1.Establishment of osteosarcoma cell models:effects of NS-398 on invasive ability of MG-63 cells under hypoxic condition
Chinese Journal of Tissue Engineering Research 2015;(40):6408-6412
BACKGROUND:In anoxic environment, invasive ability of tumor cels significantly increased. The increase in above invasive ability is possibly associated with the changes in urokinase or matrix metaloproteinases. Studies on hypoxia and tumor therapy especialy the effect on chemotherapy have made great progress, and found that hypoxia-inducible factor 1 is a key nuclear factor mediating celular hypoxia response, and also an important factor for regulating cel apoptosis during hypoxia. OBJECTIVE:To observe the effects of cyclooxygenase-2 inhibitor NS-398 on invasive ability of MG-63 cels under hypoxic condition in cel models of hypoxia osteosarcoma. METHODS: Chemical hypoxia inducer cobalt chloride was used to establish chemical hypoxia environment of concentration gradient of 0, 100, 200, 400 μmol/L. Under 200 μmol/L cobalt chloride, concentration gradient of 30, 60, 90 μmol/L of NS-398 were set. Osteosarcoma cels were determined after 48 hours of culture. Quantitative PCR was used to detect the expression of matrix metaloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA. Western blot assay was applied to measure hypoxia-inducible factor 1α expression. Flow cytometry was utilized to examine cel apoptosis in each group. RESULTS AND CONCLUSION:PCR and western blot assay results demonstrated that with increased concentration of cobalt chloride, matrix metaloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA and protein expression increased (P < 0.05), showing a concentration-dependent manner. Under cobalt chloride concentration of 200 μmol/L and hypoxia, with increased concentration of NS-398, matrix metaloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA and protein expression gradualy reduced, showing a concentration-dependent manner (P < 0.05). Flow cytometry results showed that under 400 μmol/L cobalt chloride concentration, cel apoptosis noticeably reduced. Under cobalt chloride concentration of 200 μmol/L and hypoxia, with increased concentration of NS-398, cel apoptosis increased (P < 0.05). Results confirm that anaerobic environment is an important factor for increased invasive ability of osteosarcoma. Under the anaerobic environment, NS-398 could effectively suppress the invasive ability of osteosarcoma and promote cel apoptosis.
2.EXPERIMENTAL STUDY OF BONE REPAIR INDUCED BY CRYOPRESERVED ALLOGRAFT PERIOSTEUM AND FETAL BONE COMPOSITION IN BONE DEFECT
Yongtao XU ; Jiefu GU ; Ping SHANG
Chinese Journal of Reparative and Reconstructive Surgery 2001;15(3):183-187
Objective To investigate the repairing effect of transplantation of allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft to bone defect. Methods Twenty Long-eared white male rabbits were chosen as experimental model of bilateral 12 mm combined bony and periosteal radial defect. Cryopreserved allograft periosteum with allogeneic fetal bone were implanted in the left defect as experimental side and fetal bone was simply transplanted in the right defect as control side. Bone repair process in the two groups were compared by macroscopy, microscopy, roentgenograms and the contents of calcium and phosphate in the defect area at 2、4、8 and 12 weeks after transplantation. Results There was significant statistic difference in the contents of calcium and phosphate between the experimental and control sides at 4、8 and 12 weeks after transplantation (P<0.05). With time passing by, the contents of calcium and phosphate have the increasing trends. In the experimental group, lamella bone was seen and medullary canal recanalized at 8 weeks postoperatively. The histological section showed the bone lacuna and lamella bone were formed. Conclusion It suggests that allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft can promote bone repair, and allogeneic fetal bone is excellent bone substitute.
3.Single iliac screw and dual iliac screws and titanium mesh cage fixation in the reconstruction of lumbosacral defects with finite element analysis
Liang MA ; Weichun GUO ; Yongtao XU
Chinese Journal of Tissue Engineering Research 2016;20(39):5859-5866
BACKGROUND:The surgical management to reconstruct the stability in lumbosacral region is very chal enging. There are many techniques to reconstruct the stability of the lumbosacral region, but the internal fixation loosening, fracture and pseudoarthosis are not uncommon. OBJECTIVE:Finite element analysis was used to analyze the stability of two types of reconstruction procedures and the stress of the internal fixation system. METHODS:A 64-slice spiral CT scan was used in a subject from L1 to pelvis. The scan data were imported into the Mimics 15.0 software to generate a three-dimensional surface model. The three-dimensional solid model was established using in the SolidWorks software. The solid geometry model of the L3, 4 pedicle screws combined with single iliac screw fixation and L3,4 pedicle screws combined with dual iliac screws and anterior titanium mesh cage support fixation were constructed by the Solidworks software. Two kinds of reconstruction procedures were simulated. The models were given material properties and analyzed by using the ABAQUS software. RESULTS AND CONCLUSION:(1) The maximum stress was 195.3 MPa in the complete model, 189.5 MPa in single iliac screw model, and 149.2 MPa in dual iliac screw+titanium cage model when constrained the roof of double acetabulum and applied a vertical load of 1 000 N. (2) The axial compression rigidity was 551.572 N/mm in the complete model, 613.87 N/mm in the single iliac screw model and 1 683.50 N/mm in the dual iliac screw+titanium cage model. (3) The bending rigidity of the dual iliac screw+titanium cage model was bigger than other models at 6 directions when applied 7 N?m bending loads. The maximum stress of single iliac screw model was bigger than other models. (4) The results suggest that the dual iliac screw+titanium mesh cage reconstruction can effectively restore the stability of the lumbosacral area. The stress of internal fixation system is smal er and more dispersed. There is obvious stress concentration in the connecting rod and the tail of the single iliac screw. The single iliac screw internal fixation system is easy to fatigue fracture and loosening.
4.Effect of single bundle anatomic reconstruction of the anterior cruciate ligament with the hamstring autograft through anteromedial approach
Liang MA ; Yongtao XU ; Yuanju SHE
China Journal of Endoscopy 2016;22(3):51-56
Objective To investigate the effect of single bundle anatomic reconstruction of the anterior cruciate ligament (ACL) with the hamstring autograft through anteromedial approach. Methods From January 2012 to Jan-uary 2014, 39 patients with ACL tore had been treated with single bundle anatomic ACL reconstruction with the hamstring autograft. In this group, 20 males and 19 females, 16 cases of the left knee and 23 cases of the right knee;including 27 cases of sports injuries, 5 cases of traffic accident injury and 7 cases of fall injury.8 cases were acute injury the others were old injury. 22 cases were had the meniscus injury. All patients were treated with single bundle anatomic ACL reconstruction with the hamstring autograft. The femoral tunnel was establish through the anteromedial approach. The medial meniscus suture was performed in 10 cases, partial resection in 2 cases, lateral meniscus su-ture in 9 cases, partial resection in 1 cases. The hamstring autograft was fixed with the RIGFIX system in the femoral tunnel and the BIOSURE SYNC in the tibia tunnel. The Lysholm score was used to evaluate the recovery of knee function, and the MRI score was used to evaluate the integraty of the grafts. Results All cases were followedup for 15 to 39 months with an average of 21.5 months. All cases had no vascular nerve injury, joint stiffness, infec-tion and other complications. All cases had good stability after 1 year and recover to normal sports after six months and intense sports after half past a year postoperation. The anterior drawer test and the Pivot shift tests were negative in all patients. The Lacheman test wasⅠin 1 patient and others were negative. The mean Lysholm score was (93.85± 4.33) which significantly increased compared with preoperative scores which was (36.84 ± 5.60) after 15 months (P <0.05). The MRI showed good maturity and normal morphology of the ACL autograft, the MRI score was (9.80 ±1.70) after 1 year postoperation. Conclusion The arthroscopic single bundle individual anatomic ACL reconstruction with hamstring autograft through anteromedial approach is effective, simple to perform and gives a graft the same as nor-mal anatomy. The grafts revascularization is good and effectively restore the stability of the knee.
5.Retroperitoneal Laparoscopic Ureterolithotomy
Weihua SHEN ; Qingkang XU ; Yongtao YU ; Weifei LI
Journal of Medical Research 2006;0(10):-
Objective To investigate standardization of technique of retroperitoned laparoscopic ureterolithotomy.Methods Retroperitoneal laparoscopic ureterolithotomy was performed on 85 cases of ureterolithiasis(the stone size ranging from 0.8~2.5cm,40 cases in the left,45 cases in the right;and 25 had undergone unsuccessful ESWL).Results 84 cases succeeded,1 case opened becase of play D-J tube difficultly,the mean operation time was 75 minutes(48~220min),and the estimated blood loss 20~100ml,the drain tube was removed 5~12 days(mean 7 days),1 case experienced urinary leakage,1 case infective in the retroperitoneal.Conclusion Retroperitoneal laparoscopic ureterolithotomy was a safe operation,it can take the place of open surgery.
6.Directional differentiation of bone marrow mesenchymal stem cells into oligodendrocytes induced by the combination of various neurotrophic factors
Yongtao XU ; Feng LI ; Tie LIU ; Hongbo YOU ; Zhong FANG
Chinese Journal of Tissue Engineering Research 2009;13(14):2793-2796
BACKGROUND: Myelination following axonal regeneration is a key factor affecting the recovery of spinal cord injury. Oligodendrocyte survival directly affects the myelination following axonal regeneration. OBJECTIVE: To investigate the feasibility of differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into oligodendrocytes induced by neurotrophic factors. DESIGN, TIME AND SETTING: The cell molecular biology in vitro study was performed at the Laboratory of Department of Orthopaedics, Tongji Hospital from September 2006 to June 2007. MATERIALS: A total of 5 Sprague Dawley rats aged 2-4 weeks, of both gender were selected. Bilateral femur and tibia bone marrow was obtained to harvest BMSCs. METHODS: At passage 4, BMSCs were incubated in serum-free medium, supplemented with N2, 20 ng/mL basic fibroblast growth factor, 20 ng/mL epidermal growth factor for 48 hours, and incubated in medium containing 500 ng/mL insulin-like growth factor I and N2 for 3 days. MAIN OUTCOME MEASURES: Morphological changes were observed using an phase contrast microscope. Semiquantitative RT-PCR was utilized to detect specific marker mRNA expression of oligodendrocytes. Using neuron marker anti-microtubule-associated protein, astrocyte marker anti-glial fibrillary acidic protein, oligodendrocyte marker anti-galactocerebroside, anti-myelin basic protein antibody, immunocytochemical staining was performed to detect the positive rate of the differentiation of BMSCs into oligodendrocytes. RESULTS: Morphological changes in BMSCs during the differentiation into oligodendrocytes: After the induction, a majority of BMSCs presented the morphological characteristics of oligodendrocytes. Cytoplasm retraction towards nucleus, cell process extension towards outwards, and strong refraction were found. With the prolongation of time, several cell processes connected and formed a typical net-shape structure. Specific marker mRNA expression of oligodendrocytes: Following induction, specific strap of myelin basic protein mRNA and galactocerebroside mRNA could be detected. Positive rate of oligodendrocytes: During induction, the positive rates of galactocarebroside, myelin basic protein and microtubule-associated protein were 65%, 45% and 10%, respectively. CONCLUSION: The combination of epidermal growth factor, basic fibroblast growth factor and insulin-like growth factor can effectively promote the directional differentiation of BMSCs into oligodendrocytes.
7.Clinical study of characteristics of large cerebral infarction: 68 case report
Shiping SONG ; Ying LV ; Mingjie XU ; Wei LI ; Yongtao BAI
Chinese Journal of Rehabilitation Theory and Practice 2003;9(10):626-627
ObjectiveTo explore clinic features of large cerebral infarction and the relationship between infarcted area and prognosis.MethodsClinic materials such as dangerous factors, onset, course, treatment and prognosis, brain CT changes and internal carotid artery ultrasonic examinations of 68 patients with large cerebral infarction were analyzed retrospectively.ResultsThere were 28 cases caused by cerebral embolism, and 40 cases caused by cerebral thrombosis. 6 cases got recovery, and 34 cases, progress, and 11 cases,no progress, and 17 cases, death. ConclusionThe infarcted area was significantly related to prognosis (P<0.05). Internal carotid artery frequently occurring atheromas and senile non-valve atrial fibrillation were major embolic origins for large cerebral infarction.11 out of the 17 deaths could be attributed to brain herniation and secondary infection. The early abnormal brain CT showed that the rate of mortality was 62.5%, and the rate of mid-line dislocation was 42.9%. Either of them indicated that prognosis was bad.
8.Percutaneous nephrolithotomy combined with endoscopic balloon dilation in the treatment of upper ureterostenosis with recurrent renal calculi
Qingkang XU ; Weihua SHEN ; Zhefeng XU ; Feng CHEN ; Yue DUAN ; Tianqiang YU ; Yongtao YU
Chinese Journal of Urology 2012;33(5):340-343
ObjectiveTo analyze the technique and clinical effect of percutaneous nephrolithotomy combined with endoscopic balloon dilation in the treatment of upper ureterostenosis with recurrent renal calculi. MethodsFrom June 2008 to June 2011,18 ureteral stenosis patients with the history of ureteral open surgery,postoperative residual or recurrent kidney stones were treated.There were 8 males and 10 females with the age of 27 -48 years.Fourteen cases were with hydronephrosis of 2 -4 cm,3 cases were with hydronephrosis of 5 -6 cm and 1 case was with hydronephrosis >6 cm.Subsequent stone size < 1 cm was found in 15 cases,1 -3 cm in 3 cases,>3 cm in 1 case.All patients were treated with percutaneous nephrolithotomy ultrasonic lithotripsy combined with balloon dilatation.The stone clearance rate,hydronephrosis changes,complications and IVP situation before and after surgery were analyzed.ResultsAll the 18 cases were completed surgery successfully.There was 1 (6%) case with renal hemorrhage 3 days after the surgery and controlled with DSA hemostasis.There was 1 case accepted adjusting double-J tube by ureteroscopy.Sixteen (89%) patient's stones were completed removed.One case with residual calyceal stones size <5 mm was not further treated.There was 1 case treated with nephrectomy because of renal stone with infection.The patients were followed up for 6 to 36 months.Fourteen cases with hydmnephrosis improved significantly; 3 cases with no significant changes but improved following balloon dilation.All patients achieved significant improvement in imaging study comparing of preoperative and postoperative data.ConclusionThe use of percutaneous nephrolithotomy combined with endoscopic balloon dilation is a safe and efffective treatment option in the treatment of kidney stones with ureteral stenosis.
9.Colonic mucosa urethroplasty in the treatment of complex long urethral stricture: a three case report with literature review
Qingkang XU ; Yue DUAN ; Tianqiang YU ; Feng CHEN ; Yongtao YU ; Xiang HONG ; Zhefeng XU ; Weihua SHEN
Chinese Journal of Urology 2011;32(10):700-703
Objective To investigate the feasibility of urethral reconstruction with colonic mucosa graft in the treatment of complex long urethral stricture.Methods The clinical data of three cases with complex long urethral stricture were reported and analyzed.Patient ages were 71,64 and 48 yrs and the course of disease was three months,six months and six yrs,respectively.The length of urethral stricture was 13,18 and 12 cm.Removing the narrow urethral segment and intercepting the length from 12 to 18 cm sigmoid colon and stripping colonic mucosa were performed.Urethral reconstruction was done with a free graft of colonic mucosa.Follow-up included urethrography,uroflowmetry,and urethroscopy.Results The urethral reconstructions were completed successfully.The urinary peak flows of the patients were 16.7 ml/s,19.6 ml/s and 26.4 ml/s at six weeks post operation.Urethrography revealed the graft urethral lumens were bulky three months after the operation.In urethroscopy,the colonic mucosa was found to be of good color and the anastomotic site healed well.Patients were followed-up 28,16,and three months,respectively,and were all voiding well.Conclusions Colonic mucosa graft urethroplasty is a feasible procedure for the treatment of complex long urethral stricture.
10.Locking plate and hemiarthroplasty for treatment of Neer three- and four-part proximal humeral fractures
Wei XU ; Ming CHEN ; Zonggang XIE ; Yongtao MAO ; Haibin ZHOU ; Youjia XU ; Qirong DONG
Chinese Journal of Trauma 2012;28(5):402-406
ObjectiveTo analyze the clinical outcome of locking plate and hemiarthroplasty in treatment of Neer three- and four-part proximal humeral fractures.Methods A totalof 63 consecutive patients aged over 55 years with Neer three- and four-part proximal humeral fractures were treated surgically from June 2006 to June 2010.In the retrospective study,36 patients were treated by open reduction and locking plate fixation ( locking plate fixation group) and 27 patients treated by hemiarthroplasty (hemiarthroplasty group).Clinical outcomes including operation time,blood loss,visual analog scale ( VAS),Constant-Murley score and complications were evaluated.ResultsThe average 19.7 months follow-up showed complication rate of 28% in the locking plate group,including one patient with varss malunion,one with axillary nerve injury,one with humeral head necrosis,one with heterotopie calcification and four with impingement syndrome.The complication rate was 24% in the hemiarthroplasty group,including two patients with tuberosity nonunion,one with tuberosity migration,one with impingement syndrome and one with joint semiluxation.The mean Constant-Murley score of three-part fractures in the locking plate group was ( 76.5 ±5.8) points,better than (69.2 ±7.2) points in the hemiarthroplasty group (P <0.05 ).While the two groups showed no significant differences with regards to operation time,blood loss and visual analog scale (VAS).As for the four-part fractures,the mean operation time was shorter and the mean blood loss was less in the hemiarthroplasty group compared with the locking plate group (P <0.05),while there were no obvious differences in VAS score and Constant-Murley score between the two groups. Conclusions Both locking plate and hemiarthroplasty are the reliable methods for the three- and four-part proximal humeral fractures.The patients' age,bone quality,fracture type,comminution degree and evaluation of osteonecrosis possibility,are critical for surgery decision.