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Objective To confirm that rat bone marrow mesenchymal stem cells (MSC) transfected with nerve growth factor (NGF) gene in the bladder tissue of diabetic rats bladder tissues can survive and stably express NGF. Methods A diabetic rat model was constructed. The BrdU-labelled MSC transfected with NGF gene were transplanted into the diabetic rats bladder tissues. BrdUlabelled immunohistochemistry was used to observe the growth of MSC transfected with NGF gene in the diabetic rats bladder tissues. The expression of NGF mRNA and protein were checked by RT-PCR and ELISA. Results A diabetic rat model was successfully built by a single intraperitoneal injectionof STZ. The blood glucose was still high after 8 weeks. NGF gene modified MSC could be detected in the bladder of diabetic rats by BrdU-labelled immunohistochemistry. The concentration of NGF in the control group, disease group and treatment group were ( 114 ± 3), ( 70 ± 2), ( 110 ± 2) pg/ml by ELISA and mRNA quantity by RT-PCR were 0. 183±0. 004, 0. 032±0. 139, 0. 130±0. 165, respectively. Compared with the control group, the expression of NGF gene was decreased (P＜0. 05) in the incidence group. The expression of NGF gene was increased (P＜0. 05) in the treatment group compared with the disease group. Conclusions The NGF gene-modified MSC could survive in diabetic rats bladder tissues. The NGF gene in MSC could stably express in diabetic rats bladder tissues.
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BACKGROUND:There is few mesenchymal stem cells(MSCs) in the bone marrow,about one BMSC in 1?104-1?105 monocytes.Following in vitro isolation,purification and amplification,it is satisfactory for in vivo requirement.OBJECTIVE:To observe biological characteristics and potentiality of multi-directional differentiation of BMSCs in vitro.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Institute of Urinary Surgery,Union Hospital Affiliated to Fujian Medical University from October 2007 to December 2008.MATERIALS:A total of 30 clean male Sprague Dawley rats of inbreeding line were supplied by Silaike,Shanghai,China.METHODS:BMSCs from rats were isolated and cultured in vitro by using the whole bone marrow adherence method.When 80%-90% confluency,BMSCs were digested by trypsin.At the third passage,BMSCs were induced to differentiate into osteoblasts and adipocytes.MAIN OUTCOME MEASURES:Cell morphology was observed under the inverted phase contrast microscope.BMSCs surface markers were detected using flow cytometry.Osteogenic ability was examined by alizarin red staining.Adipogenic ability was measured by Oil red O staining.RESULTS:The primary cells and the passage cells were mostly fusiform in shape,to be similar to fibroblasts.Cell still kept a high potential of growth and amplification following over 10 subcultures.At the third passage,BMSCs were positive for CD44,CD90,CD106,but negative for CD34,CD45,CD11b.Following 21 days of osteogenic induction,cell alizarin red staining showed that alizarin red was positive in osteoblasts.Following 2 weeks of adipogenic induction,oil red O staining showed that red lipid droplet existed in adipocytes.CONCLUSION:Whole bone marrow adherence method can isolate,purify and amplify BMSCs in vitro.The obtained cells have general biological characteristics of MSCs,and also have potentiality of multi-directional differentiation.
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Objective To evaluate the clinical value of laparoscopic renalcyst unroofing as compared with open surgery.Methods The clinical data of 30 cases(group A) receiving glaparoscopic renalcyst unroofing from February 2002 to March 2005 and 15 cases(group B) receiving open renalcyst unroofing from February 2002 to February 2004 were retrospectively analyzed.The rating of depression and anxiety before operation,operating time,amount of bleeding during operation,recovery of intestinal function after operation,ambulation and normal activities after operation,the time of using intravenous after operation,the times of using analgesic after operation,the operation cost,total cost of hospitalization,the hospital stay and complications were compared between group A and group B.Results In group A,the amount of bleeding during renalcyst unroofing operation,the recovery of intestinal function after operation,ambulation after operation,the hospital stay,the resume of normal activities after operation and the occurrence of complications the time of using intravenous after operation,the times of using analgesic after operation were all better than those in group B(P
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Objective To assess the cause of iatrogentic urethral stricture,prevention methods.Methods A total of 21 cases of iatrogentic urethral stricture patients were retrospectively analyzed. Results 2 cases of corpus glandulare prostatae extraction on ospectinis,2 cases of urethrolithotomy, 12 cases of transurethral resection of the prostate foe the treatment of symptomatic BPH, 5 cases of exelcymosic UCL all gained urethral sounding and urethra sliver and electrotomy. Conclusion Iatrogentic urethral stricutre can be prevented by iatrogentic urethral catheterization operation technical,reconstruction the time size quality of urethral catheterization, using intravenous. By urethral sounding and intracavitary operation,iatrogentic urethral stricture all gained satisfaction.
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Objective To evaluate the effectiveness of spiral stone dislodger combined with pneumatic lithotriptor under ureteroscopy for managing upper ureteral stones. Methods 106 patients with upper ureteral stones with spiral stone dislodger combined with pneumatic lithotriptor under ureteroscopy were treated. The transverse diameter of the stones was 4～12mm and the vertical length 6～15mm. Results 87.7% (93/106) stones were fragmented for one session of treatment. 8 cases were pushed upward then undergoing extracorporeal shock wave lithotripsy(ESWL) after surgical operations. Operations in 5 cases had to be changed to open surgery,and ureteral perforations happened in 1 case. Conclusion Successful entrance of the uretersocop is a prerequisite for crushing the stone,appropriate fixation of the stone is the key to their crushing,spiral stone dislodger is an effective means of fixing ureteral stones.
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Objective:To observe the biological behaviors of colorectal cancer LS174 cells before and after pcDNA3.0-hugl-1 transfection,so as to investigate the association of hugl-1 with colorectal cancer.Methods:The eukaryotic expression vector pcDNA3.0-hugl-1 was constructed and transfected into LS174 cells.RT-PCR and Western blotting methods were used to analyze the expression of hugl-1 mRNA and protein in LS174 cells before and after transfection.Soft agar colony formation assay,wound-healing experiment,adhesion assay and Matrigel invasion assays were used to study the effects of hugl-1 expression on the proliferation,adhesion,movement and invasion in LS174 cells.Results:The recombinant plasmid pcDNA3.0-hugl-1 was successfully constructed.RT-PCR and Western blotting showed that the hugl-1 expression was higher in cells transfected with pcDNA3.0-hugl-1 than in those un-transfected or empty vector-transfected cells (P