Effect of IL-8 on the Immune Function of Patients with Acute -Lymphoblastic Leukemia and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of interleukin -8 (IL-8) on immune function in acute lymphoblastic leukemia patients and its related mechanisms.@*METHODS@#Forty-five ALL patients were selected from January 2014 to September 2017 in our hospital. Out of them, 32 relieved patients were included in group A, 13 patients did not relieved patients after treatment and were included in the group B. The serum IL-8 level was detected by ELISA.Th1 and Th2 cells were measured by flow cytometry. After Th cells were treated with different concentration of IL-8, the Western blot was used to detect the translation levels of p-STAT3 and JAK in cells.@*RESULTS@#The difference of white blood cell count and clinical risk level between the 2 groups was statistically significant (P0.05).@*CONCLUSION@#IL-8 can interfere the balance of Th1/Th2 through STAT3 signaling pathway, and has effect on the immune function of ALL patients.

Construction of recombinant GST-RCAS1 fusion gene and its expression in E. Coli / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the recombinant plasmid of RCAS1, to express and purify its fusion protein GST-RCAS1, and to investigate its biological function.</p><p><b>METHODS</b>RCAS1 encoding gene was amplified by RT-PCR from total RNA extract of MCF-7 cells and was ligated with expression plasmid vector pGEX-2T by T4 DNA ligase after digested by the restricted endonucleases BamH I and EcoR I. Then the ligated products were inserted into competence JM109 E. Coli and the positive recombinants were identified by restriction endonuclease digestion assay and DNA sequencing. The GST-RCAS1 fusion protein expression was induced by IPTG in BL21 E. Coli and was purified with GST column and identified by SDS-PAGE and Western blotting with anti-GST monoclonal antibody, anti-RCAS1 (N-18) and anti-RCAS1 (C-20) polyclonal antibody. The apoptosis of activated T cells induced by GST-RCAS1 fusion protein was detected by flow cytometry with Annexin V and propidium iodide (PI) staining.</p><p><b>RESULT</b>A 642 bp product was cloned by RT-PCR and the recombinant plasmid was constructed successfully. The GST-RCAS1 fusion protein was recognized by GST monoclonal antibody and RCAS1(N-18 and C-20) polyclonal antibody. FACS analysis showed that GST-RCAS1 fusion protein induced apoptosis in activated T cells.</p><p><b>CONCLUSION</b>The recombinant plasmid of RCAS1 has been successfully constructed and the GST-RCAS1 fusion protein expressed and purified. The apoptosis inducing effect of GST-RCAS1 fusion protein on activated T cells is demonstrated.</p>