Objective By studying the expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats′ cardiac muscle after acute myocardial infarction, we try to find out the relation between the expression of these two factors and the formation of new blood vessels under ischemia. Methods Wistar rats were divided into control group and infarction group (3, 7, 14, 28, 42, 56 d). The expression of VEGF and bFGF in cardiac muscle and endothelial cells is detected by means of immunohisto-chemial staining while the density of newly formed ressels is detected by marking the endothelial cells with antigens associated with factor Ⅷ. Results After ligating the left anterior descending coronary artery of the rats, the expression of VEGF and bFGF increased along with the prolongation of myocardial ischemia and reached the peak on the 7th day. The expression of the 2 factors began to decrease on the 28th day and the most significant decrease happened on the 42th and 56th day. The density of the newly formed capillaries is directly propontimal to the levels of the 2 factors. Less expression of VEGF and bFGF was observed in the control group. Conclusion The up-regulation of VEGF and bFGF expression might play important roles in neovascularization. Dicrectly intramyocardial injection of bFGF and VEGF gene at the time when the expression of bFGF and VEGF began to decrease maybe optimal.

Objective To establish a method for TLC identification of Guangdongzizhu in Kanggongyan tablets. Method Developer was chloroform-methyl alcoho-Formic acid (20∶4∶0.8), spraying with the test solution of chloride ferric. Result The same color spots in the TLC graphs of sample existed at the corresponding position compared with the control. Conclusion The method is proved to be sensitive and precise, and can be used for the quality control of Kanggongyan tablets.

Objective:To compare the effects of levosimendan injection at single dose and multiple doses on the left ventricular function , biomarkers and neuroho-rmonal activity in the patients with acute exacerbation of advanced heart failure .Methods:Totally 39 patients with chronic heart failure at acute exacerbation were divided into two groups , single dose group and multiple doses group .All the patients were given loading dose of 6μg· kg-1 levosimendan at baseline , and then given 0.1μg· kg-1 · min-1 continuous 24h in-fusion pump.The above regimen was administered once again after one month and six months in the patients of multiple doses group . Left ventricular end systolic volume ( LVESV ) , left ventricular end diastolic volume ( LVEDV ) , left ventricular ejection fraction (LVEF), mitral annulus systolic velocity (SM), myocardial performance index (MOI), blood brain natriuretic peptide (NT-proB-NP), tumor necrosis factor alpha (TNF-α) and interleukin 6(IL-6) were compared between the two groups before the treatment , and after tge three-day and 6-month treatment.Results:Compared with those before the treatment , LVESF, LVEF, Sm, MPI, NT-proB-NP, IL-6 and TNF-αin in the two groups were significantly improved after the three-day treatment (P<0.05).The indices were sig-nificantly improved in multiple doses group after the 6-month treatment when compared with those before the treatment and in single dose group, and the differences were statistically significant (P<0.05).Conclusion: Repeated intravenous administration of levosi-mendan is superior to the single dose administration in improving hemodynamics and inflammatory factors .

To isolate and culture adipose stromal cells (ASCs), and study the effect of cytokines secreted by ASCs on endothelial cells, human adipose tissue was digested with collagenase type I solution and ASCs were derived by culture. The cells surface phenotype was examined by flow cytometry. ELISA was used to detect the secretion of VEGF, HGF, SDF-1 alpha and RT-PCR was employed to detect the expression of their mRNA. Then the ASC medium was utilized to culture human umbilical vein endothelial cells ECV304. Cells were counted by hemacytometer to determine the proliferation and Annexin V/ PI was employed for the examination of the apoptosis rate of ECV304. ASCs were derived by culture and expressed CD34, CD105 while they did not express CD31 or CD45. ASCs secreted cytokines such as VEGF, HGF and SDF-1 alpha so the ASC medium could stimulate proliferation and counteract apoptosis of endothelial cells (P < 0.05). Bcl-2 mRNA was also found to be up-regulated in the endothelial cells. It is concluded that ASCs can secrete cytokines and has significant effect on the proliferation of endothelial cells and apoptosis.

BACKGROUND: The studies about cellular cardiomyoplasty (CCM) of bone marrow stromal cell (BMSC) are centered on modeling autologous cell transplantation while the study of myocardium transplantation of allogeneic BMSC is seldom reported domestically.OBJECTIVE: To study the hypothesis that the allogeneic BMSC, after transplanted into the myocardial infarction (MI) regions, can survive, further proliferate, differentiate, and its effects on host hearts.DESIGN: A randomized and controlled trial with experimental animals as subjects.SETTING: Laboratory of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Ten Wistar rats of one-month old, body mass of 100-120 g and unconfined sex were used to culture BMSC while eighty female Wister rats of three-month old, body mass of 200-250 g were used for animal models.METHODS: The experiment was completed in the Laboratory of of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College,Huazhong University of Science and Technology from June to December 2004. ①Eighty rats were used to establish the acute myocardial infarction (AMI) models by ligating the left anterior descending branch of coronary artery, and 45 of them were successful. ② After 4 weeks, the models were divided into 2 groups at random. In the experiment group (n=25), the passaged and uninduced BMSC cultured in vitro were injected into the MI region of the recipients while only medium was injected in the control group (n=20). ③At 4 weeks after implantation,the hemodynamic indexes of recipients' hearts were examined. Then the samples were obtained to detect the survival, differentiation and angiogenesis status of the removal cells.MAIN OUTCOME MEASURES: ①Comparison of hemodynamic indexes in rats of two groups②Results of the implantation cells③Changes of angiogenesis in rats of two groups.RESULTS: Totally 13 rats in the experiment group and 11 in the control group were involved in the result analysis. ①The hemodynamic indexes of rats were significantly improved in the experiment groups compared with the control group [Left ventricle systolic blood pressure (LVSBP): (88.61±5.99),(76.93±4.75) mm Hg, left ventricle end-diastolic pressure(LVEDP): (7.72±1.36),(12.77±2.76) mm Hg, P ＜ 0.05;The maximum changing velocity of LVSBP:(2 365.26±266.31),(2 025.04±230.25) mm Hg/s; The maximum changing velocity of LVDBP:(2 313.26±159.30),(2 140.12±191.03) mm Hg/s]. ②After implanted in the MI region, the allogeneic BMSC passed the acute inflammation period and did not induce the remarkable reject reaction of transplantation. The BMSC in the infarcted region were mainly differentiated into fibroblast. Some cells around the infarcted region were differentiated into endothelial cells, and improved the angiogenesis. ③The number of angiogenesis in and around the transplantation regions was significantly higher in the experiment group than in the control group (P ＜ 0.01).CONCLUSION: The allogeneic BMSC can not form cardiomyocyte in the infarcted region after cell transplantation, and the engendered endothelial cells of blood vessels may promote the angiogenesis after AMI and ameliorate the cardiac function.

AIM: To develop a method for determining the contents of peoniflorin,ferulaic acid and paeonol in Shi'er Wuji Baifeng Pills(Gallus Domesticus,Radix Astragali,Rhizoma Dioscoreae,Radix Codonopsis,etc.) METHODS: The analytical column was an ODS column.The mobile phase consisted of acetonitrile-0.1% phosphoric acid aqueous solution using a gradient elution.The detection wavelengths of peoniflorin,ferulaic acid and paeonol were at 230,320 and 274 nm,respectively. RESULTS: The calibration curves of peoniflorin,ferulaic acid and paeonol showed good linearity at the ranges of 21.19-211.90,1.01-10.06,10.63-106.30 ?g/mL and the average recoveries were 98.63%,98.35%,99.12%,respectively. CONCLUSION: The method is simple,rapid,reliable and can be used for the drug control.

Objective To observe the effects of preconditioning with pioglitazone on infarct size and mito-chondrial ATP-sensitive potassium channel in rats with ischemia-repedusion, and to explore its possible mecha-nism. Method The whole experiment was divided into experiment Ⅰ and Ⅱ. In experiment Ⅰ, 24 rats were ran-domly divided into four groups (6 rats in each group): (1)Sham-operated (SO) group: the coronary artery of rat was threading without hgation, and the heart was removed by cutting immediately 4 hours later; (2) Isehemia-reperfusion (I/R) group: the rats were administered with 0.9% saline intravenously via caudal vein at 24 hours before iigating the left anterior descending branch of coronary artery for 30 minutes, and followed by reperfusion for 4 hours; (3)5-hydroxydecanoate plus pioglitazone(5HD+Pio) group: the rats were injected with 10 mg/kg 5-hy-droxydecanoate (the blocker of mitochondrial ATP-sensitive potassium channels,) at 24 hours before ligation, and 30 minutes later, 3 mg/kg pioglitazone was given in 5 minutes, and then the rats were subjected to ischemia for 30 minutes, followed by reperfusion for4 hours; (4)pioglitazone treatment group (Pio): the mrs were given 3 mg/kg pioglitazone at 24 hours before occlusion, and then they were treated as done in the 5HD+Pio group. In I/R, 5HD+Pio and Pio group, the hearts were removed by cutting after reperfusion. Western blotting was used to detect the protein expression of P38MAPK, .INK and NFκB P65. In experiment Ⅱ, 30 rats were randomly divided into five groups: SO, I/R, Pio, 5HD+Pio and 5-HD group (rats were treated as done in the rats of I/R group and were injected with 10 mg/kg 5-bydroxydecanoate 24 h before ischemia/reperfusion),and the size of myocardial in-farction and isehemia were measured after reperfusion. Statistical analyses were performed using SPSS10.0 soft-ware. Multiple comparisons were analyzed by one-way analysis of variance (SNK-q test). P<0.05 was consid-ered statistically significant. Results (1) The infarct size in i/R group was(34.93±5.55)%, while pioglita-zone reduced the infarct size to(20.24±3.93)% (P<0.05). There was no significant difference between I/R and 5-HD±Pio or 5-HD groups (P>0.05). Compared with the sham-operated group, the expression of P38MAPKmRNA, JNKmRNA and protein of P38MAPK, JNK and NFκB P65 in I/R increased (P<0.05). Com-pared with the I/R group, pioglitazone inhibited these undue expressions (P<0.05). Conclusions Pioglitazone could protect the heart from ischemia-reperfusion injury evidenced by reducing infarct size. These protective effects of pioglitazone may be related to opening mitochondrial ATP-seusitive potassium channels or downregulation of JNK and p38 MAPK signaling, leading to the overexpression of NFκB p65 activation.

Objective To predict a value of ischemia modified albumin (IMA) levels for assessing secondary cerebral infarction in patients with transient ischemic attack (TIA) in anterior circulation.Methods 105 patients with TIA in anterior circulation admitted to the hospital within 3 hours were retrospectively studied.Combined with ABCD2 score,the correlations of IMA levels at 3 h,6 h and 12 h with secondary cerebral infarction after anterior circulation TIA were analyzed.Results IMA level was 75.28 u/L within 3h after TIA,and the sensitivity and specificity of TIA in anterior circulation were 66.7％ and 76.2％ respectively.In the total of 105 patients,16 cases (15.2％) suffered from secondary cerebral infarction within 7d,and 21 cases (20.0％) within 8～30d.The serum IMA levels were (87.43±19.89)U/L,(63.88±12.51)U/L and (61.21±12.28)U/L at 3h,6h and 12h after TIA,respectively.A simple analysis showed that there was a linear correlation between the IMA level and ABCD2 scores (P=0.000,r=0.666).Kaplan-Meier survival curve analysis showed that the increased IMA level within 3h,and moderate to high ABCD2 score were the risk factors for secondary cerebral infarction after TIA in anterior circulation (P=0.012,0.041).Conclusions Early detection of IMA has a clinical value similar to ABCD2 score to predict secondary cerebral infarction in patients with TIA in anterior circulation.

Objective To investigate the effects of changes of miR-126 and spouty related EVH,domain containing proteinl(SPRED1) after transient ischemic attack(TIA)on prognostic value for pathogenesis of secondary cerebral infarction.Methods Retrospective analysis of the clinical data of 106 patients with TIA was performed.The expression levels of miR-126,SPRED1 and vascular endothelial growth factor(VEGF)in peripheral blood were detected at 3 h,6 h and 12 h after TIA onset respectively.The specificity and sensitivity of miR 126 and SPRED1 in the diagnosis of TIA were analyzed.The miR-126 and SPRED1 levels versus ABCD2 score were compared for evaluating their predictive value in the diagnosis of secondary cerebral infarction within 30 days after TIA onset.Results The miR-126 level was declined after TIA onset at 3 h(9.41±1.04),especially at 12 h(6.59 ±2.78),versus in healthy control (9.35±1.76)(t =-7.764,P=0.000).The SPRED1 level after TIA onset was increased at 3 h(58.05 ± 17.53)pg/L,12 h(82.64 ± 18.60)pg/L versus in healthy control(52.38 ± 13.24)pg/L(t=12.374,P =0.000).A closely negative correlation was found between levels of miR 126 and SPRED1 at 12 h point but not at 3 h and 6 h(r=-0.278,P=0.004).Both miR-126 and SPRED1 levels at 12 h after TIA were implied to sensitivity and specificity evaluation.Additionally,VEGF was significantly increased at 3 h (345.61 ± 76.76) pg/L,6 h (461.65 ±103.87)pg/L and 12 h (519.22 ± 103.55)pg/L after TIA onset as compared with healthy control (107.77± 26.04) pg/L(t =26.569,29.756,34.699,all P =0.000).The decrease of miR-126 and increase of SPRED1 at 12h after TIA indicated high incidences of cerebral infarction but their significance was less than ABCD2 score.Combination of miR 126,SPRED1 and ABCD2 score significantly improved the prediction for cerebral infarction(Z=2.105,P =0.035).Conclusions After the onset of TIA,levels of miR-126 and SPRED1 expression in combination of ABCD2 score can improve predictive value for cerebral infarction development.

BACKGROUND: Transplantation of bone marrow mesenchymal stem cell and vascular endothelial growth factor(VEGF) can promote vascular regeneration and improve heart function. However, whether the combined application is superior to single application or not is still unclear.OBJECTIVE: To observe the effect of allogenetic bone manow stem cells transplantation combined with VEGF transfection on vascular regeneration and heart function of rats with acute myocardial infarction.DESIGN: Simple sample observation was used in culturing bone marrow mesenchymal stem cell of rats; Randomized controlled animal experiment was used in cell transplantation and gene transfection.SETTING: Department of Cardiology, Union Hospital of Huazhong University of Science and Technology; Cardiovascular Institute of Tongji Medical College MATERIALS: Totally 94 healthy male Wistar rats and expression vector PAdTrack/VEGF165 were used in this experiment.METHODS: This experiment was carried out at Cardiovascular Institute of Tongji Medical College between June 2004 and June 2005. ①Bone marrow mesenchymal stem cells of rats were isolated, purified and cultured in vitro, then labeled with bromodeoxyuridine(BrdU). ② Preparation , extraction, purification and identification of plasmid PAdTrack/VEGF165. ③Two weeks after coronary artery was ligated to create acute myocardial infarction model, rats were randomly divided into 4 groups (n=12 in each group): stem cell + plasmid group(50 μL BrdU-labeled bone marrow mesenchymal stem cell solution and 100 μL plasmid PAdTrack/VEGF165 were injected into the rats through multiple sites), stem cell group (50 μL bone marrow mesenchymal stem cell solution was injected through multiple sites), plasmid group (100 μL plasmid PAdTrack/VEGF165 was injected through multiple sites) , control group(100 μL serum-free DMEM was injected through multiple sites). ④ Immunohistochemistry andechocardiography were performed 4 weeks later.MAIN OUTCOME MEASURES: ①Immunohistochemical and haematoxylin-eosin stainings were conducted in the infarcted and ischemic areas of rats in each group; ② Blood vessel counts; ③Echocardiography.RESULTS: Totally 48 rats entered the stage of result analysis. ① BrdUlabeled transplanted cells could be seen at the infarcted and ischemic myocardium in the stem cell+plasmid group and stem cell group. Some transplanted cells at ischemic myocardium differentiated into vascular endothelial cells and formed newborn blood capillary. ②Density of Ⅷ factor positively-stained newborn blood capillary took stem cell +plasmid group ＞ plasmid group ＞ stem cell group ＞ control group in order (all P＜ 0.01).③Wall thickness and wall motion range improved after cell transplantation and gene transfection therapy. The increased range of ejection fraction took stem cell +plasmid group ＞ stem cell group ＞ plasmid group ＞ control group in order (all P ＜ 0.01) .CONCLUSION: Allogenic bone marrow mesenchymal stem cell transplantation and VEGF gene transfection could further boost vascular regeneration of infarcted ischemic area and improve wall thickness and heart function of rats.