1.Improvement for indwelling healer tube in prevention and cure the conglutination for children patiens after the surgery operation Flexor tendon tenosynovitis
Jimin PEI ; Huisheng HE ; Dan XU
International Journal of Surgery 2009;36(8):534-536
Objective Evaluate the safety and efficacy of indwelling healer tube in prevention and cure the conglutination after the surgurey of flexor tendon tenosynovitis. Methods During the year of 2001~2006, We have 38 young patients treated with indwelling of healer tube into the local or partial of the recovering muscle and sinew, then anesthetic and sodium hyaluronate were injected in the tube in certain intervals to lubicate and prevent conglutination after the operations of joint of children flexor tendon tenosynovitis. Then let the young patients do some healing training attne earlystage after surgery. Groups of patient were set up to make comparative analysis and evaluate the effectiveness of indwelling of the healer tubes according to the recovery status of the function of arthrosis and grasp after surgery. Results The result is that the rate of choiceness of 46 sinew is 89.1% in 34 cases with indwelling healer tube after the observing period from 6 months to 2 years, whereas the other group of 44 sinew in 30 cases has the rate of choiceness of 63.6%. The comparison has the significant conclusion of statistics (P<0.05). Conclusion It is convenient and safe to use indwelling healer tube to prevent the conglutination after the operation of joint the broken finger muscle and sinew of children. Therefore it is worth popularizing and promoting.
2.New bone formation by bone marrow stromal cell combined with the bioactive glass ceramic using tissue-engineering methods
Dan JIN ; Guoxian PEI ; Qian WANG
Chinese Journal of Trauma 2001;17(3):151-154
Objective To study the biocompatibility of bioactive glass ceramic (BGC) materials with bone marrow stromal cell (BMSc) and the osteogenic capability of BMSc using tissue-engineering methods. Methods The osteogenic potential in vitro of cultured BMSc in a conditional medium was examined by histochemistry stains technique. The BMSc was cultured in combination with BGC. The attaching and extending speed of the cells to the materials, the proliferation and alkaline phosphatase activity were tested. Then the composite was implanted into the skeletal muscle beds in rabbits. All implants were examined by gross observation and histological examination. Results The BMSc showed a similar property to those of osteoblasts. BMSc can attach to and extend on BGC materials. No inhibition to celluar proliferation and ALP activity were observed by the materials. New bone can be observed in the composites of the BMSc and BGC materials. Conclusions BMSc may provide a rich cellular resource in tissue-engineered bone formation. New bone tissue can be formed by tissue engineering methods.
3.Changes of CGRP and NPY expression in tissue-engineered bone with fascia flap
Jiande CUI ; Guoxian PEI ; Dan JIN
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
Objective To evaluate the expressive levels of calcitonin gene related peptide (CGRP) and neuropeptide Y (NPY) in tissue-engineered bone with fascia flap in vivo. Method A segmental bone defect 1.5cm in length was made at the both radius of 12 healthy New Zealand rabbits. The defects were repaired by implantation in two ways: for left radius, engineered bone with fascia flap was implanted (served as experiment group), and for right radius only engineered bone was implanted (served as control group). 3, 6 and 12 months after implantation, the expressions of CGRP and NPY in the new bone were determined with immunohistochemistry and semi-quantified using image analysis software. Results CGRP and NPY expressions in the both groups were significantly increased in a time-dependent manner (P
4.Bone formation by bone marrow stromal cell using tissue engineering methods in vitro and in vivo
Dan JIN ; Guoxian PEI ; Qian WANG
Chinese Journal of Microsurgery 2000;0(03):-
Objective To study the osteogenic capability of bone marrow stromal cell (BMSc) using tissue engineering methods Methods The osteogenic potential in vitro of cultured BMSc in a conditional medium were examined by phase contrast microscopy,histochemistry stains technique The BMSc were cultured in combination with bioactive glass ceramic (BGC) materials Then the composite were implanted into the skeletal muscle beds in rabbits All implants were exmined by gross observation and histological examination Results The BMSc showed a similar property to those of osteoblasts and could synthesized mineralized new bone tissue in vitro New bone tissue can be observed in the composites of the BMSc and BGC materials Conclusions New bone tissue can be formed by tissue engineering methods
5.HUMAN BONE MORPHOGENETIC PROTEIN 7 EXPRESSION IN RABBIT BONE MARROW STEM CELL TRANSFERRED BY RETROVIRAL VECTOR MEDIATED hBMP_7 GENE
Dan JIN ; Weisen ZENG ; Guoxia PEI
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
The human bone morphogenetic protein 7 (hBMP 7) gene was reconstructed in retroviral vector and transferred into incasing cells PT67 by liposome mediated method.The clones of the cells transferred with BMP 7 were selected by G418, and targeted rabbit bone marrow stem cells were infected with the virus granules which secreted from PT67 cells and also selected by G418. The mRNA and protein of BMP 7 gene in transferred cells were analyzed with hybridization in situ and immunohistochemistry. BMP 7 retrovirus vetor was successfully reconstructed. Cells transferred by PLNCX 2 hBMP 7 expressed abundant human BMP 7 mRNA and protein in the cytoplasm. However positive findings were not found in those cells that were not transferred. It may be used to increase the osteogenic capability of BMSc in the study of bone tissue engineering.
6.Feasibility of chitosan-beta-tricalcium phosphate as an injectable bone tissue engineering scaffold material
Wenjun CHENG ; Dan JIN ; Guoxian PEI
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
0.05).Under SEM,the BMSCs showed good adhesion to beta-TCP with obvious proliferation.Conclusion BMSCs can grow and proliferate well on the compound BMSCs/beta-TCP and beta-TCP has good biocompatibility with BMSCs in vitro,which may be used as a good scaffold material for injectable tissue engineering bone.
7.Association of the degradation of the hybrid layer and the durability of dentin adhesion.
Dan-Dan PEI ; Xue-Qing HUANG ; Cui HUANG
Chinese Journal of Stomatology 2011;46(1):58-60
Acid Etching, Dental
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adverse effects
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Chondroitin ABC Lyase
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chemistry
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Dental Bonding
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Dentin
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chemistry
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ultrastructure
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Dentin-Bonding Agents
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chemistry
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Humans
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Hydrolysis
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Matrix Metalloproteinase Inhibitors
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pharmacology
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Matrix Metalloproteinases
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metabolism
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Microscopy, Electron, Transmission
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Surface Properties
8.Immunological evaluation into the bone allograft and selection of experimental animal model on im-munogenicity
Mingdong LI ; Tingfei XI ; Dan JIN ; Guoxian PEI
Chinese Journal of Orthopaedic Trauma 2009;11(7):671-674
Objective To explore the effect of immunogenicity of freeze-dried bone allograft on different in vitro experimental models. Methods The lymphocytes were obtained respectively from 10 healthy young human volunteers, 10 Balb/c and 10 C57 mice and 10 New Zealand rabbits. The experiment was carried out in 6 groups: positive control group (PHA/ConA+lymphocyte), negative control group (Hydroxyapatite powder + lymphocyte), allogeneic bone group A (Freeze-dried bone powder 2. 0 g/L + lym-phocyte), allogeneic bone group B (Freeze-dried bone powder 1.0 g/L + lymphocyte), allogeneic bone group C (Freeze-dried bone powder 0.5 g/L + lymphocyte), and negative control group (culture solution + lym-phocyte). Lymphocyte transformation test (Alamarblue) was conducted to culture the 6 kinds of experimental materials in vitro. After 72 hours, samples were scanned with ELISA muhiscan at wave lengths 570 nm and 600 nm to fetal the light absorption value. Pearson analyses were performed 10 determine the relationships a-mong the 3 animals and 1 human groups and find out which animal would be highly correlated to human. Results In the human and Balb/c mice lymphocyte transformation tests, there was no significant difference (P > 0.05) between allogeneic bone groups A, B, C and negative control group (HA) ; but there was sig-nificant difference (P < 0.001) between allogeneic bone groups A, B, C and positive control group (PHA/ConA); there was no significant difference between the 3 allogeneic bone groups (P > 0.05). There was no significant difference among the 6 groups of C57 mice and New Zealand rabbits (P > 0.05). The coefficient r between Balb/c mice and human groups was 0.959, P = 0.003, showing a highly positive correlation. The coefficient r between C57 mice and human groups was 0.527, P = 0.283, while the coefficient r between New Zealand rabbits and human groups was 0.866, P =0.026. Conclusions The immunogenicity of freeze-dried bone powder in this experiment may not be sufficient enough to induce significanrt immunologic response. Balb/c mice may be preferable for immunogenicity related experiments.
9.Designation and silencing function of the small interfering RNA of HLA-A2
Jiansheng LIU ; Gang ZHAO ; Dan PEI ; Deyu ZHENG
Chinese Journal of Tissue Engineering Research 2014;(29):4605-4610
BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants.
OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2.
METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis.
RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.
10.Human bone morphogenetic protein 7 gene transfection for the proliferation and osteogenetic differentiation of the bone marrow mesenchymal stem cells
Dan JIN ; Guoxian PEI ; Ke WANG ; Kuanhai WEI ; Bin CHEN
Chinese Journal of Tissue Engineering Research 2005;9(14):206-208
BACKGROUND: The main aspect of the study in the bone histological engineering is how to maintain and improve theosteogenesis of the osteoblasts in vivo and in vitro. The gene transference may provide a new effective method to deal with theproblem.OBJECTIVE: To discuss the effect of the reverse transcription virus mediated human bone morphogenetic protein7(hBMP-7) gene transfection on the proliferation and osteogenetic differentiation of the bone marrow mesenehymal stemcells (BMSCs) of the rabbits.DESIGN:Cells taken as the study object, grouping control, repeat observation andmeasurement.SETTING: Traumatological and othopaedic lab of a medical university hospital.PARTICIPANTS: The study wascompleted in the Traumatological and Othopaedic Lab in the Affiliated Nanfang Hospital of the Southern Medical University from July 2001 to July 2003. Four New Zealand rabbits,whose weights varied from 1.0 to 1.5 kg, were provided without sexlimit by the Animal Experiment Center of the First Military Medical University of Chinese PLA.METHODS:The reverse transcription virus carriersof the hBMP-7 were constructed,and then the BMSCs were transfected by the virus containing target genes. The expression of the hBMP-7 protein was detected with the immunohistochemical method. The cell proliferation, cycle and ALP synthesis were respectively detected with the MTT method,flow cytometer and NPP method.MAIN OUTCOME MEASURES: Primary results: ① the detection results of the cell proliferation. ② the detection results of the ALP.Secondary results: ① the expression of the hBMP-7 protein in the transfected BMSCs. ② the detection results of the cell cycle.RESULTS: After the BMP-7 gene transfection, there was hBMP-7 positive expression in the BMSCs of the rabbits,using the immunohistochemical detection. There was no significant change in the BMSCs proliferation of the rabbits after the hBMP-7 gene transfection ( P > 0.05). Compared with the ALP synthesis of the transfected BMSCs(294. 592 ± 86. 567) nkat/L, there was significant difference in the ALP synthesis of the empty carrier transfected BMSCs(155. 231 ±86.567) nkat/L and the un-transfected BMSCs (160. 866 ±91. 585)nkat/L( F =5. 660, P < 0. 05).CONCLUSION: After the BMP-7 gene transfection, the BMSCs can synthesize and express the extragenous BMP-7. The hBMP gene transfection can promote the differentiation of the BMSCs cultured in vitro into the osteoblasts and can be used as the seed cells in the construction of the histological en gineering bone tissues and in further application.