Objective To evaluate the potential protective effects of erythropoietin (EPO) in combination with granulocyte-colony stimulating factor (G-CSF) on hypoxia cardiomyocytes. Methods After left ventricular cardiomyocytes were isolated from neonate rat, the cells were inoculated in a cultural atmosphere of 95%N2 and 5%CO2 for 24 h to establish hypoxia cardiomyocytes model. The protective effects of EPO and G-CSF at different concentrations were evaluated for the optimal concentration. Then the protective effects of EPO in combination with G-CSF were investigated under the optimized concentrations. The survival, apoptotic and necrotic rates of cardiomyocytes were assessed by flow cytometry. Results The mortality and ratio of apoptotic cells to total necrotic cell were higher in hypoxia cardiomyocytes than normal cells significantly (26.73% vs 5.63%,70.05% vs 37.83%, P0.05), and the combination group was much better than EPO and G-CSF alone groups (P

Objective: To investigate the effects of bone marrow-derived mononuclear cells (BMC) transplantation for the therapy of myocardial infarction (MI) in rabbits. Methods: 20 rabbits were randomly divided into 2 groups. MI was induced by ligation of the left anterior descending artery.In transplantation group(T,n=10), BMC transplantation was performed on 5-7 days after MI . Bone marrow (3-5 ml) was obtained from iliac crest and labeled with bromodeoxyuridine (Brdu) for 24-48 hours, BMC were transplanted into infracted myocardium through intramyocardial injection. Control animals (C,n=10) didn′t receive any treatment after MI. Echocardiography was performed for evaluating the cardiac function in 1 week and 5 weeks after MI. Hemodynamic and histological studies were performed in the 5 th weeks after MI. Results: LV ejection fraction of group T had no change, but group C decreased in the 1st week and 5th weeks after MI. The results of Group T having higher LV max +dP/dt and max-dp/dt, lower LV end-diastolic pressure showed comparing with that of group C in the 5th weeks after MI. Histological studies revealed that there were Brdu positive cells in the infarcted area in group T, and the vascular density of group T in the infarcted area was significantly greater in comparision with group C. No regeneration of smooth muscle cell and cardiomyocyte were found in the infarcted area. Conclusion: Transplantation of BMC may avoid the deterioration of cardiac function through vasculogenesis in the infarcted area,but the efficacy in amelioration of cardiac function is limited.

AIM: To compare bone marrow stem cell mobilization with bone marrow-derived mononuclear cells (BMCs) transplantation for the therapy of myocardial infarction (MI) in rabbits, and to explore more effective and practical stem cell therapeutic strategy for MI. METHODS: In mobilization group (M, n=10), granulocyte-colony stimulating factor (G-CSF) (30 ?g?kg~ -1 ?d~ -1 ) was injected subcutaneously 3 hours after MI and every 24 hours for 5 days. On the 5th day, the BMCs from 10 mL peripheral blood were labeled with bromodeoxyuridine (BrdU) for 24-48 hours, then reinjected intravenously. In transplantation group (T, n=10), BMCs transplantation was performed 5-7 days after MI. After being obtained from bone marrow (3- 5 mL ) of iliac crest and labeled with BrdU for 24-48 hours, BMCs were transplanted into infracted myocardium through intramyocardial injection. Control animals (C, n=10) did not receive any treatment after MI. Echocardiography were performed for the evaluation of cardiac function 1 week and 5 weeks after MI. Hemodynamic studies and histological study were performed 5 weeks after MI. RESULTS: LV ejection fraction increased significantly in group M, had no change in group T, and decreased 1 week and 5 weeks after MI in group C. Group M and group T had higher LV max +dp/dt and max -dp/dt, lower LV end-diastolic pressure compared with group C 5 weeks after MI. Histological studies revealed that there were BrdU positive cells in the infarcted area in group M and group T. The vascular density of group M and group T in the infarcted area was significantly greater in comparison with group C. No regeneration of smooth muscle cells and cardiomyocytes were found in the infarcted area. CONCLUSION: Bone marrow stem cell mobilization with G-CSF and transplantation of BMCs both significantly improve the cardiac function for the therapy of MI through vascular genesis in the infarcted area. Bone marrow stem cell mobilization may offer a new and non-invasive therapeutic strategy for MI.

OBJECTIVETo investigate whether direct administration of adenoviral vectors (Ad) containing the complementary deoxyribonucleic acid (cDNA) of vascular endothelial growth factor 165 (Ad-VEGF165) induces porcine coronary collateral vessel formation, improves regional myocardial perfusion and function and is safe.

METHODSThree weeks after miniature swine underwent left thoracotomy and placement of an Ameroid constrictor on the left circumflex coronary artery (LCX), Ad-VEGF165 (n = 6) or the control, Ad expressing beta-galactosidase cDNA (Ad-Gal, n = 6), was directly administered into the ischemic myocardium in the circumflex distribution. All animals were sacrificed 4 wk after the second surgery. Myocardial perfusion and function were assessed by electrocardiogram-gated single photon emission computed tomography (GSPECT) imaging. Ex vivo coronary angiography was performed to examine collateral vessels. Toxicity was assessed by blood analyses on the day just before (day 0) and on day 1, 3, 7, 28 after vector delivery and by vascular, myocardial and liver histology after sacrifice.

RESULTSGSPECT imaging 4 wk after administration of Ad-VEGF165 demonstrated significant reduction in ischemic area (P < 0.01) and rest ischemic severity (P < 0.01) and significant improvement in the left ventricular ejection fraction (P < 0.01) and regional wall motion (P < 0.05) compared with that of Ad-Gal and before administration of Ad-VEGF165. Collateral vessel development assessed by coronary angiography was significantly greater in the Ad-VEGF165 group than in the Ad-Gal group (P < 0.05). General safety parameters, including routine blood parameters, liver and kidney function and cardiac specific parameters demonstrated no difference between Ad-VEGF165 and Ad-Gal animals except for the red blood cell count on day 28 (P < 0.05) and blood urea nitrogen on day 7 (P < 0.05). Only transient elevations in creatine phosphokinase (P < 0.05) and aspartate transaminase (P < 0.05) on day 1 were revealed compared with that before vector administration in both groups. Histologically, no atherosclerotic lesion in the circumflex and no inflammation in liver were revealed and only a small myocardial necrosis was observed in one Ad-VEGF165 animal (area < or = 20%) and one Ad-Gal animal (area < 10%).

CONCLUSIONSAd-VEGF165 can induce coronary collateral vessel formation, improve regional myocardial perfusion and function and is safe by means of direct injection, which suggesting that this strategy may be useful in treating human ischemic heart disease.

Adenoviridae ; genetics ; Animals ; Collateral Circulation ; Coronary Angiography ; Coronary Vessels ; physiopathology ; DNA, Complementary ; administration & dosage ; genetics ; Electrocardiography ; Endothelial Growth Factors ; genetics ; physiology ; Female ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; Lymphokines ; genetics ; physiology ; Male ; Myocardial Ischemia ; diagnostic imaging ; genetics ; therapy ; Neovascularization, Physiologic ; physiology ; Swine ; Swine, Miniature ; Tomography, Emission-Computed, Single-Photon ; methods ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo evaluate the effects of erythropoietin (EPO) combined with granulocyte-colony stimulating factor (G-CSF) on left ventricular function and ventricular remodeling after acute myocardial infarction (AMI) and investigate the possible mechanism.

METHODSThe experimental design consisted of 5 groups of rats, namely the sham, myocardial infarction (MI) model, MI with EPO treatment, MI with G-CSF treatment, and MI with EPO plus G-CSF treatment groups. Apoptosis of the cardiomyocytes was detected by TUNEL staining, and HE staining, Masson trichrome staining, scarlatinum staining, and VIII agent staining were used to evaluate the survival, scar collagen deposition, and angiogenic effects. The cardiac structure and function of the rats after the treatments were assessed by echocardiography and hemodynamic examination.

RESULTSEchocardiography indicated that LVEF and FS were improved in all the intervention groups 7 days after MI, and the rats in EPO plus G-CSF treatment group showed the most obvious reduction of LVESD and LVESV (P<0.01). On day 28 after MI, all the intervention groups showed improvements in LVEF, FS, LVESD, LVEDD, LVESV and LVEDV, which were especially obvious in the combined treatment group; the interventions, especially the combined treatment, also resulted in decreased LVEDP and increased LVSP and +dP/dtmax. On day 1 after MI, the number of apoptotic cells was significantly greater in the MI model group than in EPO and G-CSF groups, and was the fewest in the combined treatment group (P<0.01). On day 28, the number of new vessels increased and the scar and collagen deposition reduced in the EPO and G-CSF groups, and these changes were more obvious in the combined treatment group.

CONCLUSIONSEPO combined with G-CSF can prevent left ventricular remodeling and improve cardiac systolic and diastolic functions by inhibiting cardiomyocyte apoptosis, reducing tissue collagen deposition and inducing neovascularisation.

Animals ; Drug Therapy, Combination ; Erythropoietin ; therapeutic use ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Myocardial Infarction ; drug therapy ; physiopathology ; Rats ; Rats, Wistar ; Ventricular Function, Left ; physiology ; Ventricular Remodeling ; drug effects

Bone Marrow Cells ; metabolism ; pathology ; CD28 Antigens ; blood ; metabolism ; CD4 Antigens ; blood ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; Dendritic Cells, Follicular ; metabolism ; pathology ; Humans ; Immunoblastic Lymphadenopathy ; metabolism ; pathology ; Immunophenotyping ; methods ; Lymphoma, T-Cell ; metabolism ; pathology ; Neprilysin ; blood ; metabolism ; Receptors, Complement 3d ; blood ; metabolism ; fas Receptor ; blood ; metabolism

OBJECTIVETo investigate the biocompatibility of new bone tissue engineering scaffolds, A:D, L-polylactic acid (PDLLA)/polylactic acid-polyethylene glycol-polylactic acid-polylactic acid (PLA-PEG-PLA)/Tricalcium phosphate and B: PDLLA/PLA-PEG-PLA in vivo, compared with PDLLA in repair of a rabbit mandibular body defect.

METHODS24 New Zealand adult rabbits were divided into 4 groups randomly. 15 mm x 6 mm defects were made surgically in the bilateral mandibular bodies and each hemi-mandible was assigned as an experimental unit. The defects were randomly repaired with scaffold materials in each group. Specimens obtained were evaluated with general observation, X-ray, histomorphology and computerized graphical analysis at 2, 4 , 8, 12 weeks after surgery.

RESULTSCompared with PDLLA, the new scaffold materials B showed biocompatibility. At the same time the quantity of new bone produced was much more than that in control group (P<0.05). The new scaffold materials A showed the clear chronic granulomatous inflammation.

CONCLUSIONNew scaffold material B had sound biocompatibility. It was much better than PDLLA. So it may be an ideal bone tissue engineering scaffold material. A is not adapted to be used as scaffold material.

Animals ; Biocompatible Materials ; Bone and Bones ; Calcium Phosphates ; Lactates ; Lactic Acid ; Polyesters ; Polyethylene Glycols ; Polymers ; Rabbits ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo investigate whether short interfering RNAs(siRNAs) of beta-site APP cleaving enzyme (BACE) can inhibit the expression of BACE in mammalian cells.

METHODSThe gene of EGFP, U6 promoter and beta-secretase targeting siRNA were cloned by PCR, respectively. The PCR products were inserted into plasmid pLXSN. The interfering vector pLXSN/EGFP-U6-siBACE was transferred into SK-N-SH cells to express BACE. The inhibition effect of BACE siRNA on BACE expression was investigated by fluoroscopy and immunohistochemistry method.

RESULTThe interfering vector pLXSN/EGFP-U6-siBACE was constructed successfully. The BACE siRNA inhibited the expression of BACE in the SK-N-SH cells specifically and effectively, and the production of A beta was reduced.

CONCLUSIONBACE siRNA can inhibit the expression of BACE gene of mammalian cells.

Amyloid Precursor Protein Secretases ; genetics ; metabolism ; Animals ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Microscopy, Fluorescence ; NIH 3T3 Cells ; Neuroblastoma ; genetics ; metabolism ; pathology ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Tumor Cells, Cultured

Objective To explore the risk factors of cardiovascular disease in adolescents with a family history of hypertension.Methods A cross-sectional study was conducted in 3874 adolescents ages 13-18 years,with normal BP in 3724 people.Based on family history of hypertension (FH),the cohort of adolescents were dichoto- mized as postive family history (FH~+,n=1145) and negative (FH~-,n=2579).Height,weight,waist circum- ference,hip circumference,blood pressure and fasting plasma glucose(FPG),total cholesterol(TC),triglyceride (TG) and high-density lipoprotein cholesterol(HDL-C) were determined.Results FH~+ adolescents had signifi- cantly higher levels of body mass index(BMI),waist circumference,WHR,FPG,TC and LDL-C(P

Objective To delineate the relationship between body mass index(BMI),waist-to-height ratio (WHtR)and hypertension in adolescents.Methods A cross-sectional study was conducted in 3874 adolescents ages 13-18 years.They were classified into four groups according to BMI and WHtR,group A:normal BMI and WHtR