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Tan J.,Chongqing Medical University | Peng X.,Chongqing Medical University | Luo G.,Chongqing Medical University | Luo G.,Chongqing Key Laboratory for Disease Proteomics | And 9 more authors.
PLoS ONE | Year: 2010

The mechanisms of hypertrophic scar formation are not fully understood. We previously screened the differentially expressed genes of human hypertrophic scar tissue and identified P311 gene as upregulated. As the activities of P311 in human fibroblast function are unknown, we examined the distribution of it and the effects of forced expression or silencing of expression of P311. P311 expression was detected in fibroblast-like cells from the hypertrophic scar of burn injury patients but not in peripheral blood mononuclear cells, bone marrow mesenchymal stem cells, epidermal cells or normal skin dermal cells. Transfection of fibroblasts with P311 gene stimulated the expression of alpha-smooth muscle actin (α-SMA), TGF-β1 and α1(I) collagen (COL1A1), and enhanced the contraction of fibroblast populated collagen lattices (FPCL). In contrast, interference of fibroblast P311 gene expression decreased the TGF-β1 mRNA expression and reduced the contraction of fibroblasts in FPCL. These results suggest that P311 may be involved in the pathogenesis of hypertrophic scar via induction of a myofibroblastic phenotype and of functions such as TGF-β1 expression. P311 could be a novel target for the control of hypertrophic scar development. © 2010 Tan et al.


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.


Liu Z.,Sun Yat Sen University | Xu Y.,Sun Yat Sen University | Chen L.,Sun Yat Sen University | Xie J.,Sun Yat Sen University | And 15 more authors.
American Journal of Translational Research | Year: 2016

The impairment of skin repair in diabetic patients can lead to increased morbidity and mortality. Proper proliferation, apoptosis and migration in keratinocytes are vital for skin repair, but in diabetic patients, hyperglycemia impairs this process. Dendritic epidermal T cells (DETCs) are an important part of the resident cutaneous immunosurveillance program. We observed a reduction in the number of DETCs in a streptozotocin-induced diabetic mouse model. This reduction in DETCs resulted in decreased IGF-1 and KGF production in the epidermis, which is closely associated with diabetic delayed wound closure. DETCs ameliorated the poor wound-healing conditions in diabetic mice by increasing keratinocyte migration and proliferation and decreasing keratinocyte apoptosis in diabetes-like microenvironments. Our results elucidate a new mechanism for diabetic delayed wound closure and point to a new strategy for the treatment of wounds in diabetic patients. © 2016, E-Century Publishing Corporation. All Rights Reserved.


Liu Z.,Sun Yat Sen University | Xu Y.,Sun Yat Sen University | Zhang X.,Chongqing Medical University | Zhang X.,Chongqing Key Laboratory for Disease Proteomics | And 17 more authors.
American Journal of Translational Research | Year: 2016

The skin serves as a physical and chemical barrier to provide an initial line of defense against environmental threats; however, this function is impaired in diabetes. Vy4 y δ T cells in the dermis are an important part of the resident cutaneous immunosurveillance program, but these cells have yet to be explored in the context of diabetes. In this study, we observed that the impaired maintenance of dermal Vy4 y δ T cells is caused by reduced production of IL-7 in the skin of diabetic mice, which was closely associated with weakened activation of the mTOR pathway in the epidermis of diabetic mice. Weakened CCL20/CCR6 chemokine signaling resulted in the impaired recruitment of dermal Vy4 y δ T cells following wounding in diabetic mice. Meanwhile, reduced levels of IL-23 and IL-1β in the dermis around the wounds of diabetic mice resulted in the impaired production of IL-17 by dermal Vy4 y δ T cells. Therefore, diminished dermal Vy4 y δ T cells and impaired IL-17 production by these cells were important factors in the markedly reduced IL-17 levels in the skin around the wounds of diabetic mice. Because reduced IL-17 levels at the wound edge have been closely associated with delayed wound closure in diabetic mice, defects in dermal Vy4 y δ T cells may be an important mechanism underlying delayed wound healing in diabetic mice. © 2016, E-Century Publishing Corporation. 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.


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.


Yang S.-S.,Chongqing Medical University | Yang S.-S.,Chongqing Key Laboratory for Disease Proteomics | Tan J.-L.,Chongqing Medical University | Tan J.-L.,Chongqing Key Laboratory for Disease Proteomics | And 14 more authors.
Journal of Cell Science | Year: 2015

Eukaryotic initiation factor 6 (eIF6) is a pivotal regulator of ribosomal function, participating in translational control. Previously our data suggested that eIF6 acts as a key binding protein of P311 (a hypertrophic scar-related protein; also known as NREP). However, a comprehensive investigation of its functional role and the underlying mechanisms in modulation of myofibroblast (a key effector of hypertrophic scar formation) differentiation remains unclear. Here, we identified that eIF6 is a novel regulator of transforming growth factor-β1 (TGF-β1) expression at transcription level, which plays a key role in myofibroblast differentiation. Mechanistically, this effect is associated with eIF6 altering the occupancy of the TGF-β1 promoter by H2A.Z (Swiss-Prot P0C0S6) and Sp1. Accordingly, modulation of eIF6 expression in myofibroblasts significantly affects their differentiation via the TGF-β/Smad signaling pathway, which was verified in vivo by the observation that heterozygote eIF6+/- mice exhibited enhanced TGF-β1 production coupled with increased a- smooth muscle actin (α-SMA)+ myofibroblasts after skin injury. Overall, our data reveal a novel transcriptional regulatory mechanism of eIF6 that acts on facilitating Sp1 recruitment to TGF- β1 promoter via H2A.Z depletion and thus results in increased TGF-β1 transcription, which contributes to myofibroblast differentiation. © 2015.


Wang Y.,Chongqing Medical University | Wang Y.,Chongqing Key Laboratory for Disease Proteomics | Chen Z.,Chongqing Medical University | Chen Z.,Chongqing Key Laboratory for Disease Proteomics | And 16 more authors.
ACS Applied Materials and Interfaces | Year: 2016

Vasoactive intestinal peptide (VIP) was reported to promote angiogenesis. Electrospun nanofibers lead to idea wound dressing substrates. Here we report a convenient and novel method to produce VIP loaded microspheres in polycaprolactone (PCL) nanofibrous membrane without complicated processes. We first coated mussel-inspired dopamine (DA) to nanofibers, then used strong adhesive DA to absorb the functional peptide. PCL membrane was then immersed into acetone to generate microspheres with VIP loading. We employed high pressure liquid chromatography to record encapsulation efficiency of (31.8 ± 2.2)% and loading capacity of (1.71 ± 0.16)%. The release profile of VIP from nanosheets showed a prolonged release. The results of laser scanning confocal microscope, scanning electron microscope and cell counting kit-8 proliferation assays showed that cell adhesion and proliferation were promoted. In order to verify the efficacy on wound healing, in vivo implantation was applied in the full-thickness defect wounds of BALB/c mice. Results showed that the wound healing was significantly promoted via favoring the growth of granulation tissue and angiogenesis. However, we found wound re-epithelialization was not significantly improved. The resulting VIP-DA-coated PCL (PCL-DA-VIP) nanosheets with spatiotemporal delivery of VIP could be a potential application in wound treatment and vascular tissue engineering. © 2016 American Chemical Society.


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.


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.

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