ABSTRACT
Objective:To develop a method for the rapid determination of 15 elements such as Al,Ca,Cr,Mg,Zn,K,B,Sr, Cd,Ba,Pb,As,Sb,Na and Li in ibuprofen injection by ICP-MS. Methods:After diluted by 2% nitric acid solution,the samples was determined directly by inductively coupled plasma mass spectrometry(ICP-MS)with the online internal standard to correct matrix interference and signal drifting. Results:The detection limits of the method were ranged from 0. 031 to 5. 1 μg·L-1 . The average recoveries were within the range of 82. 6% -107% and RSDs were all less than 5% . Conclusion:The method is rapid, accurate and highly sensitive,which can provide technical support for the quality control of ibuprofen injection.
ABSTRACT
Objective By detecting the mutations spectrum of phenylalanine hydroxylase(PAH)gene in phe-nylketonuria(PKU)patients and their parents. The researchers analyzed the gene mutation features and high - frequency mutations and determined the relationship between the genotype and the phenotype,which would provide a theoretical basis for the early diagnosis and genetic consultation of PKU children in the region. Methods In this study,13 exons and their flanking introns of the PAH gene in 32 PKU patients and their parents from Wuxi and Suqian in Jiangsu province were sequenced by using the next - generation sequencing(NGS)technology. Results Sixty - one mutant sites and 32 mutant genes were detected in 32 PKU patients,and the mutation detection rate was 95. 31%(61 / 64 cases). The variants at c. 721C ﹥ T,c. 1068C ﹥ A,c. 611A ﹥ G,c. 1197A ﹥ T,c. 728G ﹥ A,c. 331C ﹥ T and c. 442 -1G ﹥ A were common mutations in the region with mutation frequency over 5% . What's more,4 novel variants of c. 699C ﹥ G,c. 265C ﹥ T,c. 722G ﹥ A and c. 1194A ﹥ G were found. Of those,c. 699C ﹥ G was not recorded in the PAH variant database and HGMD database and. c. 265C ﹥ T,c. 722G ﹥ A,and c. 1194A ﹥ G were first reported in the Chi-nese population. Genotype - accurate biochemical phenotype correlation by using the Guldberg AV system revealed con-sistency rate of 38. 0%(8 / 21 cases),which the consistency rate between accurate biochemical phenotype and predic-tive phenotype of moderate to severe genotype was 92. 3%(12 / 13 cases),and mild genotype was 50. 0%(4 / 8 cases). Conclusions The PAH gene variants of PKU patients in Jiangsu province are distributed mainly in exons 7,of which the highest frequency gene mutation is c. 721 c ﹥ T. Moreover,one novel variant c. 699C ﹥ G was reported for the first time. The PKU children inherit the PAH mutation gene mainly from both parents. There are definite correlation between the genotypes and phenotypes.
ABSTRACT
Objective To observe the regulation of sry-related high-mobility-group box-containing 2 (Sox2) on epidermal growth factor receptor(EGFR)and on the sphere formation rate of glioma stem cells .Methods Promoter reporter plasmids of epidermal growth factor homologous receptor ErbB2 ,ErbB3 ,ErbB4 were respectively constructed .Sox2 expression plasmid was co-transfected together with the reporter plasmid into U251 cells .Then the luciferase activity was analyzed by dual-luciferase reporter assay sys-tem to test the regulation of Sox2 on ErbB2 ,ErbB3 ,ErbB4 promoters .Sphere formation assay was used to observe selfrenew of the cancer stem cells after transfection of Sox 2 .The expression of EGFR and ErbB2 proteins in the spheres was examined by Western blot .Results Sox2 could dose-dependently increase the ErbB2 ,ErbB3 ,ErbB4 promoter drived luciferase activity .Sox2 promotes the sphere-forming rate of U251 cells and upregulates the expression of EGFR and ErbB2 in the spheres .Conclusion Sox2 could up-regulate the expression of EGFR and promote selfrenew of glioma stem cells .
ABSTRACT
Objective To compare the proteome difference between multiple myeloma cell line U266 cells treated and untreated with PS-341, to investigate the potential drug targets, and to provide theoretical evidence for clinical therapy of multiple myeloma. Methods Two-dimensional gel electrophoresis (2-DE) was performed to separate proteins from treated and untreated U266 cells with proteasome inhibitor PS-341. ImageMaster 2D Platinum software was used to analyze 2-DE image, and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) was used to identify the differentially expressed proteins. The expression levels of differential protein BAG-2 in the 2 groups of U266 cells lines were detected by Western blot. Results The 2-DE reference pattern of treated and untreated U266 cells with PS-341 was established. A total of 31 differential proteins were identified by MALDI-TOF-MS, 27 of which were down-regulated after PS-341 treatment. The differential expression level of BAG-2 in the 2 groups of U266 cells was confirmed by Western blot. Conclusion Some down-regulated proteins may be the potential drug targets of proteasome inhibitor PS-341.