ObjectiveTo discuss the clinical classification and treatment protocols of complete duplication of kidney and ureter in children. MethodsBetween March 2000 and February 2015, 106 children with complete duplication of kidney and ureter were treated, and the clinical data were retrospectively analyzed. Of them, there were 11 boys and 95 girls, aged from 1 month to 11 years (mean, 3.5 years); one side was involved in 88 cases and two sides in 18 cases. They were divided into 4 types according to image examinations and clinical presentations:14 patients who needed no special treatment were classified into the first type, 15 patients who underwent reconstruction into the second type, 74 patients who underwent segment removal of renal dysplasia and subtotal excision of abnormal duplicated ureter into the third type, and 3 patients who underwent removal of the whole affected kidney and subtotal excision of whole ureter into the forth type. ResultsThe patients were followed up 2 months to 14 years (median, 23 months). There was no deteriorating case in the first type. There was no complication such as leakage of urine, discomfort over the back and loins, ureterocele, reproductive tract infection, or hematuresis in the other types. The results of white blood cell count, renal function, and electrolyte presented no abnormality. One patient in the second type and 6 patients in the third type had ureteral stump syndrome; 1 patient in the second type and 3 patients in the third type had urinary tract infection; and 3 patients in the second type had mild hydronephrosis after operation. ConclusionIt can obtain good clinical outcome to choose individualized treatment according to clinical classification of complete duplication of kidney and ureter, which can reserve effective renal units as much as possible and improves the patients' quality of life.
To establ ish a simple and stable cervical ectopic renal transplantation rat model that increase surgical successful rate. Methods A total of 208 male inbred Wistar rats (weighing 220-260 g) were randomly served as donors and recipients. The graft consisting of kidney, renal vein (RV) and renal artery (RA) was obtained, and perfused in situ. The donor RA was end-to-end anastomosed to the recipient left common carotid artery (CCA) by using of “sleeve” anastomosis,and the donor RV to the recipient right external jugular vein by using of “cuff” technique. The distal end of the ureter wasbrought out to form cervical cutaneous stomas. Results A total of 104 ectopic renal transplantations were performed in rats, including stages of the pre-experiment (62 operations) and experiment (42 operations). The success rates of the two stages were 80.6% and 95.2%, respectively. The causes of failure in the pre-experimental stage were anesthesia accidents, thrombosis of the arterial anastomosis, massive hemorrhage, air embol ism and phlebemphraxis. In the experimental stage, 2 rats died due to late anastomotic hemorrhage and thrombosis. The remaining 40 transplanted kidney survived more than 6 months. The time for surgery was (40 ± 6) minutes, the average time for donor surgery was (20 ± 5) minutes, the preparation time for the graft was (8 ± 2) minutes, the operative time for the recipient was (18 ± 3) minutes, including the time for the arterial anastomosis (5 ± 2) minutes and venous anastomosis (2 ± 1) minutes, the cold ischemia time of graft was (15 ± 3) minutes. Conclusion The cervical ectopic renal transplantation technique has the advantages of easy-and fast-to-perform, shorter operation and cold ischemia time, higher successful rate.
Objective To study the effect of motor nerve implantation after ectopic transplantation of skeletal muscle on nerve regeneration in rat. Methods Sixty Sprague-Dewley male 8 monthold rats were randomly divided into 3 groups: control group,in situ implantation group and ectopic transplantation group. In control group, obturator nerve controlling right gracilis was cut off. In in situ implantation group, the right gracilis was cut off and replanted to its original site, and the obturator nerve was implanted to the muscle. In ectopic transplantation group, the right gracilis was cut off and transplanted to the muscle of the left leg, and the obturator nerve was implanted to the muscle. After 25 weeks, the neurophysiological information was collected through electromyography and the weight of the muscle was measured. Results The potentialwithout control of the nerve existed in control group. There were no significant differences in latency, amplitude and conduct velocity betweenin situ implantation group and ectopic transplantation group(Pgt;0.05).The atrophy of gracilis was dominant incontrol group, the weight of the muscle was 158.0±19.3 mg. The weights of the muscle were 509.6±14.5 mg in ectopic transplantation group and 516.8±12.7 mg in in situ mplantation group, showing no significant difference (P＞0.05). The weights of the muscle in in situ implantation and ectopic transplantation group were larger than that in control group, showing significant difference(P＜0.05). Conclusion Motor nerve implantation after ectopic transplantation of skeletal muscle could prevent the atrophy of the muscle and resume partial function of nerve.
Objective To observe the release pattern of the microcysts and the effect of ectopic osteogenesis of combined micromorselized bone by optimized preparation of microcysts. Methods Optimized poly-DLlactide-co-glycolide (PLGA) microcysts manufacturing method was performed with the orthogonal design, and the accumulated release amount of microcysts was calculated at 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, 48 h, 60 h, 72 h, 84 h, 96 h, 120 h, 144 h, 168 h, 192 h, 216 h, 240 h and 264 h. Twentyfour Wistar rats were divided into 4 groups (n=6) and 1 cm length incision was cut in their bilateral thighs skin, forming 48 gluteus maximus muscle sackmodels. In group A,collagen was implanted to bilateral muscle sacks respectively. In group B, collagen and autologous morselized bone were implanted to bilateral muscle sacks. Ingroup C, collagen and rhBMP-2/PLGA delayed release microcysts were implanted to bilateralmuscle sacks respectively. In group D, collagen and morselized bone/rhBMP-2/PLGA delayed release microcysts were implanted to bilateral muscle sacks. Gross and histologic observations were made at 3, 4 and 5 weeks postoperatively.Results Every optimized variance had an effect on particle diameter of microcyst and its encapsulating rate. The microcyst’s surface was smooth and had a fine spheroplast, which released slowly within 11 days in vitro. In thethird week postoperatively, the graft in group A could not be touched, while the graft in all other 3 groups was still found. After 3 weeks, collagen was absorbed completely in group A, the residual collagen could be seen in groups B, C andD. After 4 weeks, collagen could be seen in group A; micromorselized bone continued to be absorbed and became smaller in group B; microsphere became smaller, osteoblasts increased in group C; micromorselized bone and microsphere continuedto be absorbed, oteoblasts and chondroblasts increased. After 5 weeks, implantsbecame small, microsphere was absorbed, osteoblasts and chondroblasts became more in groups B, C and D. Microcysts presented with white granuloshape and were packaged in tissue pieces. Histologic observation showed that the PLGA microcysts in 3 weeks and 4 weeks could be absorbed gradually as the time in vivo, if combining with morselzed bone they could produce abundant induced osteoblasts and chondroblasts. Conclusion Optimizing the preparation technology of microcysts has delayed their release during a long period in vitro. Autologous micromorselized bone can be ectopicly induced to produce large amount of osteoblasts in gluteus maximus muscle sack, where PLGA microcysts can combine organically and bring about the bone formation with less amount of growth factors.
ObjectiveTo evaluate the ectopic osteogenesis of recombinant human bone morphogenetic protein 2 (rhBMP-2) loaded chitosan (CS)/dextran sulfate (DS) by micro-CT. MethodsrhBMP-2/CS/DS microspheres were prepared by the ionic crosslinking and its shape was observed under the scanning electron microscope. The release of rhBMP-2 was determined from resultant microspheres by ELISA assay. Forty-eight Sprague Dawley male rats were randomly divided into 4 groups (n=12), quadriceps muscle bag model was made, gelatin sponge (group A), CS/DS microspheres (group B), rhBMP-2 (group C), and CS/DS/rhBMP-2 microspheres (group D) were implanted into the bags respectively. The tissue samples with heterotopic ossification were harvested for micro-CT scanning at 4, 8, 12, and 16 weeks. The tissue mineral density (TMD), bone volume fraction (BVF), trabecular thickness (Tb.Th), trabecular number (Tb.N), bone mineral density (BMD), and tissue mineral content (TMC) were measured. ResultsThe prepared rhBMP-2/CS/DS microspheres with smooth surfaces were spherical and evenly disperses without obvious agglomeration. At 2 hours, microsphere started a sudden release period in vitro; the release reached a peak at 2 days; and the release cycle lasted about 20 days. The rats survived to the end of the experiment. At each time point after operation, no radiation developed and no osteogenesis was observed by three dimensional reconstruction in groups A and B. However, radioactive strength and reconstructed bone tissue gradually increased in groups C and D, and group D had more radioautography and more bone tissues than group C. At each time point, TMD, BVF, Tb.Th, Tb.N, BMD, and TMC of groups A and B were zero. Ectopic bone formed with time, the other parameters showed an increasing trend except Tb.N in groups C and D, showing significant difference when compared with groups A and B at each time point (P < 0.05). There was no significant difference between groups C and D at 4 weeks (P>0.05); the parameters of group D were significantly higher than those of group C at 8-16 weeks (P < 0.05). ConclusionrhBMP-2/CS/DS microspheres have stronger ability of ectopic bone formation than single rhBMP-2.
To study the recovery method and effect of amputated foot after temporary ectopic implantation. Methods Two male patients with amputated foot were treated with temporary ectopic implantation in July 2001 and January 2002. Amputated foot was caused by mechanical injury and crush injury. After 6 hours, temporary ectopic implantation of amputated foot was given and replantation was done 3 months after primary operation. The recovery methods were as follows: automatic and passive motion, high pressure oxygen, massage, protective and positional feel ing training, etc. The effects of recovery was observed. Results All amputated foots survived after operation, the time of follow-up was 6 years,and 5 years and 7 months. Extension degree of first metatarsal digital joint was 12º and 15º, flex degree of first metatarsal digital joint was 15º and 13º, and extension degree of other metatarsal digital joints was 8º and 9º. Force degree of extension muscle was 4, force degree of flex muscle was 4, and two-point discrimination was 20 mm and 18 mm. Patients recovered their superficial sensibil ity, touch sense, deep pain sense and topognosis. The skin color and temperature were normal. And the patients could do some housework. Conclusion Temporary ectopic implantation of amputated foot can recover the function of amputated foot by motor and sensitive recovery methods.
Objective To study the method of reinnervation after ectopic transplantation of the gracilis muscle in rats. Methods Sixty healthy male rats (age, 8 months; weight, 400-500 g) were randomly divided into 3 groups: the control group, the motor reinnervation group, and the sensory reinnervation group. The right gracilis of the rat was cut off, and the muscle was transplanted to the left leg. In the control group, no reinnervation was performed on the obturator nerve; in the sensory reinnervation group, the obturator nerve was coapted with the recipient saphenous nerve; in the motor reinnervation group, the obturator nerve was coapted with the femoral nerve motor branch. After 25 weeks, the weight of the muscle was measured, and the histological examination was performed. Results Atrophy of the gracilis was found to be a dominant effect in the control group, where the weight of the muscle was 204.0±15.3 mg. In the motor reinnervation group, the weight ofthemuscle was 394.8±12.9 mg, and in the sensory reinnervation group, it was 389.2±13.5 mg, with no significant difference between the two groups (P＞0.05). The weight of the muscle in the motor reinnervation group and in the sensory reinnervation group was significantly greater than that in the control group (P＜0.05).The tissue observation revealed that the nerve axon was diffusedin the motor reinnervated group, with no nerve endplates found. The motor nervereinnervated flaps showed the viable axons out to the motor endplates. The histological examination revealed evidence of reinnervation. Conclusion The motor or sensory nerve anastomosis after the ectopic transplantation of the skeletal muscle can prevent the atrophy of the muscle and restorepart of the nerve function.
Objective Tissue engineered bone (TEB) lacks of an effective and feasible method of storage and transportation. To evaluate the activity of osteogenesis and capabil ity of ectopic osteogenesis for TEB after freeze-dried treatment in vitro and in vivo and to explore a new method of preserving and transporting TEB. Methods Human bone marrow mesenchymal stem cells (hBMSCs) and decalcified bone matrix (DBM) were harvested from bone marrow and bone tissue of the healthy donators. TEB was fabricated with the 3rd passage hBMSCs and DBM, and they were frozen and dried at extremely low temperatures after 3, 5, 7, 9, 12, and 15 days of culture in vitro to obtain freeze-dried tissue engineered bone (FTEB). TEB and FTEB were observed by gross view and scanning electron microscope (SEM). Western blot was used to detect the changes of relative osteogenic cytokines, including bone morphogenetic protein 2 (BMP-2), transforming growth factor β1 (TGF-β1), and insul in-l ike growth factor 1 (IGF-1) between TEB and FTEB. The ectopic osteogenesis was evaluated by the methods of X-ray, CT score, and HE staining after TEB and FTEB were transplanted into hypodermatic space in athymic mouse. Results SEM showed that the cells had normal shape in TEB, and secretion of extracellular matrix increased with culture time; in FTEB, seeding cells were killed by the freeze-dried process, and considerable extracellular matrix were formed in the pore of DBM scaffold. The osteogenic cytokines (BMP-2, TGF-β1, and IGF-1) in TEB were not decreased after freeze-dried procedure, showing no significant difference between TEB and FTEB (P gt; 0.05) except TGF-β1 15 days after culture (P lt; 0.05). The ectopic osteogenesis was observed in TEB and FTEB groups 8 and 12 weeks after transplantation, there was no significant difference in the calcified level of grafts between TEB and FTEB groups by the analysis of X-ray and CT score. On the contrary, there was no ectopic osteogenesis in group DBM 12 weeks after operation. HE staining showed that DBM scaffold degraded and disappeared 12 weeks after operation. Conclusion The osteogenic activity of TEB and FTEB is similar, which provides a new strategy to preserve and transport TEB.
Objective To analyze the effectiveness of conservative medical treatments for ectopic pregnancy (EP): methotrexate (MTX) + mifepristone + Ectopic Pregnancy II decoction (EP-II) vs. methotrexate + mifepristone. Methods A total of 95 patients with EP in Shenzhen Shajing Affiliated Hospital of Guangzhou Medical University from January 2009 to January 2011 were randomly divided into two groups: 45 patients in the experimental group were treated with MTX, mifepristone and EP II decoction, while the other 50 patients in the control group were treated with MTX and mifepristone. The effectiveness of the two groups was analyzed with SPSS 13.0 software. Results There were significant differences in the time of serum β-HCG return to normal (16.13±8.13 ds vs. 22.05±7.15 ds, Plt;0.05), time of EP mass absorption (30.46±7.56 ds vs. 39.99±18.26 ds, Plt;0.05) and tubal patency rate (80% vs. 75%, Plt;0.05) between the two groups. But there were no significant differences in effective rate (95.56%, 43/45 vs. 94%, 47/50, χ2=0.0809, Pgt;0.05) and side effects. Conclusion The combination of methotrexate, mifepristone and EP II decoction for ectopic pregnancy is more effective than mifepristone and methotrexate in coordinately killing the embryo, shortening the time of serum β-HCG return to normal and the time of EP mass absorption, and improving the function of oviducts.
Objective To investigate the ectopic bone formation of the chitosan/phosphonic chitosan sponge combined with human umbil ical cord mesenchymal stem cells (hUCMSCs) in vitro. Methods Phosphorous groups were introduced in chitosan molecules to prepare the phosphonic chitosan; 2% chitosan and phosphonic chitosan solutions were mixed at a volume ratio of 1 ∶ 1 and freeze-dried to build the complex sponge, and then was put in the simulated body fluid for biomimetic mineral ization in situ. The hUCMSCs were isolated by enzyme digestion method from human umbil ical cord and were cultured. The chitosan/phosphonic chitosan sponge was cultured with hUCMSCs at passage 3, and the cell-scaffoldcomposite was cultured in osteogenic medium. The growth and adhesion of the cells on the scaffolds were observed by l ight microscope and scanning electron microscope (SEM) at 1 and 2 weeks after culturing, respectively. The cell prol iferation was detected by MTT assay at 1, 2, 3, 4, 5, and 6 days, respectively. Bilateral back muscles defects were created on 40 New Zealand rabbits (3-4 months old, weighing 2.1-3.2 kg, male or female), which were divided into groups A, B, and C. In group A, cellscaffold composites were implanted into 40 right defects; in group B, the complex sponge was implanted into 20 left defects; and in group C, none was implanted into other 20 left defects. The gross and histological observations were made at 4 weeks postoperatively. Results The analysis results of phosphonic chitosan showed that the phosphorylation occurred mainly in the hydroxyl, and the proton type and chemical shifts intensity were conform to its chemical structure. The SEM results showed that the pores of the chitosan/phosphonic chitosan sponge were homogeneous, and the wall of the pore was thinner; the coating of calcium and phosphorus could be observed on the surface of the pore wall after mineral ized with crystal particles; the cells grew well on the surface of the chitosan/phosphonic chitosan sponge. The MTT assay showed that the chitosan/phosphonic chitosan sponge could not inhibit the prol iferation of hUCMSCs. The gross observation showed that the size and shape of the cell-scaffold composite remained intact and texture was toughened in group A, the size of the complex sponge gradually reducedin group B, and the muscle defects wound healed with a l ittle scar tissue in group C. The histological observation showed that part of the scaffold was absorbed and new blood vessels and new bone trabeculae formed in group A, the circular cavity and residual chitosan scaffolds were observed in group B, and the wound almost healed with a small amount of lymphocytes in group C. Conclusion The chitosan/phosphonic chitosan sponge has good biocompatibil ity, the tissue engineered bone by combining the hUCMSCs with chitosan/phosphonic chitosan sponge has the potential of the ectopic bone formation in rabbit.