p300/p53/Smad3 pathway involved in aging-related atrial fibrosis in human atrial fibroblasts

AIM To investigate the role of p300 in aging-related atrial fibrosis in human atrial fibroblasts （HAFs） and its potential mechanism. METHODS HAFs were obtained from human left atrial tissue, and the senescence model was established by cell passage. Senescence-associated β-galactosidase （SA-β-Gal） staining was used to detect the cell senescence, and Western blot was used to determine the protein levels of p300, p53, Smad3 and other senescence and fibrosis associated proteins in HAFs. RESULTS Compared to passage 3 HAFs, the proportion of senescent cells, and the protein levels of p300, p53, p-Smad3 and other senescence and fibrosis associated proteins were increased in HAFs at passage 7 and 11 （P<0.05）. After treated with curcumin （a p300 inhibitor） or transfection with p300 small-hairpin （sh） RNA plasmid, the protein levels of p300, and the senescence and fibrosis associated proteins were decreased in HAFs at passage 7（P<0.05）. Up-regulation of p300 by transfection with p300 over-expression plasmid increased the protein levels of p53, Smad3 and MMP-2 in HAFs at passage 3 （P<0.05）. CONCLUSION p300/p53/Smad3 signaling pathway plays an important role in aging-related atrial fibrosis in HAFs.     

Correlation between progression of atherosclerosis accelerated by emotional stimulation and imbalance of SDF-1/CXCR4 in mice

AIM To observe effects of emotional stimulation on expression of stromal cell-derived factor-1（SDF-1） and CXC chemokine receptor 4 （CXCR4） in plasma, platelets, aortas, hippocampus and bone marrow of apolipoprotein E gene knockout （ApoE^-/-） mice, and to reveal the possible mechanism of the aggravated atherosclerotic plaque vulnerability by emotional stimulation. METHODS Thirty 8-week-old male ApoE^-/- mice were randomly divided into normal control group, high fat group, and emotional stimulation group. Ten 8-week-old inbred C57BL/6J mice served as blank control group. After 12 weeks of intervention, the serum levels of SDF-1 and CXCR4 were investigated by ELISA. The protein levels of SDF-1 and CXCR4 in platelets, aortas, hippocampus and bone marrow were determined by Western blot. The pathological damage of aortas was observed by oil red O staining. RESULTS Compared with blank control group, normal control group and high fat group, the mice subjected to emotional stimulation showed more serious atherosclerosis in aortas detected by oil red O staining, and increased levels of SDF-1 and CXCR4 in the plasma and aortas were also observed （P<0.05）. The results of Western blot showed that the protein levels of SDF-1 and CXCR4 in platelets, aortas and hippocampus were increased in the mice subjected emotional stimulation, but the expression of SDF-1 and CXCR4 in the bone marrow was decreased （P<0.05）. CONCLUSION Imbalance of SDF-1/CXCR4 may be the key target by which emotional stimulation accelerates the progression of atherosclerosis.     

不同有机酸对小白菜镉污染毒性和品质的影响

以小白菜为试材,以贵州典型黄壤为盆栽试验基质,在黄土中共设置5个不同处理,1)原始无污染土壤(CK);2)单一镉污染土壤处理(Cd);3)镉污染土壤加草酸处理(Cd+OA);4)镉污染土壤加柠檬酸处理(Cd+CA);5)镉污染土壤加酒石酸处理(Cd+TA),探究镉胁迫下不同有机酸对提高小白菜品质的效果,以期为改善蔬菜品质提供参考依据。结果表明:外源添加镉导致小白菜单株鲜质量降低了41.1%,植株体内镉大量积累,活性氧类物质、丙二醛(MDA)和脯氨酸含量升高。与对照组相比,镉污染小白菜叶片内硝酸盐含量增加了2.9倍,维生素C和可溶性糖含量分别降低了19.3%和9.9%。在镉污染土壤上施入草酸、柠檬酸和酒石酸后均显著改善了小白菜的生长状态。与镉污染处理小白菜相比,柠檬酸对小白菜的促生效果最显著,生物量相对增加了48.5%,叶片和根的镉含量分别降低了45.3%和42.6%。酒石酸对超氧阴离子的影响大于草酸和柠檬酸,对过氧化氢和脯氨酸的影响小于草酸和柠檬酸,但草酸抑制硝酸盐积累和提高可溶性糖含量的作用大于其它有机酸。上述结果说明,在镉污染土壤施用柠檬酸可以有效缓解镉对小白菜的毒害作用,提高小白菜的产量和品质。     

藁本属药用植物分子生物学研究进展

藁本属内大多植物为我国传统中药材,具有活血行气、散寒、除湿、祛风止痛的作用.然而目前对藁本属植物的研究大多集中在栽培种植技术、药用成分价值等方面,分子技术研究较少,研究面比较窄,导致对其更进一步的分子育种、药效成分的基因定位等研究进展缓慢.该研究归纳国内外藁本属植物的遗传多样性、DNA基因鉴别、功能基因克隆、转录组等研究进展,分析了藁本属植物分子生物学研究的不足,并对未来研究应关注的方向进行了展望,以期为藁本属植物的分子生物学研究提供参考依据.     

鸡白细胞介素-6原核表达及双抗夹心ELISA方法的建立

应用分子生物学技术原核表达ChIL-6，以ChIL-6重组蛋白为免疫原，按免疫程序分别制备兔抗和鼠抗IL-6重组蛋白的多克隆抗体。应用此抗体建立双抗夹心EL-ISA方法后，为了优化此方法本试验对该方法的最佳试验条件、标准曲线、重复性和初步应用进行确定。结果显示，经SDS-PAGE和Western-blot分析，表明ChIL-6在大肠杆菌中正确表达，为了建立检测ChIL6的双抗夹心ELISA方法，本试验应用表达的重组蛋白制备兔抗和鼠抗多克隆抗体。建立的双抗夹心ELISA方法最佳反应条件为包被抗体的质量浓度为50mg／L，4℃过夜；酶标二抗稀释度为1：400，37。C1h；应用该方法检测感染金黄色葡萄球菌后的ChIL-6，其结果与本实验室之前应用人的ELISA试剂盒检测的结果相似。结果表明，本试验建立的检测ChlL-6的双抗夹心ELISA方法可用于临床。     

农杆菌浸润瞬时表达分析番木瓜miR162a的研究

农杆菌浸润瞬时表达方法不仅在蛋白互作、鉴定基因沉默抑制子及基因功能研究方面得到广泛应用,目前还应用在小RNA的研究中.在对植物microRNA的研究中,农杆菌浸润方法通过将将转基因导入农杆菌浸润本生烟瞬时表达,可以快速、稳定地对植物microRNA进行检测和鉴定,样品在几天内就可分析完成,比获得稳定遗传的转基因有优势.笔者借鉴国外拟南芥microRNA瞬时表达分析的方法,克隆了番木瓜的miR62a前体序列连接到双元表达载体pSuper1300上构建成35S::cpa-miR162a质粒,导人农杆菌GV3101后浸润本生烟,浸润后48 h提取本生烟叶片总RNA后通过miRNA northern blot检测到miR162a,而本生烟和番木瓜自身则无法检测到低丰度的miR162a.本文介绍的农杆菌浸润瞬时表达方法能够在体外加工产生成熟的miRNA,该技术有助于对microRNA的结构-功能的相关性及靶标验证进一步研究.     

生物质燃料常温变径成型锥头研究

针对生物质常温闭式成型机的不足,本文引入了开式成型的概念,并在原有闭式成型机液压驱动设备的基础上,改进了成型腔.通过对生物质原料的力学分析,设计了开式成型的变径成型锥头.并使用小刨花、锯末等材料进行验证,探讨了对生物质常温变径成型机成型效果的影响因素,同时确定了较高成型质量时成型腔的压缩比范围.     

乡村旅游开发项目的社会评价

分析了乡村旅游开发项目开展社会评价的必要性,总结了开展社会评价的方法原则;在综合现有研究的基础上,提出了从社会经济影响、社会环境影响、社会生态影响、项目与当地社会互适性、社会风险五个方面开展乡村旅游开发项目的社会评价,并构建了一整套的评价指标体系;最后针对指标体系的完善和应用提出了一些思考和讨论。     

“紫科1号”红豆杉微枝繁殖技术研究

以"紫科1号"红豆杉的6种半木质化枝条为扦插繁殖材料,经3种处理液预处理,在8种基质上进行扦插繁殖试验。结果表明:3~12 cm长的枝条均可用作插穗,选用长5 cm左右的枝条进行微枝繁殖较为节约材料;用IBA∶NAA=3∶2,总溶质浓度为1 000 mg/L的处理液速蘸基部后,插于黄心土∶河沙=3∶2的基质中,其后每周用稀释1 000倍的处理液喷洒1次,生根率平均可达97.36%。