1.Isolation and purification of human coagulation factor Ⅶ from Cohn fraction Ⅲ paste
Shizhou XU ; Qingrong ZHAO ; Fangzhao LIN ; Ling XIAO ; Xiaopu XIAO ;
Chinese Journal of Blood Transfusion 2002;0(05):-
Objective To isolate and purify human coagulation factor Ⅶ from Cohn fraction Ⅲ precipitate.Methods The purification procedure of human factor Ⅶ from Cohn fraction Ⅲ precipitate involves dissolving fraction Ⅲ,absorbing factor Ⅶ onto barium citrate and eluting,ammonium sulfate fractionation and DEAE-Sepharose Fast Flow ion exchange chromatography,and Sephadex G-100 gel filtration chromatography.Results 10.1mg purified FⅦ was obtained from 400g Cohn fraction Ⅲ precipitate.The purified FⅦ has a specific clotting activity of 1775.8U/mg and the overall yield of FⅦ specific clotting activity is 17.6% of the starting material.The purity of FⅦ was judged by SDS-PAGE and there was only one protein band on the gel.Conclusion The procedure of purifying Ⅶ from Cohn fraction Ⅲprecipitate is established with satisfactory purity.
2.TNFR-1 on tumor cells contributes to the sensitivity of fibrosarcoma to chemotherapy.
Jingjing DENG ; Xiaopu ZHAO ; Lijie RONG ; Xiao LI ; Xiaoman LIU ; Zhihai QIN
Protein & Cell 2013;4(5):393-401
Impaired tumor necrosis factor receptor-1 (TNFR-1) signaling has been found in some malignant tumors with poor prognosis. However, the exact role of TNFR-1 signaling in fibrosarcoma remains unclear. Here, we explored the question by comparing the growth of TNFR-1 deficient (Tnfr1 (-)) and TNFR-1 competent (Tnfr1 (+)) fibrosarcoma FB61 cells (FB61-m and FB61-R1) in mice. TNFR-1 expression on fibrosarcoma cells delayed their growth in vivo but not in vitro. Moreover, reduced FB61-R1 tumor growth was also obtained in TNFR-1 knockout mice. The mechanism relies mainly on the TNFR-1-mediated downregulation of vascular endothelial growth factor (VEGF) production by tumor cells. Importantly, treatment of FB61-m tumors with melphalan resulted in a short delay of tumor growth, followed by a quick remission. However, when FB61-R1 tumors were treated with melphalan, tumor growth was similarly delayed at first and then completely rejected. Our results reveal evidence for TNFR-1 on tumor cells as a prerequisite in chemotherapy for fibrosarcoma, and provide novel insight into the therapeutic approach against some types of tumors using TNFR-1 angonist.
Animals
;
Down-Regulation
;
drug effects
;
Fibrosarcoma
;
drug therapy
;
genetics
;
pathology
;
Gene Expression Regulation, Neoplastic
;
drug effects
;
Humans
;
Melphalan
;
administration & dosage
;
Mice
;
Molecular Targeted Therapy
;
Receptors, Tumor Necrosis Factor, Type I
;
genetics
;
Signal Transduction
;
drug effects
;
Vascular Endothelial Growth Factor A
;
biosynthesis