To study the characteristics of acute rejection after limb allotransplantation, 29 male Sprague-Dawley rats were randomly divided into 2 groups, with 15 rats in control group and 14 rats in experimental group. Each rat in control group underwent limb replantation. Each rat in experimental group received limb transplantation from Wistar rat. No immunosuppressive drugs were used after operation. The circulation of the transplanted limb, time and signs of rejection, histopathological changes in the tissues of the limb graft when rejected and survival time of limb grafts were evaluated. In the control group, no signs of rejection were observed, the circulation of each replanted limb was normal, it could survive for a longer time. The experimental group showed clinical signs of rejection (sub dermal edema and erythema) after a mean time of 3.36+/-1.15 days, and the mean survival time of the allografts was only 7+/-0.78 days. Histopathological examination showed most violent rejection reaction in skin. It is concluded that with Wistar-to-SD limb transplantation without use of immunosuppression, rejection of the grafts would occur after a mean time of 3.36+/-1.15 days; the earliest signs of rejection were edema and erythema of the skin, skin being the most representative component of limb graft rejection.
Acute Disease ; Extremities/*transplantation ; *Graft Rejection ; Graft Survival ; Random Allocation ; Rats, Sprague-Dawley ; Rats, Wistar ; Skin/immunology ; Transplantation, Homologous

Protective effect of interleukin-1beta (IL-1beta) on motor neurons was studied after peripheral nerve injury. Twenty Wistar rats were divided into 2 groups randomly. The right sciatic nerve of each rat was resected. After silicon tubulization of sciatic nerve in rat, 15 microl 1 ng/ml IL-1beta and PBS solution were injected into the silicon capsule respectively. Enzyme histochemistry was performed to show acetyle cholesterase (AchE) and nitric oxide staining (NOS) activity of spinal alpha motor neurons in spinal segments 2 weeks later. Neurons were counted and the diameter and cross sectional (c/s) area of neurons were analyzed by using computer image analysis system. The results showed that as compared with the normal side, both enzyme activities significantly changed in motor neurons in PBS group. The diameter and c/s area of both neurons changed significantly too (P < 0.01). These results suggest that exogenous IL-1beta protects alpha-motor neurons from degeneration and necrosis after peripheral nerve injury.
Interleukin-1/*pharmacology ; Motor Neurons/*pathology ; Neuroprotective Agents/*pharmacology ; Random Allocation ; Rats, Wistar ; Sciatic Nerve/*injuries ; Spinal Cord/pathology

Objective To investigate the clinical manifestation,therapeutic methods and treatmenteffectiveness of hand contracture due to extravasation injury from peripheral intravenous therapy in children.Methods Data of 6 children who had received treatment for hand contracture in our hospital from March 2004 to September 2011 were retrospectively analyzed.There were 3 boys and 3 girls,aged from 24 to 51 months (average,38 months).Two children underwent extensor tenolysis and dorsal capsular release at the metacarpophalangeal (MCP) joints.The other 4 children received conservative treatment,including physical therapy (ultrasound and medium frequency pulse) to reduce the hand swelling and peritendinous adhesion,active and passive functional exercise of digital joints and night splint protection.Results The MCP joint could be passively flexed to 90° intraoperatively in 2 children who underwent operation; however the contracture recurred at 2 months and 6 months after operation,respectively,and then the MCP joints were fixed at 0° position.For 4 children who received conservative treatment,although the active flexion of their MCP joints ranged from 30° to 45° during the rehabilitative procedure,recurrence of hand contracture was observed after the cessation of the rehabilitation in all children,and the range of active MCP joints flexion varied from 0° to 15°.Conclusion The mechanism underlying hand contracture due to extravasation injury from peripheral intravenous therapy remains elusive.Either simple conservative or surgical management results in poor functional outcomes.Combined therapy based on different indications for conservative and surgical management may improve the outcomes.

BACKGROUND: In repair of nerve defect with allogenic nerve graft, to reduce immune rejection is one of the key problems. At present, the main approach is to reduce antigenicity of grafted nerve segment and apply generally immune inhibitor.OBJECTIVE: To observe the effects of freeze/thaw treatment and local application of transforming growth factor beta 1 (TGF-β1) plasmid on frozen nerve allograft.DESIGN: Randomized controlled animal experiment was designed.SETTING: Department of Hand Surgery, Union Hospital, Tongji Medical College Affiliated to Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Tongji Medical College of Huazhong University of Science and Technology from January 2003 to December 2004, in which 40 Wistar healthy and adult rats were employed,from different delivery and were randomized into experimental group and control, 20 rats in each one.METHODS: Transforming growth factor-β1 (TGF-β1) plasmid and frozen allogenic sciatic nerve were prepared. In experimental group and control,sciatic nerve was cut off 2.0 cm in length, in the foramen 0.5 cm beneath piriformis. The nerve defect was repaired with pre-frozen allogenic nerve 2.0 cm in length. In experimental group, TGF-β1 plasmid was injected in local muscle and two broken ends of nerve. In the control group, physiological saline of equal volume was injected. In the 6th and 12th weeks, the samples were collected from 10 rats in each group for sectioning, staining,axonal counting and statistical analysis.RESULTS: No any animal was died in experiment and all of animals entered result analysis. In the 6th weeks, in the control group, mild edema appeared among axons on the grafted segment of nerve and in the experimental group, there was no edema among axons and the regenerated nerve numbers were close to the normal. In the 12th week, in the experimental group, the entire grafted nerve segment was basically filled up by the regenerated axons;myelinated nerve fiber was arranged in order and both axons and myelins were developed well. The regenerated axonal count in experimental group was more significantly than the control, indicating extremely significant difference [(98.6±4.8), (75.8±5.1) counts/μm2, t=2.962, P ＜ 0.01].CONCLUSION: Freeze/thaw treatment can decrease antigenicity of allogenic nerve, which provides the possibility of repair of nerve defect. Local application of TGF-β1 plasmid can provide immune inhibition locally and reduce immune rejection in the host.

BACKGROUND: Auto-neural transplantation is used widely on peripheral neurological defect, but it also has some difficulties. So some scholars try to use xenoma-neural transplantation; however, it is hard todeal with immunological rejection.OBJECTIVE: To study the effect of transforming growth factor-β1 (TGFβ1) used in local area on neural regeneration after transplantation of fresh nerve allograft.DESIGN: Randomized controlled study.SETTING: Hand Surgery Department of Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and TechnologY.MATERIALS: The experiment was conducted in the Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology between August 2001 and October 2002. Totally 60healthy adult Wistar rats from different confinements were randomly divided into three groups including experimental group, blank group and control group with 20 in each group.METHODS: TGF-β1 plasmid was prepared for using. Establishment of animal model: Sciatic nerve at the 0.5 cm deep of piriformis muscle of rats in the two groups was cut with disinfectant razor into chip regularly about 2.0 cm. The excisional nerve segment was exchanged to transplant plerosis neurological defect. TGF-31 was injected into the local muscles and bisection of nerve in the experimental group, and equal volume of saline was injected into rats in the blank group and the control group. In addition, rats in the experimental group and the blank group were not treated with any drugs, but cyclosporine A (15 mg/kg) was used to feed rats in the control group. Ten rats from each group were taken for section and staining at the 6th and the 12th week: ① Glees-luxot fast blue staining method; ② myelin sheath fast blue staining method. Axonal amount: Fields were randomly taken from the middle staining samples 12 weeks later and 1.0 mm2 interaxis-cylinder was counted under light microscope of 400 times. Comparisons among groups were analyzed with i test.MAIN OUTCOME MEASURES: Morphological observation and axonal amount of transplanted area in each group.RESULTS: Quantitative analysis of the experimental animals: Totally 60rats entered the final analysis without any loss. ① Infiltration of monocytes was observed widely in various areas of graft in the blank group;meanwhile, desiccation of myelin sheath and plenty of vacuolations were also observed, especially at the sixth week. The whole graft was infiltrated by monocyte with severe rejection. Few axis-cylinders were regenerated in the transplanted segment. At the 12th week, graft was slender, plenty of scar tissues were proliferated, edema was observed obviously, few Schwann cells and regenerated axis-cylinders were observed, and lots of regenerated axis-cylinders did not pass the whole graft. A few infiltrative monocytes were observed, and edema was observed obviously, but new vessel was formed in transplanted nerve, and regenerated axis-cylinders passed the whole graft in the experimental group and the control group.Lots of Schwann cells were observed at the 6th week; meanwhile, regenerated axis-cylinders passed the whole graft at the 12th week, a quantitative myelinization was formed, Schwann cells proliferated obviously, and edema between axis-cylinder was relieved. Numbers of peripherally regener ated axis-cylinder of nerve and remyelination in each ransplanted area were more than those in the central area, and edema between peripheral axis-cylinder was milder than that in the central area in the experimental group. ② Twelve weeks after operation, 5 rats in each group were selected to observe their fields, which were taken randomly from neural graft,under the microscope of 400 times to count 1.0 mm2 inter-axis-cylinders.Number of axis-cylinder was higher in the experimental group and the control group than that in the blank group, and the differences were significant [(78.3±4.6), (76.1±4.2) , (15.0±3.5) ,t=3.056, t=2.948, P ＜ 0.01];however, number in the experimental group was similar to that in the control group, and differences were not significant [(78.3±4.6), (76.1±4.2),t=1.982 P ＞ 0.05].CONCLUSION: TGF-β1 used in local area plays an immunosuppressive action locally, decreases host immunological rejection, increases the number of axis-cylinder, and accelerates growth of nerve.

BACKGROUND: At present, the repair by means of suture is still commonly used to repair the peripheral nerve injury and rupture, while the adhesion of the fibrin glue repairing peripheral nerve injury has been considered as a new topic of study.OBJECTIVE: To study the countertraction intensity of peripheral nerve and its dynamic changes after repaired with the adhesion of fibrin glue.DESIGN: A randomized controlled experimental study.SETTING and MATERIALS: The study was completed in the Laboratory of Biodynamics, Department of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology. The healthy adult male Wistar rats weighing 250- 300 g were selected for the experiment.INTERVENTIONS: Totally 96 Wistar rats were completely randomized into the suture group and the adhesion group. Their sciatic nerves were cut, and the incisions were well lined. The fibrin glue was adopted in the adhesion group, while 11 -0 suture was adopted in the suture group. On the very day and 3, 7, 14, 21, 28 days after the operation, 8 rats were respectively taken each from the suture group and the adhesion group. The free sciatic nerves of them were detected immediately by the biodynamic test.MAIN OUTCOME MEASURES: The peak load and the power consumption were measured when the nerves ruptured and the nerve stress-strain curve was described.RESULTS: In normal countertraction intensity curve of the nerve, the elastic peculiarity can be manifested. Between the suture group and the adhesion group, there were no notable significances of the maximal countertraction intensity and power consumption on the very day and 14, 21, 28 days after the operation( P ＞ 0.05). While 3 days after the operation, the maximal countertraction intensity of the two groups was(1.35± 0. 27),( 1.97 ± 023) N/mm2 respectively, the power consumption was (0. 028 ± 0.007), (0.040 ± 0.003) J/mm2 respectively. Seven days after the operation, the maximal countertraction intensity was( 1.93 ± 0.26), (2.74± 0.30) N/mm2 respectively, the power consumption was(0.047±0.009), (0.063±0.007) J/mm2 respectively. The differences both had the notable significance ( P ＜ 0.05).CONCLUSION: The fibrin glue has enough countertraction intensity and can gratify the need of such nerve repairs.

Objective To explore the diagnostic value of post processing techniques of MSCT for diagnosing congenital heart disease associated with tracheobronchial stenosis in children. Methods Thirty four patients with congenital heart disease complicated by tracheobronchial stenosis were evaluated with MSCT. MPR, CPR, MinIP and VR were performed to show the tracheobronchial morphology. Findings in 43 segments of 32 cases were compared with the findings of surgical operation. Mann-Whitney test was employed to assess the significance of measurement between the post processing techniques and the operation. Results The lenghth of stenotic segments were variable seen at operation, with values between 4-39 mm in trachea, 4-33 mm in main bronchi and 3-12 mm in lobe bronchi, respectively. The biggest difference between the measurement of operation and MSCT was 4 mm (3 segments). Other differences ofstenotic segments were within 3 mm. Six segments exhibitee mild, 16 moderate and 21 severe stenosis at surgery. By contrast, five segments exhibited mild, 17 moderate and 21 severe stenosis by MPR or CPR,2 mild,9 moderate and 32 severe stenosis by MinIP and 4 mild, 11 moderate and 28 sever stenosis by VR,respectively. There was no significant difference in measuring the degree of tracheobronchial stenosis between MPR or CPR, VR and the surgery (Z =-0. 105,- 1. 479;P ＞0.05), while MinIP frequently overrated the degree of stenosis compared with the measurement at surgery (Z=-2.484,P =0. 013). Conclusion The integrated three dimensional reformations of MSCT scan can accurately evaluate the degree and extent of congenital heart disease associated with tracheobronchial stenosis in children and provide valuable information for clinical management.

Objective To investigate the clinical application of flow-through latissimus dorsi musculocutaneous flaps for functional reconstruction of musculus flexor digitorum defects.Methods 9 forearm traumatic patients were selected admitted from March 2011 to February 2013,aged 22 to 57 years old (average age 39.2),including 5 male and 4 female.All cases were combined injuries.The flexion of the fingers was constrained.The injury areas ranged from 15 cm×6 cm to 21 cm×9 cm while the flaps were 17 cm×7 cm at least and 24 cm×10 cm at most.For traumatic musculus flexor digitorum,skin,soft tissue and radial artery defects were all well repaired with functional flow-through latissimus dorsi musculocutaneous flaps.Functional exercise and physical therapy were done 3 weeks later.Electromyography,grip strength,and pinch strength evaluation were performed at the 6,12months after operating.Enneking evaluation was scored during follow-up periods from 12 to 24 months.All the results were measured and evaluated.Results All flaps survived completely in 9 patients and recipient radial artery were bypassed well.The flaps had good appearance and high aesthetic satisfactory.Seven cases had more than 12 months' follow-up periods.Muscle contraction can be touched 3 months after transplantion.Diagram of EMG tracing showed that the maximum voluntary contraction action potentials presented simple phase at 3 months,intermix phase at 6 months,and interference phase at 12 months.The average grip strength of the second to fifth finger was 8.97±2.91 g/mm2 at 6 months,11.69±3.71 g/mm2 at 12 months.The average pinch strength of thumb and the index finger was 1.39±0.22 g/mm2 at 6 months,1.67±0.34 g/rmm2 at 12 months.For the muscle power recovery and motor nerve function scale,4 cases were M3+,3 cases were M3.The average ratio of Enneking evaluation score with nomal was 65.7％± 15.9％.Conclusion As functional musculocutaneous flaps,flow-through latissimus dorsi musculocutaneous flaps are effective for reconstruction of musculus flexor digitorum and wound defects.The vascular status of the extremities are protected and maintained,which is extremely useful in forearm-salvage procedures from both functional and structural viewpoints.

ATP was added to the cultured sensory neurons obtained from the dorsal root ganglia of the neonatal rats and PBS was added to serve as control. MTT assays were conducted to evaluate the survival and activity of the cultured neurons. And the silicone regenerative chamber was used after the sciatic nerve incision of the mature SD rat. 1 mmol/L ATP was injected into the left chamber and 0.09 % natrium chloride was injected into the right chamber as controls. The changes of nitric oxide synthase (NOS) activity in the corresponding dorsal root ganglia were measured histochemically and image analysis was also performed 4 days after the sciatic nerve injury. The results showed that extracellular ATP could enhance the survival of the neurons and the number of NOS positive neurons were significantly different between the ATP and control groups (P<0.05). It was suggested that extracellular ATP had neurotrophic effect on neurons survival and could inhibit the NOS activity of the sensory neurons after the peripheral nerve incision, hence exerting the protective effect on the neurons, which was valuable for nerve regeneration after nerve injury.

ATP was added to the cultured sensory neurons obtained from the dorsal root ganglia of the neonatal rats and PBS was added to serve as control. MTT assays were conducted to evaluate the survival and activity of the cultured neurons. And the silicone regenerative chamber was used after the sciatic nerve incision of the mature SD rat. 1 mmol/L ATP was injected into the left chamber and 0.09 % natrium chloride was injected into the right chamber as controls. The changes of nitric oxide synthase (NOS) activity in the corresponding dorsal root ganglia were measured histochemically and image analysis was also performed 4 days after the sciatic nerve injury. The results showed that extracellular ATP could enhance the survival of the neurons and the number of NOS positive neurons were significantly different between the ATP and control groups (P<0.05). It was suggested that extracellular ATP had neurotrophic effect on neurons survival and could inhibit the NOS activity of the sensory neurons after the peripheral nerve incision, hence exerting the protective effect on the neurons, which was valuable for nerve regeneration after nerve injury.