Fan Yang

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

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Publications (4)0 Total impact

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    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.
    Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 06/2010; 26(6):569-71.
  • Academic Journal of Second Military Medical University 04/2010; 30(4):385-389. DOI:10.3724/SP.J.1008.2010.00385
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    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.
    Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 01/2010; 26(1):44-6.
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    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.
    Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials 09/2009; 32(9):1418-21.