Fan Yang

Shenzhen Second People's Hospital, Shen-ch’üan-shih, Zhejiang Sheng, China

Are you Fan Yang?

Claim your profile

Publications (4)0 Total impact

  • Xiao-lin Qin · Chao-qi Liu · Fan Yang · Bai-rui Lu · Bin Li
    [Show abstract] [Hide abstract]
    ABSTRACT: To prepare mouse PDL-1 membrane extracellular region (mPDL-1) and its antibody for further study of the biological activity of PDL-1. The extracellular region gene fragment of PDL-1 was amplified by RT-PCR and then was cloned into pET28a(+) prokaryotic expressing vector. The recombinant protein mPDL-1 was induced by IPTG in E.coli BL21 (DE3) and the expressed protein was detected by Western blot. The purified protein was used to immune rabbits to prepare polyclonal antibody and the specificity and the titer of the antibody were detected with ELISA, immunofluorescence assay and FCM. The plasmid pET28a(+)/mPDL-1 was successfully constructed and mPDL-1 protein was expressed in E.coli BL21(DE3) with high efficiency. Western blot showed that the recombinant protein was characterized with His antibody. Rabbit immunized with the purified protein produced high titer of antibody. Immunofluorescence assay displayed that the PDL-1 protein highly expressed in B16 melanoma cells was specifically combined with the antibody. Recombinant mPDL-1 is expressed and purified with high antigenicity. The preparation of recombinant mPDL-1 and its polyclonal antibody lay the foundation for further research on mPDL-1 bioactivities.
    No preview · Article · Jun 2010 · Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
  • [Show abstract] [Hide abstract]
    ABSTRACT: Objective: To establish a molecule binding model of mouse PD-1/PD-L1 protein in vitro, so as to lay a foundation for studying the biological activities of the recombinant protein and to establish a high throughput drug screening model. Methods: Prokaryotic expression plasmid pET28 a (+) /mPDL-1 and pGEX-4T-1/mPD-1 were transformed to E. coli BL21 (DE3), which was then induced with IPTG. The expression products were purified by fast affinity chromatography with FPLC Protein Purification Instrument or gel separation. ELISA and GST pull-down were used to detect the interaction between mPD-1 and mPD-L1. In addition, Alamar blue was used to test the role of protein mixtures in the mixed lymphocyte proliferation. Results: SDS-PAGE and Western blotting analysis showed that the recombinant proteins were successfully expressed. Then purified mPD-1 and mPD-L1 protein was obtained by affinity chromatography, Gel seperation and refolding, etc. ELISA and GST pull-down showed that mPD-1 and mPD-L1 protein had a specific binding activity in vitro. The mPD-1/ mPD-L1 protein had a significantly decreased influence on the proliferation of lymphocytes(P<0. 05). Conclusion: Mouse PD-1/PD-L1 recombinant protein model has been successfully established using purified mPD-1 and mPD-L1 protein expressed prokaryotically, which lays a foundation for high-throughput drug screening.
    No preview · Article · Apr 2010 · Academic Journal of Second Military Medical University
  • [Show abstract] [Hide abstract]
    ABSTRACT: To explore the effect of HIV-1 Nef on the ICAM-1 expression of endothelial cells ECV-304 and its signaling pathways. ICAM-1 expression on the endothelial cells ECV-304 was detected by Western blot and FCM assay. Kinase inhibitor PD98059 was used in the cells to analyze the ERK signaling pathway. Western blot showed that ICAM-1 and p-ERK protein expression increased in Nef expressed ECV304 cells (ECV304-Nef) and FCM results showed that the percentage of ICAM-1 positive cells in ECV304-Nef and control cells was (35.3+/-2.2)% and (12.5+/-0.8)% respectively (P>0.01). p-ERK inhibitor PD98059 almost completely blocked the Nef up-regulation of the p-ERK and ICAM-1. When p-ERK inhibitor was added, the percentage of ICAM-1 positive cells in ECV304-Nef (11.4+/-1.1)% was reduced to the level of the control cells (10.4+/-1.5)% (P>0.05). Erk/Mapk signaling pathway may contribute to the over-expression of adhesion molecules ICAM-1 gene in HIV-1 Nef positive cells. These findings may provide the basis for further research on the mechanism and treatment of HIV-1 infection.
    No preview · Article · Jan 2010 · Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the effects of Chaenomeles speciosa broth on immunoregulation for anti-tumor chemotherapy. Immunosuppressive model was induced by cyclophosphamide (CTX) in mice. The mice were treated with the broth for 15 days. The serum hemolysin was observed in mouse sera. Spleen lymphocyte transformation and gene transcription related to the immunoregulation in spleen lymphocytes were detected. After administrated the broth, the serum hemolysin and lymphocyte transformation rates significantly increased and the mRNA expression of foxp3, TGF-beta, PD1, Fas, Bax were downregulated compared with CTX-group. Chaenomeles speciosa broth has protective effects on the immunosuppressive mouse induce by CTX.
    No preview · Article · Sep 2009 · Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials