1.Human bone marrow mesenchymal stem cells into vascular endothelial cells before and after differentiation induced differential expression of eNOS/activity and its metabolites
Xuelan LUO ; Wei CHEN ; Guojun MO ; Peng YANG ; Hesheng OU
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(3):386-391
Objective To investigate the differences in eNOS gene expression,activity and its metabolites before and after human bone marrow mesenchymal stem cells (hBMSCs) are induced into vascular endothelial cells.Methods hBMSCs were induced into vascular endothelial cells.The morphological changes of the cells were observed under inverted microscope.Transwell assay was used to detect the cells' migration ability.The protein expression of eNOS was detected by immunofluorescence and Western blot.The activity of eNOS was detected by ELISA and the content of NO in cell culture supernatant was determined by nitrate reduction method.Results Compared with those in the undifferentiated group,the morphological changes of the differentiated cells were obvious.Cell migration ability increased by 238.10% (73.000±7.002 vs.21.000±4.359,P<0.05).The expression of eNOS protein increased by 114.72% (0.423±0.011 vs.0.197±0.079,P<0.05).The activity of eNOS was enhanced by 157.49% (4.967±0.073 vs.1.929±±0.103,P<0.05).The synthesis and release of NO increased by 155.67% (184.909±1.853 vs.72.323±0.426,P<0.05).Conclusion After hBMSCs are induced into endothelial cells,the expression of eNOS gene increases,their activities increase,synthesis and release of the metabolite NO increase.It may provide a basis for prevention and treatment of cardiovascular diseases with stem cells.
2.Effects of miR-24 on Endothelial Nitric Oxide Synthase Gene Expression and Tube Formation in Human Umbilical Vein Endothelial Cells
Wei CHEN ; Guojun MO ; Xuelan LUO ; Hui WANG ; Peng YANG ; Hesheng OU
Chinese Circulation Journal 2016;31(8):797-801
Objective: To investigate the effects of miR-24 on endothelial nitric oxide synthase (eNOS) gene expression with regulation and endothelial cell proliferation, migration, tube formation in human umbilical vein endothelial cells (HUVECs). Methods: Constructed high expression plasmid of miR-24 and miR-24 antisense sequence were introduced into HUVECs and the cells included in 3 groups: Control group, miR-24 group and miR-24 inhibitor group. HUVEC proliferation was detected by MTT test, migration was measured by Scratching and Transwell methods, tube formation was examined by Matrigel assay; mRNA and protein expressions of eNOS and Sp1were determined by RT-PCR and Western blot analysis respectively. Results:①Compared with Control group, miR-24 group had decreased cell proliferation by 45.45% as (0.36 ± 0.04) vs (0.66 ± 0.08),P<0.05; miR-24 group had lower speed of cell migration, decreased number of cell migration by 74.75% as (30.25±3.78) vs (119.80±10.94),P<0.01 and there was no obvious tube formation.②Compared with Control group, miR-24 group showed reduced eNOS mRNA expression by 46.2% as (0.49±0.02) vs (0.91±0.01),P<0.05, reduced protein expression by 49.07% as (0.55±0.05) vs (1.08±0.05),P<0.05; meanwhile, decreased Sp1 mRNA expression by 44.9% as (0.49±0. 01) vs (0. 89±0.02)P<0.05, decreased protein expression by 54.90% as (0.46±0.02) vs (1.02±0.04),P<0.05. In miR-24 inhibitor group, the above indexes were lower than Control group but higher than miR-24 group, the amount of tube formation and the length of tubes were similar between Control group and miR-24 inhibitor group. Conclusion: MiR-24 may inhibit HUVECs proliferation, migration, tube formation and suppress eNOS expression; Sp1 might be one of the important regulators.
3.Establishment of UPLC Characteristic Chromatograms and Study on the Differences of Chromaticity Values among Cornus officinalis and Its Different Wine-processed Products
Yao HUANG ; Xuelan ZHANG ; Yuqin LUO ; Lihong DENG ; Chaozuan FANG ; Mei WEI
China Pharmacy 2021;32(2):206-212
OBJECTIVE:To establish characteristic chromatogram of Cornus officinalis and its different wine-processedproducts,investigate the differences of chromaticity values,and analyze them with chemical pattern recognition technology.METHODS:UPLC method was adopted. Using loganin as reference,UPLC characteristic chromatograms were drawn for 10batches of C. officinalis and 20 batches of different wine-processed products (stewing with wine,steaming with wine). TCMFingerprint Similarity Evaluation System(2012A edition)was used for similarity evaluation,and common peaks were confirmed.The chromaticity values [lightness(L),red and green tone value(a),yellow and blue tone value(b),color difference value(ΔE)]were determined by spectrophotometer. SPSS 20.0 and SIMCA 14.0 software were used for cluster analysis,principal componentanalysis and partial least squares-discriminant analysis;taking the area of characteristic peak and chromaticity value as indexes,andthe variable importance projection greater than 1 as the standard,the difference markers affecting its quality were screened.RESULTS:There were 6 common peaks in the chromatograms for decoction piece of C. officinalis,7 common peaks forwine-processed C. officinalis(stewing with wine)and wine-processed C. officinalis(steaming with wine). Four components wereidentified as gallic acid,5-hydroxymethylfurfural,morroniside,loganin. 5-hydroxymethylfurfural was produced after processing.The similarity between C. officinalis and different wine-processed products (stewing and steaming with wine) was low(0.869-0.937,0.845-0.944),but the similarity between different wine-processed products was higher than 0.99. ΔL,Δa,Δb and ΔEof C. officinalis decoction pieces and wine-processed C. officinalis decoction pieces(stewing in wine)were -9.42--3.58,-24.92- -15.00,-11.33- -7.00 and 17.01-28.12,respectively. ΔL,Δa,Δb and ΔE of C. officinalis decoction pieces and wine-processed C. officinalis(steaming in wine)decoction pieces were -8.58--2.42,-25.08--13.83,-10.92--6.08,15.58-28.67. ΔL,Δa,Δb and ΔE of wine-processed C. officinalis decoction pieces(stewing and steaming with wine)were -2.17-3.00,-0.75-2.50, 0.25-1.42 and 1.25-3.83,respectively. Results of cluster analysis showed that 30 batches of sample were clustered into two categories,S1-S10 were clustered into one category,and S11-S30 were clustered into other category. Principal component analysis showed that cumulative contribution rate of former two main components was 83.147%. Results of partial least squares-discriminant analysis showed that morroniside,No.5 peak and chromaticity values(L,a,b)were the difference markers affecting its quality. CONCLUSIONS:Established UPLC characteristic chromatogram is stable and feasible,and can be used to rapidly identify C. officinalis and its different wine-processed products. Established chemical mode can be used to identify different wine-processed products.
4.The value of three-dimensional color power angiography in the prenatal diagnosis of placenta accreta
Baoxia JIA ; Yuqing LIU ; Tingting LUO ; Binyue LIU ; Jinhua CHEN ; Xuelan HUANG
Chinese Journal of Medical Ultrasound (Electronic Edition) 2017;14(2):117-121
Objective To explore the value of two dimentional colour Doppler flow image (2D-CDFI) combined with three-dimensional color power angiography (3D-CPA) in diagnosis of placenta accreta.Methods A total of 43 pregnant women at risk of placenta accreta selected from September 2010 to August 2015 were enrolled,and underwent 2D-CDFI and 3D-CPA to scan entire placenta.Taking the results of clinical outcome and delivery pathology of the placenta as standard,the ultrasound characteristics of 2D-CDFI and 3D-CPA were analyzed.Results Taking the results of clinical outcome and delivery pathology of the placenta as standard,24 were proved with placenta increta,3 patients with adherent placenta,2 patients with placenta percreta,14 patients with no placenta implantation.Out of 43 cases,29 cases displayed the placental thickening and rich blood vessels in placenta,and at interface of placenta and bladder wall in 2D-CDFI.For 2D-CDFI,19 cases were correctly diagnosed with placenta accrete,while 6 cases were mis-diagnosed and 4 cases missed diagnosed,the diagnosis coincidence rate by 2D-CDFI was 65.5% (19/29).The ultrasound characteristics displayed irregular arranged myometrial arcuate artery,rich blood vessels at interface of placenta and bladder wall in 3D-CPA.For 3D-CPA,23 cases were correctly diagnosed with placenta accrete,3 cases were misdiagnosed,the diagnosis coincidence rate by 3D-CPA was 79.3% (23/29).For 3D-CPA combined 2D-CDFI,1 case missed diagnosed,the diagnosis coincidence rate by combination 2D-CDFI with 3D-CPA was 96.6% (28/29).Conclusions Placenta accrete can all be prenatally diagnosed by characteristic ultrasonic features of 2D-CDFI and 3D-CPA.But 3D-CPA can clearly display the range of placenta accrete lesions and the depth of the blood vessels diffused,has more advantage than two-gray scale ultrasound and 2D-CDFI and has broad application in clinic.
5.Practice and discussion on evaluation system of exploratory experiment's teaching
Haidan LUO ; Yunling XIE ; Donghui WANG ; Hua ZHOU ; Kaiyu TANG ; Xuelan WANG ; Huiling YANG
Chinese Journal of Medical Education Research 2018;17(4):325-329
Exploratory experiment is the key to the experimental physiology,which is universally applicable training of innovative talents.More importantly,it is an important way to cultivate innovative talents of medicine.On the basis of the original multi-subject evaluation system,we have formed a comprehensive evaluation system through refining the scoring item for teachers and students,enriching the forms of student evaluation,and building the student self-assessment.This comprehensive evaluation system is set to promote the development of experimental physiology,especially the exploratory experiments.The practice of this evaluation system effectively improve the objectivity of the evaluation,provide a basis for reform of exploratory experiment,and then promote the training of innovative talents.