Objective To review the current status and problems in developing cardiac biological pacemaker(CBP) by cell transplantation. Methods The l iterature over the past decade concerning CBP constructed through celltransplantation was reviewed and summarized. Results Experiments in vivo testified that the cell transplantation was feasible for CBP construction, and the transplantation of sinus atrial node cell and stem cell was still the predominant method for constructing CBP. However, such problems as difficult ampl ification of transferred cardio muscle cell, low success rate of CBP construction as well as unstable function of CBP make it lag behind the tremendous cl inical demands. The gene transfection technology might be one of the approaches to resolve these issues. Conclusion As one feasible method for CBP construction, the cell transplantation has a bright future in the cl inical appl ication and is worthy of further study.
Objective To observe the change of sino-atrial nodal tissue structure and ectopic pacing function after xenogenic sino-atrial nodal tissue transplanted into left ventricular wall, so as to provide new ideas for the treatment of sick sinus syndrome and severe atrioventricular block. Methods Seventy healthy rabbits were selected, male or female, and weighing 1.5-2.0 kg. Of them, 42 were used as reci pient animals and randomly divided into sham operation group, warm ischemia transplantation group, and cold ischemia transplantation group (n=14), the other 28 were used as donors of warm ischemia and cold ischemia transplantation groups, which were sibl ing of the recipients. In recipients, a 6-mm-long and about 2-mm-deep incision was made in the vascular sparse area of left ventricular free wall near the apex. In sham operation group, the incision was sutrued directly by 7-0 Prolene suture; in cold ischemia transplantation group, after the aortic roots cross-clamping, 4 ℃ cold crystalloid perfusion fluid infusion to cardiac arrest, then sinoatrial node were cut 5 mm × 3 mm for transplantation; in warm ischemia transplantation group, the same size of the sinus node tissue was captured for transplantation. After 1, 2, 3, and 4 weeks, 3 rabbits of each group were harvested to make bradycardia by stimulating bilateral vagus nerve and the cardiac electrical activity was observed; the transplanted sinus node histology and ultrastructural changes were observed. Results Thirty-six recipient rabbits survived (12 rabbits each group). At 1, 2, 3, and 4 weeks after bilateral vagus nerve stimulation, the cardiac electrical activity in each group was significantly slower, and showed sinus bradycardia. Four weeks after operation the heart rates of sham operation group, warm ischemia, and cold ischemia transplantation group were (81.17 ± 5.67), (82.42 ± 7.97), and (80.83 ± 6.95) beats/ minute, respectively; showing no significant difference among groups (P gt; 0.05). And no ectopic rhythm of ventricular pacing occurred. Sino-atrial nodal tissue survived in 6 of warm ischemic transplantation group and in 8 of cold ischemia transplantation group; showing no significant difference between two groups (P gt; 0.05). Two adjacent sinoatrial node cells, vacuole-l ike structure in the cytoplasm, a few scattered muscle microfilaments, and gap junctions between adjacent cells were found in transplanted sinus node. Conclusion The allograft sinus node can survive, but can not play a role in ectopic pacing.
ObjectiveThrough comparing the efficacy of levosimendan with dopamine for severe valvular disease patients with atrial fibrillation surgery to explore the efficacy and safety of levosimendan used in cardiac surgery. MethodsWe allocated 48 severe valvular disease patients with atrial fibrillation surgery into a dopamine group (24 patients with 15 males and 9 females at age of 55.0 ± 17.4 years) and a levosimendan group (24 patients with 18 males and 6 females at age of 52.3 ± 16.2 years) by random digital table in the Affiliated Hospital of Luzhou Medical College between February and June 2014. The effects of the two groups were compared. ResultsHospitalization time (18.7±8.6 d vs 20.6±7.5 d, t=11.52, P=0.02) and the incidence of acute kidney injury(1/24 vs 5/24, χ2=25.30, P=0.01) in the levosimendan group were lower than those in the dopamine group. There was no statistical difference between the two groups in other early clinical outcomes. At each postoperative time point, there was no statistical difference in creatine kinase isoenzyme (CK-MB) between the two groups. While 6 to 48 hours after operation, there were significant differences in cardiac troponin (cTnI) and brain natriuretic peptide(BNP) level between the two groups (P < 0.05). Five days after operation, the left ventricular ejection fraction(LVEF) in the levosimendan group was higher than that in the dopamine group with statistical difference. ConclusionLevosimendan used for severe valvular disease with atrial fibrillation surgery is safe and effective, and has certain myocardial protection and renal protection effect, while its mechanism still needs further study.
Objective To study the influence of ischemia-reperfusion on the expression of the hyperpolarization activated cycl icnucleotide gated cation channel 4 (HCN4) and to discuss the mechanism of functional disturbance of sinoatrial node tissue (SANT) after ischemia reperfusion injury (IRI). Methods Eighty five healthy adult rabbits, weighing 2-3 kg, were randomly divided into 3 groups: control group [a suture passed under the root section of right coronary artery (RCA) without l igation, n=5], experimental group A (occluding the root section of RCA for 30 minutes, then loosening the root 2,4, 8 and 16 hours, n=10), experimental group B (occluding the root section of RCA for 1 hour, then loosening the root 2, 4,8 and 16 hours, n=10). At the end of the reperfusion, the SANT was cut off to do histopathological, transmission electronmicroscopical and immunohistochemical examinations and semi-quantitative analysis. Results The result of HE stainingshowed that patho-injure of sinoatrial node cell (SANC) happened in experimental groups A and B after 2 hours of reperfusion, the longer the reperfusion time was, the more serious patho-injure of SANC was after 4 and 8 hours of reperfusion, SANC reached peak of damage after 8 to 16 hours of reperfusion; patho-injure of SANC was more serious in experimental group B than in experimental group A at the same reperfusion time. Immunohistochemical staining showed that the expression of HCN4 located in cellular membrane and cytoplasm in the central area of SANC and gradually decreased from the center to borderl ine. The integral absorbance values of HCN4 expression in the control group (397.40 ± 34.11) was significantly higher than those in the experimental group A (306.20 ± 35.77, 216.60 ± 18.59, 155.40 ± 19.11 and 135.00 ± 12.30) and in the experimental group B (253.70 ± 35.66, 138.70 ± 13.28, 79.10 ± 9.60 and 69.20 ± 8.42) after 2, 4, 8 and 16 hours of reperfusion (P lt; 0.05). With reperfusion time, the expression of HCN4 of SANC decreased, which was lowest after 8 hours of reperfusion; showing significant difference among 2, 4 and 8 hours after reperfusion (P lt; 0.05) and no significant difference between 8 and 16 hours after reperfusion (P gt; 0.05). At the same reperfusion time, the expression of HCN4 was higher in the experimental group A than in the experimental group B. The result of transmission electron microscope showed that ultramicrostructure of SANC was damaged after reperfusion in experimental groups A and B. The longer the reperfusion time was, the more serious ultramicrostructure damage of SANC was, and reached the peak of damage after 8 hours of reperfusion. Ultramicrostructure of SANC was not different between 8 and 16 hours of reperfusion. At the same reperfusion time, the ultramicrostructure damage of SANC was moreserious in experimental group B than in experimental group A. Conclusion IRI is harmful to the morphous and structure ofSANC, and effects the expression of HCN4 of SANC, which is concerned with functional disturbance and arrhythmia.
ObjectiveTo investigate whether Akt1 gene transfection mediated by recombinant lentivirus (LVs) in the bone marrow mesenchymal stem cells (BMSCs) could enhance the ability of hypoxia tolerance so as to provide a theoretical basis for improving the effectiveness of stem cells transplantation. MethodLVs was used as transfection vector, enhanced green fluorescent protein (EGFP) was used as markers to construct the pLVX-EGFP-3FLAG virus vector carrying the Akt1 gene. The 3rd generation BMSCs from 3-5 weeks old Sprague Dawley rats were transfected with pLVX-EGFP virus solution as group B and with pLVX-EGFP-3FLAG virus solution as group C; and untransfected BMSCs served as control group (group A). At 2-3 days after transfection, the expression of green fluorescent was observed by fluorescence microscope; and at 48 hours after transfection, Western blot method was used to detect the expression of Akt1 protein in groups B and C. BMSCs of groups B and C were given hypoxia intervention with 94%N2, 1%O2, and 5%CO2 for 0, 3, 6, 9, and 12 hours (group B1 and group C1) . The flow cytometry was used to analyze the cell apoptosis rate and cell death rate, and the MTT method to analyze the cell proliferation, and Western blot to detect the expression of apoptosis related gene Caspase-3. ResultsAfter transfection, obvious green fluorescence was observed in BMSCs under fluorescence microscopy in groups B and C, the transfection efficiency was about 60%. Akt1 expression of group C was significantly higher than that of group B (t=17.525, P=0.013) . The apoptosis rate and cell death rate of group B1 increased gradually with time, and difference was significant (P<0.05) . In group C1, the apoptosis rate and cell death rate decreased temporarily at 3 hours after hypoxia intervention, then increased gradually, and difference was significant (P<0.05) . The apoptosis rate and cell death rate of group C1 were significantly lower than those of group B1 at each time point (P<0.05) except at 0 hour. MTT assay showed that absorbance (A) values of groups B and C were significantly higher than those of groups B1 and C1 at each time point (P<0.05) ; the A value of group B was significantly lower than that of group C at each time point (P<0.05) . The A value of group B1 was significantly lower than that of group C1 at 6, 9, and 12 hours after hypoxia intervention (P<0.05) . Western blot results showed that the Caspase-3 expression of group C1 significantly reduced when compared with group B1 at each time point (P<0.05) . ConclusionsAkt1 gene transfection mediated by recombinant LVs could significantly improve hypoxia tolerance of BMSCs by inhibiting the apoptosis, which could provide new ideas for improving the effectiveness of stem cells transplantation.