1.Cadherin-11 gene and joint formation
Guanzhi WANG ; Jianbao GAO ; Shiwu DONG
Orthopedic Journal of China 2006;0(15):-
Cadherin-11 is a kind of classical cadherin subfamliy.The function of cadherin-11 involves embryonic development,tissue morphogenesis,tumor's invasion and metastasis,signal transduction and so on.This review summarizes the function of cadherin-11 in joint formation,including the relation of cadherin-11 with the bone and cartilage,growth plate,synovial and tendon formation.
2.CULTIVATION AND OSTEOBLASTIC DIFFERENTIATION OF MESENCHYMAL STEM CELLS ON GFP-EXPRESSING MOUSE IN VITRO
Shiwu DONG ; Dajun YING ; Jianhong MI ; Tinghua WANG ; Huijun YANG
Acta Anatomica Sinica 2002;0(05):-
Objective To isolate the bone marrow mesenchymal stem cells(MSCs) from the GFP-expressing mouse, and to study the osteoblastic differentation of the cells. Methods MSCs were isolated by density gradient centrifugation, then the clutrued cells were induced to osteoblastic differentiation using the conditional medium. We detectd the expression of GFP and MSCs differentiation into osteoblasts by histochemistry and immunochemistry. Results The MSCs maintained the expression of GFP during expanded and induced process. After induced for 10 days, lots of alkaline phosphatase and osteoclcin staining positive cells were observed.Conclusion The MSCs of GFP-expressing mouse were successfully isolated and differentiated into ostoblasts. It may be valuable for tracing the seeding cells in tissue engineering bone.
3.Construction and identification of mouse Osf2/Cbfal gene recombinant adenovirus
Shiwu DONG ; Dajun YING ; Guangjiu LIU ; Chuhong ZHU ; Wei ZHANG
Journal of Third Military Medical University 2003;0(21):-
Objective To construct the recombinant adenovirus of mouse Osf2/Cbfal gene and to observe its ability to infect NIH3T3 fibroblasts. Methods The Osf2/Cbfal gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into NIH3T3 cells using Lipofectine DOTAP, The target gene was detected by poly-merase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. Results Restriction endonuclease and PCR analyses confirmed that the Osf2/Cbfal gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.6?1012 pfu/ml. The adenovirus had a strong effect on NIH3T3 cells. Conclusion The recombinant adenovirus containing Osf2/Cbfal gene was successfully constructed by the method of homogenous recombination in bacteria.
4.Cultivating the Teachers' Quality in the process of body donation
Yang LOGN ; Shiwu DONG ; Jiansen SUN ; Xiaosong LI ; Dajun YING
Chinese Medical Ethics 1996;0(01):-
The development of body donation remains relatively slow in China.One of its main influencing factors restricted to the development of donation is the anatomy teachers' quality.Lots of volunteers to donate and their families are not satisfied with it,which eventually leads to the failure of the donation.Therefore,enhancing anatomy teachers' quality has become one of the key steps in the donated work.It needs to improve the idea awareness of teachers,to standardize the behavior of teachers,to enhance the role models of teachers,and to carry out a regular humanities education and training to truly realize "people-oriented" goals.
5.Effects of heparanase and E-cadherin on the invasion and metastasis of gastric cancer
Xiaojun CHEN ; Zhichao ZHENG ; Zhiwei LIAO ; Shiwu XU ; Jian DONG ; Qi HUANG ; Shubao WANG
Chinese Journal of Digestive Surgery 2009;8(5):344-346
Objective To investigate the effects of heparanase and E-cadherin on the invasion and metastasis of gastric cancer. Methods Fifty specimens of gastric cancer which had been resected at Cancer Hospital of Liaoning Province from February 2005 to May 2007 were collected. The expression of heparanase mRNA and E-cadherin mRNA in these gastric cancer specimens was detected by RT-PCR, and the expression of E-cadherin in these gastric cancer specimens was detected by immunohistochemistry. Data were analyzed by t-test and variance analysis, and the enumeration data analyzed by chi-square test. Results There were significant differences in the expression of heparanase and E-cadherin between gastric cancer cells with high and low differentiation, presence and absence of metastasis, and TNM stages Ⅰ and Ⅱ versus Ⅲ and Ⅳ (t = 1.999, 4.258, 1.735 ; 1.286, 6.794, 3.091; χ~2 =6.273, 9.397, 5.640, P <0.05). The co-expression of heparanase (+) and E-cadherin (-) was correlated with tumor undifferentiation, lymph node metastasis and advanced TNM staging (χ~2 =11.306, 10.208, 8.420, P <0.05). Conclusions Heparanasc shows high expression while E-cadherin shows low expression in gastric cancer tissue. There is a synergistic effect between the abnormal expression of heparanase and E-cadherin, and the gastric cancer cells with coexpression of heparanase and E-cadherin have more malignant potential.
6.CXCR4 gene transfection enhancing the distribution of dMSCs to the wounded skin of rats with combined wound and irradiation injury
Zhaowen ZONG ; Tianmin CHENG ; Xinze RAN ; Yongping SU ; Shiwu DONG ; Nan LI ; Junping WANG ; Guoping AI ; Zheng LI
Chinese Journal of Radiological Medicine and Protection 2009;29(4):351-354
Objective To observe whether the transplanted dermal multipotent stem cells(dMSCs)transfected by adenovirus vector of CXCR4(Adv-CXCR4)can distribute more frequently to the wound of rats with combined wound and irradiation injury.Methods dMSCs transfected by Adv-CXCR4(group A),or transfected by adenovirus vector of green fluorescent protein(group B),and non-transfected dMSCs were labeled with 3H-TdR and then transplanted into combine-injured rats.The amount of dMSCs in wound were determined by liquid scintillation,and wounds healing process was observed by measuring the remaining wound area.Results From the 5th day after transplantation,the amount of dMSCs in the wound of group A accounted for 1.95%-3.85% of the total transplanted dMSCs,significantly greater than those in group B and group C,which accounted for 1.07%-1.86% of the total transplanted dMSCs.The remaining wound area in group A was smaller than those in group B and group C from day 12 after injury,and the healing time of group A was 1.5 day ahead than group B and group C.Conclusions dMSCs transfected by Adv-CXCR4 distributes more frequently to the wound of combine-injured rats and could accelerate wound healing.
7.Transdifferentiation of allotype BMSCs into hepatocytes in bone marrow chimeric mice
Lianyou WANG ; Hui XU ; Shiwu DONG ; Yongping SU ; Xueli PANG ; Dengqun LIU ; Junping WANG ; Xinze RAN ; Fengchao WANG
Journal of Third Military Medical University 2003;0(11):-
Objective To detect whether mice bone marrow mesenchymal stem cells(BMSCs)can contribute to the regeneration of hepatocytes in bone marrow chimeric mice.Methods Female recipient mice(C57BL/6J)underwent whole body gamma-ray irradiation with a dose of 10 Gy to ablate their bone marrow,followed by immediate tail vein injection of BMSCs isolated from male GFP transgenic mice.Animals were killed at different phase points:1 week,1 month,and 3 months.Using fluorescence microscope we directly observed GFP-positive cells in the liver frozen sections,and we also prepared the parafilm sections to detect the GFP-positive cells and the coexpression of GFP and Alb,CK18 by immunohistochemistry and immunofluorescence respectively.Results We found numerous GFP-positive cells in recipient mice liver at 1 week after BMSCs transplantation,some at 1 month and seldom at 3 months.There were some cells coexpressing GFP and Alb,CK18 at all the phase points.Conclusion Allotype BMSCs can differentiate into Alb and CK18 positive hepatocytes in bone marrow chimeric mice,which will become an ideal cell resource for liver tissue project.
8.Feasibility of MSCs mobilization by G-CSF and its prosthetic effect in traumatic brain injury
Jun DENG ; Guoping AI ; Taoli ZHOU ; Junping WANG ; Hui XU ; Zhongmin ZOU ; Shiwu DONG ; Lei HAO ; Xinze RAN ; Yongping SU
Journal of Third Military Medical University 2002;0(12):-
Objective To explore the feasibility of mobilization circulating MSCs by G-CSF and observe the repairing effect of G-CSF mobilization in severe mouse traumatic brain injury(TBI) model.Methods MSCs-derived bone marrow and peripheral blood(PB) were cultured and its CFU-F were counted after mobilization by G-CSF.At 2,24,48,96,120,144,192,264,336 h after severe TBI in mice was establish,the neurobehavior of mice was measured by neurological examination and motor functional test,and mortality rate and pathologic changes were analyzed.Results MSCs-derived PB were successfully cultured.The CFU-F of mobilization group increased significantly than that of control group(P
9.Effects of hPDGF-A/hBD_2 genes transfection on rat bone marrow mesenchymal stem cells
Yimei FENG ; Hui XU ; Zhongmin ZOU ; Lei HAO ; Tao WANG ; Jun DENG ; Shiwu DONG ; Yongping SU ; Tianmin CHENG
Journal of Third Military Medical University 1988;0(06):-
Objective To study the biological characteristics of platelet-derived growth factor A and human beta defensin 2 (hPDGF-A/hBD2) gene-modified rat bone marrow mesenchymal stem cells (BMSCs). Methods By using liposome transfection technique, recombinant adenovirus vector expressing hPDGF-A/hBD2 (Adv-hPDGF-A-IRES-hBD2) labeled with GFP was transfected into 293T cells for virus packaging and amplification. BMSCs were isolated, cultured and infected by adenovirus-containing supernatant. The exogenous gene-modified BMSCs were comprehensively studied on their biological features, in terms of morphology, cell growth curve, cell cycle, and adipogenic, osteogenic and myogenic differentiation ability. Results hPDGF-A-IRES-hBD2 gene-modified BMSCs did not show obvious changes in cell viability, proliferation, cell cycle distribution or cell differentiation. Conclusion BMSCs were not only good carriers for exogenous hPDGF-A and hBD2 genes but also seed cells for cell therapy even after hPDGF-A/hBD2 modification.
10.Expression of recombinant human ?-defensin 2 in dermal multipotent stem cells and its antiseptic activity
Nan LI ; Taoyuan XIAO ; Yongping SU ; Hui XU ; Junping WANG ; Zhaowen ZONG ; Xinze RAN ; Shiwu DONG ; Zhijun LIU
Journal of Third Military Medical University 1983;0(04):-
Objective To examine the expression of human ?-defensin 2 (hBD_ 2 ) recombinant adenovirus expression vector in rat dermal multipotent stem cells (dMSCs) and to observe the antiseptic activity of recombinant hBD_ 2 . Methods The expression of hBD_ 2 in dMSCs was examined by RT-PR, fluorescent immunochemistry and Western blotting, and the concentration of recombinant hBD_ 2 in supernate was measured by ELISA. The antiseptic activity of recombinant hBD_ 2 was assessed by K-B disc agar diffusion test. Results hBD_ 2 could be effectively expressed in dMSCs, and the concentration of recombinant hBD_ 2 in supernate was about 743.6 ng/ml . Recombinant hBD_ 2 in supernate showed antiseptic activity. Conclusion Recombinant adenovirus expression vector of hBD_ 2 could be effectively expressed in dMSCs, and the recombinant hBD_ 2 in supernate showed obvious antiseptic effects toward some standard bacteria lines.