating and cooling process, by monitoring the loss of solvent molecules by TGA, the shifting of the spin transition curve is found to be linearly dependent on the fraction of the residual solvent content. Additionally, the desolvated sample can re-solvate with CH3CN and recover the magnetic response reproducibly. Furthermore, after losing the acetonitrile molecules, the single-crystal-to-single-crystal transformation occurred to give 1.
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http://dx.doi.org/10.1039/c8dt03584aDOI Listing
December 2018

Comparison of wound healing effects between -derived polydeoxyribonucleotide (PDRN) and -derived PDRN.

Arch Craniofac Surg 2018 Mar 20;19(1):20-34. Epub 2018 Mar 20.

Eulji Medi-Bio Research Institute, Eulji University, Seoul, Korea.

Background: Polydeoxyribonucleotide (PDRN) influencing cellular growth and differentiation is recognized to promote wound healing by stimulating tissue repair. Although PDRN can be extracted from human placentas, PDRN medications have recently been extracted from the semen of trout () and salmon (). The present study was designed to evaluate the wound healing effects of -derived PDRN for injection (Rejuvenex) and PDRN cream (Rejuvenex Cream) in comparison with those of -derived PDRN injection (Placentex).

Methods: Full-thickness skin defects were made on the back of mice (n=60). The mice were divided into the following four groups according to the dressing used for the wounds: -derived PDRN injection group, -derived PDRN injection group, -derived PDRN cream group, and normal saline soaked dressing group (control group). We analyzed the gross findings, wound sizes, histological findings, immunohistochemistry and enzyme-linked immunosorbent assays for the groups immediately after the treatment, and again after 4, 7, and 10 days of treatment.

Results: The wound healing effects were the greatest in the -derived PDRN injection and -derived PDRN injection groups, which showed similar scores, followed by the -derived cream and normal saline soaked dressing groups.

Conclusion: The injection of PDRN extracted from was found to be as effective at healing full-thickness skin defects as the -derived PDRN injection, which is currently used in the clinic. Moreover, the -derived PDRN injection was also found to reduce the time required for wound healing.
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http://dx.doi.org/10.7181/acfs.2018.19.1.20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5894545PMC
March 2018

Effects of corticotropin-releasing hormone on the expression of adenosine triphosphate-sensitive potassium channels (Kir6.1/SUR2B) in human term pregnant myometrium.

Obstet Gynecol Sci 2018 Jan 11;61(1):14-22. Epub 2017 Dec 11.

Department of Obstetrics and Gynecology, Eulji General Hospital, Eulji University, Seoul, Korea.

Objective: Corticotropin-releasing hormone (CRH) is a crucial regulator of human pregnancy and parturition. Adenosine triphosphate (ATP)-sensitive potassium (KATP) channels are important for regulating myometrial quiescence during pregnancy. We investigated regulatory effects of different concentrations of CRH on KATP channel expression in human myometrial smooth muscle cells (HSMCs) in conditions.

Methods: After treating HSMCs with different concentrations of CRH (1, 10, 10, 10, 10 pmol/L), mRNA and protein expression of K channel subunits (Kir6.1 and SUR2B) was analyzed by reverse transcription-polymerase chain reaction and western blot. We investigated which CRH receptor was involved in the reaction and measured the effects of CRH on intracellular Ca concentration when oxytocin was administered in HSMCs using Fluo-8 AM ester.

Results: When HSMCs were treated with low (1 pmol/L) and high (10, 10 pmol/L) CRH concentrations, K channel expression significantly increased and decreased, respectively. SUR2B mRNA expression at low and high CRH concentrations was significantly antagonized by antalarmin (CRH receptor-1 antagonist) and astressin 2b (CRH receptor-2 antagonist), respectively; however, Kir6.1 mRNA expression was not affected. After oxytocin treatment, the intracellular Ca concentration in CRH-treated HSMCs was significantly lowered in low concentration of CRH (1 pmol/L), but not in high concentration of CRH (10 pmol/L), compared to control.

Conclusion: Our data demonstrated the regulatory effect was different when HSMCs were treated with low (early pregnancy-like) and high (labor-like) CRH concentrations and the KATP channel expression showed significant increase and decrease. This could cause inhibition and activation, respectively, of uterine muscle contraction, demonstrating opposite dual actions of CRH.
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http://dx.doi.org/10.5468/ogs.2018.61.1.14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5780309PMC
January 2018

Fermented Soymilk Alleviates Lipid Accumulation by Inhibition of SREBP-1 and Activation of NRF-2 in the Hepatocellular Steatosis Model.

J Microbiol Biotechnol 2018 Feb;28(2):236-245

Eulji Medi-Bio Research Institute (EMBRI), Eulji University, Daejeon 34824, Republic of Korea.

Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free fatty acid-induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with palmitic acid (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free fatty acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.
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http://dx.doi.org/10.4014/jmb.1707.07061DOI Listing
February 2018

The toothless pterosaur Jidapterus edentus (Pterodactyloidea: Azhdarchoidea) from the Early Cretaceous Jehol Biota and its paleoecological implications.

PLoS One 2017 26;12(9):e0185486. Epub 2017 Sep 26.

Department and Earth and Environmental Science, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

Background: In the Early Cretaceous Jehol Biota, the toothless pterosaurs flourished with the chaoyangopterids and tapejarids playing a key role in understanding the early diversity and evolution of the Azhdarchoidea. Unlike the more diverse tapejarids, the rarer chaoyangopterids are characterized by a long and low rostrum, supporting a close relationship with the huge azhdarchids. Unfortunately, our knowledge is still limited in the osteology, paleoecology, and taxonomy of the Chaoyangopteridae. As one of the best preserved skeletons, the type and only specimen of Jidapterus edentus provides an opportunity to understand the morphology and paleoecology of the chaoyangopterids.

Results: Our study of the osteology of Jidapterus edentus reveals valuable information about the morphology of the Chaoyangopteridae such as a rostrum with a curved dorsal profile, high Rostral Index (RI), larger angle between the dorsal and postorbital processes of the jugal, sequentially shorter fourth to seventh cervical vertebrae, sternum with a plate wider than long, contact of the metacarpal I with the distal syncarpal, pneumatic foramen on first wing phalanx, hatchet-like postacetabular process with unconstricted neck and small dorsal process, distinctly concave anterior margin of pubis, subrectangular pubic plate with nearly parallel anterior and posterior margins, longer proximal phalanges of pedal digits III and IV, as well as reduced and less curved pedal unguals. These features further support the validity of Jidapterus edentus as a distinct species and the close relationship of the chaoyangopterids with the azhdarchids. Paleoecologically, the chaoyangopterids are probably like the azhdarchids, more terrestrial than the contemporaneous and putatively arboreal tapejarids, which may have been limited to the forest-dominated ecosystem of the Jehol Biota.

Discussion: The osteology of Jidapterus edentus further supports the close relationship of the Chaoyangopteridae with the Azhdarchidae in sharing a high RI value and reduced and mildly-curved pedal unguals, and it also implies a possible paleoecological similarity in their terrestrial capability. Combined with the putatively arboreal and herbivorous tapejarids, this distinct lifestyle of the chaoyangopterids provides new insights into the diversity of pterosaurs in the ecosystem of the Jehol Biota.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0185486PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5614613PMC
October 2017

Galectin-1 from conditioned medium of three-dimensional culture of adipose-derived stem cells accelerates migration and proliferation of human keratinocytes and fibroblasts.

Wound Repair Regen 2018 12 8;26 Suppl 1:S9-S18. Epub 2017 Nov 8.

Eulji Medi-Bio Research Institute, Eulji University, Seoul, Republic of Korea.

Keratinocytes and fibroblasts cells play important roles in the skin-wound healing process and are the cell types activated by trauma. Activated cells participate in epithelialization, granulation, scar tissue formation, wound remodeling, and angiogenesis via a series of cellular activities including migration and proliferation. Previous studies reported that the conditioned medium (CM) of adipose-derived stem cells (ADSCs) stimulated the migration and proliferation of cell types involved in the skin wound healing process; however, these studies only show ADSC-CM effects that were obtained using 2-dimensional (2D) culture. Recently, 3-dimensional (3D) culture has been considered as a more physiologically appropriate system than 2D culture for ADSC cultures; therefore, ADSC-CM was collected from 3D culture (ADSC-CM-3D) and compared with ADSC-CM from 2D culture (ADSC-CM-2D) to investigate the effects on the migration and proliferation of human keratinocytes (HaCaTs) and fibroblasts. The migrations of the HaCaT cells and fibroblasts were significantly higher with ADSC-CM-3D compared with the 2D culture; similarly, the proliferation of HaCaT cells was also highly stimulated by ADSC-CM-3D. Proteomic analyses of the ADSC-CM revealed that collagens and actins were highly expressed in the 3D-culture system. Chitinase 3-like 1 (CHI3L1), tissue inhibitor of metalloproteinases (TIMP), and galectin-1 were specifically expressed only in ADSC-CM-3D. Especially, through antibody neutralization, galectin-1 in ADSC-CM-3D was found to be an important factor for the migration of human keratinocytes. Therefore, these results suggest that ADSC-CM-3D was more effective in the wound healing than ADSC-CM-2D, and galectin-1 in ADSC-CM-3D was could be a promising option for skin-wound healing. Furthermore, the differential expressions of several ADSC-CM proteins between the 2D- and 3D-culture systems may be used as basic information for the development of efficient wound-healing strategies.
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http://dx.doi.org/10.1111/wrr.12579DOI Listing
December 2018

Conditioned medium from the three-dimensional culture of human umbilical cord perivascular cells accelerate the migration and proliferation of human keratinocyte and fibroblast.

J Biomater Sci Polym Ed 2018 May - Jun;29(7-9):1066-1080. Epub 2017 Jun 18.

a Eulji Medi-Bio Research Institute, Eulji University , Seoul , South Korea.

Previous studies have reported that the conditioned medium (CM) of bone marrow-mesenchymal stem cells (BM-MSCs) stimulate the migration and proliferation of cell types involved in the wound healing process. However, these studies only show MSC-CM effects that were obtained using a two-dimensional (2D) culture. Recently, a three-dimensional (3D) culture has been considered to be a more physiologically appropriate system than the 2D culture. In addition, it has been shown that the procurement of BM-MSC is invasive, and other sources of MSC are thus being explored. Recently, perivascular cells (PVCs) have been considered as an alternative source of cells for dermal wound healing. Therefore, in this study, a PVC-conditioned medium (CM) was collected from a 3D culture (PVC-CM-3D) using highly porous polystyrene-based membranes and compared with PVC-CM from a 2D culture (PVC-CM-2D) to investigate the effects on the migration and proliferation of human keratinocytes and fibroblasts. Moreover, the PVC-CM components from the 2D and 3D cultures were identified using 2D gel electrophoresis. The migrations of the keratinocytes cells and fibroblasts were significantly higher with PVC-CM-3D than with the 2D culture; similarly, the proliferation of keratinocytes was also highly stimulated by PVC-CM-3D. Proteomic analyses of the PVC-CM revealed that type I collagen was highly expressed in the 3D-culture system. Microtubule-actin cross-linked factor 1 (KIAA0465), nebulin-related anchoring protein, and thioredoxin were specifically expressed only in PVC-CM-3D. In addition, more EVs could be isolated from the PVC-CM-3D, and EVs were found to stimulate keratinocyte migration. Taken together, 3D-culture using a polystyrene scaffold is demonstrated to be a better system for providing better physiological conditions; therefore, PVC-CM-3D could be a promising option for skin-wound healing.
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http://dx.doi.org/10.1080/09205063.2017.1340045DOI Listing
June 2019

Topology Engineering of Proteins Using Genetically Encoded, Mechanically Interlocking SpyX Modules for Enhanced Stability.

ACS Cent Sci 2017 May 10;3(5):473-481. Epub 2017 May 10.

Key Laboratory of Polymer Chemistry & Physics of Ministry of Education, Center for Soft Matter Science and Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China.

Recombinant proteins are traditionally limited to linear configuration. Herein, we report protein topology engineering using highly efficient, mechanically interlocking SpyX modules named AXB and BXA. SpyX modules are protein domains composed of p53dim (X), SpyTag (A), and SpyCatcher (B). The p53dim guides the intertwining of the two nascent protein chains followed by autocatalytic isopeptide bond formation between SpyTag and SpyCatcher to fulfill the interlocking, leading to a variety of backbone topologies. Direct expression of AXB or BXA produces protein catenanes with distinct ring sizes. Recombinant proteins containing SpyX modules are obtained either as mechanically interlocked obligate dimers if the protein of interest is fused to the N- or C-terminus of SpyX modules, or as star proteins if the protein is fused to both N- and C-termini. As examples, cellular syntheses of dimers of (GB1) (where GB1 stands for immunoglobulin-binding domain B1 of streptococcal protein G) and of four-arm elastin-like star proteins were demonstrated. Comparison of the catenation efficiencies in different constructs reveals that BXA is generally much more effective than AXB, which is rationalized by the arrangement of three domains in space. Mechanical interlocking induces considerable stability enhancement. Both AXB and BXA have a melting point ∼20 °C higher than the linear controls and the BXA catenane has a melting point ~2 °C higher than the cyclic control BX'A. Notably, four-arm elastin-like star proteins demonstrate remarkable tolerance against trypsin digestion. The SpyX modules provide a convenient and versatile approach to construct unconventional protein topologies via the "assembly-reaction" synergy, which opens a new horizon in protein science for stability enhancement and function reinforcement via topology engineering.
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http://dx.doi.org/10.1021/acscentsci.7b00104DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5445526PMC
May 2017

Serum leptin and adiponectin levels in atopic dermatitis (AD) and their relation to disease severity.

J Am Acad Dermatol 2016 Sep;75(3):629-631

Department of Dermatology, Eulji Medical Center, College of Medicine, Eulji University, Seoul, Korea. Electronic address:

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http://dx.doi.org/10.1016/j.jaad.2016.04.036DOI Listing
September 2016

Brainstem gangliogliomas: prognostic factors, surgical indications and functional outcomes.

J Neurooncol 2016 07 25;128(3):445-53. Epub 2016 Apr 25.

Medical Center, Tsinghua University, Haidian District, Beijing, 100084, China.

To explore the prognostic factors and discuss the surgical indications of brainstem gangliogliomas. Twenty-one patients with brainstem ganglioglioma were surgically treated at our hospital between 2006 and 2014. The clinical, radiological, operative, and pathological findings of these patients were retrospectively reviewed. The 3-years overall survival and event-free survival (EFS) rates were 90.5 % and 68.4 %, respectively. Four patients (4/18, 22 %) experienced a recurrence with a mean recurrence-free survival of 5.5 months and a mean follow-up of 37 months. Three patients died of surgery-related complications. Three growth patterns were identified: exophytic (6/21), intrinsic (2/21), and endo-exophytic (13/21). Eight patients (8/15, 53 %) harbored a BRAF V600E mutation. All recurrent tumors were endo-exophytic, and except the one without molecular information, were BRAF V600E mutants. A Cox hazard proportion ratio model was used to identify factors influencing EFS, including sex, age, location, growth patterns, extent of resection (EOR), and BRAF V600E mutation status. On univariate analysis, none of these factors reached statistical significance. Among them, EOR and growth patterns were strongly associated with each other (Fisher's exact test, P < 0.01). A multivariate analysis demonstrated that growth patterns were the only factor associated with EFS (P = 0.02; HR 49.05; 95 % CI 1.76-1365.13). Growth patterns may be useful to select surgery candidates and predict prognosis for patients with brainstem gangliogliomas. BRAF V600E was frequently present and appeared to be associated with shorter recurrence-free survival. Studies on BRAF V600E-targeted therapy for patients with high surgical risks are needed.
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http://dx.doi.org/10.1007/s11060-016-2131-zDOI Listing
July 2016

Design, synthesis and evaluation of novel tacrine-multialkoxybenzene hybrids as multi-targeted compounds against Alzheimer's disease.

Eur J Med Chem 2016 Jun 29;116:200-209. Epub 2016 Mar 29.

School of Pharmaceutical Sciences, Guangzhou Medical University, Xinzao, Panyu District, Guangzhou, 511436, PR China.

A series of benzoates (or phenylacetates or cinnamates) - tacrine hybrids (7a-o) were designed, synthesized and evaluated as multi-potent anti-Alzheimer drug candidates. The screening results showed that most of them exhibited a significant ability to inhibit ChEs, certain selectivity for AChE over BuChE and strong potency inhibitory of self-induced β-amyloid (Aβ) aggregation. All IC50 values of biological activity were at the nanomolar range. Especially, compound 7c displayed the greatest ability to inhibit AChE with an IC50 value of 5.63 nM and the highest selectivity with ratio of BuChE/AChE value of 64.6. Moreover, it also exhibited a potent inhibitory of Aβ aggregation with an IC50 value of 51.81 nM. A Lineweaver-Burk plot and molecular modeling study showed that compound 7c targeted both the CAS and PAS of ChEs. A structure-activity relationship analysis suggested that the electron density of aromatic ring which was linked with tacrine through acetyl group played a significant role in determining the inhibitory activity.
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http://dx.doi.org/10.1016/j.ejmech.2016.03.077DOI Listing
June 2016

Proposition of a Silica Nanoparticle-Enhanced Hybrid Spin-Microcantilever Sensor Using Nonlinear Optics for Detection of DNA in Liquid.

Sensors (Basel) 2015 Sep 25;15(10):24848-61. Epub 2015 Sep 25.

Key Laboratory of Artificial Structures and Quantum Control (Ministry of Education), Department of Physics and Astronomy, Shanghai Jiao Tong University, Shanghai 200240, China.

We theoretically propose a method based on the combination of a nonlinear optical mass sensor using a hybrid spin-microcantilever and the nanoparticle-enhanced technique, to detect and monitor DNA mutations. The technique theoretically allows the mass of external particles (ssDNA) landing on the surface of a hybrid spin-microcantilever to be detected directly and accurately at 300 K with a mass responsivity 0.137 Hz/ag in situ in liquid. Moreover, combined with the nanoparticle-enhanced technique, even only one base pair mutation in the target DNA sequence can be identified in real time accurately, and the DNA hybridization reactions can be monitored quantitatively. Furthermore, in situ detection in liquid and measurement of the proposed nonlinear optical spin resonance spectra will minimize the experimental errors.
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http://dx.doi.org/10.3390/s151024848DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4634466PMC
September 2015

Sensitive detection of Majorana fermions based on a hybrid spin-microcantilever via enhanced spin resonance spectrum.

Nanotechnology 2015 May 21;26(19):195501. Epub 2015 Apr 21.

Key Laboratory of Artificial Structures and Quantum Control (Ministry of Education), Department of Physics and Astronomy, Shanghai Jiao Tong University, 800 Dong Chuan Road, Shanghai 200240, People's Republic of China.

Motivated by recent experimental progress towards the detection and manipulation of Majorana fermions in ferromagnetic atomic chains on a superconductor, we present a novel proposal based on a single-crystal diamond (SCD) microcantilever with a single nitrogen-vacancy (NV) center spin embedded in ultrapure diamond substrate to probe Majorana fermions in an all-optical domain. With this scheme, a possible distinct Majorana signature is investigated via the electron spin resonance spectrum. In the proposal, the SCD microcantilever behaves as a phonon cavity and is robust for detecting of Majorana fermions, while the NV center spin can be considered as a sensitive probe. Further, the vibration of the microcantilever will enhance the coupling effect, which makes the Majorana fermions more sensitive to detection and the well-established optical NV spin readout technology will certainly promote the detection. This proposed method may provide a potential supplement for the detection of Majorana fermions.
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http://dx.doi.org/10.1088/0957-4484/26/19/195501DOI Listing
May 2015

A biologically inspired lung-on-a-chip device for the study of protein-induced lung inflammation.

Integr Biol (Camb) 2015 Feb;7(2):162-9

Institute of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.

This study reports a biomimetic microsystem that reconstitutes the lung microenvironment for monitoring the role of eosinophil cationic protein (ECP) in lung inflammation. ECP induces the airway epithelial cell expression of CXCL-12, which in turn stimulates the migration of fibrocytes towards the epithelium. This two-layered microfluidic system provides a feasible platform for perfusion culture, and was used in this study to reveal that the CXCL12-CXCR4 axis mediates ECP induced fibrocyte extravasation in lung inflammation. This 'lung-on-a-chip' microdevice serves as a dynamic transwell system by introducing a flow that can reconstitute the blood vessel-tissue interface for in vitro assays, enhancing pre-clinical studies. We made an attempt to develop a new microfluidic model which could not only simulate the transwell for studying cell migration, but could also study the migration in the presence of a flow mimicking the physiological conditions in the body. As blood vessels are the integral part of our body, this model gives an opportunity to study more realistic in vitro models of organs where the blood vessel i.e. flow based migration is involved.
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http://dx.doi.org/10.1039/c4ib00239cDOI Listing
February 2015

Hybrid spin-microcantilever sensor for environmental, chemical, and biological detection.

Nanotechnology 2015 Jan 8;26(1):015501. Epub 2014 Dec 8.

Key Laboratory of Artificial Structures and Quantum Control (Ministry of Education), Department of Physics and Astronomy, Shanghai Jiao Tong University, 800 Dong Chuan Road, Shanghai 200240, People's Republic of China.

Nowadays hybrid spin-micro/nanomechanical systems are being actively explored for potential quantum sensing applications. In combination with the pump-probe technique or the spin resonance spectrum, we theoretically propose a realistic, feasible, and an exact way to measure the cantilever frequency in a hybrid spin-micromechanical cantilever system which has a strong coherent coupling of a single nitrogen vacancy center in the single-crystal diamond cantilever with the microcantilever. The probe absorption spectrum which exhibits new features such as mechanically induced three-photon resonance and ac Stark effect is obtained. Simultaneously, we further develop this hybrid spin-micromechanical system to be an ultrasensitive mass sensor, which can be operated at 300 K with a mass responsivity 0.137 Hz ag(-1), for accurate sensing of gaseous or aqueous environments, chemical vapors, and biomolecules. And the best performance on the minimum detectable mass can be [Formula: see text] in vacuum. Finally, we illustrate an in situ measurement to detect Angiopoietin-1, a marker of tumor angiogenesis, accurately with this hybrid microcantilever at room temperature.
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http://dx.doi.org/10.1088/0957-4484/26/1/015501DOI Listing
January 2015

Fibrocyte trafficking in patients with chronic obstructive asthma and during an acute asthma exacerbation.

J Allergy Clin Immunol 2015 May 24;135(5):1154-62.e1-5. Epub 2014 Oct 24.

Department of Thoracic Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan; Department of Medicine, Chang Gung University, Taoyuan, Taiwan. Electronic address:

Background: Fibrocytes express several chemokine receptors (CCR7 and CXCR4) that regulate their recruitment and trafficking into tissue-damage sites in response to specific chemokine gradients (CCL19 and CXCL12).

Objective: We investigated whether these chemoattractants and S100A9, through the receptor for advanced glycation end-products (RAGE; ie, its receptor), are involved in fibrocyte trafficking in patients with chronic obstructive asthma (COA) and during an acute exacerbation (AE) in patients without airflow obstruction (Asthma AE group).

Methods: We collected peripheral blood from 14 asthmatic patients with normal pulmonary function, 14 patients with COA, 11 patients in the Asthma AE group, and 14 healthy subjects. Isolated circulating fibrocytes were used for migration assay. Expression of CCR7, CXCR4, S100A9, and RAGE in fibrocytes was measured by using flow cytometry. CCL19 and CXCL12 expression in bronchial tissues was determined by using immunohistochemistry and RT-PCR.

Results: There were higher numbers of circulating fibrocytes in patients in the Asthma AE group and patients with COA. The expression of CXCL12 in bronchial tissues and CXCR4 in circulating fibrocytes was higher in the Asthma AE group and, to a lesser extent, in patients with COA. The expression of CCL19 in bronchial tissues and CCR7 in fibrocytes was higher in patients with COA. CXCL12/CXCR4 and CCL19/CCR7 enhanced fibrocyte transmigration in the Asthma AE group and in patients with COA, respectively. The upregulated expression of S100A9 and RAGE in fibrocytes of patients in the Asthma AE group and those with COA contributes to the enhanced basal migratory motility of fibrocytes.

Conclusion: The CXCR4/CXCL12 axis contributes to chemotaxis of fibrocytes in patients in the Asthma AE group, whereas the CCR7/CCL19 axis plays an important role in patients with COA. S100A9 enhances the basal migratory motility of fibrocytes from patients in the Asthma AE group and patients with COA.
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http://dx.doi.org/10.1016/j.jaci.2014.09.011DOI Listing
May 2015

Fabrication and properties of irradiation-cross-linked poly(vinyl alcohol)/clay aerogel composites.

ACS Appl Mater Interfaces 2014 Sep 8;6(18):16227-36. Epub 2014 Sep 8.

Institute of Nuclear Physics and Chemistry, Chinese Academy of Engineering Physics , Mianyang, Sichuan 621000, China.

Poly(vinyl alcohol) (PVOH)/clay aerogel composites were fabricated by an environmentally friendly freeze-drying of the aqueous precursor suspensions, followed by cross-linking induced by gamma irradiation without chemical additives. The influences of cross-linking conditions, i.e., absorbed dose and polymer loading as well as density on the aerogel structure and properties, were investigated. The absorbed dose of 30 kGy was found to be the optimum dose for fabricating strong PVOH composites; the compressive modulus of an aerogel prepared from an aqueous suspension containing 2 wt % PVOH/8 wt % clay increased 10-fold, and that containing 1 wt % PVOH/9 wt % clay increased 12 times upon cross-linking with a dose of 30 kGy. Increasing the solids concentration led to an increase in the mechanical strength, in accordance with the changes in microstructure from layered structure to network structure. The increase of absorbed dose also led to decreased porous size of the network structure. Cross-linking and the increase of the PVOH lead to decreased thermal stability. The strengthened PVOH/clay aerogels possess very low flammability, as measured by cone calorimetry, with heat, smoke, and volatile products release value decreasing as increasing clay content. The mechanism of flame retardation in these materials was investigated with weight loss, FTIR, WAXD, and SEM of the burned residues. The proposed mechanism is that with decreasing fuel content (increasing clay content), increased heat and mass transport barriers are developed; simultaneously low levels of thermal conductivity are maintained during the burning.
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http://dx.doi.org/10.1021/am504418wDOI Listing
September 2014

Dermal clusters of mature dendritic cells and T cells are associated with the CCL20/CCR6 chemokine system in chronic psoriasis.

J Invest Dermatol 2014 May 13;134(5):1462-1465. Epub 2013 Dec 13.

Department of Dermatology, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, South Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, South Korea. Electronic address:

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http://dx.doi.org/10.1038/jid.2013.534DOI Listing
May 2014

Molecular epidemiology of enterovirus 71 infection in the central region of Taiwan from 2002 to 2012.

PLoS One 2013 31;8(12):e83711. Epub 2013 Dec 31.

Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung, Taiwan ; Institute of Microbiology and Immunology, School of Life Sciences, National Yang-Ming University, Taipei, Taiwan ; Department of Microbiology and Institute of Medical Research, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.

Enterovirus 71 (EV71), a causative agent of hand, foot, and mouth disease can be classified into three genotypes and many subtypes. The objectives of this study were to conduct a molecular epidemiological study of EV71 in the central region of Taiwan from 2002-2012 and to test the hypothesis that whether the alternative appearance of different EV71 subtypes in Taiwan is due to transmission from neighboring countries or from re-emergence of pre-existing local strains. We selected 174 EV71 isolates and used reverse transcription-polymerase chain reaction to amplify their VP1 region for DNA sequencing. Phylogenetic analyses were conducted using Neighbor-Joining, Maximum Likelihood and Bayesian methods. We found that the major subtypes of EV71 in Taiwan were B4 for 2002 epidemic, C4 for 2004-2005 epidemic, B5 for 2008-2009 epidemic, C4 for 2010 epidemic and B5 for 2011-2012 epidemic. Phylogenetic analysis demonstrated that the 2002 and 2008 epidemics were associated with EV71 from Malaysia and Singapore; while both 2010 and 2011-2012 epidemics originated from different regions of mainland China including Shanghai, Henan, Xiamen and Gong-Dong. Furthermore, minor strains have been identified in each epidemic and some of them were correlated with the subsequent outbreaks. Therefore, the EV71 infection in Taiwan may originate from pre-existing minor strains or from other regions in Asia including mainland China. In addition, 101 EV71 isolates were selected for the detection of new recombinant strains using the nucleotide sequences spanning the VP1-2A-2B region. No new recombinant strain was found. Analysis of clinical manifestations showed that patients infected with C4 had significantly higher rates of pharyngeal vesicles or ulcers than patients infected with B5. This is the first study demonstrating that different EV 71 genotypes may have different clinical manifestations and the association of EV71 infections between Taiwan and mainland China.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0083711PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3877097PMC
September 2014

Role of the cAMP-dependent carbon catabolite repression in capsular polysaccharide biosynthesis in Klebsiella pneumoniae.

PLoS One 2013 11;8(2):e54430. Epub 2013 Feb 11.

School of Chinese Medicine, China Medical University, Taichung, Taiwan. Republic of China.

K. pneumoniae is the predominant pathogen isolated from liver abscesses of diabetic patients in Asian countries. Although elevated blood glucose levels cause various immune problems, its effects on K. pneumoniae virulence are unknown. This study investigated the regulation of capsular polysaccharide (CPS) biosynthesis, a major determinant for K. pneumoniae virulence, in response to exogenous glucose. We found that K. pneumoniae produce more CPS in glucose-rich medium via reduction in cyclic AMP (cAMP) levels. Individual deletion of cyaA or crp, which respectively encode adenylate cyclase and cAMP receptor protein in K. pneumoniae, markedly increased CPS production, while deletion of cpdA, which encodes cAMP phosphodiesterase, decreased CPS production. These results indicate that K. pneumoniae CPS biosynthesis is controlled by the cAMP-dependent carbon catabolite repression (CCR). To investigate the underlying mechanism, quantitative real-time PCR and promoter-reporter assays were used to verify that the transcription of CPS biosynthesis genes, which are organized into 3 transcription units (orf1-2, orf3-15, and orf16-17), were activated by the deletion of crp. Sequence analysis revealed putative CRP binding sites located on P(orf3-15) and P(orf16-17), suggesting direct CRP-cAMP regulation on the promoters. These results were then confirmed by electrophoretic mobility shift assay. In addition, we found putative CRP binding sites located in the promoter region of rcsA, which encodes a cps transcriptional activator, demonstrating a direct repression of CRP-cAMP and P(rcsA). The deletion of rcsA in mutation of crp partially reduced CPS biosynthesis and the transcription of orf1-2 but not of orf3-15 or orf16-17. These results suggest that RcsA participates in the CRP-cAMP regulation of orf1-2 transcription and influences CPS biosynthesis. Finally, the effect of glucose and CCR proteins on CPS biosynthesis also reflects bacterial resistance to serum killing. We here provide evidence that K. pneumoniae increases CPS biosynthesis for successful infection in response to exogenous glucose via cAMP-dependent CCR.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0054430PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3569464PMC
September 2013

Attenuation of contact hypersensitivity by cell-permeable heat shock protein 70 in BALB/c mouse model.

Exp Dermatol 2012 Dec;21(12):969-71

 In contact hypersensitivity (CHS), multiple cells, inflammatory mediators and cytokines are known to be involved in the regulation of the immune response. Previously, we revealed the reactive oxygen species generation by 2, 4, 6-trinitrobenzene sulphonic acid (TNBS) in vivo, followed by heat shock protein 70 (Hsp70) carbonylation and the exogenous antioxidant role of cell-permeable Hsp70. Here, we demonstrate the role of Hsp70 using cell-permeable Hsp70 in the mouse CHS model. Pretreatment of cell-permeable Hsp70: (i) suppressed ear swelling; (ii) down-regulated phosphorylated p38, but up-regulated phosphorylated extracellular signal-regulated kinase; (iii) increased population of CD4(+) CD25(+) Foxp3(+) T cells; (iv) decreased secretion of tumor necrosis factor-α (TNF-α), IL-12, interferon-γ and IL-2 and (v) but up-regulated IL-4 and transforming growth factor beta (TGF-β) in the lymph nodes. In conclusion, cell-permeable Hsp70 attenuates CHS through modulation of MAPK pathway and regulation of Th1, Th2 and regulatory T cells.
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http://dx.doi.org/10.1111/exd.12044DOI Listing
December 2012

Assessment of an imiquimod-induced psoriatic mouse model in relation to oxidative stress.

Arch Dermatol Res 2012 Nov 5;304(9):699-706. Epub 2012 Aug 5.

Department of Dermatology, Gachon University, Seongnam, Korea.

Psoriasis is a chronic inflammatory skin disease that is thought to be related to oxidative stress. Much progress has been made in understanding the pathophysiology of psoriasis in relation to the immunologic and antioxidant systems. However, this progress has been hindered by the lack of an appropriate animal model for psoriasis. Recently, imiquimod (IQM)-induced psoriasis-like cutaneous inflammation has been reported in mice and humans. We verified the usefulness of an IQM-induced mouse model in relation to the antioxidant system. BALB/C female mice at 8-10 weeks of age were treated with IQM cream in this study. We analyzed clinical and histopathological changes. Increased reactive oxygen species production was measured by glutathione assay. Levels of myeloperoxidase (MPO) and superoxide dismutase-1 (SOD1) were determined by western blotting and immunohistochemical analyses. The activity of SOD was measured by a SOD activity assay kit. Application of IQM-induced skin inflammation similar to psoriasis in clinical and histopathological aspects. Accumulation of immune cells was confirmed. Oxidative stress was increased, the antioxidant enzyme MPO levels were increased, and both SOD levels and activity were decreased. In conclusion, the IQM-induced mouse model showed an aberrant antioxidant system. Levels of MPO and oxidative stress were increased, and the level and activity of SOD were decreased. Since this model seemed to be an appropriate model for psoriasis, it can be used to further study the pathogenic role of redox imbalance in psoriasis.
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http://dx.doi.org/10.1007/s00403-012-1272-yDOI Listing
November 2012

Corticotropin-releasing hormone downregulates IL-10 production by adaptive forkhead box protein 3-negative regulatory T cells in patients with atopic dermatitis.

J Allergy Clin Immunol 2012 Jan 13;129(1):151-9.e1-6. Epub 2011 Oct 13.

Department of Dermatology and Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea.

Background: Corticotropin-releasing hormone (CRH) is the central regulating hormone of the hypothalamic-pituitary-adrenal axis. CRH also has diverse functional effects in the periphery and is related to the aggravation of several cutaneous diseases; however, the effect of CRH on T cells in patients with atopic dermatitis (AD) has not been well evaluated.

Objective: We investigated whether CRH directly affects peripheral T(H)1, T(H)2, and regulatory T (Treg) cells in patients with AD.

Methods: We assessed whether T cells express the CRH receptor protein and mRNA by using flow cytometry, Western blotting, immunofluorescence, immunohistochemistry, and RT-PCR. We evaluated cytokine expression using ELISA after treating the T cells extracted from patients with AD and healthy control subjects (HCs) with CRH. Flow cytometry was then used to evaluate any direct effects of CRH on T(H)1, T(H)2, and Treg cells from patients with AD and HCs.

Results: T cells from patients with AD expressed significantly lower CRH receptor 1/2 mRNA levels than T cells from HCs. T cells from HCs reacted with different IL-4 and IFN-γ secretions to CRH treatment, whereas T cells from patients with AD did not. IL-10 production was significantly decreased in the supernatants from both the HCs and patients with AD after CRH treatment. CRH upregulated IL-4 production by T(H)2 cells and downregulated IFN-γ production by T(H)1 cells in HCs. CRH also suppressed the production of IL-10 by forkhead box protein 3-negative Treg cells in both groups, but the difference was only significant in patients with AD.

Conclusions: CRH-mediated suppression of IL-10 secretion from Treg cells might explain stress-related exacerbations in patients with AD.
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http://dx.doi.org/10.1016/j.jaci.2011.09.008DOI Listing
January 2012