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Objective:To explore the relationship between the Spike and the Gamma rhythm of the local field potential (LFP) in Alzheimer's disease (AD) transgenic mice during fear memory activity.Methods:Six-month-old APP/PS1/tau three transgenic (3xTg) mice and wild-type (WT) mice were divided into 3xTg group and WT group, with 10 mice in each group. The electrodes were embedded into the hippocampus of mice under sterile conditions, and the behavioral experiment of conditioned fear box test was carried out two weeks later. The changes of Gamma rhythm, Spike and Burst firing were recorded and analyzed by the wireless telemetry device which embedded in the mouse head. Finally, the correlation between Gamma rhythm and Spike was calculated by entropy value.Results:(1) In behavioral experiments, the freezing ratio caused by conditioned stimulation (CS) in 3xTg mice was ((54.07±2.32)%), which was significantly lower than that of WT mice ((76.21±2.88)%) ( t=4.796, P<0.01). (2) Simultaneously recorded the average power of the Gamma oscillation in the Pre-CS period of the WT mice was ((11.574±1.147) dB), which increased to ((18.108±1.177) dB) after CS ( t=3.386, P<0.01). After CS administration, the average power of Gamma in 3xTg group((12.346±1.345) dB) was significantly lower than that of WT group ( t=3.423, P<0.01). (3) The frequency of Spike release in WT mice during the Pre-CS period was ((5.667±1.475)times/s), significantly increased to ((11.008±1.335) times/s) after CS ( t=3.542, P<0.01). The frequency of Spike release of 3xTg mice after CS ((5.249±1.033) times/s) was significantly lower than that of WT group ( t=4.788, P<0.01). (4) The Burst duration of WT group in pre-CS and CS period were ((0.550±0.043)s) and ((1.075±0.034)s), respectively. It suggested that the Burst firing frequency of WT group increased significantly after conditional stimulation ( t=5.188, P<0.01). However, the release interval of 3xTg group after CS ((0.619±0.033)s) was significantly lower than that of WT group ( t=3.352, P<0.01). (5) After CS, the Spike-Gamma entropy curve of WT mice was always higher than that of 3xTg mice. The maximum correlation of WT group and 3xTG group were (0.403±0.031) and (0.314±0.028), respectively. The Spike-Gamma correlation of the 3xTg group was significantly lower than WT mice ( t=3.372, P<0.01). Conclusion:The defect of fear memory in Alzheimer's disease may be caused by the disharmony of Spike-LFP (Gamma) distribution.
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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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[Abstract ] Objective Cardiac shock wave therapy (CSWT) can promote arteriogenesis in ischemic myocardia , but the mo-lecular mechanism remains unclear .The study aimed to explore the effect of CSWT on arteriogenesis in human cardiac microvascular endothelial cells ( HCMEC ) and the role of focal adhesion kinase (FAK) and Calcium-activated potassium channels (KCa) in the sig-nal conduction pathway of CSWT arteriogenesis . Methods HC-MEC cells cultured in vitro were randomly divided into control group , CSWT group , CSWT +T ( FAK inhibitor PF-573228 ) group and CSWT+F( SCa inhibitor iberiotoxin ) group.Each group received one CSWT(0.09 mJ/mm2, 200Times) 48 h after added stimulant.24 hours'conventional culture later , tests were made on the levels of endothelial nitric oxide synthase ( eNOS ) and vascular endothelial growth factor ( VEGF) mRNA as well as the changes of related protein expression . Results ①QPCR test showed that eNOS , VEGF mRNA expressions increased in CSWT group compared with control group (4.61 ±0.19 vs 3.99 ±0.17, P<0.05), while compared with CSWT group, eNOS, VEGF mRNA expressions in CSWT +T group were decreased (0.62 ±0.10 vs 0.40 ±0.02, P<0.05), eNOS, VEGF mRNA expressions in CSWT +F group were also decreased (0.53 ±0.02 vs 0.64 ±0.02, P<0.05), all the differ-ences were of statistical significance .②Western blot showed that eNOS , VEGF protein expressions increased in CSWT group compared with control group(0.63 ±0.02 vs 0.43 ±0.02, P<0.05), while compared with CSWT group , eNOS, VEGF protein expressions in CSWT+T group were decreased (0.36 ±0.01 vs 0.29 ±0.02, P<0.05), eNOS, VEGF protein expressions in CSWT +F group were also decreased (0.37 ±0.02 vs 0.30 ±0.02, P<0.05), all the differences were of statistical significance . Conclusion CSWT can improve eNOS , VEGF mRNA and protein expressions in HCMEC cells and FAK and KCa may participate in the signal conduction pathway of CSWT arteriogenesis .
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Objective: To observe the effect of extracorporeal shock wave therapy (ESWT) on proliferation, cell cycle and intercellular adhesion molecule-1 (ICAM-1) expression in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were culturedin vitro at the concentration of (1×105/ml) and the cells were divided into 2 sets of groups:CSWT group, the cells were treated by different energy of (0.03, 0.09, 0.18, 0.24) mJ/mm2 respectively and corresponding Control group, in which the cells had no CSWT. HUVEC proliferation was detected by CCK colorimetric method, cell cycle was measured by lfow cytometry, mRNA and protein expressions of ICAM-1 were examined by RT-PCR and Western blot analysis respectively. Results: Compared with Control group, (0.09 mJ/mm2) CSWT group had promoted HUVECs proliferation,P0.05; (0.09 mJ/mm2) CSWT group showed decreased proportion of G0/G1 stage and increased S and G2/M stages, allP0.05. Compared with Control group, (0.09 mJ/mm2 ) and (0.03mJ/mm2) CSWT groups showed increased mRNA expression of ICAM-1 (9.27±0.95) vs (1.02±0.27),P<0.001 and (7.08±0.60) vs (1.02±0.27),P<0.01; (0.09 mJ /mm2) CSWT group had elevated protein expression of ICAM-1,P<0.05. Conclusion: ESWT especially at (0.09 mJ/mm2) may accelerate cell cycle transition from G0/G1 stage to S and G2/M stages, promote HUVECs proliferation and increase ICAM-1 expression which may play important roles in ESWT facilitated angiogenesis in vitro.
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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:Studies have shown that extracorporeal shock wave therapy is an effective, safe, and non-invasive treatment for ischemic heart disease, which can improve angiogenesis in the ischemic myocardium. OBJECTIVE:To summarize the research advances in promotion of angiogenesis for ischemic myocardium by extracorporeal shock wave therapy. METHODS:A computer-based online search of PubMed database and CNKI database was performed for relevant articles published between 1998 and 2014 with key words of “shock wave, ischemic heart disease, angiogenesis, cytokine, stem cel” in English and Chinese, respectively. Articles related to the promotion of angiogenesis for ischemic cardiovascular disease by extracorporeal shock wave were selected. Repetitive articles were excluded. According to inclusion criteria, 51 literatures were selected in result analysis. RESULTS AND CONCLUSION: Extracorporeal shock wave therapy can improve angiogenesis in the ischemic myocardium by mobilizing proliferation and differentiation of stem cels into vascular endothelial cels, and by enhancing the expression of growth factors such as vascular endothelial growth factor and basic fibroblast growth factor. Moreover, the extracorporeal shock wave therapy can create a local favorable microenvironment for angiogenesis by inhibiting inflammation, oxidative stress and apoptosis and by regulating components of the extracelular matrix. Extracorporeal shock wave therapy plays an important role in the angiogenesis of ischemic myocardium and displays a good clinical prospect in the treatment of ischemic heart disease. However, the specific mechanism requires further studies.
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Objective To observe the effect of integrated therapy of Chinese medicine and chemical drugs on adverse reaction and curative effect of initial treatment of secondary pulmonary tuberculosis. Methods Totally 1404 patients with secondary pulmonary tuberculosis and TCM lung consumption diagnostic criteria (syndrome of lung yin deficiency, qi-yin deficiency, yin-deficiency caused excessive fire) were chosen for single blind, randomized, controlled, multicenter clinical trials. Trial group was given 2HRZE/4HR, 1 time/day with Chinese medicine 2 or 3 times/day, and control group was given 2HRZE/4HR only for six months. The adverse reactions and clinical symptoms were observed to evaluate clinical efficacy and safety. Results In terms of reducing liver damage and other adverse reactions, the ratio of trial group had statistical difference with that of control group (P<0.001). In symptom scores of lung yin deficiency syndrome treated for 2, 4, 6 months, yin-deficiency caused excessive fire syndrome treated for 6 months, qi-yin deficiency syndrome treated for 4, 6 months, the differences between the two groups were significant (P<0.001). TCM syndrome curative effect between the two groups was statistical different (P<0.001). Safety evaluation result between the two groups was statistical different by tratified analysis (P<0.001). Conclusion Integrated therapy of Chinese medicine and chemical drugs can improve the symptoms and reduce adverse reactions caused by chemical drugs. It can enhance the curative effect and safety.
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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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To evaluate the safety and efficacy of andrographolide drop-pill in treatment of acute upper respiratory tract infection with external wind-heat syndrome.
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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.