Decreased small arterial compliance with increased serum vascular endothelial growth factor-A and circulating endothelial progenitor cell in dilated cardiomyopathy

Background Evidence showed that both myocardium and blood vessels were damaged in dilated cardiomyopathy （DCM）. However, the changes in arterial compliance, serum cytokines and circulating endothelial progenitor cells （EPC）, and their correlations remain unknown.
Methods Sixty-five DCM patients and 49 healthy volunteers were studied. Both large artery compliance （C1） and small artery compliance （C2） were measured with the CVProfUor DO-2020. Quantitative enzyme-linked immunosorbent assays （ELISAs） were used to measure the levels of vascular endothelial growth factor-A （VEGF-A） and VEGF receptor 2 （VEGF-R2）. Circulating EPC was assessed by EPC colony-forming assays and flow cytometry （CD133^＋/CD34^＋cells）. Phagocytized Dil-acLDL and binded FITC-UEA-I were used to analyze endothelial lineage marker expression by immunofluorescence.
Results Although C2 was markedly lower in DCM patients than in control group （（3.8±1.8） ml/mmHg × 100 vs （5.0±2.2） ml/mmHg × 100, P〈0.0001）, there was no statistically significant difference in C1 between the two groups （P〉0.05）. Levels of VEGF-A, the numbers of colony-forming units （CFU） and the fractions of EPC were obviously higher in DCM patients than in control group （（127.6±139.5） pg/ml vs （58.8±42.9） pg/ml, P〈0.0001; （2.5±1.5）% vs （0.5±0.3）%, P〈0.05; 23.5±12.8 vs 10.8±7.4, P〈0.01, respectively） and however, there was no significant difference in VEGF-R2 between two groups （P〉0.05）. LgVEGF-A was positively correlated with the number of EPC-CFU （r=-0.435; P〈0.05） and inversely correlated with C2 （r=-0.543; P〈0.001） in DCM patients. Conclusions The reduction of C2, a sensitive marker reflecting endothelial dysfunction, was observed in DCM patients and closely related to the increase in serum VEGF-A.     

Combined treatment with erythropoietin and granulocyte colony-stimulating factor enhances neovascularization and improves cardiac function after myocardial infarction

Background Erythropoietin （EPO） and granulocyte colony-stimulating factor （G-CSF） are both potential novel therapeutics for use after myocardial infarction （MI）.However,their underlying mechanisms remain unclear and the efficacy of monotherapy with EPO or G-CSF is also controversial.Therefore,we investigated the effects of combined treatment with EPO and G-CSF on neovascularization and cardiac function in post-infarction rats and explored the potential mechanisms.Methods Four groups of rats were used：control （saline injection after MI,i.h.）,EPO （a single dose of 5 000 IU/kg after MI,i.h.）,G-CSF （a dose of 50 μg· kg-1· d-1 for 5 days after MI,i.h.）,and both EPO and G-CSF （EPO＋G-CSF,using the same regiment as above）.Cardiac function was assessed by echocardiography before and 1 day,7 days,14 days and 21 days after MI.CD34＋/Flk-1＋ cells in the peripheral blood were evaluated by flow cytometry before and 3 days,5 days and 7 days after MI.The infarct area and angiogenesis in the peri-infarct area were analyzed.The mRNA and protein expression of vascular endothelial growth factor （VEGF） and stromal-derived factor-1α （SDF-1α） in the peri-infarct area were detected by real-time quantitative RT-PCR and Western blotting.Results Compared with the control and monotherapy groups,the EPO＋G-CSF group had significantly increased CD34＋/ Flk-1＋ endothelial progenitor calls （EPCs）in the peripheral blood （P ＜0.05）,up-regulated VEGF and SDF-1α levels in the peri-infarct region （P ＜0.05）,enhanced capillary density （P ＜0.05）,reduced infarct size （P ＜0.05） and improved cardiac structure and function （P ＜0.05）.G-CSF alone did not dramatically increase EPCs in the peripheral blood,enhance capillary density in the peri-infarct area or reduce infarct size compared with the control group.Conclusions Combined treatment with EPO and G-CSF increased EPCs mobilization,up-regulated VEGF and SDF-1α levels in the post-infarction microenvironment,subsequently enhanced neovascularization in the peri-infarct region and reduced infarct size.All factors contributed to its beneficial effects on cardiac function in post-infarction rats.     

Inhibitory Effects of Parthenolide on the Angiogenesis Induced by Human Multiple Myeloma Cells and the Mechanism

The inhibitory effects of parthenolide （PTL） on angiogenesis induced by multiple myeloma （MM） cells in vitro and the mechanism were investigated. Human MM line RPMI8226 cells were cultured in vitro. The effects of MM culture supernatant on the migration and tubule formation ability of human umbilical vein endothelial cells （HUVECs） treated with PTL were observed. By using Western blot, the expression of p65 and IкB-α in MM cells was detected. RT-PCR was used to assay the expression of VEGF, IL-6, MMP2 and MMP9 mRNA in MM cells. ELISA was used to measure the levels of VEGF and IL-6 in MM cell culture supernatant. The expression of MMP2 and MMP9 in MM cells was examined by immunohistochemistry. （1） In 3.5, 5.0, 7.5 and 10 μmol/L PTL groups the number of migrated cells was 310±56, 207±28, 127±21 and 49±10 respectively, which was significantly different from that in positive control group （598±47） （P〈0.01）. In 3.5 and 5.0 μmol/L PTL groups the areas of capillary-like structures were 0.092±0.003 and 0.063±0.002 mm2, significantly less than in positive control group （0.262±0.012 mm2） （P〈0.01）, but in 7.5 and 10 μmol/L PTL groups no capillary-like structures were found; （2） After treatment with different concentrations of PTL for 48 h, the expression of p65 protein was gradually decreased, while that of IкB-α was gradually enhanced with the increased concentration of PTL; （3） After treatment with 3.5, 5.0, 7.5 and 10 μmol/L PTL for 48 h, the VEGF levels in the supernatant were 2373.4±392.2, 1982.3±293.3, 1247.0±338.4 and 936.5±168.5 pg/mL respectively, significantly different from those in positive control group （2729±440.0 pg/mL） （P〈0.05）. After treatment with 7.5 and 10 μmol/L PTL, the IL-6 levels in the culture supernatant were 59.6±2.8 and 41.4±9.8 pg/mL respectively, signifi- cantly lower than in positive control group （1287.3±43.5 pg/mL） （P〈0.05）; （4） RT-PCR revealed that PTL could significantly inhibit the expression of V     

Clinical study of electro-acupuncture treatment with different intensities for functional constipation patients

Functional constipation （FC） is a common functional bowel disorder disease that affects life quality of a large number of people. This study aimed to explore the impact of different intensities of electro-acupuncture （EA） treatment for FC patients. Totally, 111 patients with FC meeting the Rome III criteria were randomly assigned to different intensities of EA groups （low and high intensity of EA groups） and medicine-controlled （MC） group. In EA groups, patients were treated with EA at quchi （Llll） and shangjuxu （ST37） bilaterally for 4 weeks, 5 times/week in the first 2 weeks, and 3 times/week in the last 2 weeks. In MC group, 5 mg mosapride citrate was administered orally 3 times/day for 4 weeks. Spontaneous bowel movement frequency each day was recorded using a consti- pation diary. Self-rating anxiety scale （SAS） and self-rating depression scale （SDS） were used to assess the patients＇ psychological state. Cortisol （CORT）, substance P （SP）, and vasoactive intestinal polypep- tide （VIP） were evaluated at baseline and at the end of 4 weeks after treatment. As compared with the baseline, there was statistically significant increase in stool frequency every week （P〈0.01）, but there was no statistically significant difference among the three groups. As compared with the baseline, after 4 weeks of EA therapy, the scores of SDS and serum levels of CORT were decreased significantly in low intensity of EA group （P〈0.01）, and the serum levels of SP and VIP were increased significantly （P〈0.05）; the scores of SAS and SDS and serum levels of CORT were decreased significantly in high intensity of EA group （P〈0.05）, and the serum levels of SP and VIP were increased significantly （P〈0.05）; the serum levels of CORT and VIP were increased significantly in MC group （P〈0.05）. As compared with MC group, after 4 weeks of treatment, the serum levels of SP were signifcicantly in- creased in low intensity of EA group （P〈0.01）. Low and high intensi     

Influences of Refined Konjac Meal on the Levels of Tissue Lipids and the Absorption of Four Minerals in Rats

This paper reports a study on the hypocholesterolemic effect of the refined konjac meal (RKM) containing about 80% glucomannan prepared from the tubers of Amorphophallus konjac K.Koch.Male and female Sprague-Dawley rats, 5 weeks old, were divided into five groups and fed a normal basal diet, a hypercholesterolemic diet (control diet), and three test diets (RKM added to the control diet at levels of 2.5.5, and 10%, respectively) for 12 weeks.The results obtained indicate that RKM could markedly lower the cholesterol levels in the serum and the liver of rats eating hypercholesterolemic diets.At the end of the 4th week of the feeding experiment, the serum cholesterol levels in the 5 and the 10% RKM groups and the liver cholesterol level in the 10% RKM group were significantly lower than those in the control groups.At the end of the 12th week, the serum cholesterol levels of all the three RKM groups were lowered to a normal level as was the liver cholesterol level of the 10% RKM group.The lipolropic effect of RKM was also confirmed by histopathologic examination of the livers.In addition to the hypocholesterolemic effects, RKM diets also increased stool bulk.Minor effects on the absorption and utilization of Ca, Fe, Zn, and Cu were found. 1990 Academic Press.inc.     

Abdominal Fat Accumulation with Hyperuricemia and Hypercholesterolemia Quail Model Induced by High Fat Diet

Objective To establish abdominal fat accumulation with hyperuricemia and hypercholesterolemia quail model fed with high fat diet. And then to investigate the pathological characteristics of this quail model. Methods Thirty Longcheng quails were randomly divided into two groups： control group and model group （n=15）. The control group quails were fed with normal diet and model group quails were fed with high fat diet for 14 days. After a 12-hour overnight fast, liver and abdominal fat at euthanasia as well as serum were collected. The levels of serum uric acid, total cholesterol, high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C）, triglyceride, free fatty acid （FFA）, and blood glucose were assayed. The activity changes of adenosine deaminase （ADA）, xanthine oxidase （XOD）, lipoprotein lipase （LPL）, hepatic lipase （HL）, and fatty acid synthetase （FAS） were analyzed. Results Compared with control group, the abdominal fat content （0.74±0.63 vs. 1.36±0.65 g, P〈0.05） and abdominal fat index （0.44%±0.30% vs. 0.85%±0.30%, P〈0.01） as well as live lipid index （3.61%±0.65% vs. 11.33%±2.14%, P〈0.01） in model group significantly increased; the levels of serum uric acid （210.61±94,76 vs. 304.25±141.94 /amol/L, P〈0.05）, total cholesterol （4.20±0.51 vs. 20.10±11.25 mmol/L, P〈0.01）, LDL-C （1.16±0.29 vs. 10.78±6.48 mmol/L, P〈0.01）, and FFA （0.39±0.14 vs. 0.55±0.15 mmol/L, P〈0.01） in model group significantly increased; HDL-C （5.85±0.95 vs. 4.14±2.03 mmol/L, P〈0.05） significantly decreased; the levels of triglyceride and blood glucose had no significant changes （P〉0.05）; the activities of ADA （9.71±3.05 vs. 17.19±5.10 U/ml, P〈0.01） and XOD （10.58±6,88 vs. 19.22＋9.44 U/L, P〈0.01） in model group significantly increased; and FAS, LPL, HL had no significant changes （P〉0.05）. Conclusions High fat diet can induce abdominal fat accumulation with hyperuricemia and hypercholesterolemia quail model. The changes of uric acid and lipid metabolic enzyme activities may he the pathological mechanism of abdominal fat accumulation with hyperuricemia and hypercholesterolemia.     

Acute hyperenhancement on delayed contrast-enhanced magnetic resonance imaging is the characteristic sign after coronary microembolization

Background Detection of coronary microembolization is of clinical importance for patient management and prediction of long-term outcome. However, there are few studies of the changes of magnetic resonance imaging after coronary microembolization. This study was designed to investigate the imaging of the left ventricle using delayed contrast enhanced magnetic resonance imaging as well as the left ventricular ejection fraction after coronary microembolization in animal models.
Methods Eight miniswine, of either sex （body weight 21-25 kg）, were used to make the coronary microembolization model. After coronary angiography, a 2.8F infusion catheter was placed in the left anterior descending artery with the tip located between the second and third diagonal branches. Microspheres with the diameter of 42μm and mean dosage of 1.2×10^5 were selectively infused into the left anterior descending artery. First pass and stressed first pass perfusion scan were performed after cine images were acquired. Then a second bolus of 0.15 mmol/kg gadolinium DTPA was given at a rate of 2 ml/s. Ten minutes later, delayed contrast enhanced magnetic resonance images of the left ventricular wall were evaluated. Serum changes of tumor necrosis factor a （TNF-α） were evaluated by enzyme-linked immunosorbent assay （ELISA）.
Results Hypoenhancement was not observed at first pass perfusion at the anterior wall of the left ventricle. Hyperenhancements of the anterior-septal and anterior wall of the left ventricle was in evidence on delayed enhancement images 6 hours after microembolization and disappeared one week later. The characteristic change of coronary microembolization on delayed contrast enhanced magnetic imaging was non-enhanced regions within the hyperenhancement zone. Left ventricular ejection fraction measured by magnetic resonance imaging decreased significantly from 0.451±0.063 at baseline to 0.362±0.070 at the sixth hour （P 〈0.01）, and recovered to 0.431±0.053 one week later （P 〈0.01 vs 6th hour）. Compared with baseline values, the left ventricular end systolic volume enlarged significantly at 6th hour and at one week after microembolization （P 〈0.05 and P 〈0.01 respectively）. Serum TNF-α increased significantly at 6th hour （22.62±6.96） pg/ml compared with baseline （16.83±3.45） pg/ml （P 〈0.05） and it further increased to （27.44±3.97） pg/ml at one week after coronary microembolization and was significantly higher than that at baseline （P 〈0.01）.
Conclusions On delayed contrast enhanced magnetic resonance imaging, hyperenhancement of the anterior-septal and anterior wall of the left ventricle show at 6th hour but not at one week after coronary microembolization. This might represent the characteristic imaging after coronary microembolization. The left ventricular ejection fraction decreased at 6th hour and recovered one week later after coronary microembolization. Although impairment of left ventricular function could be recovered at 1 week after coronary microembolization, the left ventricular remodeling process still continued in concert with continuously elevation of serum TNF-α.     

Vascular endothelial growth factor expression in serum of patients with hepatocellular carcinoma

Objectives To determine the pre-therapeutic serum level of vascular endothelial growth factor (VEGF) in patients with hepatocellular carcinoma (HCC) and to elucidate the relation between the serum level and clinical characteristics and metastasis of HCC. Methods One-hundred and fifteen HCC patients, 40 patients with benign liver lesions, and 30 healthy control subjects were included in this study. The serum VEGF level was measured with the quantitative sandwich enzyme linked immunosorbent assay (ELISA, R&D systems). Results The serum VEGF levels in the HCC group (465.62±336.24 pg/ml) was significantly elevated as compared with those in patients with benign liver lesions (159.54±120.58 pg/ml) and those in normal controls (123.53±51.84 pg/ml). The VEGF levels were not significantly different between the patients with benign liver lesions and the normal controls. The serum VEGF concentration showed a positive rate of 77.4%, 25%, and 3.3% in the HCC patients, benign liver lesion patients and normal controls, respectively. In the 115 HCC patients, the serum VEGF levels in patients with portal vein (PV) emboli (n=26, 582.76±441.89 pg/ml), with metastasis (n=43, 548.29±438.57 pg/ml) or with large HCC lesions (≥5 cm in diameter) (n=69, 554.43±369.99 pg/ml) were significantly higher than those without PV-emboli (n=89, 431.39±292.84 pg/ml), without metastasis (n=72, 416.24±247.27 pg/ml) or with small HCC lesions (n=42, 328.67±227.47 pg/ml). The serum VEGF levels in stage Ⅰ, Ⅱ, Ⅲ, Ⅳa and Ⅳb HCC patients were 340.6 pg/ml, 451.55±307.84 pg/ml, 397.44±257.18 pg/ml, 486.10±397.73 pg/ml and 647.93±344.56 pg/ml, respectively. Conclusion The pre-therapeutic serum VEGF levels in HCC patients appear to reflect the disease’s potential activity of vascular invasion and metastasis.     

Effect of cholesterol lowering on stiffness of aortic and femoral arterial walls in rabbits on a high fat diet

Background Researches in arterial elasticity have increased over the past few years. We investigated the effects of simvastatin on vascular stiffness in fat fed rabbits by ultrasonography. Methods Thirty rabbits were assigned randomly to 3 groups： normal control group （A）, the cholesterol group （B）, simvastatin group （C： high fat diet for 4 weeks and high fat diet ＋ simvastatin for further 4 weeks）. Stiffness coefficient, pressure strain elastic modulus and velocity of pulse waves in abdominal aorta and femoral artery were measured by ultrasonographic echo tracking at the end of the 4th and the 8th weeks. Results At the end of the 4th week, stiffness coefficient, pressure strain elastic modulus and pulse wave velocity of femoral artery were significantly increased in group B compared with those in group A. Similarly, at the end of the 8th week, the same parameters of abdominal aorta were significantly increased in group B compared with those in group A. In contrast, stiffness coefficient, pressure strain elastic modulus and pulse wave velocity of femoral artery were significantly decreased in group C compared with those in group B, however, there was no significant difference in parameters of abdominal aorta between groups B and C. Conclusion Short term administration of simvastatin can improve the elasticity of femoral artery but not abdominal aorta.     

Expression of stromal cell-derived factor-1 in diabetic retinopathy

Background Neovascularization can cause vision loss in proliferative diabetic retinopathy （PDR） and may be affected by many factors. Stromal cell-derived factor-1 （SDF-1） is a potent stimulator of angiogenesis. The study was aimed to investigate the expression of SDF-1 and its correlation with vascular endothelial growth factor （VEGF） in the eyes with diabetic retinopathy. Methods The levels of SDF-1 and VEGF were measured by enzyme-linked immunosorbent assay in the vitreous of 41 eyes of 41 patients with PDR and 12 eyes of 12 patients with idiopathic macular hole （IMH）. Vitreous fluid samples and fibrovascular preretinal membranes were obtained at vitrectomy. SDF-1 and VEGF were localized using immunohistochemistry. Results The vitreous concentration of VEGF was significantly higher in eyes with PDR （（2143.7~1685.21） pg/ml） than in eyes with IMH （（142.42~72.83） pg/ml, P 〈0.001）. The vitreous level of SDF-1 was also significantly higher in eyes with PDR （（306.37~134.25） pg/ml） than in eyes with IMH （（86.91~55.05） pg/ml, P 〈0.001）. The concentrations of both VEGF and SDF-1 were higher in eyes with active PDR than in eyes with inactive PDR. Panretinal photocoagulation （PRP） could decrease the SDF-1 levels in the vitreous of PDR patients. The vitreous concentration of SDF-1 correlated with that of VEGF in eyes with PDR （r=0.61, P 〈0.001）. The costaining of SDF-1 and VEGF was confined to the vascular components in preretinal membranes. Conclusions SDF-1 protein is highly expressed in both the vitreous and preretinal membranes of PDR patients; SDF-1 may be correlated with VEGF in angiogenesis in PDR.     

SDF-1α与G-CSF动员造血干细胞的相关性研究

目的 :探讨趋化因子基质细胞衍生因子 (SDF 1α)与造血干细胞移植中G CSF动员造血干细胞的关系。方法 :应用酶联免疫吸附 (ELISA)试验检测 8例自体外周血造血干细胞移植 (auto PBSCT)病人及 9例异基因外周血干细胞移植 (allo PBSCT)供者应用G CSF前 1天、后第 5天血浆SDF 1α水平的变化。结果 :应用G CSF前 1天 ,allo PBSCT供者及auto PBSCT病人SDF 1α的血浆水平分别为 ( 1918.0 1± 73 .2 2 )pg/ml;( 14 43 .42± 175 .3 7)pg/ml ,应用G CSF后第 5天 ,SDF 1α血浆水平分别降至为 ( 1841.94± 60 .70 )pg/ml;( 13 12 .2 8±111.3 3 )pg/ml,二者比较有统计学意义。allo PBSCT供者在G CSF动员前 1天、后第 5天的SDF 1α血浆水平均高于auto PBSCT病人。结论 :SDF 1α不参与G CSF诱导造血干细胞的动员过程 ,auto PBSCT病人的SDF 1α血浆水平的降低可能是骨髓基质细胞功能受损的表现。     

不同胎龄新生儿血清血管内皮生长因子水平变化及与早产儿视网膜病变的关系

目的 测定不同胎龄新生儿和早产儿视网膜病变(ROP)患儿血管内皮生长因子(VEGF)血清水平变化,并探讨其与ROP的关系.方法 复旦大学附属儿科医院新生儿科入院新生儿184例,根据胎龄分为3组:<32周早产儿组、32～36周早产儿组、足月儿组,各组根据是否吸氧再分为未吸氧和吸氧2个亚组;ROP组为出生体重<2 kg,且在眼科筛查中确诊为ROP的早产儿.各组新生儿分别在生后第1、3、5、7周用酶联免疫吸附法检测血清VEGF水平.结果 未吸氧早产儿组血清VEGF水平随着日龄增加而下降,足月组血清VEGF水平较平稳.32～36周早产儿组血清VEGF水平(ng/L)在第1、3周显著高于足月儿组,差异有统计学意义(522±224比247±123,P=0.000;393±220比247±177,P=0.022).<32周早产儿组血清VEGF水平在第1周稍高于早产儿组,差异无统计学意义(P=0.071).32～36周早产儿组中,吸氧和未吸氧亚组血清VEGF水平(ng/L)差异有统计学意义(第1周:324±148比522±224,P=0.000;第3周:264 ±106比393±220,P=0.005).ROP组血清VEGF水平在第5周反常性升高,与<32周早产儿组相比,差异无统计学意义.ROP组血清VEGF水平(ng/L)在纠正胎龄31周高于非ROP早产儿组,差异有统计学意义(515±154比347±161,P=0.047).结论 早产儿血清VEGF水平异常增高时,可能与ROP的发生有关.动态监测早产儿血清VEGF水平可能有助于预测ROP的发生.     

Microvessel angiogenesis: a possible cardioprotective mechanism of external counterpulsation for canine myocardial infarction

Enhancedexternalcounterpulsation(EECP)isanoninvasivetherapyforpatientswithcoronaryarterydisease(CAD).Itconsistsofapplyingthreepairsofair filledcuffstothelowerextremitiesofthepatients.Compressivepressureofupto200-220mmHgis appliedattheonsetofdiastole,sequentiallyfromthe distaltotheproximalcuffinsynchronizationwiththe cardiaccycle,usingtheR waveofthepatient’s electrocardiogramasthetriggersignal.Attheonsetof systole,theexternalpressureinthecuffsisreleased.EECPproduceshemodynamiceffectssimil…     

急性心肌梗死患者血浆VEGF、SDF-1和外周血CD34+细胞的动态改变及其意义

目的研究急性心肌梗死时血管内皮细胞生长因子(VEGF)、基质细胞衍生因子(SDF-1)以及外周血CD34 细胞的动态改变,探讨其在心肌梗死(心梗)中的作用。方法采集急性心梗患者发病第1、3、7、10、14天外周静脉血,应用酶联免疫方法检测心梗患者以及对照组患者血VEGF和SDF-1的水平。应用流式细胞仪检测心梗患者第1、7、14天外周血CD34 细胞的水平。同时对心梗患者进行心肌酶、肌钙蛋白、心电图及心脏超声等常规检查。结果(1)心梗后第7天外周血CD34 细胞(个/μl)明显增高(2.35±0.72vs1.48±0.49,P<0.05);(2)心梗后血VEGF(pg/ml)明显升高,于第14天达到高峰(197.56±39.87vs53.79±18.12,P<0.01);(3)心梗第1天血SDF-1(pg/ml)明显降低(1683.12±224.79vs2178.67±265.34,P<0.01),以后渐恢复至与对照组相同水平;(4)心梗第7天VEGF水平与外周血CD34 细胞高峰值之间有明显相关性;(5)VEGF高峰值与CK-MB高峰值和肌钙蛋白I之间均有明显相关性。结论心肌梗死本身即可动员干细胞至外周血;心梗后VEGF明显升高至少可持续2周以上,SDF-1短暂降低;两者的动态变化可能与干细胞动员及促使更多干细胞归巢至损伤心肌有关。     

培哚普利对急性心肌梗死伴2型糖尿病患者内皮祖细胞动员及预后的影响

目的观察应用培哚普利治疗急性心肌梗死(AMI)伴2型糖尿病(T2DM)患者的预后,血浆血管内皮生长因子(VEGF)、基质细胞衍生因子(SDF-1α)水平,骨髓内皮祖细胞(EPC)动员情况。方法将20例AMI伴T2DM患者随机分为培哚普利组(10例,予口服培哚普利4mg/d)及对照组(10例,未予培哚普利)。采用密度梯度离心法分离外周血单个核细胞,应用流式细胞仪检测AMI不同时间点[急诊经皮冠状动脉介入治疗(PCI)前,PCI后1、3、5、7、14和28d]外周静脉血中CD45-/low+/CD34+/CD133+/KDR+早期EPC数量。采用酶联免疫吸附法检测血浆中VEGF、SDF-1α及超敏C反应蛋白(hs-CRP)水平。结果培哚普利组在PCI后7d的外周静脉血中EPC数量为(211±78)个/1×106,显著高于对照组的(147±57)个/1×106(P<0.05),且在PCI后5d较PCI前显著升高(P<0.05)。培哚普利组在PCI后5、7d的血浆中VEGF水平[(172±72)、(183±63)ng/L]均显著高于同时间点的对照组[分别为(105±63)、(77±37)ng/L,P值均<0.05]。培哚普利组在PCI后7d(高峰点)的SDF-1α水平为(3296±680)ng/L,显著高于同时间点的对照组[(2115±570)ng/L,P<0.05]。培哚普利组在PCI后3d(高峰点)的hs-CRP水平为(26.2±6.5)mg/L,显著低于同时间点的对照组[(33.8±10.6)mg/L,P<0.05]。随访6个月时培哚普利组的左心室射血分数(LVEF)为0.491±0.059,显著高于入院时的0.469±0.051(P<0.05);对照组LVEF分别为0.482±0.054、0.479±0.052,差异无统计学意义(P>0.05)。结论使用培哚普利治疗AMI伴T2DM患者,可改善其组织缺血后SDF-1α、VEGF-EPC动员通路障碍,并改善其预后。     

心包腔内注入腺病毒-血管内皮生长因子165基因治疗心肌缺血的实验研究

目的:探讨经心包腔途径转染血管新生基因对缺血心肌的血管生成及舒缩功能的影响。方法:第一部分:随机将12头中国小型猪分为实验组和对照组,每组6头。两组猪均采用球囊堵塞前降支第一对角支远端以建立心肌梗死模型,心肌梗死模型建立后即刻,采用经皮剑突下穿刺方法,将中心静脉导管插入心包腔内转染Ad-LacZ。以胶原酶1200 u及透明质酸酶3000u预处理心包后,在心包腔内注射含Ad-LacZ基因2.0×109pfu。对照组心包腔内注射生理盐水。分别于注射后3天、7天及28天处死动物,对缺血心肌进行染色及病理观察。第二部分:随机将20头中国小型猪分为实验组和对照组,每组10头,每组又分3天(n=2)、7天(n=6)及28天(n=6)三个亚组。注射后3天、7天及28天分别用免疫组化、超声心动图对缺血心肌血管新生情况进行检测,并以酶联免疫吸附试验(ELISA)检测血浆、心包及心肌组织中Ad-VEGF165的表达。第三部分:20头小型猪随机分为心包转染组(心包组)和冠脉转染组(冠脉组)。心包组和冠脉组均注射Ad-VEGF1651.0 ml(2×109pfu),于注射前及其后3、7、28天分别测定组织内VEGF水平、微血管密度(MVD)、心功能。结果:①实验组注射Ad-LacZ基因后第3天、第7天及28天后X-gal染色有阳性细胞,以第7天最明显,对照组无阳性细胞。②Ad-VEGF165基因经心包腔转染缺血心肌组织后,在心包及组织中成高表达,于7天达到高峰,28天降至基线水平,血浆中无目的基因的表达;28天时,实验组缺血心肌微血管密度(MVD)、心功能均明显高于对照组[MVD,517.0±75.7/mm2vs 226.5±54.1/mm2,P=0.009;LVEF72.11±5.2%vs 55.14±4.37%,P=0.005]。③心包组和冠脉组的心脏均表达有VEGF165基因,组织内VEGF水平在7天时达高峰,28天时降至基线水平,前组高于后组(702±85pg/ml vs 592±59 pg/ml,P=0.026)。而两组的MVD、心功能随转染时间延长均明显增加,但心包组优于冠脉组(28d,MVD,517.0±75.7/mm2vs 326.4±24.1/mm2,P=0.001;FS,32.9±2.2%vs 30.6±2.1%,P=0.049;LVEF,72.11±5.2%vs 65.87±2.16%,P=0.034)。结论:①应用球囊堵塞法可成功建立猪急性心肌梗死模型,胶原酶及透明质酸酶预处理心包后,腺病毒载体可转染缺血心肌,并持续表达4周。②用胶原酶及透明质酸酶预处理心包腔后,经其转染Ad-VEGF165可以诱导急性心肌梗死模型局部VEGF蛋白表达,促进缺血心肌组织血管新生并能改善心功能。③导管介导的心包腔与冠脉转染Ad-VEGF165基因治疗心肌缺血是有效的、切实可行的,而前者可能是更有前途的新方法。     

骨髓基质细胞衍生因子1及其受体在骨髓增生异常综合征患者中的表达

目的 探讨基质细胞衍生因子1(SDF-1)与其受体趋化因子受体CXCR4在骨髓增生异常综合征(MDS)发病中的可能作用,为MDS的治疗寻找有意义的靶点.方法 收集2006年10月至2010年6月59例MDS病例,根据国际预后积分系统(IPSS)分为低危和高危两组,分别为33例和26例,以10份正常的骨髓标本作为对照.采集骨髓标本,检测骨髓血浆中SDF-1的含量、CD34+细胞CXCR4的表达率、CD34+细胞的凋亡率及血管内皮生长因子(VEGF)在骨髓血浆中的表达.结果 SDF-1在低危组和高危组患者骨髓血浆中的含量[(2301±413)、(1173 ±501)ng/L]显著高于对照组[(689±190)ng/L,P＜0.05],低危组显著高于高危组(P＜0.05).CD34+细胞CXCR4的表达在高危组(68.1％±18.8％)显著高于低危组(21.0％±9.7％)和对照组(19.4％±5.3％)(P＜0.05),在后两组的表达率差异无统计学意义(P＞0.05).CD34+细胞的凋亡率在低危组、高危组和对照组分别为54.8％±10.2％,24.3％±7.9％,l8.5％±8.7％,前组显著高于后两组(P＜0.05).骨髓血浆中VEGF的含量在低危组、高危组、对照组分别为(286±97)、(407±168)、(157±46)ng/L,差异有统计学意义(P＜0.05).相关分析显示:低危组CD34+细胞的凋亡率与骨髓血浆SDF-1的含量呈正相关(r =0.805,P＜0.05);高危组血浆VEGF的含量与CD34+细胞CXCR4的表达呈正相关(r=0.683,P＜0.05).结论 SDF-1/CXCR4在MDS中存在异常表达,且与骨髓细胞的凋亡和血管新成具有相关性,针对该生物轴的干预可为该病的治疗提供新的靶点.     

肾缺血大鼠心肌血管活性物质的变化

目的 研究肾缺血时心肌组织内皮素-1(ET-1)、降钙素基因相关肽(CCRP)和一氧化氮(NO)含量与心功能变化的关系.方法 将Wistar大鼠随机分为狭窄组(n=30)和假手术组(n=10),其中狭窄组在左右肾动脉之间结扎腹主动脉(狭窄程度50%).于术后16周测定大鼠血液动力学指标,分析心肌组织中ET-1、GCRP和NO含量变化以及与心功能的关系.结果 大鼠左右肾动脉之间结扎腹主动脉使狭窄组单侧肾脏萎缩、血压持续升高,左室收缩及舒张功能下降、左室重量指数增加.术后16周时,与假手术组相比,狭窄组心肌组织血管活性物质ET-1含量显著增高(P<0.01),CGRP含量显著下降(P<0.01),NO含量差异无显著性(P>0.05);心肌组织ET-1含量与左心室内压最大上升速率呈负相关(r=-0.37,P<0.05).结论 大鼠腹主动脉狭窄导致的慢性肾缺血可使心肌组织ET-1含量增高、GCRP含量下降,其中ET-1含量与大鼠左室收缩功能呈负相关.     

内皮祖细胞对慢性脑缺血大鼠空间学习记忆影响的机制研究

目的 探讨自体移植内皮祖细胞对慢性脑缺血大鼠行为学习功能的影响.方法 制作慢性脑缺血大鼠模型,静脉自体移植内皮祖细胞,进行水迷宫检测学习记忆功能,并从脑缺血区新生细胞、脑部血管、血管内皮生长因子(VEGF)等方面探讨可能的机制.结果 ①Morris水迷宫实验中,训练第2～5天的逃避潜伏期实验组(EPC组)[分别为(44.45 ±9.44)s,(38.32±1.51)s,(34.95±6.76)s,(24.46±5.47)s明显短于对照组(PBS组)[对应为(52.79 ±6.47)s,(43.15±11.21)s,(50.29±11.41)s,(53.75±7.35)s](P＜0.01);第一象限游泳时间EPC组(26.76±3.79)s明显长于PBS组[(14.28±2.40)s],(P＜0.0).②缺血脑组织内的BrdU阳性细胞数PBS组为(12.17±3.49)个,实验组为(26.8±5.76)个,差异有显著性(P＜0.05).③脑血管共聚焦检测结果显示:EPC组较PBS组比较毛细血管内径显著性变小(分别为(2.8±0.2)μm与(3.4±0.24)μm,同源组织缺血边界地区的分支点数目显著增加[(分别为:(210.1±13.80)与(164.2±12.3)],EPC组可以显著增加微血管总面积(分别为:(84365±3897)与(74568±4626)μm2/0.002mm3),P＜0.05].④大鼠血浆VEGF检测[分别为(63.91±6.71)pg/ml;(21.81 ±4.25)pg/ml,P＜0.05]显示干预组明显增加(P＜0.05).结论 内皮祖细胞可以显著改善慢性脑缺血大鼠的学习记忆行为能力,其可能的机制是VEGF相关联的神经保护和血管再生,在治疗慢性脑缺血疾病中有着很大的应用前景.