Objective To construct human acidic fibroblast growth factor (aFGF) recombinant eu karyotic expression vector and transfect it into muscle satellite cells(MSCs) of rat,in purpose of further study the method to set up cell bank.Methods The aFGF gene was cloned from human total RNA which was obtained from human skeletal muscle tissue by RT-PCR method.Human interleukin 2 (IL-2) signal peptide sequence (SPS) was obtained by direct chemosynthesis method.Then aFGF and SPS were fused to obtain SPS-aFGF.Finally,directional cloning SPS-aFGF into pEGFP-N1,the recombinant (pEGFP-N1-SPS-aFGF) was obtained.The recombinant was confirmed by endonuclease digestion and DNA sequencing.MSCs were purified by difference-speed adherence method and were ideontified by immunofluorescence assay.The correct cells were divided into 3 groups:Experimental group (aFGF +N 1),control group (N 1),blank group (blank).All the groups were transfected by Lipofectamine 2000TM Reagent,and pEGFP-N1-SPS-aFGF,pEGFP-N1 were respectively added in experimental group and control group while blank group was added none plasmid.Fluorescence microscope was employed to detect transfection efficiency tendency along with time changes.The expression of target gene was detected by fluorescent quantitation PCR and Western blot.Results (1) The sequencing of pEGFP-N1-SPS-aFGF was completely correct and the outcome of endonuclease was equal to actual ban s-ize.(2)The expression of GFP in transfected cells were observed by fluorescencemicroscope and transfection efficiency reached the peak at 72 h.(3)Real-time fluorescent quantitation PCR proved strong aFGF mRNA expression in transfected cells (the average relative expression of experimental group was 1464.95)with aFGF gene,while it was detected a little in the other groups (the average relative expression of control group was 1.016 and blank group was 1.000) (P ＜ 0.05).Western blot also proved strong expression in Experimental group then the other two groups.Conclusion aFGF eukaryotic expression vector was successfully constructed and transfected into MSCs.This study may be expected to obtain some specific functions cells.

Objective To compare the clinical effects of dynamic hip screw(DHS),Gamma interlocking intramedullary nail and proximal femoral nail antirotation (PFNA) in the treatment of intertrochanteric femoral fractures in elderly patients.Methods From January 2008 to June 2015,103 elderly patients with intertrochanteric femoral fracture were treated with DHS (DHS group,33 cases),Gamma nails (Gamma group,30 cases),or PFNA (PFNA group,40 cases).By the AO classification,there were 44 cases of type 31-Al,30 cases of type 31-A2 and 29 cases of type 31-A3.The 3 groups were compared in terms of incision length,operation time,intraoperative blood loss,fracture healing time,postoperative weight-bearing time,Harris scoring,and incidence of postoperative complications.Results PFNA group incurred significantly shorter incision length (5.4 ±0.5 cm) and operation time (70.8 ± 16.2 min) than DHS group (12.6 ±2.7 cm and 102.6±17.4min) and Gamma group (7.5±0.8 cmand93.0±35.9 min) (P ＜0.05).The intraoperative blood loss in PFNA group (163.2 ± 60.6 mL) was significantly less than in DHS group (280.5 ±89.8 mL) and in Gamma group (204.9 ±62.2 mL),and that in Gamma group was also significantly less than in DHS group (P ＜ O.05).PFNA group had significantly shorter weight-beating time (11.0 ± 0.8 weeks),fracture healing time(13.6 ± 1.5 weeks) and significantly higher Harris good to excellent rate (92.5％) than DHSgroup (13.3±1.0weeks,15.8 ± 1.2 weeks and 84.8％) and Gamma group (12.5±1.3 weeks,14.2 ± 1.0 weeks and 86.7％) (P ＜ 0.05).The incidence of postoperative complications in DHS group (21.2％)was significantly higher than in Gamma group(10.0％) and in PFNA group (7.5％) (P ＜ 0.05).Conclusions DHS,Gamma nail and PFNA are effective means for the treatment of intertrochanteric femoral fractures in the elderly.Intramedullary fixation,especially by PFNA,shows superiority in the clinical outcomes.