【Abstract】Objective To provide experimental evidence for gene therapy of thrombophilia diseases by constructing eukaryotic expression plasmid of human thrombomodulin(hTM) gene and transfecting the plasmid into COS7 cell and human umbilical vein endothelial cells(HUVECs). Methods The coding fragment of hTM gene was amplified by PCR. Both hTM gene and pcDNA3.1(+)/neo empty vector were digested with HindⅢ and EcoRⅠ. Two digested fragments were combined into pcDNA3.1/hTM with T4DNA ligase. After identification, the pcDNA3.1/hTM was transfected into COS7 cell and HUVECs using cation liposome. The expression of hTM mRNA and protein on the COS7 cell and HUVECs was detected by RTPCR and immunohistochemistry respectively. Results The hTM recombinant plasmid was confirmed by double endonuclease digesting and sequencing. It was transfected into COS7 cell and HUVECs successfully with liposome.Conclusion The pcDNA3.1/hTM plasmid can be successfully constructed and highlevel hTM can be expressed in eukaryotic cells. All of this provides us experimental evidence for gene therapy and further study of TM anticoagulant mechanism.
Objective To investigate the appl ication and cl inical result of flap in the repair of wounds with Achilles tendon exposure. Methods Between May 2006 and May 2010, 21 patients with Achilles tendon skin defects were treated with microsurgical reconstruction. There were 15 males and 6 females, aged 7-63 years with a median of 34 years. The defect causesincluded sport injury in 4 cases, wheel twist injury in 7 cases, crush injury in 5 cases, chronic ulcer in 3 cases, and Achilles tendon lengthening in 2 cases. The areas of wounds with Achilles tendon exposure ranged from 2 cm × 2 cm to 10 cm × 8 cm. After debridement, wounds were repaired with the medial malleolus fasciocutaneous flap (5 cases), sural neurocutaneous vascular flap (8 cases), foot lateral flap (2 cases), foot medial flap (2 cases), and peroneal artery perforator flap (4 cases). The size of the flaps ranged from 3 cm × 3 cm to 12 cm × 10 cm. The donor sites were either sutured directly or covered with intermediate spl it thickness skin grafts. The Achilles tendon rupture was sutured directly (2 cases) or reconstructed by the way of Abraham (2 cases). Results All flaps survived and wounds healed by first intention except 2 flaps with edge necrosis. Twenty-one patients were followed up 6-18 months (mean, 12 months). The flaps had good appearance and texture without abrasion or ulceration. The walking pattern was normal, and the two point discrimination was 10-20 mm with an average of 14 mm. The Ameritan Orthopedic Foot and Ankle Society (AOFAS) Ankle-Hindfoot Scale assessment revealed that 10 patients had an excellent result, 7 had a good result, 3 had a fair result, and 1 had a poor result with an excellent and good rate of 81.0%. Fourteen cases could l ift the heels with power; 5 cases could l ift the heels without power sl ightly; and 2 cases could not l ift the heels. Conclusion The wounds with Achilles tendon exposure should be repaired as soon as possible by appropriate flap according to the condition of wound.
Objective To investigate the operative procedures and cl inical outcomes of the modified superficial peroneal neuro-fasciocutaneous flap in repairing soft tissue defect of forefoot. Methods From May 2006 to May 2009, 5 male patients (aged 40-63 years) with soft tissue defect of forefoot were treated with the modified superficial peroneal eurofasciocutaneous flap. Tendons and bones were exposed in all cases. Defect was caused by object crash (4 cases) and traffic accident (1 case). The sizes of soft tissue defects of forefoot were 4 cm × 2 cm-8 cm × 4 cm. Rotating point of the modified superficial peroneal neuro-fasciocutaneous flap pedicled with the peripheral vessels network of ankle joint was at the level of tibiotalar joint. The flaps ranging from 5 cm × 4 cm to 10 cm × 6 cm were adopted to repair soft tissue defects of forefoot. The donor sites were either sutured directly or covered with intermediate spl it thickness skin grafts. Results All flaps survived and all wounds healed by first intention. Skin graft at donor site survived completely in all cases. All patients were followed up 6-18 months (mean 11 months). The appearance, texture, and function of the flap were satisfactory. There was a protective sensibil ity in all flaps without abrasion or ulceration, and the two-point discrimination of the flaps was 10-13 mm. The walking pattern was normal. No obvious discomfort was observed at the skin-graft donor sites. Conclusion With rel iable blood supply, no sacrifice of vascular trunks, favorable texture, and thickness, the modified superficial peroneal neuro-fasciocutaneous flap pedicled with the peripheral vessels network of ankle joint is useful to repair skin soft tissue defect of the forefoot.