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BACKGROUND:Studies have shown that thymosinβ4 can improve the anti-apoptotic ability of a variety of cells, but the reports describing the changes in the anti-apoptotic ability of bone mesenchymal stem cells modified with thymosinβ4 gene in the hypoxic environment are rare. OBJECTIVE:To observe the change in the apoptotic rate of bone mesenchymal stem cells modified with thymosinβ4 gene in the hypoxic environment, and to explore whether thymosinβ4 affects the apoptosis of bone marrow mesenchymal stem cells via regulating the expression of Bax and Bcl-2. METHODS:Bone marrow mesenchymal stem cells were transfected with the recombinant lentiviral vector over-expressing the thymosinβ4 gene, and then we observed the expression of thymosinβ4 using western blot assay. cells were divided into three groups:thymosinβ4 transfection group, control virus group, and untreated group. Al three groups were placed in a hypoxic environment. The apoptotic rate of bone marrow mesenchymal stem cells was evaluated by the flow cytometry assay. The expression of Bax and Bcl-2 protein was detected by western blot. RESULTS AND CONCLUSION:Thymosinβ4 gene was expressed successful y in the bone marrow mesenchymal stem cells showed by the western blot. The apoptotic rate of bone marrow mesenchymal stem cells in the hypoxic environment was lower in the thymosinβ4 transfection group than the control virus group and untreated group;while there was no difference between the latter two groups. Western blot results showed that the expression of Bcl-2 protein was higher in the thymosinβ4 transfection group than the control virus group and untreated group, and the expression of Bax protein was lower in the thymosinβ4 transfection group than the control virus group and untreated group. No difference in the expression of Bax and Bcl-2 was found between the control virus group and untreated group. These findings indicate that thymosinβ4 overexpression can improve the anti-apoptotic ability of bone mesenchymal stem cells modified in the hypoxic environment, and its possible mechanism is through regulating the expression of Bax and Bcl-2 protein.
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Objective To summarize the diagnosis and treatment of one case of combined heart liver transplantation. Methods On November 24, 2011, one case of combined heart-liver transplantation was performed on a patient with Ebstein's anomaly and tricuspid valve replacement after 5 years,complicated with congestive cirrhosis,liver failure dccompensation,preoperative heart failure Ⅲ degree and B grade of liver function Child-Pugh score. The operation was done with the graded cardiopulmonary bypass assisted mode:first creating the vena cava-aortic bypass to complete heart transplantation, second creating the femoral vein-ascending aorta bypass to complete liver transplantation,and third stopping and neutralizing.The aortic cross-clamping time was 54 min and the an hepatic phase was 38 min.The total time of three times of cardiopulmonary bypass was 199 min and the total time-consuming of operation was 517 min. The patient was given basiliximab +methylprednisolone for immune induction therapy, and tacrolimus + mycophenolate mofetil +prednisone solution for anti-rejection. After operation, liver protecting treatment, anti-infection therapy and nutrition support therapy were given.Results The recipient died of multiple organ failure after 78 days.The mechanical ventilation treatment duration for this recipient was 78 days and ECMO adjuvant therapy for postoperative hypoxemia time lasted 63 days.Conclusion The combined heart liver transplantation is an effective measures for treatment of heart and liver failure.
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Objective To compare the results of mitral valve reconstruction and replacement as treatments for moderate to severe ischemic mitral regurgitation(IMR), and report the mid-term outcome. Methods From June 2002 to May 2008, 83 pa-tients with moderate IMR(35 cases) and severe IMR (48 cases) underwent coronary artery bypass grafting(CABG) combined with mitral valvuloplasty (MVP) (n = 43) or mitral valve replacement (MVR) (n = 40). There were 49 males and 34 females with a mean age of (59.3±7.5) years(51 -77years). The procedures of MVP included annuloplasty with a Dacron or autologous per-icardium ring in 21cases, commissural annuloplasty in 9, quadrangular resection of the posterior leaflet in 9 and using St. Jude mitral annuloplasty ring in 4. In the cases underwent MVR, 28 patients received mechanical prostheses and 12 received biopros-theses. Results 30-day mortality rate was 2.3% for MVP and 5.0% for MVR (P >0.05). The 30-day complication rate was similar for the 2 groups but mechanical ventilation time was longer for MVR patients. Mild MR ocurred in 6 patients with MVP (P <0.05). Sevonty-six patients were followed by outpatient department visit or telephone for (20.2 ± 4.9) months (3 - 60 months). During the follow-up period, 7 patients with MVP had mild insufficiency but free off etber complications. All the valve prothesis functioned well. However, 3 cases had thromboembolic complications and 7 late deaths were recorded in MVR group. Five-year complication-free survival rate was 90% for MVP group and 61% for MVR. Conclusion MVP resulted in excellent durability and provided significant mid-term survival benefit over MVR. MVP should be the first choice for patients with chronic IMR.
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The feasibility of bone marrow stromal cells autologous transplantation for rabbit model of dilated cardiomyopathy induced by adriamycin was studied. Twenty rabbits received 2 mg/kg of adriamycin intravenously once a week for 8 weeks (total dose, 16 mg/kg) to induce the cardiomyopathy model with the monitoring of cardiac function by transthoracic echocardiography. Marrow stromal cells were isolated from cell-transplanted group rabbits and were culture-expanded on the 8th week. On the 10th week, cells were labeled with 4,6-diamidino-2-phenylindole (DAPI), and then injected into the myocardium of the same rabbits. The results showed that viable cells labeled with DAPI could be identified in myocardium at 2nd week after transplantation. Histological findings showed the injury of the myocardium around the injection site was relieved with less apoptosis and more expression of bcl-2. The echocardiography found the improvement of local tissue movement from (2.12±0.51) cm/s to (3.81±0.47) cm/s (P＜0.05) around the inject site, but no improvement of heart function as whole. It was concluded bone marrow stromal cells transplantation for dilated cardiomyopathy was feasibe. The management of cells in vitro, the quantity and the pattern of the cells transplantation and the action mechanism still need further research.
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The feasibility of bone marrow stromal cells autologous transplantation for rabbit model of dilated cardiomyopathy induced by adriamycin was studied. Twenty rabbits received 2 mg/kg of adriamycin intravenously once a week for 8 weeks (total dose, 16 mg/kg) to induce the cardiomyopathy model with the monitoring of cardiac function by transthoracic echocardiography. Marrow stromal cells were isolated from cell-transplanted group rabbits and were culture-expanded on the 8th week. On the 10th week, cells were labeled with 4,6-diamidino-2-phenylindole (DAPI), and then injected into the myocardium of the same rabbits. The results showed that viable cells labeled with DAPI could be identified in myocardium at 2nd week after transplantation. Histological findings showed the injury of the myocardium around the injection site was relieved with less apoptosis and more expression of bcl-2. The echocardiography found the improvement of local tissue movement from (2.12+/-0.51) cm/s to (3.81+/-0.47) cm/s (P<0.05) around the inject site, but no improvement of heart function as whole. It was concluded bone marrow stromal cells transplantation for dilated cardiomyopathy was feasibe. The management of cells in vitro, the quantity and the pattern of the cells transplantation and the action mechanism still need further research.
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<p><b>OBJECTIVE</b>To investigate the role of anti- interleukin-2 receptor (CD25) monoclonal antibody in the regulation of cytokine mRNA expression of IL-1beta, IL-2, CD25, IL-4, IL-5, IL-6, IL-10, tumour necrosis factor-alpha (TNFalpha), and interferon-gamma (IFNgamma) in cardiac allografts to elucidate its immunological mechanism and role in rats that have undergone cardiac transplantation.</p><p><b>METHODS</b>These in vivo studies were conducted using a rat MHC mismatch SD to Wistar heterotopic cardiac transplant model. Simulect, an anti-CD25 antibody, was used to prevent allograft rejection. An increase in the rate of allograft survival was observed. Rats were sacrificed on day 1, 3, 5, 7, 9, 11, 14 post-transplantation and hearts were harvested for further study. Cytokine mRNA expression was determined by semiquantitative RT-PCR.</p><p><b>RESULTS</b>In the control group, cardiac allografts were rejected at 8.3 +/- 1.7 days after transplantation (x +/- s). The rats who received CsA rejected the cardiac allograft at 26.4 +/- 5.7 days post-transplant. Allograft survival of Simulect-treated rats was 29.2 +/- 7.1 days (P < 0.05 vs controls). Rats treated with simulect and CsA had the longest survival of 55.0 +/- 11.6 days (P < 0.001 vs controls). CD25 mRNA expression in the heart tissue samples of treated rats was undetectable or very weak. However, the untreated group, CD25 expression increased, although anti-CD25 decreased this CD25 expression in the heart graft. Furthermore, in untreated allografts, IL-2, TNFalpha and IFN-gamma were strongly expressed, an effect that markedly decreased after simulect treatment. Finally, IL-4, IL-5, IL-6 and IL-10 expression was strong in anti-CD25-treated allografts.</p><p><b>CONCLUSIONS</b>These results suggest that anti-CD25 antibody treatment may not only neutralize CD25 activity but also play a role in altering cytokine mRNA expression and prolong the survival of allografts.</p>
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Animais , Masculino , Ratos , Anticorpos Monoclonais , Usos Terapêuticos , Citocinas , Genética , Sobrevivência de Enxerto , Transplante de Coração , Alergia e Imunologia , RNA Mensageiro , Ratos Sprague-Dawley , Ratos Wistar , Receptores de Interleucina-2 , Fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transplante HomólogoRESUMO
Objective: Interleukin-10 (IL-10) is an anti-inflammatory helper T cell type 2 (Th2) cytokine that modulates Th1-type cytokine production. The purpose of present study was to detect the expression and changes of IL-10 in the rats heart transplantation, as determine the role of IL-10 in the acute rejection of rats heart transplantation. Methods: Hearts from DA rats were transplanted into the necks of Wistar recipients. Allografts was divided into five groups: group A (control, n=12), group B (DST treated, n=12), group C (IL-2 Mab treated, n=12), group D (IL-4 Mab treated, n=12), group E (IL-2 Mab treated, n=12). The local expression of IL-10 was analyzed at day 1,3,5,7,9,11,14 by reversed transcripition polymerase chain reaction (RT-PCR). Results: Allografts survival time was (8.3±1.7) days, (29.2±7.1) days, (26.4±5.7) days, (10.2±1.9) days and (55.0±10.6) days (P＜0.05， P＜0.01), respectively. When compared with group A, the level of IL-10 was upregulated in group B, group C and group E, especially in group E. The upregulation of IL-10 was related to the grafts survival time. Conclusion: IL-10 plays an important role in the development of acute transplantational rejection, suggests that Th-2 type deviation may contribute to the prolongation of graft survival.