Objective To evaluate the repair of the immense abdominal wall defect after resection of malignant tumours occurred primarily in the abdominal wall or that invaded the abdominal wall secondarily. Methods Among the 20 eases, 12 eases were of malignant tumours within abdominal wall (9 eases of rhabdomyosareoma, 3 eases of malignant fibrous histocytoma), 8 eases with the invasion of abdominal wall from malignant turnouts were of retroperitoneal or intraabdominal malignant tumors (3 eases with retroperitoneal malignant tumours, 1 ease of transverse eolon carcinoma, 3 eases with local reeurrenee in abdominal wall of ascending colon carcinoma undergoing fight hemieoleetomy 2～3 years ago, 1 ease with abdominal wall implantation metastasis of renal carcinoma after resection 5 years ago ). Abdominal wall defects left over by resection were repaired with polypropylene and expanded polytetrafluoroethylene (PPePTFE) mesh. The postoperative eomplieation and healing rate were observed. Results Wounds healed by primary intension, with no subeutaneous hydrops, no incision infeetion, dehiscence and hernia, no intestinal adhesion in all eases. The rate of successful repairing for the abdominal defect was 100%. All patients were followed up, the meshes with no rejection, inflammation and incision hernia, no reeurrenee of tumour in the repaired abdominal wall. Conclusion The PP-ePTFE mesh is a favourable repairing material for the abdominal defect after tumour resection, with the advantages of strong tensility, good bioeompatibility.

BACKGROUND:Subcutaneous fat of human body is a rich reservoir of adipose derived stromal stem cells(ADSCs).ADSCs can proliferate rapidly when being cultured in vitro,and has the capacity of multi-directional differentiation.ADSCs attracted much attention in research of tissue engineered seed cells.OBJECTIVE:To isolate and culture stromal vascular fraction(SVF)cells from rabbit subcutaneous fat in vitro,and to testify whether it has multiple differentiation capacity.METHODS:SVF cells were isolated in vitro from rabbits,and cultured under standard condition.Cellular surface antigens CD44,CD45 and CD29 of passage 3 SVF cells were examined using flow cytometry.Passage 3 SVF calls were induced to differentiate into osteoblasts,chondrocytes,and lipocytes.Oil red staining was used to examine lipocyte induction.Alkaline phosphatase(ALP)staining,alizarin red staining and von Kossa staining were used to examine osteoblast induction.Type Ⅱ collagen immunohistochemical staining and type Ⅱ collagen mRNA RT-PCR were used to examine chondrocyte differentiation.RESULTS AND CONCLUSION:Primary SVF cells were multi-angular or short spindle-shaped.Passage 3 SVF calls were long spindle-shaped.Flow cytometry showed CD44+,CD29+,CD45.Oil red staining exhibited positive reaction in lipocyte induction group.ALP staining,alizarin red staining and Von kossa staining demonstrated positive reactions in osteoblast induction group.Type Ⅱ Collagen immunohistochemical staining and alcian blue staining have suggested positive reactions at 14 days of chondrogenic induction group.RT-PCR of type Ⅱ collagen mRNA test showed that the product band had strong signal at 14 days of chondrogenic induction group compared with that before induction.Above mentioned results have indicated that SVF cells isolated from rabbit subcutaneous fat have identical surface makers of stem cells,and have the ability to differentiate into lipocytes,ostsoblasts and chondrocytes in vitro by induction,and it could be concluded that the SVF cells were ADSCs.

BACKGROUND: Small intestinal submucosa (SIS) has good compatibility with cells and tissues, and has good degradabUity. It is an ideal scaffold for tissue engineering. Inducing adipose derived mesenchymal stem cells (ADSCs) seeded on SIS can construct target tissues, which has the potential to be used in clinical treatment.OBJECTIVE: To prepare decellularized porcina SIS matrix, and testify its biocompatibility with rabbit ADSCs cultured in vitro. METHODS: SIS was processed by enzyme digestion-hypertonic saline decellularization, lyophilized at low temperature, and sterilized by gamma radiation. Paraffin sections were used to observe the effect of decellularization of SIS, and the surface structures of SIS were observed by scanning electron microscope (SEM). Rabbit ADSCs were isolated and cultured, and passage 3 ADSCs were seeded onto one side or both sides of SIS. After one weak of co-culture, the cell-scaffold composites were observed.RESULTS AND CONCLUSION: SIS was white and semi-transparent film. Paraffin sections showed no cells on SIS matrix; electron microscopy showed loose weave structure of serosal surface and dense packing structure of mucosal surface. After one week of co-cultivation, plenty of ADSCs were observed on the surface of SIS. In ADSCs seeded onto one side of SIS group, a large number of cells grew on the superior surface, and few even no cells were observed on inferior surface of SIS. When ADSCs were seeded onto both sides of SIS, cells adhered to SIS in paraffin sections. Results show that enzyme digestion-hypartonic saline decellulariation can decellularize SIS completely, and SIS can support ADSCs growth.

Acetabulum fracture, the inner fracture of the joint, has be en divided into 10 kinds. Because acetabulum is an irregular body, no single ope ration approach can solve all kinds of acetabular fracture. Appropriate selectio n of the operative approach according to the type of fracture is very important. Kocher-Langenbeck approach, extended-iliofemoral approach and ilioinguinal ap proach are often used in clinic now. Because of its complicated procedures and m any complications resulting from the approach, the application of extended-ilio femoral approach is decreasing. Because of its predominant advantage and improve ment of the operation techniques, the use of ilioinguinal approach is obviously increasing in proportion. Since the united approach can reveal clearly so that i t facilitates reduction and internal fixation, it is recommended by the surgeons at home and abroad.

ObjectiveTo study the possibility of the liver histioid construction through co-culturing hepatocytes and sinusoidal vessel endothelial cells (SVECs) on hyaluronic acid(HA) scaffold.MethodsHepatocytes and SVECs of a mouse were isolated respectively, and then were cultured successively in HA scaffold. The scaffold-cell complexes formed later were implanted onto the surfaces of liver in the mouse. After 2 weeks, the implants were taken out for HE staining and compatibility evaluation.ResultsThe isolated hepatocytes and SVECs were vigorous. The implants inosculated well with liver tissue of the host with visible growth of blood vessels in the implants. The hepatocyte aggregation grown around the vessels, the liver-like tissue, were observed.ConclusionIt is feasible to construct liver histioid through co-culturing hepatocytes and sinusoidal vessel endothelial cells on hyaluronic acid scaffold.

ObjectiveTo investigate the feasibility of construction of engineered blood vessel using chitosan tube and fibrin gel as scaffold.MethodsVascular endothelial cells and smooth muscle cells were harvested from aortas of a rat, respectively. After expansion in vitro, vascular endothelial cells were seeded onto the inner surface of chitosan tube and smooth muscle cells mixed with fibrin gel seeded onto outer surface of the scaffold to construct engineered blood vessels. Inverted microscope, immunohistochemical staining and scanning electronic microscope were used to evaluate the construct.ResultsVascular endothelial cells formed monolayer and covered the inner surface of chitosan tube. Smooth muscle cells survived in the fibrin gel and grew in a 3-dimensional manner. ConclusionChitosan-fibrin gel may be potentially used as scaffold of engineered blood vessels.

BACKGROUND: Studies demonstrated that small intestinal submucosa (SIS) had no immunogenicity, which can not lead to rejection following transplantation, thus, this is an ideal skin substitutes for natural skin.OBJECTIVE: Basic fibroblast growth factor (bFGF) gene was transfected into bone marrow mesenchymal stem cells (BMSCs),and combined the cells with SIS to construct tissue-engineered skin.METHODS: BMSCs were obtained from Japanese big-earad rabbits, and in vitro cultured. Then the subculturad BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry. In addition, the result of transfecting BMSCs with pCDNA-bFGF vector was measured by Western blot, and the structure of tissue-engineered skin was observed.RESULTS AND CONCLUSION: After passaged, BMSCs were grown quickly with Iong-fusiform shape. The cells were positive expressed CD90 and CD44, but negative expressed CD45. bFGF had been transfected into BMSCs, and stable expressed. The transfected BMSCs grew well in SIS. By this method, tissue-engineered skin can be constructed in vitro.

BACKGROUND: Basic fibroblast growth factor (bFGF) has significant promotion effects on repair in trauma, but local application cannot play a role for a long time.OBJECTIVE: To observe expression of bFGF gene in rabbit bone marrow mesenchymai stem cells (BMSCs) following transfection.DESIGN, TIME AND SETTING: The cytogene in vitro observation was performed at the College of Life Science, Yunnan University from March to August 2009.MATERIALS: One Japan flap-eared rabbit was purchased from the Department of Animal, Kunming Medical College. pCDNA3.1plasmid (Invitrogen, USA) was used in this study.METHODS: Bone marrow was extracted from rabbit anterior superior iliac spine. BMSCs were harvested by the adherent method.Cells were digested and subcultured when 80% confluent. According to GeneBank bFGF cDNA sequence, gene was designed and synthesized. Following electropherosis, the gel was retrieved using xhol I, BamH I enzyme digestion. Restriction enzyme was used to perform enzyme digestion, electropherosis and gene recovery in PcDNA Vector plasmid. bFGF DNA was connected with PcDNA Vector plasmid. PcDNA-bFGF eukaryotic expression vector was constructed. Recombinant was transfected into rabbit BMSCs using liposome infection protocol, and stable transfected line was screened.MAIN OUTCOME MEASURES: BMSC surface antigen expression was measured. Western blot was utilized to determine the expression of target protein.RESULTS: Results of flow cytometry showed that cultured cells were positive for CD90 and CD44, but negative for CD45.Results of immunohistochemistry demonstrated that vessel-derived BMSCs were negative for CD34, but positive for CD44. In cell disruption liquid of bFGF-transfected BMSCs, a significant positive zone of hybridization was visible at M, 23 000. However, no positive band was found in protein from pCDNA3.1(-) blank vector-transfected BMSCs.CONCLUSION: The bFGF gene was successfully transfected into BMSCs, and this target gene can stably express.

Objective To compare the curative effect of anatomical plate and locking plate in treatment of calcaneous intraarticular fracture. Methods 67 petiants with calcaneous intraarticular fracture were randomly divided into anatomical plate group (n=33) and locking plate group (n=34) . The Bo..hler angle, the Gissane angle, the length of calcaneal axis, the width and height of calcaneous and the Maryland grade were compared at 1 week and 6 month after operation. Results (1) week after operation, the Bo..hler angle, the Gissane angle, the height and width of calcaneous, the length of calcaneal axis, the Maryland grade had no significant difference between 2 groups . 6 months after operation, the Bo..hler angle, the Gissane angle, the height of calcaneous had significant differe nce between 2 groups. There was no significant difference in the length of calcaneal axis and the grade of Maryland between 2 groups. Conclusions The locking plate group is better than anatomical plate group in major anatomical measure indicators in 6 months follow up. The therapy of locking plate is worth of clinical promotion.

Objective To investigate the clinical characteristics of idiopathic retroperitoneal fibrosis (IRF) and its diagnosis and treatment. Methods Data of 27 IRF cases admitted into our hospital during recent 8 years were retrospectively analyzed. Result The main clinical manifestations of IRF included abdominal pain, lumbago, mass in retroperitoneal cavity and ureteral obstruction. The main diagnostic approach to IRF was the image examination. Preoperative correct diagnosis was established in 85 2% of cases. Ureterolysis and wrapping up with the omentum was performed in 21 cases of ureteral obstruction. Two cases suffering from renal artery (RA) stenosis were relieved with arteria renalis lysis. One superior mesenteric artery (SMA) stenosis was managed by arterial lysis. Twenty six cases were cured and one case died postoperatively. Conclusion IRF has nonspecific clinical manifestations. The preoperative diagnosis depends on the image characteristics. The treatment mainly consists of the relief of the obstructive symptoms.