Modeling of Acute Deep Venous Thrombosis in Rabbits / 中国康复理论与实践

@#Objective To establish a model of deep venous thrombosis (DVT) in rabbits. Methods Animal models of venous thrombosis were made by blocking venous flow with a vascular clamp temporarily, injuring the vascular wall, and injecting thrombin in the distal vein in one side of rabbits. Then fixed the hip and knee joints of the operated sides in the flection position with plaster. 48 h later, the femoral veins on both sides were examined with the ultrasonography and pathology. Results All the rabbits survived after operation. The ultrasonography showed that the femoral veins on both sides were virtually anechoic. However, the veins on the operated sides couldn't be compressed and no flow was detected, the control sides were just the reverse. The veins on the operated side were filled with thrombus which had not adhered on the wall, but no thrombosis occurred in the control side. Conclusion A model of DVT was established in rabbits.

Inducing Human Bone Marrow Mesenchymal Stem Cells to Cardiomyo-Like Cells with Supernatant of Normal or Injured Rat Cardiomyocytes in Vitro / 中国康复理论与实践

@# Objective To observe whether human bone marrow mesenchymal stem cells (hMSCs) could differentiate into cardiomyo-like cells by culturing with supernatant of normal or injured rat cardiomyocytes (CMs) in vitro. Methods hMSCs were cultured with supernatant from normal or injured rat CMs for 27~30 d. The morphologies of induced hMSCs were observed with inverted microscope and the special cardio-markers cTnI and Desmin were identified with immunocytochemisry. Results A few cells cultured with supernatant from normal CMs enlarged and expressed cTnI, but little Desmin. While more cells cultured with supernatant from injured CMs enlarged and expressed cTnI, and parts of them expressed Desmin. The incidence of cTnI or Desmin positive cells were significantly different between these two groups (P<0.01). Conclusion Supernatant from both normal and injured CMs can induce hMSCs into cardio-like cells in vitro, and that from injured CMs is more effectively.

Action Potential of Cardiomyocyte-like Cells Differentiated from Human Marrow Mesenchymal Stem Cells in Vitro / 中国康复理论与实践

@#Objective To detect whether the cardiomyocyte-like cells differentiated from human marrow mesenchymal stem cells (hMSCs) could produce action potential (AP). Methods Isolated and cultured hMSCs were induced into cardiomyocyte-like cells with 5-Azacytine in vitro. They were measured for their AP by patch clamp technique, and compared with those of hMSCs of the same generation and beating cardiomyocytes (CMs) derived from 2 day-old SD rats. Results 6/30 cardiomyocyte-like cells produced AP. The CMs produced significant AP, hMSCs appeared no AP, and cardiomyocyte-like cells appeared weak AP. Conclusion The hMSCs manifested the potential to differentiate into CMs in the electrophysiology characteristics following 5-Azacytine induction.

Using damaged cardiomyocytes onditional induction culture medium to induce the differentiation of human bone marrow mesenchymal stem cells / 国际生物医学工程杂志

Objective To investigate the possibility of conditionally induced culture medium (CICM) of oxidative damaged cardiomyocytes(CMs) derived from SD suckling rats to induce the differentiation of human bone marrow mesenchymal stem cells (hMSCs) into cardiomyo-like cells. Methods hMSCs and SD suckling rats' CMs were respectively isolated and cultured in vitro. Beating CMs were damaged by H_2O_2 and the the resultant culture medium wherein the oxidative damaged CMs were euhivated was used as CICM. Passaged hMSCs were induced by CICM for 4 weeks. The morphologies of induced hMSCs were observed by inverted microscope and the special cardio-markers-Cardiac troponin I(cTnI) and desmin were identified by immunocytochemistry method. Results After coincubation with CICM, hMSCs became larger and expressed cTnI and desmin.Conclusion CICM of oxidative injured CMs could induce hMSCs differentiation into cardiomyo-like cells.

Role of 5-azacytidine in differentiation of human mesenchymal stem cells into cardiomyocytes in vitro / 老年心脏病学杂志（英文版）

Objective 5-azacytidine could induce the diffcrentiation of stem cells into cardiomyocytes (CMs). The aim of this study was to screen the optimal condition for 5-azacytidine inducing differentiation of human mesenchumal stem cells (hMSCs) into CMs, and the effect of 5-azacytidine on adherence, cell vigor and chromosome karyotype ofhMSCs. Methods hMSCs were isolated from human bone marrow and cultured in vitro. The phenotypes ofhMSCs were identified by flow cytometric analyses. MTT test was used to investigate the effect of different concentrations of 5-azacytidine on proliferation ofhMSCs. Four weeks after 5-azacytidine induction, semi-quantitative RT-PCR, transmission electron microscopy (TEM), single-cell action potentials, detection ofcardio-cnzyme AST and LDH, cell adherence, cell viability and chromosome karyotype test were performed. Results The typical morphological features of hMSCs were fibroblast-like in shape, hMSCs expressed CD44 and CD105,and did not express CD34, CD45 and CD31. The optimal concentration of 5-azacytidine was 10μmol/L. The shape ofhMSCs treated with 5-Azacytidine changed from fusiform to polygon or astrocyte gradually, and passaged cells were evenly arranged as polarity structure. Induced-hMSCs connected with neighbouring cells, forming myotube-like structures 4 weeks later. It was confirmed that induced hMSCs shaped myotubule-like structure and had some of micro-histologic structures of CMs by TEM. RT-PCR showed that induced hMSCs expressed cardiac specific product BNNP and early cardio-myogenesis specific transcription factor NKX2.5mRNA. Besides, induced-MSCs led to the weak action potential and secreted cardio-enzyme AST and LDH. There was no significant difference in cell adherence and viability before and after induction. Both hMSCs and induced-hNSCs kept stable normal diploid nucleus. Conclusion The optimal condition for inducing effect of 5-azacytidine is 10 μ mol/L and 24-hour incubation; and under this condition, the adherence, vigor and chromosome karyotype ofhMSCs would not be affected.

Isolation and identification of hepatic stem cells from rat fetal liver in vitro / 吉林大学学报(医学版)

Objective To isolate and cultivate hepatic stem cells(HSCs) from rat fetal liver in vitro and identify their biological features.Methods Collagenase perfusion method and mechanical cutting method were used to isolate HSCs from rat fetal liver which were then cultivated by H-DMEM containing 10% fetal bovine serum.The cell surface antigen expression of HSCs was observed with immunocytochemical method under confocal laser scanning microscope.Results The isolated HSCs from rat fetal liver grew to monolayer 5 d after cultivation in vitro.They presented pykno-round cells and distinct borderline under the light microscope.After 8 d the cells grew like epithelium.These cells expressed AFP antigen,CK 18 and CK19.Conclusion The cultivated cells are proved to be HSCs and can proliferate quickly in vitro.

Rehabilitation on sequelae of epidemic encephalitis: 41 case report / 中国康复理论与实践

@#ObjectiveTo study effect of rehabilitation on the sequelae of encephalitis. Methods 41 patients were divided into two groups, Group A( only treated with traditional Chinese medicine)and group B ( treated with both traditional Chinese medicine and rehabilitation). Each patient was evaluated before and after the treatment. Result The treatment in the group B was significantly better than that in the group A (P<0.05), especially in motor function. Conclusion Comprehensive rehabilitation is an effective therapy on the sequelae of encephalitis, which shortens the course of the sequelae and improves the motor function and speech.

Culture,proliferation and differentiation of human marrow mesenchymal stem cells to cardiomyocytes in vitro / 中国免疫学杂志

95% purity were treated for 24 h with 1,5,10 and 20 ?mol/L 5-azacytidine to induce cellular differentiation.Expression of the cardiac-specific marker-troponin Ⅰ and desmin,identified by immunohistochemistry,was used to identify cardiac muscle differentiation.Results:hMSCs expressed a high level of CD44(hMSCs 93.26%?2.48% vs 3.42%?1.09% in a control group,P