Building and identification of prion gene eukaryotic expression vector of human / 重庆医学

Objective To constructan eukaryotic expression recombinant plasmid named pEGFP-N2-PRNP .Methods Total RNA was extracted from alzheimer (AD) disease peripheral blood ,and the PRNP gene was amplified by reverse transcription-poly-merase chain reaction(RT-PCR) .By using gene recombination technique ,human PRNP cDNA was inserted into retroviral vector pEGFP-N2 .The recombinant plasmid was identified by a pair of specified primers containing the restriction sites of Xho Ⅰ and EcoRⅠ .Results The PRNP gene could be obtained by RT-PCR ,the recombinant plasmid was identified by restriction endonucle-ase analysis ,PCR and sequence analysis ,and the expression vector pEGFP-N2-PRNP ,which could be stably expressed in SH-SY5Y cells .Conclusion The recombinant plasmid pEGFP-N2-PRNP is constructed successfully ,Which offers a basic for the further re-search on PRNP biological fuction .

Complications after femoral shaft fracture treated with intramedullary internocking nails and its appropriate managements / 中国基层医药

Objective To investigate the complications and appropriate managements. Methods 109 cases were treated with intramedullary interlocking nails. 81 cases were simple femoral shaft fratures and 28 cases were combined with subtrochanteric fractures. Results Distal nailing failure in 18 patients(2 distal nails unlocked in 1 cases,1 nail unlocked in 15 cases,nails were loosened and exited in 1 case,1 nail breakage in 1 case). Split of fracture ended in 3 cases. Delayed union in 16 cases,nonunion in 1 case. Flexion restriction of knee joint in 20 cases. Broken of main nail in 2 cases. Conclusion Using the intramedtllary interlocking nails to treat femoral shaft fractures had deftnite effect. To avoid the complications, choosing suitable nails preoperative, standard operation, timely and scientific rehabilitation postoperation are necessary.