1.The role of B7-H1mRNA in immune privilege of homogeneity
Xiaohuan HAUNG ; Jing WU ; Jian YU ; Linjiang CHEN ; Ming MA
Chinese Journal of Organ Transplantation 2012;33(9):567-570
Objective To clarify the role of B7-H1 in the immune privilege after corneal transplantation in homogeneity variant mice.Methods We established the experimental animal model of allograft mice by using C57BL/6 mouse as donor and Balb/c mouse as recipient.We allocaated the mice with long time survival (>50 days) corneal graft into survival group,mice with rejection occurring in 50 days into rejection group,and normal C57BL/6 mice into control group.The transplanted corneal grafts were obtained for future reference at the 8th week after transplantation in survival group,and the time of rejection in rejection group.The expression of B7-H1 mRNA was detected by using immunohistochemistry and real time quantitative PCR (RT-PCR),and the relationship between B7-H1 and the immune privilege after corneal transplantation was analyzed. Results The B7 H1 mRNA was highly expressed in epithelium and endothelium of corneal grafts both in survival and control group,in comparison to an obviously lower expression in rejection group.The relative expression level of B7-H1 mRNA was 200.0 ± 11.5 in survival group,44.7 ± 10.8 in control group,and 6.9 ± 12.0 in rejection group,respectively. There were statistically significant differences among the three groups (F=241.164,P<0.01 ).The was a significant correlation between the level of B7-H1 mRNA and occurrence rate of rejection in corneal graft (P<0.01 ).Conclusion The results suggest that the immune privilege after corneal transplantation might be mediated by B7-H1,which plays an important role in maintaining the state of corneal immune privilege.
2.Effects of Basic Fibroblast Growth Factor on Proliferation as well as Collagen and Fibronectin Expression on Fibroblasts
Yan WU ; Lan YANG ; Yufei ZHANG ; Xiaohuan YUAN ; Yanhui CHU
Herald of Medicine 2014;(11):1416-1419
Objective To explore the mechanism and effects of basic fibroblast growth factor( bFGF)on skin wound healing. Methods Fibroblasts( FB)were isolated from normal skin and hypertrophic scar and cultivated by direct adherence method. FB were then treated with different concentrations of bFGF(0,0. 1,1,10,100,1 000 ng·mL-1 )and cultivated with serum-free medium for 72 hours. The proliferation and apoptosis of FB in each group were detected by cell counting and trypan blue staining. Content and gene expression of typeⅠand type Ⅲ collagen and fibronectin were determined by ELISA and RT-PCR,respectively. Results bFGF promoted the proliferation of FB at low concentrations promoted apoptosis of FB at higher concentrations. The proliferation of FB from hypertrophic scar was slower than that from the normal skin. bFGF significantly inhibited type Ⅰ collagen production from hypertrophic scar FB but not from the normal skin. Moreover,bFGF up-regulated fibronectin expression in the normal fibroblasts,but not in the hypertrophic scar. No change in type Ⅲ collagen expression and production was observed in FB from either source. Conclusion bFGF has differential effects and mechanisms on FB of the normal skin and hypertrophic scar,suggesting that bFGF may play a role in early phase of skin wound healing and scar formation.
3.Effects of bone marrow mesenchymal stem cell conditioned medium on bioactivity of scar fibroblasts
Yan WU ; Chunlei ZHANG ; Yang LIU ; Hongzhi LI ; Jing YU ; Haihua BAO ; Ran GUO ; Xiaohuan YUAN
Chinese Journal of Tissue Engineering Research 2014;(7):1009-1014
BACKGROUND:Mesenchymal stem celltransplantation promoted skin repair in trauma via various regulatory mechanisms and inhibited scar formation. At present, many scholars believed that bioactive factors secreted by mesenchymal stem cells played an important role.
OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem cellconditioned medium on the proliferation and col agen synthesis of hypertrophic scar fibroblasts.
METHODS:Human bone marrow mesenchymal stem cells and hypertrophic scar fibroblasts were isolated and cultured, and bone marrow mesenchymal stem cellconditioned medium was prepared. Hypertrophic scar fibroblasts were cultured in vitro with 12, 24, and 48 hour-col ected conditioned medium for 24 hours, which was compared with blank control group. The proliferation of cells was determined by CCK-8. Type I and type III col agen expression in hypertrophic scar fibroblasts was detected using real-time PCR.
RESULTS AND CONCLUSION:Compared with the blank control group, 24 and 48 hour-col ected conditioned medium significantly inhibited the proliferation of hypertrophic scar fibroblasts (P<0.01), and also suppressed col agen synthesis of hypertrophic scar fibroblasts (P<0.01). Results suggested that bone marrow mesenchymal stem cellconditioned medium inhibited the proliferation and col agen synthesis of hypertrophic scar fibroblasts by secreting anti-fibrotic bioactive factors, which may provide new theoretical supports for celltherapy to reduce cutaneous scarring.
4.The regulatory effect of recombinant human fibroblast growth factor-21 on the expression of liver X receptor α and glucose transporter protein 1 in the type 2 diabetes rats
Wenping ZOU ; Yufei ZHANG ; Huiyan WANG ; Yan WU ; Xiaohuan YUAN ; Yanhui CHU
Chinese Journal of Biochemical Pharmaceutics 2009;30(6):386-389
Purpose To examine the regulatory effect of recombinant human fibroblast growth factor-21 on the expression of liver X receptor α and glucose transporter protein 1 in the type 2 diabetes mellitus rats.Methods The rat models of type 2 diabetic mellitus were divided into four groups at random, ic. rhFGF-21 every day, after eight weeks of these treatment, Inspect the fasting blood glucose (FBG), fructosamine(FA), triglyceride(TG), T-cholesterol(TC), high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol(LDL-C) of these rats, then detecting the mRNA expression of LXRα and GLUT1 by RT-PCR.Results (1) rhFGF-21 can reduce blood glucose steadily to near normal levels in diabetic rats. (2) The expression of LXRα and GLUT1 level was significantly higher in the rhFGF-21 treatment group than that in the model group. (3) rhFGF-21 megadoses and middle doses decreased FA, TG, TC,and LDL-C and elevated HDL-C.Conclusion rhFGF-21 could regulate the mRNA expression of LXRα and GLUT1 in diabetes rats, increase basal level glucose transport, then reduce blood glucose, improve lipid metabolize dysfunction.
5.Effects of mesenchymal stem cells conditioned media on the transforming growth factor-beta signaling pathway in normal and hypertrophic scar fibroblasts
Yan WU ; Xiaohuan YUAN ; Lan YANG ; Xiaojin ZHAO ; Houzhong LI ; Ying WANG ; Hong JIN
Chinese Journal of Tissue Engineering Research 2016;20(29):4349-4354
BACKGROUND:Numerous studies have shown that mesenchymal stem cel s (MSCs) can effectively attenuate the fibrosis of damaged heart, lung and kidney by secreting various bioactive factors. OBJECTIVE:To evaluate the anti-fibrotic therapeutic potential of bone marrow MSCs conditioned media in vitro. METHODS:Normal fibroblasts and hypertrophic scar fibroblasts were treated with bone marrow MSCs conditioned media, then transforming growth factor-βand col agen production were analyzed by ELISA, and mRNA expression level of Smad7 and hydroxyproline content were detected by RT-PCR and colorimetry, respectively. RESULTS AND CONCLUSION:Bone marrow MSCs conditioned media significantly inhibited the production of both transforming growth factor-βand col agen in hypertrophic scar fibroblasts (P<0. 01), and up-regulated the mRNA expression level of Smad7 (P<0. 01), a major inhibitory regulator in the SMAD family. However, the normal fibroblasts were scarcely influenced by bone marrow MSCs conditioned media. These findings indicate that bone marrow MSCs conditioned media is considered a promising candidate for the treatment of hypertrophic scars, which may provide new theoretical supports to reduce cutaneous scarring.
6.Role of B7-H1 protein in immune privilege following corneal allograft transplantation in mice.
Xiaohuan HAUNG ; Jin WU ; Jian YU ; Ke XIONG ; Ming MA
Journal of Southern Medical University 2012;32(6):901-903
OBJECTIVETo study the expression level of B7-H1 protein in the eyeball tissues of mice receiving corneal allograft transplantation and explore the role of B7-H1 protein in corneal immune privilege.
METHODSMouse models of corneal allograft transplantation were established, and the corneal opacity and angiogenesis index was evaluated according to the Sonoda method. Eight weeks later, the mice were examined for the occurrence of graft rejection, and the expression level of B7-H1 protein in the eyeball tissues were detected by immunohistochemistry, using 8 normal mice as the control group.
RESULTSB7-H1 protein was expressed highly in the corneal and the choroidal/ciliary body in the normal control mice and the survived mice, but was absent in mice with graft rejection.
CONCLUSIONB7-H1 protein may play a role in the immune privilege following corneal allograft transplantation in mice.
Animals ; B7-H1 Antigen ; immunology ; Corneal Transplantation ; Female ; Immune Tolerance ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation, Homologous ; immunology
7.The clinical significance of sCD40L and Lp-PLA2 in the assessment of coronary artery severity and risk classification in patients with non-ST-segment elevation acute coronary syndrome
Xuan GUO ; Chunyan ZHANG ; Congxia WANG ; Yan ZHANG ; Shan JIA ; Weidong MA ; Xiaohuan LIU ; Xiaohui QUAN ; Haoyu WU ; Yang ZHENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(3):322-326
Objective To investigate the clinical significance of soluble CD40 ligand (sCD40L ) and lipoprotein associated phospholipase A2 (Lp-PLA2 ) in the assessment of coronary artery severity and risk classification in patients with non-ST-segment elevation acute coronary syndrome (NSTE-ACS).Methods Of the 9 6 patients with coronary heart disease diagnosed by coronary angiography,2 8 patients had stable angina pectoris (SAP),38 patients unstable angina pectoris (UAP)and 30 patients acute non-ST-segment elevation myocardial infarction (NSTEMI).Another 30 patients with non-coronary heart disease (NC)served as controls.The sCD40L and Lp-PLA2 levels were determined by enzyme-linked immune sorbent assay (ELISA)method.The Gensini score was used to assess the severity of coronary artery and analyze the correlation with sCD40L and Lp-PLA2.The correlation of sCD40L and Lp-PLA2 with GRACE risk score was analyzed too.Results ① sCD40L was significantly higher in NSTEMI and UAP groups than in SAP and NC groups (P<0 .0 5 ),but there was no significant difference between NSTEMI and UAP groups (P>0 .0 5 )or SAP and NC groups (P>0 .0 5 ).Lp-PLA2 was significantly higher in NSTEMI group than in UAP,SAP and NC groups (P<0.05).Lp-PLA2 was significantly higher in UAP group than in SAP and NC groups (P<0.05).② We found that sCD40L had obvious correlation with Lp-PLA2 (r=0.284, P<0.01),Gensini score (r=0.213,P<0.05),and GRACE (r=0.224,P<0.05).Lp-PLA2 was significantly correlated with Gensini score (r=0.270,P<0.05),and GRACE (r=0.323,P<0.01).③ Multivariate logistic regression analysis showed that Lp-PLA2 was independently associated with NSTE-ACS (P<0.05).Conclusion The sCD40L and Lp-PLA2 which were significantly elevated in NSTE-ACS are correlated with the severity of coronary artery disease.The two indexes indicate the instability of atherosclerotic plaque;thus they can be used as predictors of risk assessment in coronary heart disease.
8.Effect of Curcumin on Non-Small Cell Lung Cancer A549 Cells
Junxiang LIU ; Jieting LIU ; Chunyin TANG ; Lingyu LI ; He BAI ; Xiaohuan YUAN ; Wei ZHANG ; Liyan SUN ; Yan WU
Herald of Medicine 2017;36(8):865-869
Objective To study the inhibitory effect of curcumin on the proliferation,migration and invasion of non-small cell lung cancer cell A549,and to discuss further if it is closely related to the expression of c-Jun N-terminal kinase (JNK) and relative protein p38.Methods A549 cells were cultured by conventional method,and then treated with different concentration of curcumin (10,20,40,80 μmol · L-1).The proliferation,migration and invasion of A549 cells were measured by real-time cellular analysis (RTCA).The expression levels of JNK,p-JNK,p38 and P-p38 were detected by real-time PCR and Western blotting.Results Curcumin showed an antiproliferation effect against A549 cells with IC50 =40 μmol · L-1,and curcumin exhibited obviously inhibitory effect on the migration and invasion of A549 cells.Additionally,compared with control group,curcumin suppressed the expression of JNK and p38 at the gene level,and significantly inhibited the expression of JNK,P-JNK,p38 and p38 (P<0.05) at the protein level.Conclusion These results demonstrated that curcumin can inhibit the proliferation,migration and invasion of A549 cells via reducing the level of JNK,p38 phosphorylation,and blocking JNK signal transduction pathway.
9.The correlation research between three ultrasonic technologies for the measurement error in breast cancer and the expression of ER, PR and VEGF
Peiwei CAI ; Teng LIN ; Xiaohuan ZHU ; Cong HUANG ; Yingbin XU ; Weizhen LIAN ; Guoliang GONG ; Qiancheng QIU ; Baoan WU
Chinese Journal of Primary Medicine and Pharmacy 2016;23(5):641-644,645
Objective To explore the three ultrasonic technologies of two -dimensional ultrasound(2D -US),ultrasonic elastography(UE) and contrast -enhanced ultrasound(CEUS) for the measurement error in breast cancer and the correlation with the expression of ER,PR,VEGF.Methods 50 patients with breast cancer were meas-ured by 2D -US,UE,CEUS preoperatively,and the pathological specimen were measured postoperatively.Then used the immunohistochemistry to detect the expression of ER,PR,VEGF in tumor,and analyzed the correlation with the measurement errors.Results The results of differences between 2D -US,UE,CEUS and pathology were respectively as follows:( -0.59 ±-0.34)cm,( -0.20 ±-0.14)cm,( -0.40 ±-0.31)cm,and the differences were statistically significant(F =20.497,P <0.001).The positive expression rate of ER and PR was high if the difference between UE and 2D -US was less than or equal to 0.44cm.And the positive expression rate of VEGF was low if the difference between CEUS and 2D -US was less than or equal to 0.19cm.Three ultrasonic technologies in the measurement of breast cancer were different,the trend of difference between UE and 2D -US was smaller if the ER and PR were positively expression,and the trend of difference between CEUS and 2D -US was bigger if the VEGF was positively expression.Conclusion There is correlation between different immunohistochemical expression of breast cancer with measurement error in three different ultrasonic imaging technologies.The results suggest that the molecular pathology difference of breast cancer can impact on ultrasonic imaging,which contributes to know the reason and regulation of measurement error in different ultrasonic imaging technology.
10.Expression and clinical significance of serum angiopoietin-2 in patients with chronic heart failure
Haoyu WU ; Chunyan ZHANG ; Lei LIANG ; Congxia WANG ; Xiling SHOU ; Yan ZHANG ; Weidong MA ; Shan JIA ; Yang ZHENG ; Xiaohuan LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(3):336-338,348
Objective To investigate the expression of serum angiopoietin-2 (Ang-2 ) and its clinical significance in patients with chronic heart failure (CHF).Methods The levels of serum Ang-2,N-terminal pro-B-type natriuretic peptide (NT-proBNP)and left ventricular ejection fraction (LVEF)were detected in 1 1 3 patients with CHF,who were divided into four groups according to New York Heart Association (NYHA:class Ⅰ,n=3 2;Ⅱ,n=30;Ⅲ,n=26;Ⅳ,n=25).Another 20 healthy volunteers for physical check-up were chosen as the control group.Results With the change of cardiac function (from Ⅰ to Ⅳ),the levels of plasma Ang-2 and NT-proBNP were increased significantly (P<0.05 ).The level of serum Ang-2 was positively correlated with NT-proBNP (r=0.774,P<0.001),but negatively correlated with left ventricular ejection fraction (r=-0.725,P<0.001).Conclusion The level of serum Ang-2 has a significant correlation with the severity of heart failure.Ang-2 is expected to be used to predict and evaluate the severity of CHF.