Objective To report the methods and clinical effect of the lateral crural flaps in repairing anterior tibal, dorsal and calcaneal softtissue defects. Methods From August 1999 to December 2004, 18cases of defects were repaired with lateral crural flap, including 15 cases of anterior tibal, dorsal and calcaneal softtissue defects with vascular pedicled island lateral crural flaps and 3 cases of dorsal pedal soft-tissue defects with free vascular lateral crural flaps.〖WTHZ〗Results All flaps survived after operation.Insufficient arterial supply of the flap occurred in 2 cases after operation, the pedicled incision sewing thread was removed and lidocain was injected around vascular pedicle, then the flap ischemia was released. Inadequate venous return and venous hyperemia occurred in 1 case because peroneal vein was injured duringoperation.The flap edge skin was cut and heparin was locally dripped for one week, the flap vascular cycle was resumed. All patients were followed up two months to one year, the flaps were not fat, and the elasticity was good. Conclusion It is safe and reliable to use lateral crural flap to repair anterior tibial, dorsal pedal and calcaneal soft-tissue defects.
Objective To investigate the method and the effectiveness of open pelvic fractures associated with perineal injury. Methods Between August 2000 and July 2010, 16 cases of open pelvic fractures associated with perineal injury weretreated. There were 12 males and 4 females with an average age of 41 years (range, 17-69 years). Injury was caused by traffic accidents in 9 cases, by falling from height in 6 cases, and by crushing in 1 case. The mean time between injury and admission was 8 minutes (range, 5-20 minutes). According to Tile classification, 2 cases were rated as type A, 6 as type B, and 8 as type C. The wound size ranged from 5 cm × 3 cm to 15 cm × 12 cm. The perineal injured location included intraperitoneal rectal injury in 2 cases and extraperitoneal anorectal injury in 14 cases. The average injury severity score (ISS) was 29 (range, 25-48). The main treatments included emergency resuscitation, colostomy, external fixation of fractures, repeated debridement with pulsatile irrigation followed by intravenous antibiotics, and vacuum seal ing drainage (VSD). Results In 5 deaths, 3 cases died of hemorrhagic shock and 2 cases died of multi ple system organ failure within 4 days of admission. The other 11 cases were followed up 6-46 months (mean, 14 months). The X-ray films showed that bone union was achieved after 2-4 months of operation. Infection in varying degree occurred at perineal wounds; second stage healing of wounds was achieved in 10 cases after debridement and VSD treatment, and wound healed in 1 case after gracil is muscle flap repair. No anal incontinence occurred in the patients having anorectal injury during follow-up. Conclusion For patients with perineal injury and open pelvic fractures, the following treatments should be carried out so as to obtain good effectiveness: early anti-shock, protection of important organ function, treatment of complications, late resistance to infection and stabil ity restoration of the pelvic ring, functional repair and reconstruction of rectum and anal canal and urinary tract.
【Abstract】 Objective To explore an effective method to cultivate esophageal mucosa epithel ial cells (EMECs)of canine in vitro, and to observe the biological characteristics of EMECs growing on SIS in order to provide an experimental basis for esophagus tissue engineering. Methods Esophageal tissues were obtained from five healthy dogs aged 2 to 5 weeks under sterile conditions. The primary EMECs were cultivated with defined keratinocyte serum free medium (DKSFM) containing 6% FBS. The morphological characteristics and the growth curve of EMECs of the 2nd generation were observed for 1 to 5 days. The expressions of the EMECs marker (cytokeratin 19, CK-19) were examined by immunocytochemistry. The 2nd generation of EMECs was seeded on SIS and observed by HE staining, immunohistochemical staining, and SEM for 4 and 8 days. Results The primary culture of canine EMECs arranged l ike slabstone. Immunohistochemical staining of CK-19 of the2nd generation EMECs showed positive broadly. The cells growth reached the peak level at 2 days by MTT method. E MECs werepolygon in shape and arranged l ike slabstone, and formed a single layer on the surface of SIS. The cells were contact ed closely with each other for 4 days. Eight days later, 2 to 3 layers stratified structure was formed. Lots of EMECs were grown on SIS, andshowed laminate arrangement. Conclusion With mixed enzymatic digestion, the culture of EMECs in DKSFM containing 6 %FBS is a simple and feasible method. SIS shows good biocompatibil ity and can be used as a good scaffold material in th e tissue engineered esophagus.
Objective To investigate the feasibil ity of inducing canine BMSCs to differentiate into epithel ial cells in vitro with epithel ial cell conditioned medium (ECCM). Methods Five mL BMSCs were obtained from il iac spine of a healthy adult male canine with weighing 10 kg, and then isolated and cultured. The oral mucosa was harvested and cut into 4 mm × 4 mm after the submucosa tissue was el iminated; ECCM was prepared. BMSCs of the 2nd passage were cultured and divided into two groups, cultured in ECCM as experimental group and in L-DMEM as control group. The cell morphological characteristics were observed and the cell growth curves of two groups were drawn by the continual cell counting. The cells were identified by immunohistochemical staining through detecting cytokeratin 19 (CK-19) and anti-cytokeratin AE1/AE3 on the21st day of induction. The ultra-structure characteristics were observed under transmission electron microscope. Results The cells of two groups showed long-fusiform in shape and distributed uniformly under inverted phase contrast microscope. The cell growth curves of two groups presented S type. The cell growth curve of the experimental group was right shifted, showing cell prol iferation inhibition in ECCM. The result of immunohistochemical staining for CK-19 and anti-cytokeratin AE1/AE3 was positive in the experimental group, confirming the epithel ial phenotype of the cells; while the result was negative in the control group. The cells were characterized by tight junction under transmission electron microscope. Conclusion The canine ECCM can induce allogenic BMSCs to differentiate into epithel ial cells in vitro.