ABSTRACT
In order to construct the recombinant retrovirus vector of bovine sox2 gene and obtain infectious retroviral particles, we successfully amplified the ORF (open reading frame) of bovine sox2 gene from the primodial genital ridges of bovine embryo by RT-PCR. The cDNA of ORF was subcloned to pMD18-T vectors and verified that its sequence was highly homologous to the GenBank counterpart (GenBank Accession No. NM-001105463) by sequencing. The correct fragment was digested by EcoR I/Bgl II from recombinant pMD18-T vector and inserted into the same restriction sites f retroviral vector pMSCVneo. We got recombinant retrovirus vector pMSCV-sox2 which was transfected into PT67 by lipofectamine 2000 with pMIG (including green fluorescence protein) as a control. Flow cytometry analysis showed that its transfected efficiency was 68.3%. Subsequently, we established the stable cell strain by G418 selection which could produce virus. Its viral titer was up to 8.16x10(7) CFU/mL. This greatly facilitates the further study of bovine induced pluripotent stem cells induced from bovine somatic cells by defined factors.
Subject(s)
Animals , Cattle , Mice , Cloning, Molecular , Genetic Vectors , Genetics , NIH 3T3 Cells , Open Reading Frames , Genetics , Recombinant Proteins , Genetics , Retroviridae , Genetics , Metabolism , SOXB1 Transcription Factors , Genetics , Stem Cells , Metabolism , TransfectionABSTRACT
Objective To analyze the features of pelvic arteriography in postpartum hemorrhage.Methods The pelvic arteriographic results of postdelivery hemorrhage caused by differeut cause in 10 patients whom were treated with artery embolization were analysed and compared with the clinical data.Results The arteriographic features included irrgeular nodose or diffuse outflow of contrast medium in bilateral upper branches of uterine artery in 5 cases with atonic uterus streaky or cloudy contrast medium in the site of uterine incision in 2 cases after caesarean.In two cases with incomplete laceration in the lower portion of soft puerperant tract,there was distinct outflow of contrast medium in one side or bilateral down branches of uterine artery but its bilateral upper branches was displayed clearly.On angiography in one case with uterine vascular malformation displayed one of upper uterine branch was enlarged and the constrast medium flowed into utrine cavity directly.Conclusion Pelvic arteriography in postpartum hemorrhage with different causes has its special findings.Arteriography in combination with the clinical data can afford more direct and accurate informations for analysing the causes of postpartum hemorrhage causes.