F9克隆胚胎癌细胞株在诱导分化过程中H-2抗原及花生凝集素受体的变化

近年来有不少研究工作表明,曾被认为是无能的胚胎癌细胞(EC)株的F9细胞,在维生素A酸(简称RA)的影响下能分化成内胚层细胞或神经样细胞等。分化细胞的纤维蛋白溶酶原激活因子和胶原样蛋白都有显著增加,而且细胞表面的抗原、糖蛋白及植物凝集素受体等随之也发生明显变化。某些抗原(如F9抗原)随着分化会逐渐消失,而另一些抗原(如H-2抗原)又会出现。在细胞分化中植物凝集素受体的变化较为显著,其中最令人感兴趣的是花生凝集素受体(简称PNA     

Ordovician conodont biogeography – reconsidered

Review of the traditional separation of global Ordovician conodont distribution into the North American Midcontinent Province (NAMP) and North Atlantic Province (NAP) reveals a confusing variety of concepts and definitions that hinder biogeographic analysis. Use of this twofold scheme and its subsequent variants should be­discontinued in favour of the more detailed divisions proposed here. Major biogeographical entities of the Shallow-Sea and Open-Sea Realms, separated by the shelf-slope break, are both further subdivisible into Tropical, Temperate and Cold Domains. In the Cold domains, faunal differences between the two Realms and their subdivisions are not easily discernible, since biofacies zones and different habitats were highly condensed. Faunal differences are amplified in the tropical regions, where the North American Midcontinent Province and North Atlantic Province were originally defined. Recognition of endemic taxa is essential for finer classification within domains of the Shallow-Sea Realm (SSR). Our preliminary analysis of Early Ordovician conodont distribution identifies the Laurentian Province (in the Tropical Domain), Australian­(Tropical Domain), North China (Tropical Domain), South China (Temperate Domain), Argentine Precordillera (Temperate Domain) and Balto-Scandian Province (in the Cold Domain). The Open-Sea Realm (OSR) is dominated by cosmopolitan and widespread taxa, and formal subdivision at provincial level is yet to be achieved. The North Atlantic Province encompasses both the Open-Sea Realm and the Temperate and Cold Domains of the Shallow-Sea Realm. The North American Midcontinent Province sensu stricto is more or less equivalent to the Laurentian Province, representing shallow-water regions fringing Laurentia; in a broader sense the North American Midcontinent Province includes all provinces of the Tropical Domain within the Shallow-Sea Realm.     

A Comparison of Nitrate-selective Microelectrodes made with Different Nitrate Sensors and the Measurement of Intracellular Nitrate Activities in Cells of Excised Barley Roots

Nitrate-selective microelectrodes based on a number of nitratesensors were compared. The electrode properties tested includedlog-linear slope of the calibration curves, detection limit,and ion selectivity. The nitrate sensor mixture described inan earlier paper performed favourably when compared with othernitrate-selective mixtures or with commercially-available macroelectrodes.This earlier mixture consisted of 6% methyltridodecylammoniumnitrate, 65% n-phenyloctyl ether, 23% poly(vinylchloride), 5%nitrocellulose, and 1% methyltriphenyl phosphonium bromide.These electrodes, even when stored backfilled remained nitrate-selectivefor several days although there was eventually some deteriorationin performance. The electrodes were used in vitro for assaying nitrate in barleyroot extracts and a linear relationship was found between resultsfrom ion chromatography and microelectrode measurements. Intracellularmeasurements made in vivo in epidermal cells of excised barleyroots identified two populations of measurements believed tobe the cytoplasm and the vacuole. Significant decreases in compartmentalnitrate activities were measured 2 to 6 h after excision. Theseresults indicate that the nitrate pool in both the cytoplasmand vacuole of root epidermal cells is sensitive to root excisionand question the physiological significance of measurementsmade on excised roots. Key words: Nitrate-selective microelectrodes, barley root, compartmentation, nitrate     

抗精氨酸加压素血清的制备及其初步应用

用戊二醛为偶联剂将精氨酸加压素(AVP)与甲状腺球蛋白连接合成AVP免疫原,免疫家兔获得了高滴度和良好特异性与亲和力的抗精氨酸加压素血清。用此抗血清进行了大鼠和豚鼠下丘脑内AVP样免疫反应物质的定位研究,测定了正常人血浆及大鼠部分脑区AVP免疫活性物质含量,结果与国外报道相近,初步应用结果表明此抗血清适用于放射免疫测定(RIA)和免疫细胞化学(ICC)研究。     

Intracellular expression of a single-chain antibody directed against type IV collagenase inhibits the growth of lung cancer xenografts in nude mice

It was documented that type IV collagenase with two subtypes of 72 ku/MMP-2 and 92 ku/MMP-9 plays an important role in tumor invasion and metastasis. The endoplasmic reticulum (ER)- retained, single chain Fv antibody fragment (scFv) was used to inhibit the function of type IV collagenase. For expression in mammalian cells, the assembled scFv M97 gene with ER retention signal encoding 6 additional amino acids (SEKDEL) was reamplified by PCR. The amplified fragments were cloned into the pcDNA3.1 vector. The resulting plasmid was sequenced and then introduced into PG cells, a highly metastatic human lung cancer cell line, by lipofectAMINE method. The result of intrabody gene therapy showed that type IV collegenase expression was down regulated significantly as measured by ELISA. The biological behavior of PG cell, such as the ability of in vitro invasion through Matrigel, colony formation on soft agar, was also inhibited by scFv M97 transfection. Animal experiments in a xenograft model of human lung cancer showed that scFv M97 transfection significantly prolonged the survival time of nude mice. The results indicate that intracellular antibody technology represents a novel and efficient way to abrogate selectively the activity of type IV collagenase.     

过量表达棉花CBF2基因提高转基因拟南芥抗旱耐盐能力

CBF/DREB是一类植物中特有的转录因子,在植物抵抗逆境胁迫过程中发挥重要功能。本研究从陆地棉(Gossypium hirsutum L.)Coker 312中克隆获得1个棉花CBF/DREB基因,命名为Gh CBF2,该基因编码一个由216个氨基酸组成的CBF蛋白。序列分析结果显示,Gh CBF2与其他植物的CBF蛋白类似,含有AP2转录因子典型的保守结构域。干旱或高盐胁迫处理明显增加了Gh CBF2基因的表达量。亚细胞定位分析结果发现Gh CBF2定位在细胞核中。将Gh CBF2基因构建到由35S启动子调控的植物表达载体p MD上并转化拟南芥(Arabidopsis thaliana L.),结果表明,在干旱和盐胁迫条件下,过量表达Gh CBF2基因拟南芥的成活率显著高于野生型,并且游离脯氨酸和可溶性糖含量也高于野生型,说明转Gh CBF2基因提高了拟南芥的耐盐抗旱能力。采用实时荧光定量PCR方法分析胁迫相关标记基因COR15A、RD29A和ERD6的表达情况,结果显示转基因株系中的表达量显著高于野生型,说明Gh CBF2参与调控拟南芥干旱和盐胁迫相关基因的表达。     

α-亚麻酸对干旱胁迫下水稻种子萌发的影响

亚麻酸属ω-3系统的多不饱和脂肪酸,是细胞膜的重要组成部分。在干旱胁迫下,植物通过释放α-亚麻酸重塑细胞膜的流动性,但外源添加α-亚麻酸能否提高植物对干旱的抗性尚未可知。该研究采用PEG6000模拟干旱胁迫,探讨了种子萌发过程中α-亚麻酸对水稻萌发和幼苗生长过程中抗旱性的影响。结果表明:在14%和16%PEG干旱胁迫下,水稻种子推迟萌发,幼苗的生长受到抑制。25μmol·L~(-1)和250μmol·L~(-1)的α-亚麻酸可缓解干旱对水稻幼根和幼苗生长的抑制,且随着干旱程度的增加,缓解作用增强。16%PEG胁迫下,25μmol·L~(-1)、250μmol·L~(-1)的α-亚麻酸分别提高幼苗的根长34.3%和29.1%、苗长67.8%和52.0%、根重43.9%和35.2%、苗重59.1%和43.6%。α-淀粉酶活力测定发现,25μmol·L~(-1)和250μmol·L~(-1)的α-亚麻酸分别提高干旱胁迫下水稻种子α-淀粉酶活力为56.7%~70.7%和36.8%~43.8%。水稻幼根活力测定显示,25μmol·L~(-1)和250μmol·L~(-1)的α-亚麻酸分别提高干旱胁迫下幼根活力为11.4%~28.4%和5.4%~22.2%。此外,还对不同干旱胁迫下水稻种子α-淀粉酶活性和幼根活力的变化进行了讨论,认为α-亚麻酸主要通过提高种子萌发过程中α-淀粉酶的活性来缓解所遭遇的干旱胁迫,而对水稻幼根活力的影响则相对有限。     

冠状动脉狭窄对血流量的影响

在22条开胸犬上观察了冠脉狭窄对血流量(CBF)的影响。用一可调节的微米缩窄器定量调节左旋支缩窄程度,测量了主动脉平均压(Pa)、冠脉远端小动脉平均压(Pc)和狭窄端压力降(ΔP)。冠脉狭窄程度与血流量变化曲线显示:在冠脉狭窄程度小于85％时,CBF相对稳定;随着狭窄程度的进一步增加,CBF急剧下降;而在狭窄程度大于95％后,CBF又缓慢下降。冠状动脉狭窄程度与CBF下降的曲线可用下列方程式表达: CBF=1.48×10~(10)e~(-27.6A)(A=冠脉狭窄程度) 冠脉狭窄程度大于50％时,狭窄程度与Pc呈负相关:Pc=159.1—1.36A(r=-0.73,P<0.01)。Pc与CBF呈正相关;Pc=16.9 1.3CBF(r=0.74,P<0.01)     

西藏八角莲与桃儿七根及根茎SSH文库的建立

以西藏八角莲(Dysosma tsayuensis Ying)和桃儿七[Sinopodophyllum hexandrum(Royle)Ying]为材料,构建其根及根茎的SSH文库,从中筛选鬼臼类植物属种间与鬼臼毒素生物合成相关的差异表达基因。从文库中随机挑取201个阳性克隆测序后得到183条ESTs。去除载体序列和冗余序列,聚类拼接得到17个西藏八角莲的unique ESTs。经BLAST同源比较和功能查寻,有功能注释的unique ESTs共12个,占70.6%,所编码的蛋白涉及光合作用、合成代谢、转录调控等功能;无功能注释和匹配结果的共5个,占29.4%。该研究成功构建了西藏八角莲和桃儿七SSH文库,为进一步揭示鬼臼毒素生物合成途径及其调控机制奠定了基础。     

基于局域模型的凉水国家自然保护区土壤全氮空间分布

以凉水国家自然保护区激光雷达数据为基础,建立数字高程模型,提取基本地形属性如坡度、坡向和复合地形属性湿度指数和相对径流强度指数等,在成土因素学说基础上,对全氮含量(TN)进行地理加权回归建模(GWR),同时运用反距离加权(IDW)、普通克里格(OK)和泛克里格(UK)对TN进行空间插值.结果表明： 对于研究区TN的预测,GWR模型预测精度(77.4%)高于其他3种空间插值方法,IDW预测精度(69.4%)高于OK(63.5%)和UK(60.6%)的预测精度.利用GWR模型预测研究区域土壤TN平均达到4.82 g·kg^-1;在高海拔、地形湿度大以及相对径流强的区域,土壤TN相对较高.对预测结果进行探讨发现,不同坡位、坡向的土壤TN也存在一定差异.因此,基于地形属性的局域模型是土壤属性空间分布预测的更为有效的方法.