Xiao W.,Second Clinical Medical College |
Xiao W.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
Wu K.,Second Clinical Medical College |
Wu K.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
And 12 more authors.
Journal of Immunotherapy | Year: 2015
Wogonin exerts effective antitumor activities through direct cytotoxicity against cancer cells and indirect immune modulation. However, the molecular mechanisms of these activities remain poorly understood and need further study. We found that wogonin could efficiently downregulate the expression of B7H1, retinoic acid early induced transcript-1ε (RAE-1ε), and vascular endothelial growth factor in gastric cancer cells. Wogonin also promoted the secretion of calreticulin and high-mobility group protein 1 by tumor cells. Apoptotic bodies from dying tumor cells treated with wogonin were susceptible for uptake by neighboring dendritic cells (DCs). With the xenograft tumor model, wogonin inhibited tumor growth and promoted the recruitment of DC, T, and NK cells into tumor tissues. Infiltrated frequencies of DC, T, and NK cells in tumors were inversely correlated with expression levels of vascular endothelial growth factor, B7H1, and RAE-1ε of tumor tissues. Wogonin directly inhibited the activation of STAT3 on tyrosine 705 in tumor cells. The dephosphorylation of STAT3 contributed to the decreased expression of B7H1 and MHC class I chain-related protein A, and the enhancement of calreticulin on the cell membrane. Our study confirmed the immune-enhancing function of wogonin, and indicated that wogonin could be used in collaboration with DC vaccine or activated lymphocytes for tumor therapy. Copyright © 2015 Wolters Kluwer Health, Inc. All rights reserved.
Yan S.,Yangzhou University |
Yan S.,Nanjing Medical University |
Yang B.,Yangzhou University |
Shang C.,First Peoples Hospital of Lianyungang |
And 9 more authors.
Molecular Medicine Reports | Year: 2016
Platelet-rich plasma (PRP) is blood plasma that has been enriched with platelets, and the number of platelets is correlated with rheumatoid activity. PRP is a concentrated source of autologous platelets, and contains several different growth factors and cytokines, including platelet-derived growth factor, transforming growth factor-β and insulin-like growth factor-1, which stimulate healing of bone and soft tissue. Rheumatoid arthritis (RA) is characterized by synovial hyperplasia, cell activation, articular inflammation and invasion of the synovium into the adjacent bone and cartilage. The adhesion of fibroblast-like synoviocytes (FLSs) onto the extracellular matrix (ECM), migration and invasion are important for the erosion and destruction of the articular cartilage of patients with RA. The aim of the present study was to investigate the effects of PRP on the adhesion, migration and invasion of RA-FLSs. Scratch and Transwell migration assays determined that PRP at a concentration of 2 and 5% significantly enhanced the migration ability of RA-FLSs. Treatment of RA-FLSs with 2 and 5% PRP promoted the adhesion and invasion of the cells. Additionally, the immunofluorescence assay revealed that PRP induced a decrease in the number of centrally located stress fibers and led to an increase in the formation of filopodia and lamellipodia in the detectable leading edge protrusions in RA-FLSs. In addition, reverse transcription-quantitative polymerase chain reaction and western blot analysis determined that PRP upregulated the protein and mRNA expression levels of matrix metalloproteinase-1 (MMP-1). In conclusion, the promotion of RA-FLS cell migration, invasion and adhesion on the ECM by PRP may be modulated through the upregulation of MMP-1 expression and the induction of actin cytoskeletal reorganization.
Qian L.,Yangzhou University |
Qian L.,Jiangsu Electric Power Company |
Qian L.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
Chen W.,Yangzhou University |
And 7 more authors.
Microbiology and Immunology | Year: 2015
Because inappropriate activation of Toll-like receptor9 (TLR9) may induce pathological damage, negative regulation of the TLR9-triggered immune response has attracted considerable attention. Nonpathogenic immune complex (IC) has been demonstrated to have beneficial therapeutic effects in some kinds of autoimmune diseases. However, the role of IC in the regulation of TLR9-triggered immune responses and the underlying mechanisms remain unclear. In this study, it was demonstrated that IC stimulation of B cells not only suppresses CpG-oligodeoxynucleotide (CpG-ODN)-induced pro-inflammatory IL-6 and IgM κ production, but also attenuates CD40 and CD80 expression. Furthermore, our results suggest that the receptor for the Fc portion of IgG (FcγR) IIb is involved in the suppressive effect of IC on TLR9-mediated CD40, CD80 and IL-6 expression. Finally, it was found that IC down-regulates TLR9 expression in CpG-ODN activated B cells. Our results provide an outline of a new pathway for the negative regulation of TLR9-triggered immune responses in B cells via FcγRIIb. A new mechanistic explanation of the therapeutic effect of nonpathogenic IC on inflammatory and autoimmune diseases is also provided. © 2015 The Societies and Wiley Publishing Asia Pty Ltd.
Li Q.,Yangzhou University |
Li X.,Yangzhou University |
Wang C.,Yangzhou University |
Wang C.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
Wang C.,Jiangsu Electric Power Company
Neoplasma | Year: 2016
Ultraconserved regions (UCRs) are non-protein coding gene sequences with strict conserved across among different species. Emerging evidence demonstrates that UCRs encoding noncoding RNAs (ncRNAs) serve as regulators of gene expression. In recent decades, increasing evidence implicates the involvement of UCRs in carcinogenesis. Previous studies showed RNA expression of uc.206 was increased in colorectal cancer. Until now, the role of uc.206 in cervical cancers remains undefined. This study revealed that uc.206 is significantly up-regulated in cervical cancer (CC) tissue and negatively correlates with the expression of the pro-apoptotic gene P53 in RNA level. We show that uc.206 specifically targets the 3’ untranslated region (3’UTR) of P53 and regulates its expression. Inhibition of uc.206 effectively delays cervical cells proliferation and promotes apoptosis, accompanied by increased expression of P53 protein. Thus, these findings suggested that uc.206 acts as a novel oncogene by targeting the P53 gene and promoting CC cell growth, which might be beneficial for cervical cancer therapy. © 2016, Cancer Research Institute Slovak Acad. of Sciences. All rights reserved.
Shen L.,Yangzhou University |
Shen L.,Jiangsu Electric Power Company |
Zhu L.,Yangzhou University |
Luo Q.,Yangzhou University |
And 5 more authors.
Chinese Journal of Natural Medicines | Year: 2015
The present study was designed to isolate and purify chemical constituents from solid culture of endophyte Aspergillus terreus LQ, using silica gel column chromatography, gel filtration with Sephadex LH-20, and HPLC. Fumigaclavine I (1), a new alkaloid, was obtained, along with seven known compounds, including fumigaclavine C (2), rhizoctonic acid (3), monomethylsulochrin (4), chaetominine (5), spirotryprostatin A (6), asperfumoid (7), and lumichrome (8). The structure of compound 1 was elucidated by various spectroscopic analyses (UV, MS, 1D and 2D NMR). The in vitro cytotoxicity of compound 1 was determined by MTT assay in human hepatocarcinoma cell line SMMC-7721, showing weaker cytotoxicity, compared with cisplatin, a clinically used cancer chemotherapeutic agent. © 2015 China Pharmaceutical University.
Yan C.,Yangzhou University |
Jie L.,Yangzhou University |
Yongqi W.,Yangzhou University |
Weiming X.,Yangzhou University |
And 8 more authors.
Biochemical and Biophysical Research Communications | Year: 2015
Nanoparticles are becoming promising carriers for gene delivery because of their high capacity in gene loading and low cell cytotoxicity. In this study, a chitosan-based nanoparticle encapsulated within a recombinant pcDNA3.1-dsNKG2D-IL-15 plasmid was generated. The fused dsNKG2D-IL-15 gene fragment consisted of double extracellular domains of NKG2D with IL-15 gene at downstream. The average diameter of the gene nanoparticles ranged from 200 nm to 400 nm, with mean zeta potential value of 53.8 ± 6.56 mV. The nanoparticles which were loaded with the dsNKG2D-IL-15 gene were uptaken by tumor cells with low cytotoxicity. Tumor cells pre-transfected by gene nanopartilces stimulated NK and T cells in vitro. Intramuscular injection of gene nanoparticles suppressed tumor growth and prolonged survival of tumor-bearing mice through activation of NK and CD8+ T cells. Thus, chitosan-based nanoparticle delivery of dsNKG2D-IL-15 gene vaccine can be potentially used for tumor therapy. © 2015 Elsevier Inc. All rights reserved.
Wan Q.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
Wan Q.,Jiangsu Electric Power Company |
Wan Q.,Yangzhou University |
Bennett B.C.,University of Virginia |
And 5 more authors.
Proceedings of the National Academy of Sciences of the United States of America | Year: 2014
Dihydrofolate reductase (DHFR) catalyzes the NADPH-dependent reduction of dihydrofolate (DHF) to tetrahydrofolate (THF). An important step in the mechanism involves proton donation to the N5 atom of DHF. The inability to determine the protonation states of active site residues and substrate has led to a lack of consensus regarding the catalytic mechanism involved. To resolve this ambiguity, we conducted neutron and ultrahigh-resolution X-ray crystallographic studies of the pseudo-Michaelis ternary complex of Escherichia coli DHFR with folate and NADP+. The neutron data were collected to 2.0-A˚ resolution using a 3.6-mm3 crystal with the quasi-Laue technique. The structure reveals that the N3 atom of folate is protonated, whereas Asp27 is negatively charged. Previous mechanisms have proposed a keto-to-enol tautomerization of the substrate to facilitate protonation of the N5 atom. The structure supports the existence of the keto tautomer owing to protonation of the N3 atom, suggesting that tautomerization is unnecessary for catalysis. In the 1.05-A˚ resolution X-ray structure of the ternary complex, conformational disorder of the Met20 side chain is coupled to electron density for a partially occupied water within hydrogen-bonding distance of the N5 atom of folate; this suggests direct protonation of substrate by solvent. We propose a catalytic mechanism for DHFR that involves stabilization of the keto tautomer of the substrate, elevation of the pKa value of the N5 atom of DHF by Asp27, and protonation of N5 by water that gains access to the active site through fluctuation of the Met20 side chain even though the Met20 loop is closed. © 2014, National Academy of Sciences. All rights reserved.
Li G.,Yangzhou University |
Li G.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
Shi W.,U.S. Center for Disease Control and Prevention |
Chen G.,Yangzhou University |
And 9 more authors.
Plasmid | Year: 2014
The authors created a mammary gland-specific expression vector p205C3 based on the beta lactoglobulin gene and an intron of the beta casein gene from Chinese dairy goat. We utilized this vector to express human lysozyme in mice and characterized expression levels. The present work indicated the feasibility that p205C3 could be used to develop big animal (. e.g., dairy goat or cow) mammary gland bioreactors expressing hLYZ. A mammary gland-specific expression vector p205C3 was constructed with the 5'- and 3'-flanking regions of β-lactoglobulin gene and the first intron of β-casein gene of Chinese dairy goat as regulatory sequences. Human lysozyme (hLYZ) cDNA from mammary gland was cloned into p205C3 and the recombinant vector was used to generate transgenic mice by microinjection. Based on the lysoplate assay, four female offspring of one male founder were detected expressing recombinant hLYZ in their milk at the levels of 5-200 mg/l, and the expressed protein had the same molecular weight as that of normal hLYZ. Besides mammary glands, ectopic expressions were also found in the spleens and the small intestines of the transgenic mice. Among the offspring, the female transgenic mice maintained and expressed the transgene stably with a highest expression level of 750 mg/l. Therefore, p205C3 could be used to develop animal mammary gland bioreactors expressing hLYZ. © 2014 Elsevier Inc.
Zhang Y.,Yangzhou University |
Zhang Y.,Jiangsu Key Laboratory of Integrated Traditional Chinese |
Li Y.,Yangzhou University |
Li Y.,Jiangsu Coinnovation Center for Prevention |
And 6 more authors.
Journal of Neurogastroenterology and Motility | Year: 2016
Background/Aims The functional variant (rs56109847) in the 3'-untranslated regions (3'-UTR) of the serotonin receptor 3E (HTR3E) gene is associated with female diarrhea predominant irritable bowel syndrome (IBS-D) in British populations. However, the relationship of the polymorphism both to HTR3E expression in the intestine and to the occurrence of Chinese functional gastrointestinal disorders has yet to be examined. Methods Polymerase chain reaction amplification and restriction fragment length polymorphism analyses were employed to detect polymorphisms among Chinese Han women, particularly 107 patients with IBS-D, 99 patients with functional dyspepsia (FD), 115 patients with mixed IBS and 69 patients with IBS-D + FD. We also assessed microRNA-510 (miR-510) and HTR3E expression in human colonic mucosal tissues with immunohistochemistry and other methods. Dual-luciferase reporter assays were conducted to examine the binding ability of miR-510 and HTR3E 3'-UTR. Results Genotyping data showed the variant rs56109847 was significantly associated with IBS-D, but not with FD, mixed-IBS, or FD + IBS-D. HTR3E was abundantly expressed around the colonic mucosal glands but less expressed in the stroma. miR-510 expression decreased, whereas HTR3E expression increased in the colonic mucosal tissue of patients with IBS-D compared with those in controls. HTR3E expression was significantly higher in patients with the GA genotype than that in patients with the GG genotype. The single-nucleotide polymorphisms disrupted the binding site of miR-510 and significantly upregulated luciferase expression in HEK293 and HT-29 cells. Conclusions The single-nucleotide polymorphisms rs56109847 led to reduced microRNA binding and overexpression of the target gene in intestinal cells, thereby increasing IBS-D risk in the Chinese Han population. The decreased expression of miR-510 might contribute to IBS-D. © 2016 The Korean Society of Neurogastroenterology and Motility.
Wang C.,Yangzhou University |
Wu X.,Yangzhou University |
Shen F.,Yangzhou University |
Li Y.,Yangzhou University |
And 4 more authors.
Development Growth and Differentiation | Year: 2015
Long noncoding RNAs (LncRNAs) are longer than 200 nucleotide noncoding RNAs without apparent functional coding capacity that function as regulators of cell growth and development. In recent years, increasing evidence implicates the involvement of LncRNAs in erythropoiesis. shlnc-EC6 is a LncRNA associated with erythroid differentiation but the mechanism remains undefined. In this study, we found that knockdown of shlnc-EC6 in purified mouse fetal liver erythroid progenitor and hematopoietic stem cells (FLEPHSCs) significantly blocked erythroid enucleation. We also showed that Rac1 was negatively regulated by shlnc-EC6 at the posttranscriptional level via specific binding to sites within the 3'UTR of Rac1 mRNA. Moreover, we found that knockdown of shlnc-EC6 led to upregulation of Rac1, followed by the activation of the downstream protein PIP5K, and subsequently resulted in the inhibition of enucleation in cultured mouse fetal erythroblasts. Thus, our findings suggest that shlnc-EC6 acts as a novel modulator to regulate mouse erythropoiesis via Rac1/PIP5K signaling pathway. © 2015 Japanese Society of Developmental Biologists.