Objective:The three-dimensional(3D) reconstruction method of calf perforator vessel was explored to provide the basis for clinical safe and accurate harvesting of calf chain perforator flap.Methods:Gelatin-lead oxide is automatically perfused the femoral artery of 3 fresh adult lower limbs specimens. Data were collected after the CT scan of the full limb, 3D reconstruction of bones, arteries and their perforators was processed in Mimics Research 19.0 software. Dynamic display of vascular hierarchy and adjacent relationships by adjusting CT values. And show the anatomical region of each perforator artery in different colors, measure the feeding area of the single perforator using Scion Image software. The 3D printed calf bone and vascular model can be used for direct observation. The specimen anatomy was measured according to a perforator vessel parameter with an outer diameter of 0.5 mm through the deep fascia. The number of perforators, vessle pedicle length, and outer diameter were observed and statistically analyzed.Results:The digitized 3D reconstruction images can clearly show the 3D structured of the calf artery and perforator vessels and micro vessels, as well as the course, distribution characteristics of blood vessels, and the 3D relationship with adjacent structures. The printed 3D model visually observes the hierarchy of the perforator vessels and the anastomosis relationship between the perforators. 3 calf specimen arteries with perforator artery 22±4, vessel pedicle length (46.7±18.3) mm, outer diameter (0.8±0.3) mm, the average blood supply area of the single perforator (38.2±10.7) cm 2. Conclusion:The Mimics Research 19.0 software should be used to dynamically reconstruct the 3D structure of the branch vessel by adjusting the CT value, which improves the display effect of the perforator vessels. The 3D printing model makes the source, course, diameter and distribution range of the perforator vessels visually visible, which provides experimental support for the clinical construction of digital calf chain perforator flap harvest scheme for patients.

Objective:The three-dimensional(3D) reconstruction method of calf perforator vessel was explored to provide the basis for clinical safe and accurate harvesting of calf chain perforator flap.Methods:Gelatin-lead oxide is automatically perfused the femoral artery of 3 fresh adult lower limbs specimens. Data were collected after the CT scan of the full limb, 3D reconstruction of bones, arteries and their perforators was processed in Mimics Research 19.0 software. Dynamic display of vascular hierarchy and adjacent relationships by adjusting CT values. And show the anatomical region of each perforator artery in different colors, measure the feeding area of the single perforator using Scion Image software. The 3D printed calf bone and vascular model can be used for direct observation. The specimen anatomy was measured according to a perforator vessel parameter with an outer diameter of 0.5 mm through the deep fascia. The number of perforators, vessle pedicle length, and outer diameter were observed and statistically analyzed.Results:The digitized 3D reconstruction images can clearly show the 3D structured of the calf artery and perforator vessels and micro vessels, as well as the course, distribution characteristics of blood vessels, and the 3D relationship with adjacent structures. The printed 3D model visually observes the hierarchy of the perforator vessels and the anastomosis relationship between the perforators. 3 calf specimen arteries with perforator artery 22±4, vessel pedicle length (46.7±18.3) mm, outer diameter (0.8±0.3) mm, the average blood supply area of the single perforator (38.2±10.7) cm 2. Conclusion:The Mimics Research 19.0 software should be used to dynamically reconstruct the 3D structure of the branch vessel by adjusting the CT value, which improves the display effect of the perforator vessels. The 3D printing model makes the source, course, diameter and distribution range of the perforator vessels visually visible, which provides experimental support for the clinical construction of digital calf chain perforator flap harvest scheme for patients.

Objective:To evaluate the clinical effect of fibula flap in the repair of large segment of composite tissue defect in upper limb.Methods:From April, 2015 to June, 2019, 7 patients with large composite tissue defects in upper limbs were treated. All of them with various skin, vessel, nerve, tendon and other tissue defects. Repairing was well planned before surgery according to the type, location and size of defect. While in repairing of the bone and skin defect, fibula flap was taken from the shank and to repair the defects of nerve, tendon and vessels in upper limb. Regular followed-ups were made after surgery.Results:The 7 fibula flaps all survived. Postoperative follow-up ranged between 8 to 36 (averaged of 15) months. All the reconstructed limbs were in satisfactory appearance and function recovery. All the patients were able to manage their daily activities and live independently. The shape and function of donor sites were good. According to the Enneking system, the outcomes were graded as excellent in 4 cases and good in 3, with the average score was 25.9 points.Conclusion:Free grafting of vascularised fibula flap is especially feasible to be used in the repair of large bone tissue defect of upper limb. It repairs the defects of skin, vessel, nerve and tendon with the flap from a single donor site.

Objective:To introduce an improved method for reconstruction of thumb by transplanting partial second toe combined with hallucis flap.Methods:Since April, 2015 to December, 2019, 12 thumbs were reconstructed by transplanting partial second toe combined with hallucis flap. The average age of the patients was 28.5 years old. The severity of thumb defect was grade III-V. The compound tissue flap of common vascular pedicle was obtained from the hallux and the partial second toe of the contralateral foot, according to the condition of the defect of thumb. The hallucis flap was rotated transversely and connected with the partial second toe. Make up the characteristics and defects for the reconstructed thumb. The remaining skin and soft tissue of the second toe and the donor area were tiled to cover the wound. The patients were followed-up regularly.Results:All of the 12 transplantations completely survived. Postoperative follow-up period ranged from 6 to 26 months, 14 months in average. Appearances of all reconstructed thumbs and the donor toes was satisfactory. According to the Functional Assessment Criteria of the Upper Limb set by the Hand Surgery Society of Chinese Medical Association, the outcomes were graded excellent in 9 cases and good in 3 cases. All patients were able manage daily activities and living independently. Shape and function of donor foot were good.Conclusion:Part of the second toe combined with the hallucis flap transfer for thumb reconstruction may effectively improve the appearance of the reconstructed thumb. The reconstructed thumb is symmetrical, good in shape and satisfactory in function.

Objective: To introduce the method for reconstruction of thumb defect by transplanting free flap of second toe combined with the hallucis flap. Methods: From June, 2012 to February, 2017, a total of 9 cases of thumb defect were treated. The average age of these 9 patients was 26 years. The plane of thumb defect was class II area A to class III area A. According to the condition of thumb defect, designed the incision on the hallux and the second toe of the contralateral foot, and cut the 2nd digit and big-toe nail flap with a common arterial trunk. The nail of hallucis flap was rotated 90 degree and connected to the distal end of the 2nd toe. The hallucis flap covered the narrow neck of the 2nd toe. Thus, the circumference of the reconstructed finger and the length of the toenail were increased. Regular followed-up was made after operation. Results: All 9 transplantation flaps survived, and donor sites healed primarily. Postoperative followed-up period ranged from 4 to 12 (averge,7) months. All the reconstructed thumbs survived and donor toes were in satisfactory appearances. According to the Functional Assessment Criteria of the Upper Limb Formulated by the Hand Surgery Society of Chinese Medical Association, the outcomes were graded as excellent in 6 cases and good in 3 cases. All the patients were able to manage their daily activities independently. Donor toe injury was small, and their shape and function was good. Conclusion Free flap of second toe combined with hallucis can effectively improve the appearance of the reconstructed thumb. The appearance of the reconstructed thumb is symmetrical, beautiful, and the function is good.

BACKGROUND: Mesenchymal stem cells can protect and repair the liver of rats with liver failure, but the mechanisms are not completely clear. OBJECTIVE: To explore the protective effects and related mechanisms of intravenous injection of human adipose-derived mesenchymal stem cells on acute liver failure in rats. METHODS: Thirty-six Sprague-Dawley rats (provided by Qingdao Daren Fucheng Animal Husbandry Co., Ltd. in China) were randomly divided into control group, model group and transplantation group. Animal models of acute liver failure were established by intraperitoneal injection of D-galactosamine in the model group and the transplantation group. One day after modeling, the rats in the transplantation group were injected with human adipose-derived mesenchymal stem cell suspension, and those in the model group were injected with the same amount of saline. After 1 and 3 days of cell transplantation, the serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were measured. Three days after cell transplantation, the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-10 were detected, the pathological changes of the rat liver were observed by hematoxylin-eosin staining, and the activity of glycogen synthase kinase-3β protein in the liver tissue was detected by western blot. RESULTS AND CONCLUSION: Compared with the model group, there was a significant reduction in the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, tumor necrosis factor-α, interleukin-6 and interleukin-10 in the transplantation group (P < 0.05). Inflammation and necrosis of liver tissues in the transplant group were alleviated compared with the model group. The activity of glycogen synthase kinase 3β in the liver tissue of the transplanted group was lower than that of the model group (P < 0.05). Overall, these results indicate that human adipose-derived mesenchymal stem cells can alleviate hepatic inflammation and pathological injury, and improve the liver function in rats with acute hepatic failure. Moreover, the mechanism may be related to the inhibition of glycogen synthase kinase 3β activity.

Objective To investigate the effect of Irisin on proliferation,apoptosis,migration and invasion of human cholangiocarcinoma cell line Hucct-1.Methods After treatment with Irisin,CCK-8 assay and flow cytometry assay were conducted to investigate the effect of Irisin on proliferation and apoptosis of cholangiocarcinoma cells.Scratch test and transwell invasion assay were used to studythe effect of Irisin on the migration and invasion ability of cholangiocarcinoma cells.Western blot was utilized to detect the expression of E-cadherin,N-cadherin and Vimentin in cholangiocarcinoma cells.Results CCK-8 assay showed that Irisin inhibited cholangiocarcinoma cell proliferationin a dose-dependent manner.Flow cytometry assay showed that the apoptosis rate of Irisin group [(14.8 ±0.9)％] was higher than that in the control group [(5.4±0.6)％],(P＜0.05).The scratch test showed that the rate of cell scratch healing in Irisin group [(15.0± 1.0)％] was significantly lower than that in the control group [(28.0±2.0)％] (P＜0.05).Transwell invasion test showed that the number of cells in Irisin group was (96.0±7.0),which was significantly lower than that in control group (155.0± 9.0) (P＜0.05).Western blot showed that the expression of E-cadherin increased and N-cadherin and Vimentin decreased after Irisin treatment.Conclusion Irisin inhibits proliferation,migration and invasion and promote apoptosis of cholangiocarcinoma cells.

Objective To investigate value of peripheral NLR and PLR for the survival of patients with neuroblastoma.Methods The clinical data of 41 neuroblastoma patients were analyzed by the Kaplan-Meier,Log-rank test,and multivariate COX regression.Results NLR,PLR levels of neuroblastoma patients were significantly higher than that in the healthy control group (1.81 ±0.29 vs.1.07 ±0.29,P ＜ 0.01) (169 ± 23 vs.76 ± 3,P ＜ 0.01);The elder the age,the higher the clinical stages,the higher the serum levels of NSE,and urine VMA were,the higher was the NLR (x2 =3.93,6.286,7.676,6.689,all P＜0.05) and PLR (x2 =4.111,5.707,8.019,8.922,all P ＜0.05).The higher the serum level of LDH,the higher was the NLR (x2 =7.769,P =0.02).3-year overall survival in low NLR group was 84％ and that in high NLR group was 73％ (x2 =4.002,P =0.045);3-year progression-free survival in low NLR group was 74％ and that in high NLR group was 50％ (x2 =4.082,P =0.043);3-year progression-free survival of low PLR group was 85％ and high PLR group was 38％ (x2 =9.388,P =0.002).The clinical stages,MYCN genetic expression,NLR levels were independent factors for the overall survial in patients with neuroblastoma (P ＜ 0.05).Conclusion Pretreatment NLR level can effectively predict the prognosis of neuroblastoma.

Background Researches showed that transforming growth factor-β2 (TGF-β2) promotes the activity of human Tenon capsular fibroblasts (TFs),which plays a role in the scarring of filtering blebs after antiglaucoma surgery.However,its mechanism is not fully clear.Lysyl oxidases (LOXs) are important extracellular matrix proteases which can catalyze the cross-linking of collagen and elastin.Investigating the impact of TGF-β2 on the expression of LOXs has a great significance for the understanding of the pathogenesis of filtering bleb scarring and its prevention.Objective This study was to investigate the effect of TGF-β2 on the expression of LOXs in cultured human TFs.Methods The TFs at 4-8 generations were divided into normal control group and different concentrations of TGF-β2 treated-groups,and 100,200,400,800 μ1 of TGF-β2 with the final concentration of 2,4,8 and 16 ng/ml was added into the medium to treat human TFs respectively for 24 hours.The LOXs in the cells were detected by Western blot to determine the optimal dose of TGF-β2.The 4 ng/ml TGF-β2(200 μ1) was used to treat human TFs for 6,12,24 and 48 hours respectively,and the change of LOXs expression in the cells over time was assayed by Western blot.The expression and distribution of LOX protein in the normal cells and TGF-β2-treated cells was detected by using immunofluorescence technique.This study was approved by Daping Hospital of Third Military Medical University Ethic Commission.The guardians of the patients who offered the specimen knew the purpose of the study and signed informed consent.Results Western blot assay showed that the expressions of LOX,LOXL1,LOXL2,LOXL3 and LOXL4 in the cells were gradually elevated from the normal control group and 2,4,8,16 ng/ml TGF-β2-treated groups,showing significant differences among the groups (F =37.338,13.438,31.067,11.767,15.167,all at P＜0.01).The expression of LOXL2 protein in the cells was 0.68±0.07,1.09±0.10,1.32±0.07,1.50± 0.06 and 1.89±0.12 in the normal control group and 6-,12-,24-and 48-hour groups respectively after 4 ng/ml TGF-β2 treatment,with a significant increase over time (F =82.832,P=0.000).The expression of LOX was weak in the normal cultured TFs,while the fluorescence intensity of LOX expression was evidently enhanced in the cytoplasm of the cells in the TGF-β2-treated group.Conclusions TGF-β2 upregulates the expressions of LOXs in human TFs in a dose-and time-dependent manner,which probably offers a basis for the further study on the prevention of filtering bleb scarring after glaucoma surgery.

BACKGROUND:Recent studies have shown that adipose-derivedmesenchymal stem cells (ADMSCs) not only have multilineage differentiation potential, but also exert an important role in blood sugar balance and hormone production.OBJECTIVE:To observe the differentiation potential of human ADMSCs (hADMSCs) into functional islet-like cells and the therapeutic effect of hADMSCs transplantation in diabetic rats.METHODS:PDX-1 gene was transfected into hADMSCs by adenovirus. Cell differentiation and insulin secretion were identified and detected by dithizone staining and ELISA, respectively. Twenty male Sprague-Dawley rats were randomly divided into control group (n=4), diabetes group (n=8) and transplantation group (n=8). Rats in the latter two groups were subjected to making diabetic models by 65 mg/kg streptozotocin injection. Afterwards, rats in the transplantation group were given PDX-1 transfected ADMSCs via the tail vein.RESULTS AND CONCLUSION:At 15 days after transfection, the number of insulin positive cells and insulin secretion were both increased significantly (P < 0.05). Fasting glucose levels in the transplantation group decreased significantly (P < 0.05), while the body weight increased significantly (P < 0.05). In the diabetic group, the fasting glucose level still maintained at a high level, and the body weight of rats was significantly decreased. These results implicated that PDX-1 gene could induce hADMSCs differentiating into functional islet-like cells. PDX-1 transfected ADMSCs transplantation is effective in treating diabetic rats, but the mechanism needs further study.