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Ke-yi Wang

Publications and source records attributed to Ke-yi Wang.

3 recordsLinked to original sources

[Identification of peptides binding to Pisum sativum agglutinin from a phage-displayed random peptide library].

OBJECTIVE: To obtain peptides binding specifically to Pisum sativum agglutinin (PSA) from a phage-displayed random peptide library. METHODS: (1) A phage-displayed random hexapeptide library was screened with PSA as target. (2) Dot blot was used to analyze the influence of the alpha-Met-D-mannoside on binding between PSA and phage-displayed peptides. (3) Three peptides (RMWSF, RYDYSY, LRLRQL) were selectively synthesized, and different concentrations were used to inhibit PSA and ConA binding to the HRP. RESULTS: The enrichment occurred obviously after three rounds of screening. The insert sequences of amino acids, displayed on 22 phage DNAs from the third round of screening, were divided into three groups. The binding of phage-displayed peptides to PSA was specific as shown by dot blot and could be inhibited by alpha-Met-D-mannoside. LRLRQL was not dissolved in water. ARMWSF and RYDYSY inhibited binding of PSA to HRP, but failed to inhibit binding ConA to HRP. CONCLUSION: The binding site of peptides ARMWSF and RYDYSY is different to that of alpha-Met-D-mannoside.

Binding Sites↗

[Interaction between polysaccharides and interferon-gamma using an improved ELISA approach].

OBJECTIVE: To establish an ELISA approach to study the interaction of polysaccharides with cytokine in vitro. METHODS: The heparin BSA complexes (HBC) were synthesized with a chemistry method and separated using a 1 X 90 cm column of Separose 4B. After identification of the complex via SDS-PAGE,the wells of ELISA plates were coated with HBC and the interaction of HBC with interferon-gamma (IFN-gamma) was detected. The effects of heparin, low molecular weight heparin (LMW heparin), chondroitin sulfate (CS), hyaluronic acid (HA) and carrageenans on the binding of HBC to IFN-gamma were tested in this system. RESULT: Human recombinant IFN-gamma bound to heparin in a concentration dependent manner, the binding of IFN-gamma to HBC was detected at the concentration of 0.25 ng, and saturated at around 2 ng. Free heparin, LMW heparin, CS,HA and carrageenans competed for the binding of IFN-gamma to HBC with significant different ability. The IC(50)concentrations of heparin and LMW heparin were 2.40 microg/ml and 18.60 microg/ml respectively. CONCLUSION: IFN-gamma is a cytokine with high binding affinity to heparin and carrageenans family but poor to CS-A and CS-C. ELISA is a simple, sensitive approach to detect the interaction of polysaccharides with cytokine in vitro.

Enzyme-Linked Immunosorbent Assay↗

[The expression and significance of costimulatory molecule in peripheral blood B lymphocytes in rheumatoid arthritis].

OBJECTIVE: To research the mechanisms of immune function disorder in the patients with rheumatoid arthritis (RA). METHODS: Monoclonal antibodies against CD(80), CD(86), CD(40) were used for flow cytometry and expression of costimulatory molecule CD(80), CD(86), CD(40) on peripheral blood B lymphocytes was studied in patients with RA, and healthy human were used as control groups. We also used enzyme linked immunosorbent assay to detect the cytokines level of Th1 cell including interleukin (IL)-2, interferon-gamma and those of Th2 cell including IL-6, IL-10. These samples are serum and synovial fluid of patients with RA, respectively. RESULTS: Compared with control group, the expression of CD(86) on peripheral blood B lymphocytes in patients with RA was obviously decreased (P < 0.01), the expression of CD(40) was also significantly increased (P < 0.05). No statistically significant differences in CD(80) positive peripheral blood B lymphocytes between RA group and control group were observed. Comparing with control group, the level of IL-2 and interferon-gamma in serum and synovial fluid of patients with RA are obviously increase (P < 0.01 or P < 0.05). While, the level of IL-6 and IL-10 are decrease (P < 0.01 or P < 0.05). CONCLUSIONS: The disorder expression of costimulatory molecule CD(86), CD(40) on peripheral blood B lymphocytes may be associated closely with the imbalance of Th1/Th2 cytokines level in patients with RA. These will give us a new idea on the therapeutic strategy of RA.

Adult↗