Expression of tissue factor,tissue factor pathway inhibitor in the atherosclerotic plaque on femoral artery of New Zealand rabbits

AIM: To observe the regulatory changes of tissue factor (TF),tissue factor pathway inhibitor-1 (TFPI-1) and tissue factor pathway inhibitor-2 (TFPI-2) associated with the formation of atherosclerotic plaque on femoral artery of New Zealand rabbits.METHODS: New Zealand rabbits were fed with high cholesterol diet and received femoral artery balloon injury to establish moderated atherosclerotic plaque animal model.These rabbits were classified into 8 weeks group,10 weeks group and 12 weeks group.The expressions of factor TF,TFPI-1 and TFPI-2 in femoral artery were detected by immunohistochemical methods at 8th,10th and 12th week.The positive areas of the slice were analyzed by semi-quantification system.Fluorescence quantification PCR (FQ-PCR) was used to detect the copies of mRNA of TF,TFPI-1 and TFPI-2 in each group.RESULTS: The expressions of TF,TFPI-1 and TFPI-2 determined by immunohistochemical methods were increased from 8th,10th to 12th week,respectively.In each group,the quantity of mRNA of TF and TFPI-1 at the 8th,10th and 12th week was soared,respectively.In each group,the quantity of mRNA of TFPI-2 did not change in early phase,but decreased at 12th week.CONCLUSION: In the process of the atherosclerotic plaque formation in rabbit femoral artery model,the expressions of TFPI-1 and TFPI-2 proteins are increased.However,the expression of TPFI-1 or TFPI-2 could not stave off the development of plaque formation.     

湄江翠片加工中香气与色泽的相关性初探

本文详细介绍了目前湄江翠片茶各种工艺组合，并根据制茶原理，分析了制茶因子对湄江翠片的品质影响，通过分析对比，找出了湄江翠片比较合理的工艺组合。     

应用MTT比色法评价不同牛血清促细胞生长作用

细胞培养过程中血清的选择对细胞的促生长增殖具有重要作用。本试验分别以商品新生牛血清、自制新生牛血清、自制成年牛血清培养成纤维细胞、骨髓瘤细胞、杂交瘤细胞，通过MTT比色法来判断细胞的增殖能力，进而对血清质量作出评价。结果表明，自制新生牛血清具有良好的促细胞生长作用(相对生长率＞0.96)，并且3次试验结果比较差异不显著(P＞0.05)，具有较好的重复性；而成年牛血清和无血清空白对照组促细胞作用不明显。因此应用MTT比色法能够评价血清质量。     

鸭肿头出血症病毒强毒的致病性及感染组织细胞的超微病理变化

将鸭肿头出血症病毒（DSHDV）强毒株以不同途径接种10日龄鸭胚，研究了病毒在鸭胚中的繁殖规律，并用透射电镜观察了感染鸭胚各组织器官的超微结构变化，同时用DSHDV人工感染不同日龄雏鸭，比较了不同感染途径对雏鸭的致病性和对不同日龄雏鸭的致病性。结果显示，尿囊腔和卵黄囊2种途径均能使病毒在鸭胚上繁殖并传代，对鸭胚的半数致死量分别为10^-7.24／0．2mL和10^-6.4／0．2mL。病毒感染鸭胚后，电镜下可观察到肝和尿囊膜中的病毒粒子，病毒粒子呈球形或椭圆形，大小为60～80nm，无囊膜；病毒在细胞浆内复制，可形成特征性包涵体；病毒侵害的主要靶器官为尿囊膜与肝，细胞器的变化主要表现为粗面内质网扩张以及线粒体肿胀和嵴断裂、消失。病毒经肌肉注射、口服和滴鼻均能使28日龄鸭感染发病，致死率分别为86％、89％和97％。研究表明，DSHDV能很好地在鸭胚上生长并致鸭胚各组织器官超微结构发生变化，不同感染途径均能使雏鸭感染发病且致病性强。     

一种用C18柱纯化低浓度微量寡核苷酸的新方法

通过比较4种不同的回收方法和C18硅胶4种不同的用量对回收的影响,提出了回收低浓度微量寡核苷酸的最佳方案:分别用1 mL乙腈、2 mL灭菌双蒸水和1 mL 10 mmol/L乙酸铵平衡装填10 mg硅胶的C18 Sep-Pak反相层析柱;再将寡核苷酸溶液通过C18柱;然后用1 mL 25 mmol/L碳酸氢铵洗脱C18柱,并以3000 r/m in离心2 m in;最后用1 mL 30%乙腈洗脱C18柱,将收集的溶液离心冻干后即得到纯化的寡核苷酸。该方法的回收率可达53.7%±3.0%。     

丙烯酸松香的溴化及产物的阻燃性研究

以丙烯酸松香与液溴加成,合成了溴化丙烯酸松香;研究了反应温度、原料的物质的量比、反应时间等因素对反应过程及产物性能的影响.用FT-IR、1 HNMR和热重分析(TG)等对溴化丙烯酸松香进行了表征.结果表明,当反应温度为-2 ～ 4℃、反应时间5 ～ 6h、液溴与丙烯酸松香的物质的量比值约为1.2时,收率可达93%.溴化丙烯酸松香不仅具有较好的阻燃性,同时还拓宽了松香的应用范围,提高了其附加值.     

MMP-7,MMP-10 and TIMP-4 expression in ventricular remodeling of human heart failure

AIM:To sieve matrix metalloproteinases (MMPs) and the tissue inhibitors of matrix metalloproteinases (TIMPs) closely associated with ventricular remodeling of human heart failure using antibody chip technology.METHODS:We performed cytokine-specific antibody array analysis using individual left ventricular myocardial samples from 6 patients with heart failure due to arrythmogenic right ventricular cardiomyopathy (ARVC) undergoing transplantation and matched samples from 6 non-failing subjects to screen differentially expressed MMPs and TIMPs associated with the ventricular remodeling of heart failure.The results were further validated by ELISA and immunohistochemical analysis.RESULTS:We identified high expression of MMP-7 and MMP-10 and low expression of TIMP-4 in ARVC failing hearts compared to non-failing hearts by hybridization with the cytokine-specific antibody arrays containing 17 MMPs and 4 TIMPs on the chips.ELISA and immunohistochemical analyses further confirmed that differentially expressed levels of MMP-7, MMP-10, and TIMP-4 were observed not only in ARVC failing heart, but also in failing hearts due to ischemic (ICM) and dilated cardiomyopathy (DCM).CONCLUSION:Highly expressed MMP-7 and MMP-10 and lowly expressed TIMP-4 may be involved in the ventricular remodeling of heart failure derived from cardiomyopathy of different etiology.     

Soluble expression of a CXCL10-loop3-EGF fusion protein and its anti-tumor activity

AIM: To evaluate the implication of CXCL10-loop3-EGF fusion protein for the activities of targeting tumor and anti-angiopoiesis. METHODS: RT-PCR was preformed to amplify CXCL10 coding sequence from PBMC activated by IFN-γ. CXCL10-loop3-EGF fusion gene, which was conducted by Over-Lap Extention PCR, was hinged up with plasmid pTG19-T, transfected to E. coli DH5α and processed positive colony selection. After ligated with plasmid pET32a(+), recombinant CXCL10-loop3-EGF fusion gene was then transfected to E. coli Origami B (DE3) and induced to express its coding fusion protein his-CXCL10-loop3-EGF. The recombinant fusion protein CXCL10-loop3 -EGF was purified by His-bind affinity chromatograph, enterokinase cleavage, ultrafiltration and dislysis. The transwell chemotatic test and HUVEC angiopoiesis inhibition test were performed to determine the anti-tumor responses and anti-angiopoiesis activity of CXCL10-loop3-EGF fusion protein. RESULTS: CXCL10-loop3-EGF fusion protein was successfully constructed and confirmed by SDS-PAGE analysis and Western blotting. Significant PBMC chematatic activity and HUVEC anti-angiopoiesis activity were observed. CONCLUSION: CXCL10-loop3-EGF fusion protein, which has perfect anti-tumor activity, is successfully constructed.