AIM: To determine the combined effect of transmuscle laser revascularization (TMR) and endothelial progenitor cells(EPCs) treatment on ischemic hindlimb of nude rats.METHODS: Mononuclear cells (MNCs) isolated from human umbilical cord-blood (HUCB) by density gradient centrifugation were expanded in vitro. Immunocytochemistry and flow cytometry studies were performed. EPCs were labeled with 1, 1'- dioctadecyl-1 to 3, 3, 3', 3'- tetramethyl-indocarbocyanine perchlorate (DiI) before injected into the laser induced channels or ischemic region. Acute ischemic limb was created in 4 groups of nude rats by ligating right external iliac artery. All animals were divided randomly into the following four groups: TMR+EPCs group: local transplantation of EPCs into laser channels; TMR group: transmuscular channels were created without EPCs; EPCs group: EPCs were injected into ischemic hindlimb; control group: ischemic model without TMR or EPCs. All rats underwent femoral artery ultrasonic blood flow measurements of the ischemic and nonischemic limbs to obtained a flow ratio [femoral artery flow index (FAFI): right femoral artery flow /left femoral artery flow] at baseline (after ligating artery immediately) and 28 days postoperation, and then the samples of ischemic limb muscle underwent histochemical and immunohistologic analysis. RESULTS: The attached cells expressed endothelial cell (ECs) markers (KDR, CD34, CD31, AC133 and von Willebrand factor) and exhibited function similar to that of ECs judged by Ac-LDL incorporation. Flow cytometric analysis disclosed that AT cells were positive for CD34 (62%?7%) and AC133 (57.2%?9.8%) at day 7 of culture. 28 days after therapy, FAFI was significantly higher in the TMR +EPCs (0.66?0.09, P0.05). FAFI in the control group was unchanged and no difference was found between TMR group and control group. TMR+EPCs (5.66?0.77), TMR (4.96?0.31) as well as EPCs (4.68?0.44) treatment resulted in an increased number of capillaries in the treated regional area compared with control group (2.60?0.31, P

Objective:To analyze the relationship between aquaporin 1 (AQP1) level and vascular calcification in patients with diabetes nephropathy.Methods:A total of 125 diabetic nephropathy patients admitted to Suzhou Hospital of Nanjing Medical University from March 2020 to March 2023 were retrospectively selected as case group. The case group was divided into group A (diabetes nephropathy stage Ⅰ and Ⅱ) with 31 cases, group B (diabetes nephropathy stage Ⅲ) with 32 cases, group C (diabetes nephropathy stage Ⅳ) with 39 cases, and group D (diabetes nephropathy stage V) with 23 cases. In these patients, 51 cases had vascular calcification, taken as the calcification group, and 74 cases had no vascular calcification, taken as the non calcification group. Sixty volunteers who underwent health examinations in the same hospital were selected as the control group. Receiver operating characteristic curve was used to analyze the predictive value of AQP1 on vascular calcification in diabetes nephropathy patients and to explore the related factors of vascular calcification in diabetes nephropathy patients.Results:Compared with the control group, AQP1 level and calcification rate in groups A, B, C and D were higher: 6.41 ± 1.04, 7.93 ± 1.23, 9.50 ± 1.52 and 11.37 ± 2.01 vs. 3.83 ± 0.56 ng/L, 6.45% (2/31), 28.13% (9/32), 51.28% (20/29) and 86.96% (20/23) vs. 0 ( P<0.05). Compared with group A, the level of AQP1 and calcification rate in groups B, C and D were higher ( P<0.05); compared with group B, the AQP1 level and calcification rate in groups C and D were higher ( P<0.05); compared with group C, the level of AQP1 and calcification rate in group D were higher ( P<0.05). Compared to the non calcification group, the levels of uric acid, homocysteine and cystatin C in calcification group were higher: (313.82 ± 38.72) μmol/L vs. (253.42 ± 30.14) μmol/L, (20.03 ± 3.01) μmol/L vs. (15.01 ± 2.71) μmol/L, (1.73 ± 0.26) mg/L vs. (1.30 ± 0.17) mg/L ( P<0.05). AQP1 was positively correlated with uric acid, homocysteine, and cystatin C ( P<0.05). The area under the curve of AQP1, uric acid, homocysteine and cystatin C in predicting vascular calcification in patients with diabetes nephropathy were 0.892, 0.803, 0.738 and 0.763, respectively. Taking whether vascular calcification occurs in patients with diabetes nephropathy as the dependent variable (no = 0, yes = 1), the variables of P<0.05 in the single factor analysis were selected for multivariate Logistic regression analysis. The results showed that uric acid, homocysteine, cystatin C and AQP1 were the main factors affecting vascular calcification in patients with diabetes nephropathy ( P<0.05). Conclusions:Serum AQP1 has a high predictive value for vascular calcification in diabetes nephropathy patients, and is expected to be used as a biomarker for early diagnosis of vascular calcification in diabetes nephropathy patients.