游泳对大鼠心肌碱性成纤维细胞生长因子(bFGF)表达的影响

目的:碱性成纤维细胞生长因子(bFGF)有强烈诱导血管生成和修复血管损伤的作用,故其在心肌中的表达程度可以作为判断运动对心脏侧枝血管生成影响的指标.以雄性SD大鼠为实验对象,通过施加中等强度的游泳训练,观察运动训练对SD大鼠心肌组织学结构和bFGF表达的影响,为运动医学与心脏康复的理论和实践提供依据;方法:将32只健康雄性SD大鼠按体重随机分为2组:对照组(C组);运动组(E组),每组均为16只.C组安静饲养,不进行运动训练,但需放入过膝浅水中以模拟训练组环境;E组大鼠每周训练6天,每天训练1次.从第1周开始适应性游泳训练,第一次游泳5min,此后逐日累加5min,至第3周末时每天游满2h.此后,从第4周起维持这一运动量直至第8周.第8周进行游泳训练后,即刻处死各组大鼠,取出心脏称湿重以做心脏系数统计,并切取左心室前壁组织块,做HE染色和bRGF免疫组织化学染色.用光学显微镜观察各组心肌及心肌间质的变化,并用图像采集分析系统观测各组HE染色和bFGF免疫组织化学染色变化,做统计学处理;实验结果:1)一般情况观察:心脏重量和心系数,E组较C组显著增加(P＜0.01);2)心肌组织学观察,HE染色显示C组心肌组织形态正常;而E组呈现生理性肥大变化,心脏毛细血管功能结构产生了良好的重构,心肌组织之间毛细血管的密度增加;3)bFGF免疫组织化学染色结果观察,C组呈弱阳性表达;E组呈较强的阳性表达;心肌细胞中bFGF表达呈特异性棕黄染色颗粒,以计算机显微图像采集分析系统计算bFGF染色面积,发现E组大于C组(P＜0.01);bFGF染色平均密度E组大于C组(P＜0.01);bFGF染色积分光密度,E组均大于C组(P＜0.01);结论:本实验观察到,适宜的耐力运动训练可以使心肌细胞、心肌血管内皮细胞等部位bFGF的表达增强,改善心肌细胞的分子生物学特性、心脏毛细血管功能结构产生了良好的重构,心肌组织之间毛细血管的密度增加,毛细血管腔表面积密度增高,毛细血管与心肌纤维的比例增大,使心脏产生适应性生理肥大,表明中等强度的运动可以使心脏发生生理性肥大,其机理可能与bFGF的表达增强有关.     

Basic fibroblast growth factor alleviates brain injury following global ischemia reperfusion in rabbits

The aim of this study was to explore the protective effect of basic fibroblast growth factor (bFGF) on brain injury following global ischemia reperfusion and its mechanisms. Brain injury following global ischemia was induced by four vessels occlusion and systemic hypotension. Twenty-four rabbits were randomized into three groups: group A, only dissection of vessels; group B, intravenous infusion of normal saline after reperfusion for 6 h; group C, 30 μg/kg bFGF injected intravenously at the onset of reperfusion, then infused with 10 μg/(kg·h) for 6 h. Serum neuron specific enolase (NSE), S-100B, tumor necrosis factor-α(TNF-α), interleukin-1 (IL-1), interleukin-8 (IL-8) were measured before ischemia, 30 min after ischemia, 0.5, 1, 3, 6 h after reperfusion. Brain water content was determined and cerebral histopathological damages were compared. NSE and S-100B were increased 1 h after reperfusion and reached their peaks 6 h after reperfusion, but were much higher in group B than those in group C 3, 6 h after reperfusion. In groups B and C, TNF-a was increased after ischemia and IL-1 and IL-8 were increased significantly 0.5 h after reperfusion, then reached their peaks 6 h, 3 h, 6 h after reperfusion respectively. TNF-a and IL-8 at the time points of 1 h and 3 h and IL-1 at 3 h and 6 h in group C were correspondingly lower than those in group B. These indices in group A were nearly unchanged. There were less severe cerebral histopathological damages in group C compared with group B, but no difference in brain water content. It could be concluded that bFGF alleviates brain injury following global ischemia and reperfusion by down-regulating expression of inflammatory factors and inhibiting their activities.     

Improved myocardial perfusion and cardiac function by controlled-release basic fibroblast growth factor using fibrin glue in a canine infarct model

Objective: Angiogenic therapy is emerging as a potential strategy for the treatment of ischemic heart disease but is limited by a relatively short half-life of growth factors.Fibrin glue (FG) provides a reservoir for controlledrelease of growth factors.The aim of this study was to evaluate the effects of basic fibroblast growth factor (bFGF) incorporating FG on angiogenesis and cardiac performance in a canine infarct model.Methods: Acute myocardial infarction was induced by ligation of the left anterior descending coronary artery (LAD).Group I (n=6) underwent ligation of LAD alone.In Group II,transmural channels were created in the infarct area (n=6).In Group III,nontransmural channels were created to locate FG cylinders containing bFGF (n=6).Eight weeks after operation,myocardial perfusion was assessed by single photon emission computed tomography,cardiac function by echocardiography,and vascular development by immunohistochemical staining.Results: Total vascular density and the number of large vessels (internal diameter ≥50 μm) were dramatically higher in Group III than in Groups I and II at eight weeks.Only the controlled-release group exhibited an improvement in regional myocardial perfusion associated with lower defect score.Animals in Group III presented improved cardiac regional systolic and diastolic functions as well as global systolic function in comparison with the other two groups.Conclusions: Enhanced and sustained angiogenic response can be achieved by controlled-release bFGF incorporating FG within transmyocardial laser channels,thus enabling improvement in myocardial perfusion and cardiac function.     

组织工程的一个初步研究混合多尿烷血管移植(英文)

本文的目的是研究半多孔内皮细胞的包膜(SPEUs)的效应 以PTMEG1000为基础的SPEU的多孔性为51.58%(±5.86%) 与天然脉管相比较,例如与一头6个月小牛胸部的上部下来的主动脉大血管相比较,SPEU的拉伸强度是足够强的 脉管的典型脉动压力服从静脉管压力(1.5～2%Δd/d),此压力小于动脉管压力(9～15%Δd/d) 在种植EC之前,观察了在SPEUs上培育的人皮的成纤维细胞(HDF) SPEU的DNA数量与HDF在一天之内种植;有71.9%(±11.01%)细胞被种植在SPEUs上 LDH试验表明:在有控制的和实验群数据之间没有显著的差别,说明没有细胞毒素 因此,EC在SPRUs上具有良好的粘附性,这是所期望的,而应用胶原实现了将EC种植在半微孔的SPEUs上的表面改善     

人酸性成纤维细胞生长因子cDNA的表达

目的：研究人酸性成纤维细胞生长因子（haFGF）cDNA在不同表达系统中的表达，旨在得到高效表达haFGF的表达系统。方法：采用PCR方法扩增人酸性成纤维细胞生长因子(haFGF）cDNA，将该cDNA片段分别插入表达载体pKK223-3及pET3C的表达框架中，筛选得到了重组质粒pKK223-3-haFGF及pET3C-haFGF，分别转化大肠杆菌JM109及BL21（DE3），构建了表达菌株JM109/pKK223-3-haFGF及BL21(DE3)/pET3C-haFGF,用IPTG诱导表达。结果；SDS-PAGE的结果表明，表达菌株BL21(DE3)/pET3C-haFGF能高效表达人酸性成纤维细胞生长因子，而JM109/pKK223-3-haFGF诱导后目标蛋白带不明显。结论：BL21（DE3）/pET3C表达系统更适于表达人酸性成纤维细胞生长因子。     

Human cytomegalovirus induces alteration of β-actin mRNA and microfilaments in human embryo fibroblast cells

Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV. HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time-and titer-dependent manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion: CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression of β-actin mRNA and the microfilaments. Project (No. 001103058) supported by the Key Program of Science and Technology Bureau of Zhejiang Province, China     

Differential time responses in inflammatory and oxidative stress markers after a marathon: An observational study

ABSTRACT

Acute and adaptive changes in systemic markers of oxidatively generated nucleic acid modifications (i.e., 8-oxo-7,8-dihydro-2?-deoxyguanosine (8-oxodG) and 8-oxo-7,8-dihydroguanosine (8-oxoGuo)) as well as inflammatory cytokines (i.e., C-reactive protein, interleukin-6, interleukin-10, and tumour necrosis factor alpha), a liver hormone (i.e., fibroblast growth factor 21 (FGF21)), and bone metabolism markers (sclerostin, osteocalcin, C-terminal telopeptide, and N-terminal propeptide of type 1 procollagen) were investigated following a marathon in 20 study participants. Immediate changes were observed in inflammatory cytokines, FGF21, and bone metabolism markers following the marathon. In contrast, no immediate changes in urinary excretion of 8-oxodG and 8-oxoGuo were evident. Four days after the marathon, decreased urinary excretion of 8-oxodG (-2.9 (95% CI -4.8;-1.1) nmol/24 h, P < 0.01) and 8-oxoGuo (-5.8 (95% CI -10.3;-1.3) nmol/24 h, P = 0.02) was observed. The excretion rate of 8-oxodG remained decreased 7 days after the marathon compared to baseline (-2.3 (95%CI -4.3;-0.4) nmol/24 h, P = 0.02), whereas the excretion rate of 8-oxoGuo was normalized. In conclusion marathon participation immediately induced a considerable inflammatory response, but did not increase excretion rates of oxidatively generated nucleic acid modifications. In fact, a delayed decrease in oxidatively generated nucleic acid modifications was observed suggesting adaptive antioxidative effects following exercise.     

Effect of basic fibroblast growth factor （bFGF） on the treatment of exposure of the orbital implants

Objective： To evaluate the efficacy and the indication of basic fibroblast growth factor （bFGF） in the treatment of exposure of orbital implants. Design： Retrospective and observational case series. Methods： We reviewed 41 patients （41 eyes） suffering exposure of orbital implants from Jan. 2000 to June 2006. The study group patients with mild exposure received combined treatment with bFGF and antibiotic drops, and while the control group patients with mild exposure were treated with antibiotic drops only. The study group patients with moderate and severe exposure received combined treatment with bFGF and antibiotic drops, and after 2 months they were subjected to amniotic membrane transplantation, while the control group patients with moderate and severe exposure underwent amniotic membrane transplantation after using antibiotic drops. Observation of the growth of conjunctival epithelium and comparison of the healing rate of the two groups. Results： The healing rates of the mild, moderate and severe exposure study group were 100% and 92.3%. The healing rates of the mild, moderate and severe exposure control group were 55.6% and 66.7% respectively. The difference of the healing rates of the mild exposure study group and the control group was significant （P=-0.033）. And the difference of the healing rates of the moderate and severe exposure study group and the control group was not significant （P=-0.167）. Conclusion： bFGF may promote obviously the healing of orbital implant exposure, particularly it can be the first choice for the treatment of mild degree exposure. For the moderate and severe cases, it can be administered before surgical repair to enhance neovascularization and will tend to increase the success rate of surgical repair.     

Human cytomegalovirus induces alteration of (-actin mRNA and microfilaments in human embryo fibroblast cells

Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV. HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time- and titer-dependent manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion: CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression of β-actin mRNA and the microfilaments.