1.Differentiation into osteoblasts from marrow mesenchymal stem cells of rabbits transfected by human bone morphogenetic protein-7 gene
Yan KANG ; Weiming LIAO ; Puyi SHENG ; Longjuan ZHANG
Chinese Journal of Tissue Engineering Research 2006;10(21):161-163,封三
BACKGROUND: At present, bone morphogenetic protein (BMP) is the only cytokine characterized as the ability of osteoblast, and it can promote marrow mesenchymal stem cells (MSCs) differentiating into chondroblast and osteoblast.OBJECTIVE: To investigate the differentiation into osteoblast from MSCs of rabbits transfected by human bone morphogenetic protein-7 (hBMP-7) gene with genetic engineering technique. DESIGN: Single sample observation. SETTING: Department of Orthopaedics, the First Hospital Affiliated to Sun Yat-sen University. MATERIALS: pcDNA1.1/AMP-hBMP7 plasmid.METHODS: The experiment was completed at the Surgery Laboratory of the First Hospital Affiliated to Sun Yat-sen University from May 2004 to March 2005. MSCs of rabbits were cultured with whole bone marrow technique, transfected with pcDNA1.1/AMP-hBMP7 and pcDNA1.1/AMP plasmids respectively in vitro, and left the blank controls. Transcription and expression of transfected genes were detected so as to observe form and growth of cells. Component of calcium content in cytoplasm was measured and osteogenic phenotype was identified with alkaline phosphatase (ALP) staining, calcium salts staining and transmission electron microscopy (TEM).MAIN OUTCOME MEASURES: ① Form and growth of MSCs of rabbits; ② evaluation of expressive product; ③ results of ALP staining (Ca-Co technique); ④ results of chinalizarin staining; ⑤ results of TEM; ⑥ results of scanning electron m icroscopy (SEM); ⑦ assay of osteocalcin.RESULTS: hBMP-7 gene was contained in MSCs after transfection and expressed the relevant mRNA. Cellular form changed a little after expression of amid genes, but growth curve did not changed as compared with that in non-transfected group. Expression of osteocalcin was increased as compared with that in non-transfected group, and ALP staining and calcium salts staining of transfected cells were positive.CONCLUSION: Transfection of hBMP-7 gene can promote MSCs of rabbits differentiating into osteoblast.
2.Over-expressions of macrophage migration inhibitor factor and cyclin D1 correlates with clinical pathologic characteristics of pancreatic carcinoma and it significance
Jie ZHAO ; Jingtang XIA ; Wen LI ; Longjuan ZHANG ; Hua WANG ; Zhaofeng WU ; Yuan TAN ; Cheng XU
Journal of International Oncology 2009;36(7):550-553
Objective To evaluate the expression of macrophage migration inhibitor factor (MiF) and cyclinD1 in pancreatic carcinoma and their relationships with clinical pathology characteristics. Methods The expression of MIF and eyclinD1 in 89 carcinoma and 5 normal pancreatic tissues was detected with immunohis-tochemistry methods, and the relationships among MIF and cyclinD1 expression and clinicopathological factors were studied. Results The overexpression of MIF and cyclinD1 was found in 88.8%, and 50. 6% of pancre-atic carcinoma tissues respectively. The overexpression of MIF had a significant correlation with Ⅰ,Ⅱ,Ⅲ,Ⅳ tumor stage (69. 2%, 94. 7%, 96. 4%, 100%, P <0.05), while the positive expression rate of cyclinD1 only had a significant correlation with tumor stages Ⅲ,Ⅳ (33. 3%, 68. 8%, P <0. 05). Both of the two proteins had a correlative tendency with pathological grade and lymph node metastasis. The different expression of MIF between pancreatic carcinoma with and without liver metastasis had no statistical significance, (100% ,85.9%, P >0. 05)while there was a statistically significant difference about cyclinD1 (66. 7% ,46. 5% ,P <0. 05). A significant positive correlation was also found between MIF and cyclinD1 (P < 0. 05). Conclusion The ex-pression of MIF and CyclinD1 was higher in pancreatic cancer tissues than in normal tissue, and they may be associated with the malignant stage, tumor differentiation, local lymph node and liver metastasis of this tumor.
3.Efficacy and oncological safety of bone marrow mesenchymal stem cell transplantation for colitis in inflammatory bowel disease models in mice
Xiaowen HE ; Zexian CHEN ; Longjuan ZHANG ; Xiaosheng HE ; Lei LIAN ; Jia KE ; Xutao LIN ; Xi CHEN ; Xiaojian WU ; Ping LAN
Chinese Journal of Tissue Engineering Research 2014;(23):3696-3701
BACKGROUND:Transfusion of bone marrow mesenchymal stem cells may become a novel and effective biological therapy for inflammatory bowel disease in clinical practice. Nevertheless, the oncological safety of the treatment is worrisome, and is a key to determine whether mesenchymal stem cells can be widely used in treatment of inflammatory bowel disease, and deserves further investigation. OBJECTIVE:To evaluate the therapeutic effect of bone marrow mesenchymal stem celltransfusion against inflammatory bowel disease in mouse models, and to clarify the effects of mesenchymal stem cells on tumorigenesis of inflammatory bowel disease. METHODS:Mouse model of colitis was established using Balb/c (H-2d) mice exposed to dextran sulfate sodium. Syngeneic bone marrow mesenchymal stem cells were transfused into mouse model through caudal vein. The therapeutic effect of mesenchymal stem cells was compared and observed, and pathological remission of colitis was evaluated. Mouse model of colitis-driven colon carcinogenesis was established using Balb/c (H-2d) mice exposed to dextran sulfate sodium and azoxymethane. Tumor formation within the murine colon was compared and observed after transfusion of mesenchymal stem cells. RESULTS AND CONCLUSION:In models of dextran sulfate sodium-induced colitis, weight loss and fecal occult blood were lessened in the bone marrow mesenchymal stem cellgroup compared with the phosphate buffered saline group. Histological damage score of colitis was less in the bone marrow mesenchymal stem cellgroup:mucosal structure of distal colon was almost intact under microscope, and there was smal area of epithelial defects and cryptal defects. Inflammatory cellinfiltration, proliferation of capil ary and smal vessels could be observed in mucosa and submucosa. Homing and colonization of mesenchymal stem cells in submucosa of inflamed colon could also be observed by in vivo tracing. In the dextran sulfate sodium/azoxymethane model of colitis-driven colon carcinogenesis, the number of intestinal tumors and tumor load were obviously less in the bone marrow mesenchymal stem cellgroup than in the control group. Results indicated that transfusion of bone marrow mesenchymal stem cells can apparently improve colitis lesions of mice with inflammatory bowel disease and inhibit carcinogenesis of colitis, which may provide theoretical support for the biological safety of mesenchymal stem cells transplantation for inflammatory bowel disease.
4.Effects of laminar shear stress on the expression of IL-8 receptor CXCR1 in endothelial cells.
Longjuan XUE ; Chang YU ; Xiaoheng LIU ; Yi LAI ; Ye ZENG ; Yi ZHANG
Journal of Biomedical Engineering 2007;24(5):1054-1057
This study was conducted to elucidate the effects of different fluid shear stress on the expression of IL-8 receptors CXCR1 in endothelial cells. The HUVEC cell lines, EA. Hy926 cells, were cultured in vitro, exposed to 5.56, 10.02, and 15.27 dyn/cm2 laminar shear stress, respectively, and then the shear stress grous were each investigated at the time-points of 0h, 1h, 2h, 4h and 8h. Western blotting was used for detecting the expression of IL-8 receptor CXCR1 at the time-points. The results showed that, under 5.56 dyn/cm2 shear stress, the expression of CXCR1 increased with time significantly (P < 0.01). The maximum expression of CXCR1 appeared at 4 h and was 2.2 times that of the control. When exposed to 10.02 dyn/cm2, the expression of CXCR1 increased gradually with time and finally remained at a constant higher level. When exposed to 15.27 dyn/cm2, CXCR1 expression decreased significantly with time (P < 0.01). The minimum CXCR1 expression appeared at 8 h and was 62. 59 percent of that of the control. These results indicate that the expression of CXCR1 in endothelial cell is regulated by laminar shear stress.
Cell Line
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Endothelial Cells
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cytology
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metabolism
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Humans
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RNA, Messenger
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biosynthesis
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genetics
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Receptors, Interleukin-8A
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genetics
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metabolism
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Shear Strength
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Stress, Mechanical
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Umbilical Veins
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cytology
5.Risk factor analysis of incisional infection in Crohn's disease patients after bowel resection
Huashan LIU ; Longjuan ZHANG ; Zhenxing LIANG ; Tuo HU ; Chi ZHOU ; Xiaowen HE ; Xiaojian WU ; Xianrui WU
Chinese Journal of Digestive Surgery 2018;17(9):935-942
Objective To analyze the risk factors affecting postoperative incisional infection in Crohn's disease (CD) patients after bowel resection.Methods The retrospective case-control study was conducted.The clinicopathologieal data of 239 CD patients who underwent bowel resection in the Sixth Affiliated Hospital of Sun Yat-sen University between January 2007 and December 2016 were collected.All patients underwent bowel resection.Observation indicators:(1) surgical situations;(2) follow-up;(3) risk factors analysis affecting postoperative incisional infection;(4) clinical factors affecting preoperative anemia.The follow-up using outpatient examination or ward diagnosis was performed to detect incisional infection within 30 days postoperatively up to January 2017.The normality test was done by Shapiro-Wilk.Measurement data with normal distribution were represented as x-±s,and comparison between groups was evaluated with the t test.Measurement data with skewed distribution were described as M (range),and comparison between groups was analyzed using the Wilcoxon ranksum test.The univariate analysis and multivariate analysis were done using the Logistic regression model.The P< 0.05 in univariate analysis was incorporated into multivariate analysis for analysis in the forward wald.Results (1) Surgical situations:of 239 patients,11 underwent emergency surgery and 228 underwent elective surgery;65 and 174 underwent respectively laparoscopic surgery and open surgery;179 received digestive tract reconstruction and anastomosis and 81 received enterostomy (21 combined with anastomosis and enterostomy).Among 239 patients,137,113,101,58,54 and 11 were complicated respectively with fiber stenosis,intestinal fistula,obstruction of small intestine,abscess,cellulitis and enterobrosis (some patients combined with multiple signs).(2) Follow-up:239 patients were followed up at 30 days postoperatively.During the follow-up,48 with incisional infection were improved by symptomatic treatment.(3) Risk factors analysis affecting postoperative incisional infection:① Results of univariate analysis showed that illness behavior,sedimentation rate of RBC > 20 mm/h,preoperative anemia,preoperative chronic intestinal fistula,open surgery,intraoperative fiber stenosis and intraoperative intestinal fistula were risk factors affecting occurrence of postoperative incisional infection [odds ratio (0R)=2.530,2.579,4.233,2.988,2.554,0.503,3.052,95% confidence interval (CI):1.218-2.259,1.141-5.833,1.598-11.210,1.522-5.864,1.082-6.029,0.265-0.954,1.555-5.993,P<0.05].② Results of multivariate analysis showed that preoperative anemia and intraoperative intestinal fistula were independent risk factors affecting occurrence of postoperative incisional infection (OR =3.881,2.837,95% CI:1.449-10.396,1.429-5.634,P<0.05).(4) Clinical factors affecting preoperative anemia:cases (male) with preoperative anemia,body mass index (BMI),cases with sedimentation rate of RBC > 20 mm/h,platelet (PLT) > 300x109/L,elevated C-reactive protein,albumin (Alb) <35 g/L were respectively 120,(17.4±2.9)kg/m2,130,75,139,65 in patients with preoperative anemia and 65,(18.3±2.9)kg/m2,36,12,39,10 in patients without preoperative anemia,with statistically significant differences (x2 =17.966,t =2.210,x2 =12.219,14.440,14.661,12.272,P<0.05).Conclusion The preoperative anemia and intraoperative intestinal fistula are independent risk factors affecting occurrence of postoperative incisional infection,and preoperative anemia is associated with perioperative inflammatory conditions.