| Background: Diabetes is one of the main chronic diseases worldwide,with more than 425 million people currently affected.One of the main complications of diabetes is diabetic foot(Diabetic foot,DF).The annual incidence rate in the global diabetic population is 6.3%.Diabetic foot ulcers(DFU)have a lifetime risk of 25%..Therefore,there is currently no effective treatment for severe diabetic foot,and the final outcome is often amputation or even death,which brings a heavy burden to Patients and society.Finding and researching effective methods for the treatment of severely diabetic foot is a problem that needs to be solved urgently.Peripheral arterial disease is the most prominent factor leading to the occurrence of diabetic foot ulcers,including the pathological changes of the large arteries of the lower extremities and/or the capillaries of the feet.Arteriosclerosis occlusion can cause ischemia,hypoxia,and necrosis of skin,fascia,muscle,nerve,bone and other tissues.In view of this,reconstruction of the diseased blood vessel to improve the blood supply of the lesion is one of the main surgical methods for the treatment of diabetic foot ulcers.However,studies have found that after revascularization of the sclerosed and blocked arteries of the lower extremities the foot ulcers of some Patients are still difficult to heal,because revascularization can only recanalize the large arteries of the calves,but cannot dredge the finer foot arteries and cannot repair them.Damaged blood vessels or regeneration of new blood vessels cannot completely solve the problem of ischemia in the foot.Therefore,how to promote the repair and regeneration of local capillaries in ulcers is a key issue.In the early clinical practice of this subject,the "Tibial transverse transport(TTT)" was innovatively applied to the treatment of severe diabetic foot,and fruitful results were achieved.Facing Patients with large-area soft tissue defects in diabetic foot and severe ischemic infection,successfully saved the patient’s limbs and promoted the healing of ulcers.Early imaging results showed that the patient’s microcirculation was significantly improved,with a large number of new blood vessels."Tibial transverse transport" breaks the traditional concept of soft tissue defect repair and provides new methods and concepts for the treatment of diabetic foot Patients.Then the exact mechanism of tube regeneration in the TTT process is still unclear,clarifying the TTT process The mechanism of angiogenesis,exploring the cellular and molecular mechanisms that affect angiogenesis,can provide a theoretical basis for TTT and help guide the clinical application of this technology.In this study,Enothelial progenitor cells(EPCs)were used as seed cells to verify the Ang-1/Tie-2 signal transduction pathway in EPCs and study the Ang-1/Tie-2 signal transduction pathway The series of molecular regulation events of TTT neovascularization further clarified the neovascularization mechanism in the treatment of severe diabetic foot with Tibial transverse transport,and provided a theoretical basis for“Tibial transverse transport”.Method:1.Patient enrollment criteria formulation,grouping and specimen collection: Diabetic foot Patients with Wanger level3 and above who received treatment from June 2018 to June 2020 were divided into3 groups according to the enrollment criteria: Group A:(Patients did not receive Before TTT treatment);Group B(Patients received TTT 1 month after surgery);Group C: control group(Patients received other treatments 1 month),collect peripheral blood and tissues around wounds and ulcers of the above Patients.2.Exploring the expression of peripheral blood vascular factors and microvascular regeneration in the wound surface of patients with severe diabetic foot by TTT:(1)Observe the structure of neovascularization in the tissue by HE staining;perform blood vessel by immunohistochemistry CD34 Quantitative detection of the number of production;(2)Detection of mature microvessels in wound tissue by CD31 and α-SMA immunofluorescence;(3)Detection of Ang-1,Ang-2,and VEGFA expression in wounds of each group by immunohistochemistry Situation;(4)Detect the expression of Ang-1,Ang-2and VEGFA in peripheral blood by ELISA technology.3.Exploring the mechanism of TTT and peripheral blood vascular factor expression and wound microvascular regeneration:(1)Detecting Nrf-2 in the tissues around the ulcer and peripheral blood in each group of patients through immunohistochemistry and ELISA experiments Expression;(2)Detect the expression of MMP-9 and TIMP-1 in the tissues around the ulcer and peripheral blood of each group by immunohistochemistry and ELISA;(3)Detect the ulcer of each group by immunohistochemistry and ELISA The expression of human angiopoietin receptor tyrosine kinase2(Tie-2)in peripheral tissues and peripheral blood;(4)Flow cytometry was used to detect CD34+,CD34+/CD133+,CD34+/CD133+/VEGFR+ in Patients’ peripheral blood2 positive rate of cells.Result:1.HE staining of traumatic tissues revealed that neovascularization was significantly increased in post-TTT patients compared with other groups,and the number of neovascularization and the number of mature microvessels in traumatic tissues detected by CD31 and α-SMA immunodouble fluorescence was found to be significantly higher in the post-TTT group compared with other groups(p<0.05);ELISA and immunohistochemistry tests showed that the vascular ELISA and immunohistochemistry showed that the positive rates of Ang-1 and VEGFA were significantly higher in the post-TTT group than in the other groups,and Ang-2 was lower than in the other two groups(p<0.05);2.The expression of Tie-2、Nrf-2 and TIMP-1 was significantly higher and the expression of MMP-9 was lower in the post-TTT patients’ paracolic tissue and peripheral blood than in the other groups(p<0.05).3.Tibial Transverse Transport(TTT)and severe diabetic foot Patients’ peripheral blood endothelial progenitor cell mobilization was studied: the percentage of peripheral blood endothelial progenitor cells was higher in the post-TTT group than in the remaining two groups after patients’ peripheral blood single nuclei were detected by flow cytometry,and the difference between the groups was statistically significant(p<0.05).Conclusion: 1.Tibial transverse transport can effectively promote the healing of ulcers in Patients with severe diabetic foot.2.The wounds of patients undergoing Tibial Transverse Transport l surgery have a large number of neovascularization,which confirms that the Ang-1Tie-2 signaling pathway is up-regulated in the peripheral blood and local areas of this group of patients,and promotes angiogenesis.3.Confirmation.The activation of endothelial progenitor cells in patients undergoing Tibial Transverse Transport surgery is increased,which provides a necessary condition for neovascularization. |
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