The clinical experiences in the appieation of umbilical-thoracic skin flap in the coverage of the defect of the forearm in 9 cases were reported. The flap was supplied by the branches of inferior epigastric artery.The biggest flap was 8.5×28cm,the smallest one was 7× 16cm.All flaps surviVed.The results were satisfactory. The advantages of the flap were:(1)potients felt comfortable when the upper extremity was immobilized at the side of the they;(2)the size of skin taken from the do...
Objective To explore effects of zinc on the contents of cycl in D2, cycl in-dependent kinase 4 (CDK4), and their DNA and total cellular protein in human umbil ical cord blood-drived mesenchymal stem cells (hUCBMSCs). Methods hUCBMSCs were isolated and cultured by density gradient centrifugation adherence method in vitro. At the serial subcultivation, the hUCBMSCs were randomly divided into 7 groups. In control group, hUCBMSCs were cultured with DMEM medium (containing 15%FBS). In treatment groups, hUCBMSCs were cultured with DMEM medium (containing 15%FBS plusZnSO4•7H2O). The final concentrations of zinc were 0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 mg/L, respectively. The cellular surface antigens of CD29, CD34, CD44, and CD45 at the 3rd generation of hUCBMSCs were detected by flow cytometry. MTT assay was used to detect cell activity of the 3rd generation of hUCBMSCs. The optimum concentration of zinc was selected by the results of MTT as experimental group. The cell growth curves of experimental group and control group were drown by counting cell. The cell surface antigen, reproductive cycle, and DNA content were detected by flow cytometry motheds. The contents of cycl in D2 and CDK4 were detected by Western blot method. Results The positive expression rates of CD29 and CD44 were more than 70% in hUCBMSCs. The cell activity of 2.5 mg/L treatment group was superior to other treatment groups, as experimental group. At 7, 14, and 28 days, the contents of DNA, total cellular protein, cycl in D2, and CDK4 of hUCBMSCs were significantly higher in experimental group than those in control group (P lt; 0.01). The percentage of hUCBMSCs at S stage and prol iferation index in experimental group were also significantly higher than those in control group (P lt; 0.01). Conclusion Zinc (0.5-4.5 mg/L) has the promoting effect on the hUCBMSCs activity, and 2.5 mg/L is the optimal concentration. Zinc (2.5 mg/L) can accelerate the prol iferation and DNA reproduction of hUCBMSCs and increase the contents of cycl in D2 , CDK4, and cellular total protein.
Objective To investigate the method and conditions of isolation，proliferation of multipotent mesenchymal stem cells（MSCs）from human umbilical cord blood in vitro, and to induce osteogenic and adipogenic differentiation directly for identification. Methods Human umbilical cord blood was collected in asepsis condition, isolated by density gradient centrifugation,or sedimented red cell with methylcellulose, and then the same centrifugation was done, or obtained by negative immunodepletion of CD34+. These isolated mononuclear cells were used to carry on plastic adherent culture. To obtain single cellderived colonies, these cells were proliferated clonally in medium which consists of L-DMEM orMesencultTM medium and 10% fetal calf serum（FCS） respectively, then their differentiation potentiality to osteoblasts and lipoblasts was tested. Results The mononuclear cells isolated by sedimented and centrifugated way cultured in MesencultTM medium and 10%FCS were most available. These adhesive cells could become obviously short rodshape or shuttle-shape cells after 5-7 days.The colonies form well in 3rdpassage cells. The mononuclear cells obtained by onlycentrifugalized in density gradient were hard to form colony, isolated by immunomagnetic beads were hard to culture. The surface antigens of these colonies cells presented CD29, CD59, CD71 but not CD34,CD45 and HLADR etc. The colony cells differentiating into osteoblasts that produce mineralized matrices, stained by alizarin red, and differentiating into adipocytes that accumulate lipid vacuoles, stained by oil red. Conclusion MSCs can be isolated from human umbilical cord blood and proliferate it in vitro. The way that mononuclear cells are sedimented red cell by methylcellulose and cultured by MesencultTM medium and 10% FCS is the valid method of isolation. Proliferation colonies cells present matrix cell immunophenotypes, and candifferentiate into osteoblasts and adipocytes.
Objective To investigate the effect on proliferation and apoptosis of vascular endothelial cells exposed to high glucose. Methods The cultured human umbilical vein endothelial cells (HUVEC) were treated with high glucose at concentrations of 10,20,30,40,50 mmol/L for 2,4,6,8,10,12,14 days,cpm value was studied by using tritium-labelled thymidine deoxyribose(3H-TdR)incorporation assay.Flow cytometry was used to detect apoptosis index,proliferation index and cell cycle. Results Treated with high glucose,the proliferation index and cpm were significantly reduced in a dose and time dependent manner than the control groups(Plt;0.05).The apoptosis index of HUVEC were higher compared with the control groups(Plt;0.05).The percent of the cultured cells in G1 phase in all high glucose groups were increased,the percent in S phase were largely decreased(Plt;0.05). Conclusion Exposed to high glucose,the apoptosis of HUVEC was accelerated and the suppressive effect of high glucose on the proliferation of HUVEC was observed.Endothelial cells were possibly arrested in G1/S checkpoint. (Chin J Ocul Fundus Dis,2000,16:177-180)
Objective To observe the survival of human umbilical cord derived mesenchymal stem cells (hUC-MSCs) after injection into the vitreous of rabbits,and the animal safety under those procedures.Methods Twentyseven pigmented rabbits were randomly divided into 3 groups (intravitreal injection 1 week group，2 weeks group and 4 weeks group)， each with 9 rabbits.For each animal the right eye was the experimental eye receiving hUCMSCs injection,while the left eye was the control eye receiving culture medium. The rabbit eyes were examined by slitlamp microscope, indirect ophthalmoscopy, fundus photography, fundus fluorescence angiography（FFA）and Tonopen tonometer before and after injection. hUCMSCs were labeled by CMDil in vitro, and their survival status was measured by confocal fluorescence microscopy, light microscope and transmission electron microscope at 4 weeks after injection. Results Four weeks after injection, a large number of the hUCMSCs were still alive in the vitreous cavity. The overall condition of those rabbits was good. The anterior segment and retina of experimental eyes were normal, without hyperfluorescence, hypofluorescence and leakage in the retina at 1,2 and 4 weeks after injection. There was no significant difference on IOP before and after injection at different time points (P＞0.05), and no obvious changes at cornea, anterior chamber angle,lens,retinal structure by.light microscope and transmission electron microscope examination.Conclusion hUC-MSCs can survive in the rabbit vitreous for four weeks；intravitreal injection of hUCMSCs was safe and feasible.