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Objective To investigate use of magnetic resonance diffusion tensor imaging (MR-DTI) for quantitatively evaluating the efficacy of Mailuoning Injection on non-compressive lumbar radiculitis.Methods Nine Bama mini pigs were selected and divided into group A,B and C,3 cases in each group.The non-compressive lumbar radiculitis model was established under CT-guiding.The corresponding therapeutic drugs (group A:10 mL Mailuoning;group B:10 mL normal saline;group C:10 mL diminishing inflammation fluid) were given by epidural injection on 14 d after constructing model.MR-DTI was performed before model construction,14 d after model construction and before treatment.One experimental pig in each group was taken on 3,7,14 d after treatment,performed MR-DTI and killed for taking the nerve root sample to conduct the immunohistochemical detection.The fractional anisotropy (FA) values of nerve root in MR-DTI imaging were measured.The FA values and immunohistochemical detection results were statistically analyzed.Results MR-DTI:the FA values after model construction in each group was decreased (P0.05),but the FA values increase in the group C was earlier and more rapid than other two groups (P0.05),while the FA values had statistical difference between the group B and C (P<0.05).The immunohistochemical results:TNF-α integral absorbency value(IA value) on 7 d after treatment in the group A began to decline;the TNF-α IA value on 14 d after treatment in the group A and C was significantly decreased compared to group B,the difference was statistically significant (P<0.05).Conclusion Mailuoning Injection has a certain effect on non-compressive lumbar radiculitis,which can be evaluated by using DTI.
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Objective Little research has been done on how Cx37 changes the current density of mononuclear macrophage in atherosclerosis.The purpose of this study was to detect the effects of Cx37 on the current density of mononuclear macrophage in atherosclerosis.Methods A total of 30 Wistar mice were randomly divided into Cx37+ group and Cx37-group equally.The atherosclerosis model was constructed by a high-fat diet.According to different parts of sample collection, these two groups were subdivided into Cx37+ plaque group, Cx37-plaque group, Cx37+blood group and Cx37-blood group.RT-PCR was applied to detect the expression of Cx37 in different body parts.The mononuclear macrophages were cultured after being separated from blood and plague in both groups.The current density of mononuclear macrophage was detected by the whole cell recording.Results The relative expression of Cx37 in Cx37 + plaque group was higher than that in plaque group ([1.10±0.02] vs [0.60±0.03]).Energy Spectrum CT was used to detect the carotid artery plaque in both Cx37 + and Cx37-groups, which verified the successful model construction.At 80,120 and 160ms, the current density in Cx37 + plaque group([0.61± 0.06], [0.67±0.07], [0.91±0.03]A/cm2) was significantly higher than those in Cx37 + blood group([0.49±0.02], [0.61±0.03], [0.67±0.02]A/cm2) , Cx37-plaque group([0.48±0.02], [0.60±0.02], [0.64±0.02]A/cm2) and Cx37-blood group([0.49±0.02], [0.59±0.02], [0.64±0.02]A/cm2).The same goes for those at 200, 240, 320ms(P<0.05).Conclusion Cx37 has more significant impact on the current density in the plaque of mononuclear macrophage than in the peripheral blood in promoting macrophages activation and atherosclerosis progress.
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Objective The extracorporeal cardiac shock wave therapy ( CSWT ) can promote angiogenesis in ischemic myo-cardium and improve myocardial perfusion , but its mechanisms remain to be clarified .This study aimed to explore the effects of CSWT on the endothelial progenitor cells (EPCs), vascular endothe-lial growth factor ( VEGF) and Interleukin-8 ( IL-8) as well as its re-lieving effect on angina pectoris in patients with coronary heart dis-ease. Methods After Dobutamine stress echocardiography ( DSE) and 99 mTc-MIBI myocardial perfusion imaging (MPI) at rest and un-der stress, 25 patients with coronary heart disease underwent 9 three-month cycles of CSWT .Before and after the treatment , we obtained the results of 6-min walk test, NYHA cardiac function grades , CCS angina pectoris classes, Seattle Angina Questionnaire (SAQ) scores, doses of nitroglycerin administered , left ventricular diastolic di-ameter ( LVDD) , and left ventricular ejection fraction ( LVEF) .We evaluated myocardial perfusion and myocardial contractile function using MPI and the peak systolic strain rate (PSSR) at rest and under stress, respectively. Results After CSWT, the numbers of EPCs and EPC-CFUs cultured in vitro were significantly increased as compared with the baseline (34.52±6.58 vs 19.56±4.28, P0.05).The PSSR showed no significant changes at rest (1.21±0.62 vs 1.04±0.43, P>0.05) but remark-ably increased under stress after CSWT (2.02±1.00 vs 1.35±0.66, P<0.01). Conclusion CSWT can up-regulate the expressions of VEGF and IL-8 and improve the function of EPCs in the peripheral blood , and thus plays an important role in relieving the symptoms of angina pectoris , promoting cardiac function and enhancing exercise tolerance in patients with coronary heart disease .
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Objective To investigate the effect of different time of hypoxia reoxygenation (HR) on cell autophagy and cell viability.Methods According to the different reoxygenation time,H9C2 cells were divided into five groups:normal control group (group A),hypoxia group (group B),reoxygen 2h group (group C),reoxygen 12h group (group D) and reoxygen 24h group (group E).The expression of LC3 Ⅱ/LC3 Ⅰ in each groups was detected by Western Blot,and the activity of myocardial cells was detected by MlT.Results MlT results showed that the viability of the myocardial cells after hypoxia 2h/reoxygenation 2h was significantly lower than that in the normal control group (P＜0.05).The ratio ofLC3 Ⅱ/LC3 Ⅰ of the cells in group B (hypoxia group),group C (reoxygen 2h group),group D (reoxygen 12h group),group E (reoxygen 24h group) was significantly higher than that of group A (normal control group) (P＜0.05).Among them,the ratio of LC3 Ⅱ/LC3 Ⅰ of the cells in group C and D were significantly higher than that of group B (P＜0.05),paticlularly the group D was the highest (P＜0.05).The activity of myocardial cells in group D was the lowest among all groups.Conclusion Culturing in the three gas incubator can obviously decrease the viability of H9C2cells.Hypoxia can activate autophagy,and autophagy increases significantly after reoxygenation.Autophagy may be activated most obviously at 12 hours and decrease to the level of hypoxia group at 24 hours.Cells can be damaged by hypoxia,and further increase of cellular damage may occur after oxygen inhalation.At 12 hours,the cell viability is the lowest,and the cell viability may be improved at 24h.
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Objective Extracorporeal shock wave therapy ( ESWT ) can promote angiogenesis and collateral circulation es-tablishment by introducing shock waves to ischemia myocardium , however , the specific mechanisms remain unclearly .The study aimed to explore the effects of 0.09mJ/mm2 shock wave treatment on human umbilical vein endothelial cell (HUVEC) proliferation, vascular endothelial growth factor (VEGF) and Interleukin-8 (IL-8) expressions. Methods There were an experimental group and a control group in the experiment .The tubes of the experiment group were set in shock wave devices without the treatment of shock waves .As for the experiment group, 0, 0.03, 0.09, 0.18, 0.24 mJ/mm2 shock waves were introduced , followed by the addition of CCK8 solution and the measurement of A value at the wavelength of 450nm by microplate reader .The HUVEC cell lines were performed 0.09 mJ/mm2 ultrasonic shock energy , CCK8 colorimetric method were utilized to detect the HUVEC proliferation , real time PCR and flow cytometry were applied to detect the expressions of VEGF and IL-8 mRNA respectively . Results ① CCK8 colorimetric method revealed that 0.09 mJ/mm2 shock energy markedly promoted the HU-VEC proliferation compared with the control group , with A450 value comparison (0.70 ±0.04 vs 0.54 ±0.09, P0.05 ).②RT-PCR showed that the 0.09 mJ/mm2 energy significantly enhanced the expressions of VEGF and IL-8 mRNA compared with the control group (7.93 ±0.90 vs 1.07 ±0.40, 7.34 ±1.67 vs 1.00 ±0.09, P<0.001).③Flow Cytometry showed that the expressions of VEGF and IL-8 mRNA significantly increased after 0.09 mJ/mm2 ESWT compared with the control group (39.89 ±4.79 vs 20.98 ±3.30, 31.33 ± 5.61 vs 22.60 ±3.76, P<0.05). Conclusion Low-energy ESWs can surely promote HUVEC proliferation and increase the expres-sions of VEGF and IL-8, and the up-regulation of VEGF and IL-8 may play an important role in the promotion of angiogenesis by ESWT .
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BACKGROUND:Currently, there is no uniform, standardized approach to isolate, purify and proliferate bone marrow mesenchymal stem cells. Chlormethylbenzamido-1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (CM-Dil) is a stable, reliable, high marking and simple marker. OBJECTIVE:To develop the methods for isolation, culture and identification of rat bone marrow mesenchymal stem cells in vitro. METHODS:Two male Sprague-Dawley rats, weighing 50-100 g were taken to col ect the bilateral femur and tibia bone marrow under sterile conditions, and then, primary bone marrow mesenchymal stem cells were isolated and cultured using bone marrow adherent separation and density gradient centrifugation. cells were amplified and purified through timely and repeated passage, and labeled at the third generation with fluorescent dyes CM-Dil in vitro as a source of donor cells. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells were cultured successful y in vitro using bone marrow adherent separation and density gradient centrifugation separation methods, but the former was superior to the latter in the number of cultured cells significantly, while the two methods were not different significantly in terms of cellviability and proliferation. Flow cytometry results showed that the positive rates of cultured cells were 17.5%for CD34, 97.9%for CD44, and 91%for CD90. CM-Dil can label bone marrow mesenchymal stem cells successful y, which is a stable, reliable, high marking and simple marker.
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Objective To investigate the effects of Tanshinone Ⅱ A (extracted from Chinese herb medicine Salvia miltiorrhiza Bge.) on ventricular arrhythmias of rabbit hearts induced by ischemia in order to illuminate its mechanism of anti-arrhythmia.Methods Thirty rabbits were randomly (random number)divided into normal group,ischemic group and Tanshinone Ⅱ A group.Model of wedge shaped mass of rabbit left ventricular myocardium with coronary perfusion was prepared,and then by using floating glassy microelectrode,the trans-mural ECG,QT interval,the trans-mural dispersion of re-polarization (TDR) and trans-membrane action potentials from both endocardium and epicardium were simultaneously and wholly recorded.The incidence of ventricular arrhythmia in myocardium was observed after ischemia for thirty min.Results Under the condition of acute ischemia,compared with normal group,the incidence of ventricular arrhythmia and TDR were significantly increased in ischemia group (P ＜ 0.01),while incidence of ventricular arrhythmia and TDR were significantly reduced in tanshinone ⅡA group compared with ischemia group (P ＜ 0.05).The incidences of ventricular arrhythmia in normal,ischemia and Tanshinone Ⅱ A groups were 0/10,9/10 and 2/10 respectively.Conclusions Tanshinone Ⅱ A prevents ventricular arrhythmia and reduces TDR significantly in ischemic rabbit hearts.
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BACKGROUND: It was uncertain that the migration, differentiation and survival of transplanted bone marrow mesenchymal stem cells (BMSCs) into myocardium after the acute myocardial infarction. OBJECTIVE: To investigate the migration, differentiation and survival of rabbit transplanted autologous BMSCs in myocardium after the acute myocardial infarction. METHODS: Rabbit BMSCs were isolated and labeled by DAPI in vitro. Rabbit left anterior descending branch was ligated to establish acute myocardial infarction models. Following successful model establishment, 30 New Zealand rabbits were assigned to BMSC and control groups (n = 15). In the BMSC group, autologous BMSCs were infused into the surrounding sites of the infracted region by 4 points 1 hour following coronary artery ligation. In the control group, the same region was injected with an equal volume of saline. Injection volume was 30 μL in each point. Five animals from each group were sacrificed 10 minutes, 3 days and 4 weeks following transplantation. The heart was obtained to undergo frozen sections. The distribution of DAPI-labeled BMSCs was observed using fluorescence microscope. Immunofluorescence method was used to examine the troponin Ⅰ and α-actin. RESULTS AND CONCLUSION: DAPI-labeled BMSCs with blue nuclei were distributed extensively in the myocardium of the cell transplantation group, ovoid in shape and arranged in parallel with the cardiac muscle fibers. Troponin Ⅰ and α-actin were positive immunofluorescently in the cytoplasm of the labeled BMSCs. Results indicated that transplanted BMSCs in the ischemic myocardium could differentiate into myocardial cells under stimulation of local microenvironment.
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Objective To investigate the distribution and changes of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in normal heart or heart after myocardial infarction (MI),and to explore beneficial effects of bFGF on heart after MI. Methods Eleven adult minipigs were randomly divided into 2 groups,control (n=5) and MI group (n=6). MI was induced by ligating the left anterior descending coronary artery. Sham-operation was carried out on the animals of control. bFGF-like-immunoreactivity (bFGF-ir) and FGFR-1-like-immunoreactivity (FGFR-1-ir) in heart tissues were detected by immuno-histochemistry and image analysts in 4 weeks after surgery. Results In controls,bFGF-ir and FGFR-1-ir in the atrium showed a considerable high level compared with 2 ventricles (P
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To investigate methods of treating atrial tachycardia by arrhythmias by biatrial pacing and assessments of efficacy． Left atrial pacing had been done in coronary sinus in 5 cases, including 4 c ases in specific electrodes of coronary vein sinus l case in ordinary ventricu lar electrode． Biatrial leads were con nected to atrial port of DDD pacemaker by a “Y” type connector ．The pacing mo de was AAT．Results: 4 coronary sinus leads were put in middle part and l in di stal part． We tested threshhold, sensitivity and impedance in operations． The follow-up time was 4-l 8 months． Biatrial pacing had an effect on 4 cases and had significant effects o n 3 cases． The effective rate was 80 percent．Conclusion:Biatrial pacing can treat atrial tachycardia arrhythmias effectively for intraatrial block．
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Objective To further prove the multipotentiality of rabbit bone mesenchymal stem cells (MSCs) by inducing it into cardiomyocytes with 5-azacytidine. Methods MSCs from rabbit were abstracted, isolated, cultured. Then MSCs were induced by 5-azacytidine in vitro for 24 hours. After cultured for 4 weeks, MSCs’ differentiation was studied by immunohistochemistry and electron microscopy technique. Results Some of MSCs induced by 5-azacytidine expressed troponinT . Electron microscopy showed myofilaments. Conclusion MSCs which exist in bone marrow are mutipotential stem cells, and they can be induced into cardiomyocytes by 5-azacytidine in vitro.