3.Effect of bone marrow mesenchymal stem cells combined with Shh modified nano-hydroxyapatite/collagen for femoral defect repair
Yuxing TANG ; Qing ZHAO ; Zhongmeng YANG ; Yongheng YE ; Renan HU
Chinese Journal of Tissue Engineering Research 2017;21(14):2180-2185
BACKGROUND: SSh as a Hedgehog signal protein can promote bone development, growth and remodeling.OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) combined with Shh modified nano-hydroxyapatite/collagen (nHAC) in the repair of femoral defects in rats.METHODS: Forty-eight Sprague-Dawley rats were randomly divided into four groups, and the model of femoral defects was established in these rats. At 14 days after modeling, experimental group was implanted with the BMSCs/Shh modified nHAC, scaffold group was implanted with simple nHAC, cell scaffold group was implanted with BMSCs/nHAC,and blank control group was without any implantation. At 3, 6, 9, 12 weeks after repair, X-ray examination, bone density measurement and bone biopsy in bone defect area were performed.RESULTS AND CONCLUSION: (1) X-ray examination: The Lane-Sandhu X-ray score and bone mineral density value in the experimental group at different time points after operation were significantly higher than those in the other three groups (P < 0.05). (2) Hematoxylin-eosin staining: 12 weeks after repair, a small amount of bone tissues but no bone marrow formed in the scaffold group; a small amount of bone tissues with absence of bone marrow formed in the cell scaffold group, and the residual scaffold was visible; in the experimental group, the scaffold was completely absorbed,and mature bone and medullary cavity formed with presence of bone marrow. (3) Scanning electron microscope observation: 12 weeks after repair, irregular arrangement of bone fibers and a large number of bone fossae were observed in the scaffold group; the cell scaffold group showed a large number of osteoblasts, but bone fibers still arranged irregularly; in the presence of the Haversian system, a large number of regularly arranged bone trabeculae were detective in the experimental group. These results elucidate that the Shh modified nHAC/BMSCs complex can promote the repair of bone defects.
4.Effects of polylactic acid copolymer/adipose-derived stem cells complex on the biomechanical properties after fracture healing in osteoporotic bone
Yuxing TANG ; Qing ZHAO ; Zhongmeng YANG ; Yongheng YE ; Renan HU
Chinese Journal of Tissue Engineering Research 2017;21(10):1577-1582
BACKGROUND: Polylactic acid copolymer bone scaffold has excellent biodegradability, and it is easy to be shaped and can promote the formation and growth of bone tissue and blood vessel.OBJECTIVE: To observe the effects of adipose-derived stem cells(ADSCs)/poly(lactic-co-glycolic acid) (PLGA) complex on the biomechanical properties after fracture healing in osteoporotic bone.METHODS: Sixty Sprague-Dawley rats were randomly divided into four groups: blank control group received no treatment; the bilateral tibial fracture model was made after 3 months of bilateral ovarian resection in model group; the bilateral tibial fracture model was made and ADSCs were implanted into the bone after 3 months of bilateral ovarian resection in cell therapy group; the bilateral tibial fracture model was made and the PLGA/ADSCs complex was implanted after 3 months of bilateral ovarian resection in combined treatment group.The bone mineral density, callus thickness, biomechanical parameters and the microstructure of the trabecular bone were detected.RESULTS AND CONCLUSION: (1) The bone density: The bone density of the model group was significantly lower than that of the blank control group (P < 0.05); the bone mineral density of the cell therapy group and the combined treatment group was higher than that of the model group (P < 0.05), but lower than that of the control group (P < 0.05); and the bone mineral density of the combination treatment group was higher than that of the cell therapy group (P < 0.05). (2)Thickness of the callus: The thickness of the callus in the cell therapy group and combined treatment group was higher than that of the model group and blank control group (P < 0.05); moreover, the thickness of the callus in the combined treatment group was higher than that of the cell therapy group (P < 0.05). (3) Biomechanical test: The failure load, stress and shear strength, elastic modulus were decreased in the model group compared with the blank control group (P < 0.05), while the shear strain increased (P < 0.05). Compared with the model group, the failure load, ultimate stress, shear strength, elastic modulus were increased in the cell therapy group and combined treatment group (P < 0.05), and the shear strain was decreased (P < 0.05). Moreover, the combined treatment group showed more changes in these biomechanical parameters (P < 0.05). (4) The trabecular bone microstructure: The model group presented with trabecular derangement, spacing increases, and even fracture and lacuna. After ADSCs or ADSCs/PLGA transplantation,the trabecular bones increased in number, thickness, and spacing, and the number of lacunae reduced. In conclusion,ADSCs combined with PLGA in the treatment of osteoporotic fracture can significantly improve the biomechanical parameters of bone tissue after healing.
5.Mechanism of Immune Hyporesponsiveness Induced by Recipient-Derived Immature Dendritic Cells in Rat Liver Transplantation
Li LI ; Shengning ZHANG ; Jianghua RAN ; Jing LIU ; Zhu LI ; Yongheng ZHAO ; Laibang LI
Chinese Journal of Bases and Clinics in General Surgery 2004;0(01):-
Objective To study the mechanism of immune hyporesponsiveness of allograft rejection induced by transfusion nonpufsed allopeptide syngeneic immature dendritic cell(imDC) generated from recipient bone marrow progenitors and to explore a possible strategy for liver allograft protection in clinic.Methods Forty experimental rats were randomly divided into 4 group: control group,cyclosporine A(CsA) group,mature DC(mDC) group and imDC group.In control group,Wistar rats only received liver transplantation.In CsA group,Wistar rats underwent liver transplantation plus CsA treatment(10 mg/(kg?d)).In mDC group,recipient-derived mDC 1?106 were infused intravenously through the penile vein to Wistar rats.In imDC group,ImDC with the dose of 1?106 were injected into Wistar rats via the dorsum vein of penile.In each group,five recipients were killed on the 10th day after transplantation,the other five recipients were left to observe survival time.The levels of ALT,AST,TBIL,IL-2,IFN-?,IL-4 and IL-10 were detected.The acute rejection and the expression of FasL/Fas in the grafts were detected by HE and immunohistochemical staining.Western blot was used to detect Scurfin protein expression of CD4+ CD25+ T cells.Results The median survival time of the liver allografts in CsA group and imDC group were significantly longer than that in control group and mDC group(P
6.Superiority, Confusion and Countermeasures to DCD WeChat Ethical Review
Lan LI ; Guishu CHEN ; Shaohong YU ; Yongheng ZHAO ; Chao LI ; Riuhong ZHANG ; Hongqing ZHANG ; Wenhu JIANG
Chinese Medical Ethics 2016;29(2):288-290
DCD WeChat ethical review had its necessity and superiority but problems and confusion as well. For instance, WeChat review time was urgent and limited;WeChat review discussed insufficiently even hard to car-ry out;the methods of WeChat review were limited;the authenticity of audit opinion was difficult to grasp;the re-view data privacy existed hidden danger;the review quality was difficult to guarantee;WeChat conference content file archive was difficult, and so on. As to this, the author put forward the following countermeasures:giving war-ranty on the ethical review time as far as possible;initiating the committee review by the referee and uploading the review information to WeChat group, then ethical reviewers expressing their opinions in real names to ensure data confidentiality and privacy protection; strengthening the ethics committee education for organ transplantation; im-proving the quality of the review committee members and moral cultivation;archiving the WeChat conference infor-mation.
7.Laparoscopic hepatectomy: a report of 6 cases
Shengning ZHANG ; Jing LIU ; Jianghua RAN ; Shuyuan WU ; Zhu LI ; Yongheng ZHAO ; Laibang LI ; Yizhou JIANG ; Li LI
International Journal of Surgery 2011;38(1):22-24
Objective To explore the indication and feasibility of laparoscopic hepatectomy. Methods Clinical data of 6 patients undergoing laparoscopic hepatectomy(LH) were collected and retrospectively anallyzed,5 of them with lesions located in surface or edge of Ⅱ -Ⅵ segment,one of them with lesion in Ⅷ segment. These lesions were 5 - 9.6 cm, the average diameter was (6.64 ± 2.60) cm. There were 4 cases of liver cavernous hemangioma, and 2 case of hepatic focal nodular hyperplasia. The liver functions of 6 cases w ere in Child Pugh A . Results All 6 patients were applied laparoscopic hepatetomy successfully, 5 cases were performed partial resection, 1 case underwent laparoscopic hepatic left lateral lobectomy. The average operation time was( 105.17 ± 27.97 )minutes, and the intraoperative average hemorrhage was (247. 50 ± 90.91 ) mL. All of the lesions were completely removed. There were no postoperative complications such as bile leakage or hemorrhage. All patients recovered well. The average postoperative hospitalization was (4.16 ± 1.60)days: Conclusions Laparoscopic hepatectomy is safe and feasible for lesion located in the edge or sur face of liver and left liver.
8.Construction of the recombinant adenovirus vector carrying antisense multidrug resistance (MDR1) gene.
Bo LI ; Xinghua GOU ; Lin CHEN ; Lünan YAN ; Dehua LI ; Yongheng ZHAO ; Le HAN ; Lanying ZHAO
Journal of Biomedical Engineering 2004;21(3):424-427
The fragment of MDR1 gene obtained from the plasmid pHaMDR1-1 carrying the whole human MDR1 cDNA, was cloned reversely into the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the Escherichia coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged in the 293 cells. The recombinant adenovirus MDR1 vector would introduce the antisense MDR1 gene into the human multidrug resistance hepatocellular cell line effectively, which would provide an experimental basis for studies on the multidrug resistance in human hepatocellular carcinoma.
ATP-Binding Cassette, Sub-Family B, Member 1
;
biosynthesis
;
genetics
;
Adenoviridae
;
genetics
;
metabolism
;
Carcinoma, Hepatocellular
;
drug therapy
;
genetics
;
Cell Line, Tumor
;
Drug Resistance, Neoplasm
;
genetics
;
Escherichia coli
;
genetics
;
Gene Transfer Techniques
;
Genes, MDR
;
genetics
;
Genetic Vectors
;
Humans
;
Liver Neoplasms
;
drug therapy
;
genetics
;
Multidrug Resistance-Associated Proteins
;
biosynthesis
;
genetics
;
Oligonucleotides, Antisense
;
pharmacology
;
Plasmids
;
Recombinant Proteins
;
biosynthesis
;
genetics
;
Recombination, Genetic
9.Effect of Extracorporeal Liver Alloperfusion in Treatment of Acute Liver Failure in Pigs
Guoxun LI ; Yongheng ZHAO ; Ximo WANG ; Gang LONG ; Zhongkui JIN ; Jinggang ZHU ; Jialin WANG ; Honglei WANG ; Ping WANG ; Gang LI ; Yu ZHANG ; Jin GONG ; Tao JIANG ; Shi CHEN
Chinese Journal of Bases and Clinics in General Surgery 2004;0(01):-
Objective To evaluate the feasibility and curative effect of extracorporeal liver perfusion (ECLP) in treatment of acute liver failure (ALF) in pigs. Methods The experiments were carried out in healthy pigs (weight 20-30 kg) under general anesthesia. All of the pigs were randomly divided into 3 groups. ALF model was established by liver blood supply obstructing and portal-systemic shunting. ALF group (n=5): ALF pigs were killed 8 h after establishing. ALF+ECLP group (n=5): ALF pigs were perfused with ECLP for 4 h and killed 8 h after establishing. Normal liver+ECLP group (n=4): normal pigs were dealt with just as ALF+ECLP group. The data of PT, AST, TNF, blood ammonia were collected in all groups. Pathologic changes in liver and brain were detected. Results The levels of PT, AST, TNF, blood ammonia, RBC and HCT in the ALF+ECLP group were lower than those in the ALF group (P
10.Quantitative comparison of 68Ga-NGR and 18F-FDG uptake in well-differentiated hepatocellular carcinoma bearing mice
Yongheng GAO ; Zhengjie WANG ; Fei KANG ; Xiaowei MA ; Wenhui MA ; Mingru ZHANG ; Mingxuan ZHAO ; Tianming FU ; Guoquan LI ; Shengjun WANG ; Zhe WANG ; Weidong YANG ; Jing WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(3):147-152
Objective To quantitatively compare the diagnostic capability of 68Ga-NGR and 18F-FDG in well-differentiated hepatocellular carcinoma (HCC) bearing mice by microPET/CT imaging.Methods The in vitro cellular uptake, in vivo microPET/CT imaging and biodistribution studies of 68Ga-NGR and 18F-FDG were quantitatively compared in SMMC-7721-based well-differentiated HCC.The human fibrosarcoma (HT-1080) and human colorectal adenocarcinoma (HT-29) cells/xenografts were respectively used as positive and negative reference groups for CD13.The expression of CD13 was qualitatively verified by immunohistostaining.The levels of CD13 and glucose-6-phosphatase (G6Pase) were semi-quantitatively analyzed by Western blot test for all 3 types of tumors.Two-sample t test was used for data analysis.Results The in vitro cellular uptake showed that the 68Ga-NGR uptake in SMMC-7721 and HT-1080 cells was higher than that in HT-29 cells, and the 68Ga-NGR uptake was higher than 18F-FDG uptake in SMMC-7721 cells.The in vivo microPET/CT imaging results revealed that the uptake of 68Ga-NGR in SMMC-7721 tumor was (2.17±0.21) %ID/g, remarkably higher compared to (0.73±0.26) %ID/g of 18F-FDG uptake (t=8.826, P<0.01).The tumor/liver ratio of 68Ga-NGR was 2.05±0.16, which was 2.03-fold higher than that of 18F-FDG.In the HT-1080 tumors, the uptakes of 68Ga-NGR and 18F-FDG were both high, and the values were (2.46±0.23) %ID/g, (3.47±0.31) %ID/g.The uptake of 68Ga-NGR was significantly lower than that of 18F-FDG in HT-29 tumors: (0.67±0.20) %ID/g vs (3.17±0.29) %ID/g;t=4.221, P<0.01.Western blot and immunohistostaining results were as follows: HT-1080(CD13+, G6Pase-), SMMC-7721(CD13+, G6Pase+), HT-29(CD13-, G6Pase-).Conclusions The uptake of 68Ga-NGR is higher than 18F-FDG uptake in SMMC-7721 tumor bearing mice, therefore it is worthwhile to consider the feasibility of clinical translation for PET/CT in diagnosis of HCC.Furthermore, because of the difference in 68Ga-NGR and 18F-FDG avidities in tumors with different molecular phenotypes of CD13 and G6Pase, there is an underlying potential for molecular imaging in the determination of molecular phenotypes.