ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts.</p><p><b>METHODS</b>(1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured. The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope. (2) Two batches of hAMSCs of the third passage were obtained, then the expression of vimentin of cells was observed with immunofluorescence method, and the expression of cell surface marker CD90, CD73, CD105, and CD45 was detected by flow cytometer. (3) hAMSCs-CS of the third passage at culture hour 72 were collected, and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay. (4) Human fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured. Human fibroblasts of the third passage were used in the following experiments. Cells were divided into blank control group and 10%, 30%, 50%, and 70% hAMSCs-CS groups according to the random number table (the same grouping method below), with 48 wells in each group. Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS), while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS. The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12, 24, 48, and 72, respectively, and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments. (5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4), with 4 wells in each group, at post scratch hour (PSH) 0 (immediately after scratch), 12, 24, 48, and 72, the migration distance of cells was observed and measured with inverted phase contrast microscope. (6) Human fibroblasts were grouped and treated as in (5), with 3 battles in each group, and apoptosis rate of cells was detected by flow cytometer. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, one-way analysis of variance, LSD test, and t test.</p><p><b>RESULTS</b>(1) On culture day 3, most hAMSCs were in large form, and spindle-shaped with much prominences like fibroblasts or in flat polygonal shape. hAMSCs of the third passage were spindle-shaped. The expression of vimentin of hAMSCs of the third passage was strongly positive, and the expressions of surface markers CD90, CD73, and CD105 of the cells were positive, while the expression of CD45 of the cells was negative. (2) The content of IGF-Ⅰ, VEGF, EGF, and bFGF in hAMSCs-CS were respectively (11.7±1.0), (316±68), (6.1±0.4), and (1.49±0.05) pg/mL. (3) At culture hour 12-72, the proliferation activity of human fibroblasts in each hAMSCs-CS group was significantly higher than that in blank control group (with P values below 0.01), and the proliferation activity of human fibroblasts in 50% hAMSCs-CS group was the highest. (4) The width of scratch in two groups was nearly the same at PSH 0. The migration distance of cells in 50% hAMSCs-CS group was significantly longer than that in blank control group at PSH 12-72 (with P values below 0.01). (5) The apoptosis rate of human fibroblasts in blank control group was (16.2±2.4)%, which was significantly higher than that in 50% hAMSCs-CS group [(7.4±3.6)%, t=6.710, P<0.01].</p><p><b>CONCLUSIONS</b>hAMSCs-CS can promote proliferation and migration of human fibroblasts and inhibit the apoptosis of human fibroblasts.</p>
Subject(s)
Female , Humans , Male , Pregnancy , Amnion , Cell Biology , Apoptosis , Cell Movement , Cell Proliferation , Cells, Cultured , Culture Media, Conditioned , Chemistry , Enzyme-Linked Immunosorbent Assay , Epidermal Growth Factor , Metabolism , Fibroblast Growth Factor 2 , Metabolism , Fibroblasts , Cell Biology , Flow Cytometry , Insulin-Like Growth Factor I , Metabolism , Mesenchymal Stem Cells , Chemistry , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
Objective To investigate the application of OSCE evaluation system on the medical skills examination of clinical medical students and the significance of this system on the training of their medical skills.Methods 20 teachers examed 150 students by the OSCE evaluation system with 4 test stations,by comparing the score of the students of different test stations by one-way ANOVA and evaluating the system by questionnaire survey with Likert 5 on the degree of satisfaction and Likert 3 on effects and intrinsic influencing factors of the system.Results The score of the first and forth test stations was lower than that in the other stations(P＜0.05).8/5.48％ students and 1/5％ teachers were not satisfied with the system.The OSCE evaluation system could exam the psychological diathesis,ability of communication,cooperation,and clinical thinking,practical skill of the students and its effects are moderate (the score was more than 2.0).Evaluation on the intrinsic influencing factors:Students considered the questions were more difficulty in the 2nd,3rd,1st,4th test stations order.4/20％ teachers considered the questions of the second test station was easy.8/40％ teachers considered the duration of the second test station was too long.More than 70％ students and teachers considered the other indexes were rational.Conclusion The OSCE evaluation system can play an effective role in directing the teaching and learning.It can also help to culture the comprehensive capacity of the students.We should gradually improve the design of the system by considering the intrinsic influencing factors.
ABSTRACT
Objective To investigate whether pregnant rats exposure to ketamine cause offspring changes in space cognitive abilities and exploration abilities.Methods 3-month Sprague-Dawley female rats ( n =24)were randomly divided into four groups:group N (control group),group K1 (small doses of ketamine group),group K2 ( clinical anesthesia dose of ketamine group),group K3 ( large doses of ketamine group).3-month Sprague-Dawley male rats ( n =4) and female rats were mated at the same cage by the proportion of 2∶ 1.Pregnant mice were treated at tenth day:group N were treated saline with equal-volume to ketamine vein injection; group K1,group K2,group K3 administered vein injection 3,8,20mg/kg of ketamine.Then the 20-day offspring rats'learning and memory were assessed used Open Field Test ( record the time of the offspring in the central case through the number of grid within 2 min ) and Hole Board Test ( Counting the times of offspring stretch into the hole in 5 min) at postnatal days 20.Results In the Open Field Test,the retention time in central check of group N,group K2 and group K3 were (2.45 ± 1.23)s,(6.42 ±2.50)s,(6.41 ±2.19)s.Compared with group N,the retention time in central check of group K2 and group K3 were significantly higher (F=13.42,P＜0.01 ),and group K1 were not significant different ( t =1.33,P＞0.01 ),and the locomotion of group K1,group K2,group K3 were significantly reduced( ( 15.33 ± 6.81 ),( 13.75 ± 5.93 ),( 16.92 ± 6.54 ),F =4.24,P ＜ 0.05 ).In the Hole Board Test,the times of offspring stretch into the hole were not significant different comparing with the control group(F=2.17,P ＞ 0.05 ).Conclusion The dose of ketamine that equivalented clinical anesthesia can affect offspring rats' space cognitive abilities; but the exploring cognitive ability were not significantly influenced.