Renaturation of Lysozyme by size exclusion chromatography(SEC) to improve yield as well as the initial and final protein concentration has been studied in detail, Although urea decreases the rate of proteins refolding, it can suppress protein aggregation to sustain pathway of correct refolding at high protein concentration, and there existed an optimum urea concentration in renaturation buffer. Lysozyme was successfully refolded from initial protein concentration of up to 100mg/m1 by SEC, the yield was more than 40%. And the refolding of Interferon-${\gamma}$ was further investigated.

PKB activation may contribute to resistance to antiproliferative signals and breast cancer progression in part by impairing nuclear import and action of p27. PKB transfection caused cytoplasmic p27 accumulation and cytokine resistance. The nuclear localization region of p27 contains a PKB/Akt consensus site at threonine 157 and p27 phosphorylation by PKB impaired its nuclear import in vitro. PKB/Akt phosphorylated wild type p27 but not p27T157A. PKB activation led to cytoplasmic mislocalization of p27WT but p27T157A remained nuclear. In PKB activated cells, p27WT failed to cause Gl arrest, while the antiproliferative effect of p27T157A was not impaired. Cytoplasmic p27 was seen in 41% (52/128) of primary human breast cancers in association with PKB activation. Thus, we show a novel mechanism whereby PKB impairs p27 function that is associated with an aggressive phenotype in human breast cancer.