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Objective@#To elaborate the characteristics and advantages of Whole-Mount immune fluorescence staining by observing the lymphatic vessels of mice.@*Methods@#The ear skin tissue, the hindlimb lymphatic vessels and the mesenteric lymphatic vessels were harvested from normal C57 mice. The tissue samples were subjected to whole-tissue immunofluorescence staining.These tissue samples were fixed by paraformaldehyde, blocked by bovine serum and incubated in primary and secondary antibodies. Then, the lymphatic vessels were observed and analyzed in these samples with a confocal laser-scanning microscope.@*Results@#The capillary lymphatic vessels and lymphatic endothelial cells can be clearly showed in the ear skin. The valves and smooth muscles can be clearly showed in the hindlimb and mesenteric lymphatic vessels by Whole-Mount immunofluorescence staining.@*Conclusions@#The whole-tissue immunofluorescence staining technique can observe the external morphology of lymphatic vessels clearly and stereoscopically, and can deeply observe the internal structure of lymphatic vessels. This technique can provide more accurate study on physiology and pathology of lymphatic vessels.
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Objective@#To investigate the anti-photoaging effects of human umbilical cord mesenchymal stem cells derived extracellular vesicles (hucMSC-EVs) on dermal fibroblasts.@*Methods@#Human fibroblasts were pretreated with hucMSC of different concentrations for 24 hours before ultraviolet B (UVB) irradiation. Immediately post irradiation, the intracellular reactive oxygen species (ROS) were analyzed using reactive oxygen detection kit. After 72 hours irradiation, cell proliferation was assessed using CCK-8 assay, and the percentage of senescent cells was evaluated by β-galactosidase (SA-β-gal) staining.@*Results@#The UVB irradiation increased intracellular ROS level, decreased the cell proliferation from 100% to(77.33±2.89)%, and increased the percentage of SA-β-gal positive senescent cells from (6.70±0.46)% to (17.67±1.53)%. The hucMSC-EVs pretreatment decreased the intracellular ROS level, stimulated cell proliferation from (77.33±2.89)% to (90.67±2.52)% and( 96.00±5.57)% , and decreased the positive rate of SA-β-gal positive cells from (17.67±1.53)% to (8.38±0.56)% and (7.07±1.10)%.@*Conclusions@#The hucMSC-EVs preconditioning may protect dermal fibroblasts from UVB induced photoaging, by reducing intracellular ROS.
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Objective@#To explore the relationship between protease-chymase secreted by mast cells, and activated transforming growth factor-β1(TGF-β1), in skin with secondary lymphedema(SLE)in lower extremity, so as toidentify the key factors in fibrosis of lymphedema.@*Methods@#In this study, the affected limb skin of 7 SLE patients was includedas the experimental group, and normal skin tissue of the lower limb of 7 volunteers was used as controls. The skin samples were assayed by Masson staining, and the expressions of chymase and TGF-β1 were assayed by immunohistochemistry, immunofluorescent staining and enzyme-linked immunosorbent assay.@*Results@#There was obvious fibrosis in the skin of lower extremity in patients with lymphedema. The number of MCs andthe expressions of chymase, latency-associated peptide TGF-β1 (LAP TGF-β1) and TGF-β1 were all significantly increased in fibrotic skin in lymphedema, compared with those in normal skin. At the same time, the chymase-containing mast cells accumulated in the lymphatic vessels, with higher expression of TGF-β1.@*Conclusions@#The expression of chymase and TGF-β1 was significantly increased in the fibrotic skin insecondary lower extremity lymphedema. The increased expression of chymase in the skin may activate more TGF-β1 expression, and the increased TGF-β1 may promote skin fibrosis in SLE.
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Objective@#To explore the appropriate treatment cycle of complex decongestive Therapy (CDT) for extremity lymphedema, which is the summary of the relationship between treatment time and curative effect.@*Methods@#From January to December 2015, 203 patients with limb lymphedema were divided into four stages according to the degree of edema and the degree of fibrosis, that is, four groups. All patients were given CDT treatment for a course of treatment (4 weeks). During the treatment period, the multi frequency bioelectrical resistance body composition analyzer was used to monitor the change of tissue moisture and the tape diameter was used to measure the circumference of the affected limb. SPSS software is used to analyze the data.@*Results@#203 cases were treated with CDT therapy. Water in limb tissue and limb circumference of affected side was decreased significantly (P<0.01). To the tissues water dynamic changes, in the first group there was a significant difference (P<0.01) in the first week and second week. There was no obvious difference in 2-4 weeks; In the 2, 3 and 4 groups, there was significant difference in the first week and the second week, as well as the second week and the third week (P<0.05). And there was no difference in the third and fourth week. To the circumference dynamic change, in the first group there was no significant difference in patients in the first and the second week; In the third group, there was significant difference in the first week and the second week, as well as the second week and the third week (P<0.05). There was no significant difference in the last two weeks; In the fourth group, there was significant difference in the first week and the second week, as well as the third week and the fourth week(P<0.01). There was no difference in the second week and the third week. There is a correlation between the tissue water and the circumferential diameter (r=0.643). The patient has a good subjective feeling.@*Conclusions@#CDT therapy is effective in the treatment of lymphedema. For the first stage of edema, the therapy lasted two-week at least; For the second and the third stage edema, a 3 week therapy was suitable; For the fourth stage therapy, a four-week therapy is appropriate. It is suggested that CDT therapy should be maintained for at least half a year or so.
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Objective@#To evaluate the diagnostic value of indocyanine green(ICG)lymphography in primary lymphedema of extremities.@*Methods@#61 patients with limb lymphedema were enrolled in the study. The contralateral healthy limbs were used as controls. After intradermal injection of Indocyanine Green, dynamic observation of lymphatic vessels and lymph flow was performed using PDE near-infrared fluorescent instrument. The frequency of lymphatic contraction was calculated.@*Results@#Lymphatic vessel images were clearly visualized in healthy limbs. Inguinal or axillary lymph nodes can be visualized when examined 30 minutes after injection. Lymphatic contraction frequency was 1(0.33-5.00)time per minute. 94.8% lymphedematous limb demonstrated delayed lymphatic vessel or lymph nodes. "Dermal backflow" and diffused shadow pattern can be observed in all affected limbs. Abnormal lymphatic vasculature and contraction can be visualized. The frequency of lymphatic contraction in the affected limb was higher compared to controls: 2.5(0.5-7.0) times per minute.@*Conclusions@#ICG lymphography possess high sensitivity for lymphedema diagnosis. ICG lymphography furthers our understanding of pathophysiological alterations of lymphatic disorders.