Objective To study exogenous expression and subcellular localization of wild type (WT ) and mu-tant PAX3 proteins in vitro by generating their expression plasmids for further study of pathogenesis of Waarden-burg syndrome (WS) .Methods The plasmids pECE-PAX3 and pcDNA3 .0-HA were ligased after they were cut by double enzyme digestion using molecular cloning technique to generate recombinant eukaryotic expression plasmid pcDNA3 .0-PAX3-HA ,which was as a template to generate expression plasmids pcDNA 3 .0 -H80D -HA and pcDNA3 .0-H186fs-HA of novel mutations H80D and H186fs of PAX3 gene .All constructs were verified by di-rect nucleotide sequencing .NIH3T3 cells were transfected transiently with the expression plasmids of PAX3 ,H80D and H186fs respectively .The exogenous expression of WT PAX3 protein and mutant H80D ,H186fs proteins were analysed using Western blot assay ,while their subcellular distribution were observed using immunofluorescence as-say .Results The DNA sequences of expression plasmids of PAX3 and its mutant H80D ,H186fs were correct . Both WT and mutant PAX3 proteins were detected at the expected size .WT PAX3 and H80D proteins were only lo-calized in the nucleus ,whereas H186fs protein showed aberrant localization in both cytoplasm and nucleus .Conclu-sion We successfully generated the recombinant eukaryotic expression plasmids of PAX 3 gene and its mutants and drew preliminary conlusion of gene mutation having effect on subcellular distribution of WT PAX 3 proteins in vitro , which lays experimental basis for further study of the moceluar mechanism of WS caused by PAX3 gene mutations in China .

The inhibitory effect of hydroxyapatite ultrofine powder (HAUFP) on tumor and the effect on the immunity function of body were investigated. The levels of IL-2 in the spleen cells and serum TNF levels in the tumor-bearing mice at the 7th day and 14th after peritoneal injection of HAUFP were detected by using the methods of colorimetric analysis of MTT and crystal purple decoration, respectively. The disappearance of the ascites of the mice was observed. The results showed that the levels of IL-2 and TNF in the tumor-bearing mice were higher obviously in the drug-treated group than in the control group (P<0.01), the ascites growth was inhibited. It was suggested that HAUFP could increase the levels of IL-2 and TNF of the tumor-bearing mice and improve the immune function of body.

The inhibitory effect of hydroxyapatite ultrofine powder (HAUFP) on tumor and the effect on the immunity function of body were investigated. The levels of IL-2 in the spleen cells and serum TNF levels in the tumor-bearing mice at the 7th day and 14th after peritoneal injection of HAUFP were detected by using the methods of colorimetric analysis of MTT and crystal purple decoration, respectively. The disappearance of the ascites of the mice was observed. The results showed that the levels of IL-2 and TNF in the tumor-bearing mice were higher obviously in the drug-treated group than in the control group (P<0.01), the ascites growth was inhibited. It was suggested that HAUFP could increase the levels of IL-2 and TNF of the tumor-bearing mice and improve the immune function of body.