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Objective To investigate the effect of local injection of rat adipose-derived mesenchymal stem cells (ADSCs)on the phenotype of macrophages in skin wound.Methods The cryopreserved primary SD rat ADSCs were resuscitated and then sub-cultured.ADSCs of the third generation were used in the experiments.Thirty six SD rats were divided into ADSCs group (n =18) and control group (n =18) by random numbers table method.The full thickness skin wounds were established on bothsides of the spine.After the model establishment 0.2 ml ADSCs suspension labeled by live cell stain Chloromethylbenzamido derivatives of 1,l'-dioctadecyl-3,3,3',3'-tetramethylindo-carbocyanine perchlorate (CM-Dil) with the concentration of 5 × 106/ml was subcutaneously annularly injected in the skin wound of SD rats in ADSCs group.The SD rats in control group were given 0.2 ml serum-free Dulbecco modified Eagle medium (DMEM).On 3,7,and 14 days after injury,six rats were selected from each group to measure the wound area and healing rate.The healed wound tissues were harvested to observe the morphology by HE staining.The expressions of interleukin (IL)-10 were detected by immunohistochemistry staining.The double-positive expressions of CD68 and inducible nitric oxide synthase (iNOS) (M1 type macrophages) and of CD68 and arginase-1 (Arg-1) (M2 type macrophages) were detected by immunofluorescent staining.The distribution of CM-Dil-labeled ADSCs in healed wound tissue 14 days after injury was observed by inverted fluorescence microscope.Results (1) At day 3 after injury,the wound areas in two groups were covered with crust and surrounding redness,and the wound healing rates were slightly different;at day 7 after injury,the wound area of ADSCs group was significantly smaller than that of control group,and the healing speed and rate of ADSCs group was significantly higher than that of control group (P ＜ 0.01);at day 14 after injury,the healing rate of ADSCs group was nearly 99％ (P ＜ 0.01),and the healing skin tissue texture of ADSCs group was better than that of control group.(2) At day 3 after injury,there were a large number of inflammatory cells and disorganized collagen fiber in the wound areas of two groups;at day 7 after injury,the inflammatory cells infiltration reduced in ADSCs group compared with control group,and the collagen fiber arrangement in control group was in disorder;at day 14 after injury,the inflammatory cells in both groups obviously decreased,and ADSCs group had more new vessels and more orderly arrangement of collagen fiber than control group.CM Dil labeled ADSCs were seen in the healing wound tissue in ADSCs group.(3) At day 3 after injury,there was little difference in M1 type macrophage distribution in the two groups;ADSCs group had more M2 type macrophage cells than control group significantly (P ＜0.01);the expression of IL 10 in ADSCs group was not high,which did not differ from that of control group;at days 7 and 14 after injury,ADSCs group has fewer M1 type macrophage cells,more M2 type macrophage cells,and higher expressions of IL-10 than control group (P ＜ 0.01).Conclusion The ADSCs trasplantation can promote the change from M1 type to M2 type macrophages,facilitating wound regeneration and healing.
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Objective@#To investigate the effect of local autologous dermal fibroblasts transplantation on hypertrophic scar formation and wound healing quality in early scar formation. To explore the feasibility of fibroblasts for prevention and treatment of hypertrophic scar.@*Methods@#Dermal fibroblasts were isolated from the dorsal skin tissue of New Zealand white rabbits by mechanical method combined with enzyme digestion. Passage 3 cells were induced to differentiate into osteoblasts and adipocytes. The complete epithelialization time and hypertrophic scar formation after full-thickness skin defect were confirmed by pre-experiment study. In the experiment, 6 rabbits were used, left ear as experimental group and right ear as control group. In the experimental group, the passage 4 dermal fibroblasts labeled with 5-bromodeoxyuridine (5-BrdU) were injected subcutaneously around the wound and hypertrophic scar on 20 d (day 2 after epithelialization) and 30 d (most obvious scar hyperplasia) after surgery. As a control group, physiological saline was injected following the same protocol. On 37 days after surgery, the hypertrophic scar tissue were harvested and assessed by gross view and histological examination. The transplanted cells were detected by immunofluorescence staining, transforming growth factor beta 1 (TGF-β1) and decorin(DCN) mRNA expression were assayed by real-time fluorescence polymerase chain reaction (RT-PCR), and the protein expression of TGF-β1、DCN、Collagen type Ⅰ and type Ⅲ were tested by enzyme linked immunosorbent assay(ELISA).@*Results@#Compared with the control group, the scar in the experimental group was flatter and softer, the color was slightly lighter, and the volume was reduced. The histological results showed that compared with the control group, the number of fibroblasts and inflammatory cells in the superficial dermis was reduced, the proliferation of connective tissue and collagen deposition were reduced, and the basal cells and collagen fibers were arranged in order in the experimental group. The results of RT-PCR showed that TGF-β1 mRNA expression level in the hypertrophic scar tissue reduced significantly and DCN increased significantly in the experimental group, compared with the control group (P<0.01). ELISA results showed that TGF-β1, type Ⅰ and Ⅲ collagen contents in hypertrophic scar tissue were significantly lower than that in the control group, while the DCN expression was significantly increased (P<0.01).@*Conclusions@#The rabbit dorsal dermal fibroblasts significantly inhibited the hypertrophic scar in rabbit ears. The mechanism may be related to the up-regulation of DCN expression and down-regulation of TGF-β1 expression, reduction of type Ⅰ and Ⅲ collagen synthesis and extracellular matrix deposition, in the hypertrophic scar tissue.
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Objective To propose an extraction algorithm for extraction of breast tumor boundary in its ultrasonic image. Methods A novel Global-Local Energy function was proposed in the variational Level Set formulation for the curve evolution. By enhancing the intensity of local information, the breast tumor boundary was obtained from ultrasonic images accurately. Results Experiments on a synthetic ultrasound image showed that this method was more accurate at segmenting the object from the background than the Chan-Vese algorithm based on the global optimization. Experiments on 103 ultrasonic breast tumor images also showed that the boundaries were extracted accurately with this method without any pre-processing step. Conclusion This proposed method is expected for the fast and accurate boundary extraction of ultrasonic breast tumor boundary from ultrasonic image.
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The chemical constituents of the gorgonian Isis minorbrachlasta have been studied for the first time. Th?marine specimen was collected off the Nansha Islands in the South China Sea . On the basis of spectral correlation (1H NMR,13C NMR,IR)four known compounds:gorgosterol,22 - epi-hippuristanol, ceramide and batyl alcohol, were indentified.