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Peng X.,Southwest Research Institute | Yuan S.,Southwest Research Institute | Tan J.,Southwest Research Institute | Ma B.,Southwest Research Institute | And 14 more authors.
Life Sciences | Year: 2012

Aims: P311 is an 8 kDa protein that has been shown to be of importance in the process of myofibroblast transformation, glioblastoma invasion and nerve regeneration. However, the interaction protein of P311 has yet to be found. The purpose of this study was to find the interactive protein of P311. Main methods: The yeast two-hybrid system was used for screening the potential interaction proteins of P311. Joint expression of the potential interactive protein and P311 was immunohistochemically stained. The interaction between P311 and the selected protein was further confirmed by fluorescence resonance energy transfer (FRET) in pulmonary adenocarcinoma tissue sections, and by coimmunoprecipitation in HEK293. Key findings: Integrin β4 binding protein (ITGB4BP) was confirmed as the interaction protein of P311. Co-expression and interaction of ITGB4BP and P311 were demonstrated in pulmonary adenocarcinoma by both immunohistochemistry and FRET. Moreover the interaction between P311 and ITGB4BP was demonstrated by coimmunoprecipitation in HEK293. Significance: The interactions between P311 and ITGB4BP may be very important in the process of tumor cell differentiation and metastasis. ITGB4BP may provide a potential new target for the therapy of tumors. © 2012 Elsevier Inc. All rights reserved.

He W.,Chongqing Key Laboratory for Disease Proteomics | Huang C.,Chongqing Medical University | Luo G.,Chongqing Key Laboratory for Disease Proteomics | Pra I.D.,University of Verona | And 9 more authors.
Proteomics | Year: 2012

Just as biomarkers specific for diseases, biomarkers indicative of healthy conditions are valuable for the early diagnosis, monitoring, and prognosis of diseases. Our study focused on discovering via proteomics a stable panel of urinary proteins in the human healthy population. Urine samples were collected three times during 4 months from 100 male and 100 female healthy donors and analyzed through four different fractionation techniques (i.e. in-gel, 2D-LC, OFFGEL, and mRP) coupled with HPLC-Chip-MS/MS. Thus, 1641 urinary proteins were identified with a high confidence, among which 70 exhibiting an intergender/day variation <0.25 were selected and matched with the previously published five largest urinary proteomes to get 56 candidate proteins. Next, a panel comprising 18 intact urinary proteins was constructed by comparing the urinary proteomes via SDS-PAGE and 2DE. Finally, such 18 urinary proteins were validated via enzyme-linked immunosorbent assay in eight healthy individuals. Most of these proteins had been related to multiple rather than to single diseases. Therefore, we surmise that this protein set could be used as a biomarker to assess the human health status. Further determinations of the normal fluctuations of the single urinary proteins in this series using samples from large numbers of healthy individuals are required prior to any application in clinical settings. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Wang Y.,Chongqing Key Laboratory for Disease Proteomics | Xu R.,Chongqing Key Laboratory for Disease Proteomics | Luo G.,Chongqing Key Laboratory for Disease Proteomics | Lei Q.,Chongqing Key Laboratory for Disease Proteomics | And 9 more authors.
Acta Biomaterialia | Year: 2016

The structure of dermal scaffolds greatly affects the engineered tissue's functions and the activities of seeded cells. Current strategies of dermal scaffold design tend to yield a homogeneous architecture with a uniform pore size. However, the structures of the human dermis are not homogeneous in terms of either interstitial spaces or architecture at different dermal depths. In the present study, a biomimetic fibroblasts-loaded artificial dermis composed of three-layer scaffolds with different pore sizes was prepared. The three-layer scaffolds, which look similar to a sandwich, mimic the natural structures of the human dermis, which has comparatively larger pores in the outer layers and smaller pores in the middle layer. The fibroblasts-loaded artificial dermis were shown to favor wound healing by promoting granulation tissue formation and wound re-epithelialization, as determined by a histological study and Western blotting. Our data indicated that the biomimetic fibroblasts-loaded artificial dermis with "Sandwich" structure and designed gradient pore sizes may hold promise as tissue-engineered dermis. Statement of Significance Pore size effect on wound healing had been extensively studied. However, it is still not well understood whether dermal scaffolds with a uniform pore size are better than that with varied pore sizes, which are similar to human dermis as determined by our previous work. In our study, we demonstrated that the "sandwich" collagen scaffolds mimicking the natural structures of the human dermis significantly promoted wound healing compared with the "Homogeneous" scaffolds with a uniform pore size. These results may be helpful in the design of dermal scaffolds. © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Li H.,Chongqing Medical University | Li J.,Chongqing Medical University | Wang Y.,Chongqing Key Laboratory for Disease Proteomics | Yang T.,Chongqing Medical University
Proteome Science | Year: 2012

Background: Biomarkers released from the heart at early stage of ischemia are very important to diagnosis of ischemic heart disease and salvage myocytes from death. Known specific markers for blood tests including CK-MB, cardiac troponin T (cTnT) and cardiac troponin I (cTnI) are released after the onset of significant necrosis instead of early ischemia. Thus, they are not good biomarkers to diagnose myocardial injury before necrosis happens. Therefore, in this study, we performed proteomic analysis on effluents from perfused human hearts of donors at different ischemic time.Results: After global ischemia for 0 min, 30 min and 60 min at 4°C, effluents from five perfused hearts were analyzed respectively, by High performance liquid chromatography-Chip-Mass spectrometry (HPLC-Chip-MS) system. Total 196 highly reliable proteins were identified. 107 proteins were identified at the beginning of ischemia, 174 and 175 proteins at ischemic 30 min and ischemic 60 min, respectively. With the exception of cardiac troponin I and T, all known biomarkers for myocardial ischemia were detected in our study. However, there were four glycolytic enzymes and two targets of matrix metalloproteinase released significantly from the heart when ischemic time was increasing. These proteins were L-lactate dehydrogenase B(LDHB), glyceraldehyde-3-phosphate dehydrogenase, glucose-6-phosphate isomerase (GPI), phosphoglycerate mutase 2 (PGAM2), gelsolin and isoform 8 of titin. PGAM2, LDHB and titin were measured with enzyme-linked immunosorbent assays kits. The mean concentrations of LDHB and PGAM2 in samples showed an increasing trend when ischemic time was extending. In addition, 33% identified proteins are involved in metabolism. Protein to protein interaction network analysis showed glycolytic enzymes, such as isoform alpha-enolase of alpha-enolase, isoform 1 of triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase, had more connections than other proteins in myocardial metabolism during ischemia.Conclusion: It is the first time to use effluents of human perfused heart to study the proteins released during myocardial ischemia by HPLC-Chip-MS system. There might be many potential biomarkers for mild ischemic injury in myocardium, especially isoform 8 of titin and M-type of PGAM2 that are more specific in the cardiac tissue than in the others. Furthermore, glycolysis is one of the important conversions during early ischemia in myocardium. This finding may provide new insight into pathology and biology of myocardial ischemia, and potential diagnostic and therapeutic biomarkers. © 2012 Li et al; licensee BioMed Central Ltd.

Huang Z.-M.,Chongqing Medical University | Wu J.,Chongqing Key Laboratory for Disease Proteomics | Jia Z.,Chongqing Medical University | Tian Y.,Chongqing Medical University | And 5 more authors.
BMB Reports | Year: 2012

The retinoid-related orphan nuclear receptor gamma (RORγ) plays critical roles in regulation of development, immunity and metabolism. As transcription factor usually forms a protein complex to function, thus capturing and dissecting of the RORγ protein complex will be helpful for exploring the mechanisms underlying those functions. After construction of the recombinant tandem affinity purification (TAP) plasmid, pMSCVpuro RORγ-CTAP(SG), the nuclear localization of RORγ-CTAP(SG) fusion protein was verified. Following isolation of RORγ protein complex by TAP strategy, seven candidate interacting proteins were identified. Finally, the heat shock protein 90 (HSP90) and receptor-interacting protein 140 (RIP140) were confirmed to interplay with RORγ by co-immunoprecipitation. Interference of HSP90 or/and RIP140 genes resulted in dramatically decreased expression of CYP2C8 gene, the RORγ target gene. Data from this study demonstrate that HSP90 and RIP140 proteins interact with RORγ protein in a complex format and function as co-activators in the RORγ-mediated regulatory processes of HepG2 cells. © 2012 by the The Korean Society for Biochemistry and Molecular Biology.

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