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Objective To discuss the application of ultra-thin flap technology in the thoracodorsal artery per forator flap,and appraise the effectiveness in clinical practice.Methods From May,2013 to December,2015,10 patients with soft-tissue defects of the four limbs were treated with ultra-thin thoracodorsal artery perforator flap.The classification of the perforating vessels was done by Kimura method.The flap blood supply was measured and the patients' satisfaction of the appearance of flap was investigated through questionnaires.Results The flap of 8 cases (5 cases of type Ⅰ and 3 cases of type Ⅱ) could be evenly thinned,and the flap of 2 cases (type Ⅲ) could only be thinned at the edge beyond the part with perforator as the center and a radius of 3 centimeters.The flap of 9 cases completely lived.While in 1 case (type Ⅲ),a small part of the edge of the flap was necrosed(2 cm×1 cm),and coalesced well after change dressing.The donor site was sutured directly in 7 cases and covered with skin graft in 1 case.Eight patients were followed-up from 6 to 12 months.The treatment effect was satisfied.And there were no side effects on the functioning of donor sites.Seven patients were highly satisfied with the appearance of the flap (satisfaction rate was 87.5％).Conclusion Ultra-thin flap technology can be adopted to cut thin thoracodorsal artery perforator flap,and can remarkably improve the effect in clinical practice.
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OBJECTIVE:To optimize ultrasonic extraction and purification technology of solanesol from tobacco leaf. METH-ODS:Using extraction rate and transport rate of solanesol as indexes,single factor test was used to investigate liquid-solid ratio, ultrasonic extraction temperature and time,ultrasonic power and extraction times,and the amount of soap alkali lye(volume ratio of soap alkali lye to extraction liquid),acidizing fluids (volume ratio of acidizing fluids to soap alkali lye extract),extraction times of purification technology. Optimized technology was validated,and the purity of solanesol was calculated;the amount of ex-tracted solanesol was compared between this method and traditional extraction method (spending 30 h),solvent continuous cyclic extraction (spending 5-6 h). RESULTS:Optimized extraction technology was as follows as volume ratio of soap alkali lye to ex-traction liquid 1∶14,ultrasonic extraction temperature 70 ℃,ultrasonic extraction time 60 min,ultrasonic power 120 W,extract-ing for 3 times;optimized purification technology was as follows as volume ratio of soap alkali lye to extraction liquid 2∶35,vol-ume ratio of acidizing fluids to soap alkali lye extract 2∶14,extracting for 4 times. In validation test,extraction rate,transport rate and purity were 92.45%(RSD=0.46%,n=3),79.88%(RSD=0.30%,n=3)and 55.86%(RSD=0.40%,n=3). The amount of solanesol extracted with 3 methods were 52.22,45.22 and 26.10 mg/g. CONCLUSIONS:The optimized technology is simple and stable,costs less time and saves source with high extraction amount and purity,which is suitable for production,extraction and purification of solanesol from tobacco leaf.
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Adult stem cells from skeletal muscle cells were induced to differentiate into cardiocytes to see if stem cells from another different but histologically-comparable tissues can differentiate to the target cells. Skeletal muscles-derived stem cells (MDSCs) were isolated from adult skeleton muscle tissues by differential adhesion, and immunocytochemically identified by using Sca-1. In order to induce the proliferation but not differentiation of MDSCs, the cells were cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) supplemented with 1:50 B27, 20 ng/mL basic fibroblast growth factor (bFGF), 20 ng/mL epidermal growth factor (EGF) in a suspension for 6 days. Then these stem cells were treated with 5 mumol/L 5-azacytidine for 24 h in an adherence culture. The characteristics of induced cells were examined by immunocytochemistry, quantitative real time RT-PCR and morphological observation of cell phenotype. Our results showed that the appearance of some cells gradually changed from spindle-shape into polygonal or short-column-shape. Some of these post-treated cells could contract spontaneously and rhythmically. The expression of GATA-4 and cTnT was increased 1 and 2 week(s) after the treatment. And about 16.6% of post-treated cells were cTnT-positive. Therefore, we are led to conclude that skeletal muscle-derived stem cells could differentiate into cardiocyte-like cells, which exhibited some characteristics of cardiocytes.
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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.
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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.
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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.
Subject(s)
Animals , Female , Male , Rats , Interleukin-1 , Pharmacology , Motor Neurons , Pathology , Neuroprotective Agents , Pharmacology , Random Allocation , Rats, Wistar , Sciatic Nerve , Wounds and Injuries , Spinal Cord , PathologyABSTRACT
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.
Subject(s)
Interleukin-1/pharmacology , Motor Neurons/pathology , Neuroprotective Agents/pharmacology , Random Allocation , Rats, Wistar , Sciatic Nerve/injuries , Spinal Cord/pathologyABSTRACT
Objective To explore the relationship between immunohistochemistry of LN,FN,p53 and TIMES in human brain glioma(BG). Methods Transmission electronic microscope(TEM) and immunohistochemistry were used to investigate the morphological characteristics of micrangiums in BG and the expression of LN,FN,p53 in BG and intracranial metastatic tumors. Results 1.It was found that base laminas beneath endothelial cell with locally or extensively thickened were intact and continuous in BG.The increasing thickness of BM was consistent with the staining of LN and FN and related to p53 immunostaining.BM of p53-protein positive cases grew thicker than that of p53-protein negative ones.2.Micrangiums BM in all BG were positive for LN and FN.The more malignant the BG was,the stronger the LN and FN staining became and the thicker the blood vessel walls grew(P0.05).Conclusion One of the reasons that BMs in TIMES in BG thicken may be the over expression of LN and FN of brain micrangium endothelial cells.Also,the influence of p53 on TIMES is associated with the functional state of endothelial cells.Micrangiums endothelial cells may be play a role in regulating TIMES.
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Objective To study the morphological characteristics of blood-tumor barrier(BTB)model of glioma in vitro. Methods After C 6 glioma cells/endothelia ECV 304 co-cultured mixed or in Transwell or on both sides of membrane of Transwell,the morphological characters of fenestra of endothelial cells,the junction between ECV 304 cells,the interface between C 6 cells and ECV 304 cells,and perivascular-end-feet of C 6 cells were observed under transmission electron microscope(TEM)and scanning electron microscope(SEM),and compared to BTB in 4 cases with human brain glioma. Results ECV 304 cells grown to confluence were not fenestrated,but with fomation of tight junction between ECV 304 cells after co-cultured with C 6 cells mixedly,in Transwell or on both sides of membrane of Transwell;It was not found that pseudopodia from C 6 cells as co-cultured in Transwell reaching into pore of Transwell;C 6 cells co-cultured on both sides of membrane of Transwell often sticked out to the ECV 304 cells,but with no pseudopodia from C 6 cells surrounded ECV 304 cells of penetrated the endothelia clefts.Perivascular-end-feet of C 6 cells were not integrant.These characters were similar to BTB in human brain glioma.Conclusion C 6 glioma cells/endothelia cell ECV 304 co-cultures on both sides of membrane of Transwell may simulate the morphological characters of BTB in vivo in some degree.;