Xin Xie

Northwest University, Western Springs, Illinois, United States

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Publications (12)18.1 Total impact

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    ABSTRACT: The human leukocyte-associated Ig-like receptor (LAIR) family contains two members: LAIR-1 (CD305) and LAIR-2 (CD306). Among them, LAIR-1 is a transmembrane glycoprotein bearing two intracellular immunoreceptor tyrosine-based inhibition motifs (ITIM) and LAIR-2 is soluble. Both molecules bind collagen and LAIR-2 has higher affinity than LAIR-1. LAIR-1 can mediate strong inhibitory signal but the functions of leukocytes expressing LAIR-1 are unclear because of the absence of an effective method to isolate them with resting status. In this study, we generated a monoclonal antibody (MAb) by immunizing BALB/c mice with the recombinant LAIR-2-GST fusion protein, which we termed 3G4. The subclass of 3G4 was identified as IgG1. Specificity analysis by Western blotting demonstrated 3G4 could react with both LAIR-1 and LAIR-2. Unlike another LAIR-1-specific MAb (9.1C3), 3G4 did not inhibit the lysis of target cells P815 by NK cells in a redirected cytotoxicity assay. Preincubation of LAIR-1-transfected K562 cells with 3G4 mildly prevented the binding of LAIR-1 to collagens I and III in a dose-dependent manner. Taken together, the novel MAb 3G4 provides a useful tool to isolate LAIR-1-positive cells without changing their resting state for further application.
    Monoclonal antibodies in immunodiagnosis and immunotherapy. 04/2014;
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    ABSTRACT: Substantial evidence suggests that inflammation is an important contributor to many neurodegenerative disorders. Activated microglial cells play an important role in releasing pro-inflammatory factors, including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) for inducing inflammation. Recently, some reports have suggested that glycoprotein nonmetastatic melanoma B (GPNMB) is highly expressed in microglia after LPS treatment. However, the role of GPNMB in activated microglia is not clearly understood. In this study, we used RT-PCR and Western blotting to detect GPNMB and matrix metalloproteinase-3 (MMP-3) expressions in activated microglia. GPNMB small interfering RNA (siRNA) or MMP-3 inhibitor was applied on microglial BV2 cells, and ELISA was performed to measure the expressions of TNF-α and IL-1β in BV2 cells. Levels of iNOS and NO in BV2 cells were also determined. We found that the levels of GPNMB and MMP-3 were significantly increased in BV2 cells after LPS treatment. Moreover, we found that GPNMB significantly upregulated the expression of MMP-3 in BV2 cells, and high expression of MMP-3 was dependent on the level of GPNMB. Inhibition of GPNMB or MMP-3 expression by GPNMB siRNA or MMP-3 inhibitor dramatically suppressed the expressions of TNF-α, IL-1β, iNOS, and NO in activated microglia. All of these results suggest that GPNMB is involved in the inflammatory responses of microglia.
    Journal of Molecular Neuroscience 03/2014; · 2.89 Impact Factor
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    ABSTRACT: Hepatic steatosis is the accumulation of an excess amount of triglycerides and other fats inside liver cells resulting from abnormal hepatic lipid metabolism. Mitochondrial structural and molecular defects are involved in the progression of hepatic steatosis pathogenesis. Hepatic methylation and transcriptional activity of the mitochondrial-encoded NADH dehydrogenase (MT-ND) play a critical role in the progression of non-alcoholic fatty liver disease (NAFLD). However, the expression of MT-ND3 in hepatic steatosis has not been extensively studied. In this study, liver specimens were collected from different patients, and were subjected to immunohistochemistry. Primary hepatocytes were treated with oxidative stress, hypoxia, and lipotoxicity to investigate the respective roles of these factors on MT-ND3 expression and cell apoptosis by western blotting and flow cytometry, respectively. We found that increased MT-ND3 expression in human hepatic steatosis was positively associated with histological severity of hepatic steatosis. Hypoxia, H2 O2 , and saturated fatty acid treatment induced cell apoptosis mediated by mitochondria. These three factors all had effects on MT-ND3 expression in cultured hepatocytes. Taken together, MT-ND3 may play important roles in hepatic steatosis progress. Hypoxia, oxidative stress, and lipotoxicity could all influence expression of MT-ND3 and thus may play a role in the progression of hepatic steatosis.
    Apmis 09/2013; · 2.07 Impact Factor
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    ABSTRACT: CD306, also known as soluble leukocyte-associated immunoglobulin-like receptor-2 (LAIR-2), is a member of an immunoglobulin superfamily with the shared characteristic of an immunoglobulin-like C2-type domain. CD306 is speculated to be secretory and has 84% similarity with the extracellular domain of CD305, which binds to the same ligands as CD306. However, data on its distribution are absent due to the lack of an efficient method to detect it. In this study, we successfully cloned the cDNA of CD306 from the peripheral blood mononuclear cells (PBMCs) of patients with hemorrhagic fever with renal syndrome. The fusion proteins were expressed and purified, and three strains of monoclonal antibodies (mAbs) against CD306 were prepared and characterized. The sandwich ELISA for detecting CD306 was established and optimized with sensitivity up to 15pg/ml, and the assay showed high specificity for the detection of CD306. With this method, a right skewed frequency distribution of CD306 in the sera of healthy subjects was determined. The concentrations of CD306 in sera and urine were detected in patients with different diseases. Aberrantly high levels of CD306 were found in the sera of pregnant women and patients with inflammation and rheumatic heart disease and in the urine of pregnant women. Meanwhile, there was a positive correlation between CD306 and soluble CD305 expression and secretion levels in sera and urine samples from patients, and both proteins inhibited CD305-mediated immunosuppressive functions. Our results demonstrate that CD306 represents a potentially useful predictor for disease diagnosis and that the method developed has potential for clinical application.
    Journal of immunological methods 08/2013; · 2.35 Impact Factor
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    ABSTRACT: Bone regeneration is a coordinated process involving the connection between blood vessels and bone cells. Glycoprotein non-metastatic melanoma protein B (GPNMB) is known to be vital in bone formation. However, the effect of GPNMB on bone regeneration and the underlying molecular mechanism are still undefined. Fibroblast growth factor receptor (FGFR)-mediating signaling is pivotal in bone formation and angiogenesis. Therefore, we assessed GPNMB function as a communicating molecule between osteoblasts and angiogenesis, and the possible correlation with FGFR-1 signaling. Recombinant GPNMB dose-dependently increased the differentiation of human bone marrow stromal cells (hBMSCs) into osteoblasts, as well as the mRNA levels of osteoblasts marker alkaline phosphatase (ALP) and osteocalcin (OCN). Furthermore, these increases depended on the activation of FGFR-1 signaling, as pretreatment with FGFR-1 siRNA or its inhibitor SU5402 dramatically dampened GPNMB-induced osteogenesis. Additionally, GPNMB triggered dose-dependently the proliferation and migration of human umbilical vein endothelial cells (hUVECs), FGFR-1 phosphorylation, as well as capillary tube and vessels formation in vitro and in vivo. Blocking FGFR-1 signaling dampened GPNMB-induced angiogenic activity. Following construction of a rodent cranial defect model, scaffolds delivering GPNMB resulted in an evident increase in blood vessels and new bone formation; however, combined delivery of GPNMB and SU5402 abated these increase in defect sites. Taken together, these results suggest that GPNMB stimulates bone regeneration by inducing osteogenesis and angiogenesis via regulating FGFR-1 signaling. Consequently, our findings will clarify a new explanation about how GPNMB induces bone repair, and provide a potential target for bone regeneration therapeutics and bone engineering. J. Cell. Biochem. © 2013 Wiley Periodicals, Inc.
    Journal of Cellular Biochemistry 06/2013; · 3.06 Impact Factor
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    ABSTRACT: Colon cancer is a type of malignant tumor that causes considerable mortality worldwide. Epithelial cellular adhesion molecule (EpCAM), a tumor-associated antigen of colon tumors, is a target for colon cancer therapy. EpCAM-specific monoclonal antibodies (mAbs) have been applied in human colon cancer since the 1990 s; however, the therapeutic effects are limited. EpCAM activates nuclear signaling pathways by releasing its intracellular domain (EpICD). The released EpICD stimulates the Wnt/β-catenin signaling pathway, which is also strongly associated with tumorigenesis. EpCAM is also a target gene of the Wnt/β-catenin signaling pathway. EpCAM and the Wnt/β-catenin signaling pathway form a functional interaction cycle in colon cancer. Thus, we propose a new therapeutic drug for colon cancer: an EpCAM single-chain fragment variable antibody (scFv)-truncated protamine (tp)-siRNA. EpCAM scFv can recognize and bind colon cancer cells through its EpCAM antigen activity. Furthermore, the specific siRNA transferred into colon cancer cells specifically inhibits Wnt/β-catenin signal transmission. Therefore, this new drug may efficiently interrupt the functional cycle between EpCAM and Wnt/β-catenin signaling and be an effective therapeutic strategy for colon cancer.
    Cell Biology International 04/2013; · 1.64 Impact Factor
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    ABSTRACT: Objective To detect the distribution of autoantibodies against premelanosome protein 17 (Pmel 17) in the sera of vitiligo patients with the recombinant Pmel 17 protein. Methods The cDNA encoding human Pmel 17 was amplified by RT-PCR from the primary melanocytes and was cloned into vector pMD19-T and subsequently the expression vector pGEX-4T-1. After being induced by IPTG, the recombinant Pmel 17 protein was purified by high-performance affinity chromatography. The presence of autoantibodies against Pmel 17 in the sera of vitiligo patients was determined by indirect ELISA coated with recombinant Pmel 17 protein. Results The recombinant Pmel 17 plasmids were right as we expected by DNA sequencing. The recombinant Pmel 17 protein was successfully expressed and purified. The indirect ELISA revealed that, as compared with healthy control group (0/100, 0%), positive rate of the serum autoantibodies in the progress vitiligo patients was 22% and positive rate of stable vitiligo patients was only 2%(2/100). The difference of positive rate between patients with progress and stable vitiligo was statistically significant. Conclusion The recombinant Pmel 17 protein was successfully expressed. The autoantibodies against Pmel 17 is closely related to the severity of vitiligo.
    Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 04/2013; 29(3):411-3.
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    ABSTRACT: Subarachnoid hemorrhage (SAH) is a frequent occurrence in cerebrovascular accidents, and inflammation occurs in the subarachnoid space after SAH. Arachnoid cells have the capability to present antigens and active T-lymphocytes after stimulation by cerebrospinal fluid (CSF). However, the effect of CSF on T-lymphocytes and arachnoid cell adhesion was not clearly understood. In this study, we used ELISA to detected tumor necrosis factor-α (TNF-α) content in CSF of SAH patients. CSF or recombinant TNF-α were applied on arachnoid cells and T-lymphoctes, and RT-PCR and western blotting were performed to determine the expression of intercellular adhesion molecule-1 (ICAM-1) in arachnoid cells and Lymphocyte Function-Associated Antigen-1 (LFA-1) in T-lymphocytes, respectively. Meanwhile, the Matrix Metal Proteinase-9 (MMP-9) expression in these cells was also determined. We found that the content of TNF-α in the CSF was significantly increased in the CSF of SAH patients (from 22±8pg/mL of healthy people to 436-450pg/mL of SAH patients). Treatement with CSF could increase the expression of ICAM-1in arachnoid cells and that of LFA-1 in T-lymphocytes, mainly through the increased levels of TNF-α. We also found that the co-culture of arachnoid cells and T-lymphocytes increased the expression of MMP-9 in both cells through the interaction of ICAM-1 of and LFA-1. All of these results suggested that arachnoid cells are involved in the T-lymphocytes invasion in the subarachnoid space after SAH.
    Brain research 03/2013; · 2.46 Impact Factor
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    ABSTRACT: The development of bone tissue engineering has provided new solutions for bone defects. However, the cell-scaffold-based approaches currently in use have several limitations, including low cell seeding rates and poor bone formation capacity. In the present study, we developed a novel strategy to engineer bone grafts using mesenchymal stem cell sheets and coral particles. Rabbit bone marrow mesenchymal stem cells were continuously cultured to form a cell sheet with osteogenic potential and coral particles were integrated into the sheet. The composite sheet was then wrapped around a cylindrical mandrel to fabricate a tubular construct. The resultant tubular construct was cultured in a spinner-flask bioreactor and subsequently implanted into a subcutaneous pocket in a nude mouse for assessment of its histological characteristics, radiological density and mechanical property. A similar construct assembled from a cell sheet alone acted as a control. In vitro observations demonstrated that the composite construct maintained its tubular shape, and exhibited higher radiological density, compressive strength and greater extracellular matrix deposition than did the control construct. In vivo experiments further revealed that new bone formed ectopically on the composite constructs, so that the 8-week explants of the composite sheets displayed radiological density similar to that of native bone. These results indicate that the strategy of using a combination of a cell sheet and coral particles has great potential for bone tissue engineering and repairing bone defects.
    Biochemical and Biophysical Research Communications 03/2013; · 2.41 Impact Factor
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    ABSTRACT: Parthenogenetic embryonic stem cells have pluripotent differentiation potentials, akin to fertilized embryo-derived embryonic stem cells. The aim of this study was to compare the neuronal differentiation potential of parthenogenetic and fertilized embryo-derived embryonic stem cells. Before differentiation, karyotype analysis was performed, with normal karyotypes detected in both parthenogenetic and fertilized embryo-derived embryonic stem cells. Sex chromosomes were identified as XX. Immunocytochemistry and quantitative real-time PCR detected high expression of the pluripotent gene, Oct4, at both the mRNA and protein levels, indicating pluripotent differentiation potential of the two embryonic stem cell subtypes. Embryonic stem cells were induced with retinoic acid to form embryoid bodies, and then dispersed into single cells. Single cells were differentiated in N2 differentiation medium for 9 days. Immunocytochemistry showed parthenogenetic and fertilized embryo-derived embryonic stem cells both express the neuronal cell markers nestin, βIII-tubulin and myelin basic protein. Quantitative real-time PCR found expression of neurogenesis related genes (Sox-1, Nestin, GABA, Pax6, Zic5 and Pitx1) in both types of embryonic stem cells, and Oct4 expression was significantly decreased. Nestin and Pax6 expression in parthenogenetic embryonic stem cells was significantly higher than that in fertilized embryo-derived embryonic stem cells. Thus, our experimental findings indicate that parthenogenetic embryonic stem cells have stronger neuronal differentiation potential than fertilized embryo-derived embryonic stem cells.
    Neural Regeneration Research 02/2013; 8(4):293-300. · 0.14 Impact Factor
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    Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas / Sociedade Brasileira de Biofisica ... [et al.] 10/2012; · 1.08 Impact Factor
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    ABSTRACT: The transcription factors (Oct4, Sox2, c-Myc, and Klf4) play an important role in the generation of induced pluripotent stem cells. These factors are expressed in metaphase II oocytes and embryonic stem cells (ESCs). The mechanisms responsible for the reprogramming of ooplasm during nuclear transfer are expected to be associated with the four factors. Here, we show that different paternal genetic backgrounds are able to influence the in vitro development of parthenogenetic and cloned embryos. Using real- time polymerase chain reaction (PCR) we found that the expression level of Oct4 in oocytes was less than that of ESCs, whereas oocytes from KM x C3H females showed the highest expression level of Sox2 than the other strains tested or in G1 ESCs. c-Myc mRNA levels in oocytes from KM mice were greater than those found in ESCs or oocytes of KM x C3H mice. These data demonstrate that the expression of the four transcription factors was different among the oocytes, which may be a contributing factor for the different efficiencies of parthenogenesis and the development of cloned embryos in vitro.
    Cellular reprogramming. 10/2010; 12(5):565-70.

Publication Stats

8 Citations
18.10 Total Impact Points

Institutions

  • 2013
    • Northwest University
      Western Springs, Illinois, United States
    • Shenzhen Second People's Hospital
      Shen-ch’üan-shih, Zhejiang Sheng, China
    • Shanghai Jiao Tong University
      • School of Medicine
      Shanghai, Shanghai Shi, China