Objective To assess the variation and its significance of messenger ribonucleic acid(mRNA) expression of endothelial nitric oxide synthase (eNOS) in allografts of common carotid transplantation model in white rabbits. Methods To establish an animal model of common carotid transplantation in vivo, 30 rabbits were divided into four groups with random number table. Group A (n=3): autografts; group B (n=9): allografts with the least treated; group C (n=9): allografts treated by penicillin/streptomycin and preserved under room temperature; group D (n=9): allografts treated by penicillin/streptomycin and cryopreserved in liquid nitrogen. All the transplanted grafts were harvested 1-3 weeks later, then compared and evaluated the histomorphological variation and eNOS mRNA expression. Results The vascular structures of autografts in group A were kept approximately normal, only a few infiltration of inflammatory cells could be found. The structural variations of allografts in other trial groups behaved similarly as, intima proliferation in the 1st week, intima hyperplasia in the 2nd week, and both intima and media hypertrophy in the 3rd week. And also there seemed that luminal thrombosis could be found in all the allografts. Allografts in group B were destructed utmost the worst in all the groups. The expression of eNOS mRNA in allografts of group B was significantly less than that in other groups (Plt;0.05). Conclusion The down-regulation of eNOS mRNA expression might lead to intima hyperplasia and thrombosis of allografts.
In order to seek a good method to treat the severe, complex damage of the digital flexor tendon, an anatomical study based on 30 cadavers was done, and the result showed that the source of the blood supply to the palmaris longus tendon was from the radial and ulnar arteries. Three cases of severe complex digital flexor tendon injuries were satisfactory. Its main advantages were: (1) The tendon transplant had its own blood supply and had no need to the revascularization, therefore the adhesion formed between the tendon and adjacent tisues might be less;(2) Besides reparing the tendon defect, it can simultaneously solve the problem of the defect of the soft tissues and the skin.
Objective To investigate the clinical result of treatment of bonecyst by transplantation of the autologous bone marrow combined with the allograft bone. Methods From February 2004 to March 2006, 13 patients withbone cyst were treated by transplantation of the autologous bone marrow combined the the allograft bone. Among the 13 patients, 6 were males and 7 were females, ranging in age from 5 to 16 years, averaged 11.5 years. In the patients, 5 lesions were located inthe proximal humerus, 2 in the femoral neck, 3 in the femoral shaft, 2 in the proximal tibia, and 1 in the distal tibia. Among the patients, 5 had a complication of pathologic fracture. All the patients underwent an erasion of the bone cyst, and then the transplantation of the autologous bone marrow combined with the allograft bone, and 8 of them were also given an instrument fixation. Results The follow-up for 6 months to 2 years after operation revealed that 5 of the patients had an incision healing by the first intention, 5 had an effusion in the incision site, and 3 had a delayed healing of the incision. According to the Capanne criteria, the postoperative X-ray findings indicated that 10 patients had Grade Ⅰ healing, and 3 had Grade Ⅱ healing. The complete healing took 3.5-8 months,averaged 5.2 months. There was no recurrence. When the fixation instrument was removed, no pathologic fracture occurred. The function of the upper and lower limbs recovered. Conclusion Transplantation of the autologous bone marrow combined with the allograft bone is an effective and safe procedure for treatment of bone cyst.
In order to compare the immunogenecity and biological properties of homologous tendon grafts after treatment from different methods of freezing, tendons from chickens received repeated freezing-thawing treatment or ultra-low-temperature treatment, and then, the post-treatment tendons were preserved in liquid nitrogen for 3 months before transplantation. The autogenous tendon transplantation was served as the control. It was found that in the group of repeated freezing-thawing treated tendons, the tendon cells all died and while in the ultra-low temperature treated tendons the active rate of tendon cells was 92.5% +/- 3.4%, and the histological observation showed that transplantation of frozen tendons would result in extensive infiltration of inflammatory cells in the grafted tendons and the peritendinous adhesion was serious than that of the autografts. The active flexion function, hydroxyproline levels and the biomechanical analysis showed no significant differences between the repeated freezing-thawing treated homografts and the ultra-low-temperature treated homografts, and that the autografts was definitely superior to the homografts. The conclusions were: (1) Transplantation of the homologous tendons from the two different methods of freezing could receive considerable success and there was no significant difference between them; (2) Transplantation of frozen homologous tendon graft might give successful result which was probably due to the preservation of the cellular activity of the tendon cells following freezing treatment and elimination of the antigen presenting cells in the tendon as well, and (3) Although the cellular components of the tendon were damaged and the antigenicity of the tendon was lowered, it did not necessarily mean that homologous tendon graft would always be successful in transplantation.
Objective To modify orthotopic liver transplantation method with two-cuff technique. Methods On the basis of cuff technique, the donor liver was perfused through the abdominal aorta with 20 ml cold perfusate. The anastomosis of the suprahepatic vena cave(SVC) was sutured end-to-end with 8/0 nylon line,and the continuity of infrahepatic vena cave(IVC) and portal vein(PV) were established by means of cuff method respectively.The bile duct anastomosis was performed by internal stent. Results Orthotopic liver transplantations were performed in 360 rats. The time for donor operation and liver preparation was (31.2±5.0) min and (12.0±3.0)min,and the recipient operative and anhepatic time was (45.0±5.5) min and (20.0±2.5) min, respectively. The two-day survival rate was 91.4%.In nonintervention group, one-week survival rate was 86.5%. Conclusion The modified model is easily available and highly reproducible which provides a practical and stable experimental model for the study of liver transplantation.
Objective To investigate the myogenic differentiation of mesenchymal stem cells (MSCs) after being transplanted into the local muscle tissues. Methods The serious muscleinjured model was established by the way of radiation injury, incising, and freezing injury in 36 mouses. Purified MSCs derived from bone marrow of male mouse and MSCs induced by5-azacytidine(5-Aza-CR) were transplanted into the local of normal muscle tissues and injured muscle tissues of femal mouse. The quantity of MSCs and the myogenic differentiation of implanted MSCs were detected by the method of double labeling, which included fluorescence in situ DNA hybridization (FISH) and immuno-histochemistry on the 1st, 3rd, 6th, 9th, 12th, and 15th day after transplantation. Results The quantity of implanted MSCs decreased as timepassed. MSCs’ differentiation into myoblasts and positive expression of desmin were observed on the 15th day in purified MSCs group and on the 6th day in induced MSCs groups. Conclusion MSCs could differentiate into myoblasts after being implanted into the local of muscle tissues. The differentiationoccurs earlier in the induced MSCs group than that in purified MSCs group.
ObjectiveTo study the cell apoptosis and the dynamic expression and significance of apoptosis-related genes in graft veins. MethodsA rat experimental model of autogenous graft vein was established by transplanting the right external jugular vein to infrarenal abdominal aorta in 100 Wistar rats. TUNEL and immunohistochemistry were used to detect the apoptosis, the expression of apoptosis-related genes bcl-2 and bax in vascular smooth muscle cells (VSMCs) of graft veins. ResultsWithin the 8 weeks after transplantation, the apoptotic VSMCs in the graft veins were much more than those in the control group with the apoptotic rate reaching the peak〔(28.5±16.6)%〕 on the 2nd week and dropping to (8.1±2.8)% during the 4th to 8th week. There was statistical difference compared to the control group 〔(0.5±0.2)%, P<0.01〕. From 1 to 2 weeks, the positive rate of bcl-2 was (22.1±5.4)% which was higher than that of the control group and the 4-8 week group (P<0.01); from 1 to 6 weeks, the expression of bax was higher than that of the control group 〔(5.5±2.3)%〕 and the postoperative 8th week group 〔(8.2±2.9)%, P<0.01〕.Conclusionbcl-2 and bax protein may be involved in the regulation of apoptosis of VSMCs. Apoptosis of VSMCs may be an important factor in graft remodeling and graft vein stenosis.
Objective To investigate the treatment method of ischemic necrosis of the femoral head by the transplantation of pedicled bone flap with deep iliac circumflex vessel in adult and to assess its curative effect. Methods From February 1996 to September 2003, 46 adult patients with ischemic necrosis of thefemoral head (58 coxas) were treated by transplantation of pedicled bone flap with deep iliac circumflex vessel. The locations were the left side in 34 coxas and the right side in 24 coxas(stage Ⅱ in 16 coxas, stage Ⅲ in 39 coxas and stage Ⅳ in 3 coxas).Results Thirty-seven patients (40 coxas) were followed up 6 months to 10 years after operation. According to the assess of FanQishen,the result was excellent in 19 cases (21 coxas), good in 11 cases (12 coxas), fair in 5 cases (5 coxas) and poor in 2 cases (2 coxas). The postoperative excellent and good rate was 81%. Conclusion Operative treatment of ischemic necrosis of the femoral head in adult by transplantation of bone flap pedicled with deep iliac circumflex vessel has the characteristics of restoration of blood supply ofthe femoral head, decompression of marrow cavity, elimination of necrotic bone and support of the femoral head. It is an effective and advanced method for treatment of ischemic necrosis of the femoral head in adult.
OBJECTIVE: To evaluate the therapeutic effect of amniotic membrane transplantation(AMT) for ocular burn. METHODS: Twenty patients with ocular burn(28 eyes) were treated with amniotic membrane transplantation. Of them, there were 6 cases of severe alkali burn(10 eyes), 8 cases of severe acid burn(10 eyes), and 6 cases of thermal burn(8 eyes). RESULTS: In 28 eyes, ocular inflammation was controlled after 3-7 days of surgery; no continued dissolution, perforation and iris atrophy were found. There were corneal transparency in 16 eyes, keratoleukoma in 4 eyes, and total corneal opacity in 8 eyes. All eyeballs were saved and had stable ocular surface. There was no allograft immune rejection and secondary infection. CONCLUSION: Amnitic membrane transplantation can relieve the inflammatory reaction, reduce the growth of blood vessel and restrain the proliferation of fibrous tissue. It is an effect surgical method for ocular burn in reconstruction of ocular surface and salvage of eyeball.
Objective To investigate the extent intravenously transplantation of mesenchymal stem cells (MSCs) mediated by magnetic targeting material arrive in the myocardial infarction region and its effects on the recovery of myocardial infarction. Methods Identify the phenotype of the fourth genet of ex vivo expanded MSCs, stain with DAPI after inducing with 10μmol/L 5-aza, then preserve the MSCs for transplantation. 28 SD rats were divided into three groups: group A (n=10), delivered MSCs combined with magnetic targeting material for 30 minutes to rats through tail vein,and kept on raising after placing magnets on the corresponding skin region to myocardial infarction area for 30min; group B (n=9), administration MSCs not conjuncted with magnetic targeting material through tail vein; group C (n=9), direct intramyocardial transplantation of MSCs. Two days after transplantation, evaluate the aggregation state of MSCs in the area of myocardial infarction; 30d later, estimate the functional and morphological changes in myocardial infarction region. Results We observed that each MSCs had 3-5 molecules of magnetic targeting material attached to its membrane under transmission electron microscope. The homing rates of MSCs respectively were group A 38%, group B 6%, group C 53%.The number of aggregating MSCs of group A and group C was apparently more than that of group B(Plt;0.01). After transplantation, the contraction indices of left ventricle in group A and group C had significant improvement as compared with that of pretransplantation (LVEF 46%±6% vs. 38%±8%, 51%±5% vs. 35%±4%; LVFS 28%±6% vs. 20%±7%, 32%±4% vs. 20%±5%, Plt;0.05) and administrated cells stained with DAPI could be detected in infarction region under optical microscope. After transplantation, the contraction indices of left ventricle in group B hadn’t conspicuous improvement, and the transplanted cells labeled with DAPI could not be identified in infarction region under optical microscope (homing rate of MSCs 38%). There was no statistically difference of results between group A and group C, but in experiment process, there was a high mortality in group C. Conclusion The method that intravenously delivery of MSCs mediated by magnetic targeting material could accumulate much more MSCs in infarction region, reduce infarction size, and effectively improve the cardiac function after infarction.