吉林省东部粳稻生长和产量对移栽期和温度变化的反应

以中晚熟水稻品种"吉粳811"为研究对象,于2013年和2014年在延吉市进行分期播种/移栽试验,分析了吉林省东部地区一季粳稻生长速度和产量对移栽期及温度变化的响应规律,确定该品种在研究地区的适宜播种期和移栽期,以减免低温冷害的影响。结果表明:播期/移栽期的推迟提升了生长季平均气温,水稻生长发育进程加快,有效生育期缩短,主要生长季内平均气温每升高1℃,水稻生长速率提升19%,生育期缩短5 d左右;吉林省东部地区水稻的适宜移栽温度为日均气温13.0℃,移栽过早或偏晚均导致减产;中晚熟品种水稻在4月18日前后播种、5月26日前后移栽可保证在霜前成熟,且产量高;中晚熟品种水稻移栽至成熟适宜活动积温为2280℃·d左右,活动积温每减少100℃·d,水稻产量下降1095 kg·hm~(-2)(约减产13%);积温不足导致水稻冷害发生,因而减产。     

紫穗槐叶片浸提液对长柄扁桃种子萌发和幼苗生长的影响

紫穗槐和长柄扁桃是中国西北干旱和半干旱地区的常见绿化植物.为探索紫穗槐搭配长柄扁桃进行绿化建设和生态修复的可行性,采用培养皿滤纸法和土培法测定了5种质量浓度的紫穗槐叶片浸提液(0.025、0.05、0.10、0.15和0.20 g ? mL-1)对长柄扁桃8个品种(YY1、YY3、YY4、YY5、YY6、SM6、SM7...     

α-SMA与肿瘤发展相关性的研究

以小鼠的肝、肺组织的正常细胞与肝肿瘤细胞（Hepa1-6）、肺肿瘤细胞（LLC）为研究对象,通过CTFM（cell traction force microscopy）法测定了4组细胞系的牵引力;用荧光抗体染色技术比较了小鼠细胞的α-SMA蛋白在癌变前后的变化。实验发现：与小鼠正常细胞相比,在α-SMA的表达水平上,Hepa1-6细胞α-SMA的平均光密度减小了47.9%,LLC细胞下降了52.3%;而Hepa1-6细胞牵引力的均方根值减小了53.4%,LLC细胞减小了49.7%。这说明α-SMA的表达与牵引力的变化以及细胞癌变的过程是密切相关的。     

Lectin binding to gp60 decreases specific albumin binding and transport in pulmonary artery endothelial monolayers.

The effect of albumin binding to cultured bovine pulmonary artery endothelial cell (BPAEC) monolayers on the transendothelial flux of 125I-labelled bovine serum albumin (BSA) was examined to determine its possible role on albumin transcytosis. The transport of 125I-BSA tracer across BPAEC grown on gelatin- and fibronectin-coated filters (0.8 microns pore diam.) was affected by the presence of unlabelled BSA in the medium in that transendothelial 125I-BSA permeability decreased, reaching a 40% reduction at BSA concentrations equal to or greater than 5 mg/ml. BSA binding to BPAEC monolayers was saturated at concentration of 10 mg/ml with an apparent binding affinity of 6 x 10(-7) M. In contrast, gelatin added to the medium altered neither 125I-BSA binding nor transport. Several lectins were tested for their ability to inhibit 125I-BSA binding and transport. One lectin, Ricinus communis (RCA), reduced 125I-BSA binding by 70% and transport by 40%. Other lectins, Ulex europaeus, Triticum vulgare, and Glycine max decreased neither 125I-BSA binding nor transport. The reduction of 125I-BSA transport by RCA was not observed in the presence of saturating levels of BSA, indicating that RCA influenced only the albumin-dependent component of transport. RCA, but not other lectins, precipitated a 60 kDa plasmalemmal glycoprotein from cell lysates of surface radioiodinated BPAEC monolayers. This 60 kDa glycoprotein appears to be the equivalent of gp60 identified previously as an albumin binding glycoprotein in rat microvascular endothelium. In summary, approximately 40% of albumin transport across BPAEC monolayers is dependent on albumin binding. This component of albumin transport is inhibited by 80% by the binding of RCA to gp60. These results suggest that binding of albumin to gp60 on pulmonary artery endothelial cell membrane is a critical determinant of transendothelial albumin flux involving mechanisms such as plasmalemmal vesicular transcytosis.     

Determination of diclofenac in human plasma by selected ion monitoring

A simple and rapid gas chromatographic/mass spectrometric method to determine plasma diclofenac was developed, which employs formation of the methyl ester with diazomethane. Methoxydiclofenac was used as the internal standard. Under the conditions used, the previously described partial cyclization of diclofenac to the indolone derivative was avoided. The limit of detection of plasma levels of diclofenac is 2 ng ml-1, which renders the method useful for clinical studies on oral, intravenous and rectal administration of the drug. The analysis is carried out by electron impact gas chromatography/mass spectrometry and can therefore be performed on the more common mass spectrometers. Linearity and reproducibility of the method were demonstrated by the high correlation coefficient of the calibration lines (r greater than 0.999) and from the low variation of their slopes (coefficient of variation 3%) determined on different days, respectively. Pharmacokinetic parameters (area under curve = 1.8 +/- 0.26 microgram h ml-1, tmax = 1.5 +/- 0.5 h, Cmax = 734 +/- 82 ng ml-1 and terminal half-life = 0.88 +/- 0.52 h) determined from the plasma decay of diclofenac in three healthy subjects given a single oral dose of diclofenac were in good agreement with those reported in the literature.     

东亚飞蝗羧酸酯酶基因的克隆和原核表达

羧酸酯酶是昆虫体内重要的代谢解毒酶系,其主要功能是水解和结合内源性和外源性含有酯键的有毒物质,减缓其到达靶标部位的时间。东亚飞蝗Locusta migratoria manilensis(Meyen)是我国重要的农业害虫,对其羧酸酯酶基因克隆和表达有助于深入探索杀虫剂代谢毒理机制。本研究首先对羧酸酯酶基因(CarE4)进行了克隆,并将其插入到pCold TF DNA Vector中,在大肠杆菌中进行了原核表达,最后用疏水层析和离子交换层析方法对目的蛋白进行了纯化。本文成功建立了羧酸酯酶蛋白原核表达和纯化技术体系,为进一步研究东亚飞蝗羧酸酯酶的生理功能、结构特点和作用原理提供了基础资料。     

黄土高原雨水集聚深层入渗(RWCI)系统下山地果园土壤水分时空变异特征

水资源短缺是影响黄土高原雨养农业发展的关键性因素,雨水资源开发是缓解该地区水资源短缺的有效措施.本研究利用管式 TDR 系统监测21年红富士老果园0～300 cm土层土壤含水率变化,分析了雨水集聚深层入渗(RWCI)系统下黄土高原旱作山地果园土壤水分时空分布特征.结果表明: RWCI系统能够显著增加果园土壤含水率,特别是40～80 cm土层(土壤含水率低值区)土壤含水率,在该区域,不同设计深度(40、60和80 cm)RWCI处理(RWCI₄₀、RWCI₆₀和RWCI₈₀)年均土壤含水率分别较鱼鳞坑(CK)处理提高75.3%、85.4%和62.4%,分别较裸露坡地(BS)处理提高39.2%、47.2%和29.1%.RWCI₄₀、RWCI₆₀和RWCI₈₀处理土壤水分入渗最大深度分别为80、120和180 cm,显著深于CK处理(60 cm),其中土壤水分变化幅度最大的土层分别主要发生在0～60、0～100和0～120 cm.在果树整个生育期内,RWCI处理土壤平均含水率(0～300 cm)以RWCI₈₀处理最大,其次是RWCI₄₀和RWCI₆₀处理.总体来看,RWCI系统是黄土高原实现雨水资源化和农业高效用水的有效措施.     

人Semaphorin 4D慢病毒载体的构建及鉴定

目的:构建并制备能够有效表达Semaphorin 4D的重组慢病毒。方法:从人急性T细胞白血病Jurkat细胞DNA 扩增人Semaphorin 4D基因,克隆至pWPI GW慢病毒载体上,与pVSVG及pSPAX质粒共转染人胚肾293T细胞,包装出重组慢病毒,将纯化后的重组病毒直接感染293T和HUVEC细胞,通过免疫印迹、免疫荧光染色和血管内皮细胞迁移分析等方法检测Semaphorin 4D的表达和诱导血管内皮细胞迁移的作用。结果: 重组慢病毒介导Semaphorin 4D在293T和HUVEC内获得表达,能介导血管内皮细胞迁移。结论:成功构建了表达Semaphorin 4D的重组慢病毒载体。     

慢病毒介导的GFP-Hsp90αE47A基因在HepG2细胞表达及对细胞增殖性影响的研究

目的：建立真核细胞表达的GFP-Hsp90αE47A基因重组慢病毒载体三质粒包装细胞系统,并检测其对细胞增殖性的影响,为进一步研究HSP90分子伴侣功能奠定基础。方法：制备完整的重组慢病毒载体三质粒系统：转移质粒（Hsp90αE47A/psin-GFP）,包装质粒（ΔNRF）及包膜蛋白质粒（VSV-G）。磷酸钙法将三质粒共转染293T包装细胞,48h后收集病毒上清。将制备好的慢病毒颗粒感染HepG2细胞,在荧光显微镜下观察报告基因GFP的表达情况,Westernblot检测HepG2细胞GFP-Hsp90α表达。MTT法检测细胞增殖情况。结果：转染后的293T和感染后的HepG2细胞能观察到较强的绿色荧光,培养液上清病毒滴度约为3.0×103ifu/μl,HepG2细胞中有GFP-Hsp90α蛋白的表达。内源性Hsp90α表达无明显上升（为对照组的1.05±0.15倍,P〈0.05,t检验）,有明显外源性GFP-Hsp90αE47A蛋白的表达,为对照组内源性Hsp90α的0.68±0.12倍。外源性GFP-Hsp90αE47A蛋白的表达HepG2细胞增殖活性于第4d有明显抑制。（1.051±0.03vs1.349±0.05,P〈0.05,t检验）。结论：成功建立重组慢病毒载体的三质粒包装细胞系统,并将GFP-Hsp90αE47A基因在HepG2细胞中稳定表达,且并未引起细胞明显的热休克反应而导致的内源性Hsp90α增高;且能明显抑制细胞增殖,为后期Hsp90α分子伴侣功能进行研究奠定基础。