Construction of a lentiviral vector of FoxM1 shRNA and its transfection into human prostate cancer cell lines in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation.</p><p><b>METHODS</b>Three interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay.</p><p><b>RESULTS</b>DNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability.</p><p><b>CONCLUSION</b>We have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.</p>

Repair of finger pulp defect by using arterialized venous dorsal finger flap with sensory nerves / 中华骨科杂志

Objective To explore a new microsurgical method for reconstruction of finger pulp defect.Methods From May 2008 to May 2009,10 male patients with finger pulp defect were treated in our hospital,aged from 18 to 38 years (average,26 years).The index finger was involved in 6 patients,the middle finger in 3 patients and the ring finger in 1 patient.All finger pulp defects were caused by machine injury.The defect sizes ranged from 1.5 cm×2.0 cm to 2.0 cm×3.0 cm.Six patients suffered from single skin defect,and 4 patinets suffered from skin defect combined with nail bed laceration and distal phalangeal fractures.All patients were performed emergency operations.The defects were reconstructed by using arterialized venous flap with microsurgical suture of the dorsal branch of the proper digital nerve.The fractures were fixed by Kirschner wires.The donor area was covered with skin grafts.Results All flaps survived completely.The fractures healed 8 to 10 weeks postoperatively.All patients were followed up for 4 to 6 months,all flaps presented satisfactory appearance,normal texture,with no pigmentation at the last followup.The static 2-point discrimination of the flaps ranged from 8 to 12mm.All injured fingers obtained good recovery of flexion and extension of the distal interphalangeal joints.The nails of the fingers with laceration of nail bed grew smoothly.The nail bed with laceration grew smoothly,and some new nails could be seen.The skin grafts applied to the donor area survived completely.Conclusion The arterialized venous flap with suture of the dorsal branch of the proper digital nerve is a good method for reconstruction of finger pulp defect,which had the following advantages:slight donor injury,low anesthesia risk,simple operative technique,and satisfactory postoperative function and appearance.