Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, prokaryotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS transformed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentrations in 2 h. PEP-1-P27mt of 1 micromol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological protocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.

Objective To investigate the correlation between single nucleotide polymorphisms (SNP) of gene interleukin-23 receptor (IL-23R) rs1004819, rs1495965, rs1884444, rs2201841,rs6677188, rs7517847, rs7530511, rs10489629, rs10889677 and rs11209026 with susceptibility of inflammatory bowel disease (IBD) in Han population of Jiangsu province in China. Methods The gene polymorphism in 134 healthy volunteers, 135 cases of ulcerative colitis(UC) and 43 cases of Crohn's disease(CD) were detected with SNaPshot. Experimental data were analyzed with SPSS 17.0 software. Results In UC, genotype frequency of CC and CT on rs7530511 was 99.26％ (134/135)and 0.74％(1/135), allele frequency of C and T was 99.63％(269/270)and 0. 37％(1/270). While in normal controls, which were 94.03％(126/134), 5.97％(8/134), 97.01 ％(260/268)and 2.99％(8/268)respectively. Compared genotype frequency of these two group, P value was 0. 040 (OR＝0.118、95％CI:0.014～0.953). Compared allele frequency of these two group, P value was 0. 043 (OR＝0.121、95％CI:0.015～0.973). In wild type and mutation type UC patients, the age distribution was different, more young patients in mutation type while more middle-aged patients in wild type, P value was 0.032 and 0.001 respectively. Most UC patients of rs6677188 AT type were in remission under endoscope (P＝0.032). Conclusion The mutation of IL-23R rs7530511 may be a protective factor of UC. The polymorphism of rs6677188 was associated with the age of patients and the remission under endoscope.

ObjectiveTo investigate the possible association of interleukin-23 receptor(IL-23R) polymorphisms with the susceptibility and phenotype of inflammatory bowel diseases (IBD) in Jiangsu Han population.MethodsWe genotyped 178 IBD patients including 135 patients with ulcerative colitis ( UC),43 patients with Crohn's disease (CD),and 134 headthy controls for rs11805303,rs1343151,rs11465804,rs11209032,rs17375018,rs11465788.ResultsComparing with the controls (50.4％ ),there was a significant increase in the carriage of the T allele of rs11805303 in UC (60.4％) ( P =0.020).In genotypephenotype correlation of rs17375018 in UC,clinical severity(UCDAI) was associated with the prevalence of the G allele showed a trend to mild activity.Genotype polymorphisms of rs17375018A was observed more in younger than 25 in the genotype-phenotype correlation in CD(41.7％ vs 22.0％,P =0.050,OR =2.532,95％ CI 0.988-6.494),while rs11805303 was associated with age at diagnose and disease lesion (P =O.039 and 0.044).The risk of extra intestinal manifestation in rs17375018A allele carriers was lower (23.1％ vs46.7％,P=0.040,OR =2.917,95％CI 1.027-8.283).ConclusionsWe confirmed the susceptibility of rs11805303polymorphisms with UC and first demonstrated the genotype-phenot correlation of rs11805303,rs17375018 with UC,CD in Jiangsu Han population.

Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, prokaryotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS transformed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentrations in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological protocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.