Expression of TWEAK and its receptor Fn14 in human subcutaneous adipose tissue. Relationship with other inflammatory cytokines in obesity

TWEAK, a cytokine of the TNF family, has been found to be expressed under different inflammatory conditions but no data is available concerning the expression of this cytokine and its receptor (Fn14) in human obesity. In the present work we have evaluated the expression of many pro-inflammatory TNF system cytokines (TNF-alpha, TWEAK and their respective receptors, TNFR1, TNFR2 and Fn14) in human adipose tissue of 84 subjects some with different degree of obesity and type 2 diabetes, and its relation with inflammation by also measuring the expression of macrophage marker CD68. We detected expression of TWEAK and Fn14 in isolated mature adipocytes and in the stromovascular fraction. Additionally, we found that LPS upregulates the expression of both genes on THP-1 human monocytic cell line. TWEAK was expressed in adipose tissue of all studied subjects with no differences between obesity group, and was associated with Fn14 expression in morbid obese, mainly in women with type 2 diabetes. The data obtained here also showed that TNF-alpha and TNFR2 mRNAs were significantly more expressed in subcutaneous adipose tissue of subjects with morbid obesity compared to obese and non-obese subjects. In contrast, TNFR1 gene expression was negatively associated with BMI. Our results suggest that the expression of TNF-derived pro-inflammatory cytokines are increased in severe obesity, where macrophage infiltrate could modulate the inflammatory environment through activation of its receptors.     

烟粉虱对寄主植物叶背的定向行为

烟粉虱Bemisia tabaci(Gennadius)成虫在自然条件下总是能够准确定位到寄主的叶片背面进行取食和产卵,这是一种本能的定向行为。以扶桑和油麦菜为寄主材料,通过非选择性和选择性实验,研究和评价光照、叶面物理性状和重力3个环境因子在烟粉虱成虫定向行为中的作用。结果显示,重力在定向行为中起主导作用,寄主叶片正反面的物理性状差异也能够使烟粉虱成虫定位到寄主叶片的背面,但是存在时滞效应。说明烟粉虱成虫在定向到寄主叶片背面的过程中存在2个系统,一个是以重力为线索的垂直方向感作为导航系统,另一个则是以叶面物理性状为线索的行为调节作为校正系统,二者的相互协作使得烟粉虱成虫能够精确定位到寄主叶片的背面。     

检测SDS-聚丙烯酰胺凝胶电泳中家蝇蛋白的新方法——海波银染法

介绍一种检测SDS聚丙烯酰胺凝胶电泳中家蝇幼虫蛋白的新方法-海波银染法。该方法对传统银染方法中的试剂与步骤加以改进,省略了乙醇固定与洗涤步骤,只需20 min即可完成全部染色过程,且仅在国产分析纯试剂及普通操作条件下,灵敏度可达毫微克级水平。     

基于多源数据的黄土高原陆地水循环结构变化分析

近几十年间,黄土高原的水循环进程在人类活动与气候变化的影响下已产生了剧烈的变化。为加深对水循环结构变化的了解与认识,利用1982-2010年的降水、蒸散发、径流、土壤储水量和社会经济用水等数据,运用Mann-Kendall趋势检验和线性回归分析方法,对黄土高原的水量平衡进行评估,且细化了组成水循环的12种水文变量,并分析了水循环各分量的变化趋势及其结构的演化规律。由于网格化的社会经济用水数据（1982-2010年）对本研究产生了较大的时间限制,因此本文注重于探究这29年间黄土高原各水文变量的变化趋势及水循环结构的演化规律。结果表明：在自然系统中,蒸散发以1.97 mm/a的速率上升（P<0.01）,径流、降水和土壤储水量分别以1.01 mm/a（P<0.01）、0.77 mm/a和0.46 mm/a的速率下降。在社会系统中,社会经济用水以0.50 mm/a的速率上升,其中主要由于生活、制造业、发电和采矿用水分别以0.22、0.23、0.30 mm/a和0.01 mm/a的速率增加所导致。此外,灌溉和牲畜用水分别以0.25 mm/a（P<0.05）、0.01 mm/a（P<0.01）的速率减少。就水循环结构而言,多年平均蒸散发和社会经济用水占水循环的平均比例分别为80.95%、15.27%,并以每年0.16%、0.06%的速率逐渐升高。径流、土壤储水量的变化占水循环的平均比例分别为4.00%、-0.24%,并以每年0.24%（P<0.01）、0.02%的速率逐渐下降。随着社会经济的发展和人口的增加,区域水资源供需矛盾将进一步加剧,本研究对黄土高原水资源的科学调控与可持续利用有重要参考意义。     

黄河三角洲植被空间分布特征及其环境解释

为了解黄河三角洲地区植被空间分布与环境因子之间的关系,通过局地植被样方调查、区域遥感影像提取归一化植被指数(NDVI)及地形高度、地下水位埋深、表层土壤Cl~-含量等环境数据采集,综合样地植被与环境数据进行了除趋势对应分析(DCA)和除趋势典范对应分析(DCCA),并对区域NDVI与主要环境变量进行了单因子相关性分析和多元逐步回归分析。结果显示:DCA排序可将黄河三角洲植被分为翅碱蓬、柽柳-翅碱蓬、芦苇-柽柳、芦苇4个主要群落类型(群丛),DCCA与DCA排序图总体相似,但DCCA更清晰地表明其第一轴主要代表的是潜水Cl~-浓度等关键水盐因子,且随着水土环境系统盐分含量的减小,群落由翅碱蓬逐渐向芦苇演变。区域典型植被群落和NDVI分布格局与变化趋势受地下水位埋深和潜水Cl~-浓度2个环境因素影响较大(NDVI与2个环境变量间建立的二元回归方程R~2=0.57),而土壤Cl~-含量的植被效应实际上受地下水位埋深和潜水Cl~-浓度的影响。在区域地下水普遍浅埋条件下,地下水成为影响植被生长与分布的生态环境最敏感要素,而地下水位埋深和潜水Cl~-浓度是这一要素中的2个关键因子,尤其是后者梯度变化对天然植被分布格局起重要的控制作用。     

AUXIN RESPONSE FACTOR 2 Intersects Hormonal Signals in the Regulation of Tomato Fruit Ripening

The involvement of ethylene in fruit ripening is well documented, though knowledge regarding the crosstalk between ethylene and other hormones in ripening is lacking. We discovered that AUXIN RESPONSE FACTOR 2A (ARF2A), a recognized auxin signaling component, functions in the control of ripening. ARF2A expression is ripening regulated and reduced in the rin, nor and nr ripening mutants. It is also responsive to exogenous application of ethylene, auxin and abscisic acid (ABA). Over-expressing ARF2A in tomato resulted in blotchy ripening in which certain fruit regions turn red and possess accelerated ripening. ARF2A over-expressing fruit displayed early ethylene emission and ethylene signaling inhibition delayed their ripening phenotype, suggesting ethylene dependency. Both green and red fruit regions showed the induction of ethylene signaling components and master regulators of ripening. Comprehensive hormone profiling revealed that altered ARF2A expression in fruit significantly modified abscisates, cytokinins and salicylic acid while gibberellic acid and auxin metabolites were unaffected. Silencing of ARF2A further validated these observations as reducing ARF2A expression let to retarded fruit ripening, parthenocarpy and a disturbed hormonal profile. Finally, we show that ARF2A both homodimerizes and interacts with the ABA STRESS RIPENING (ASR1) protein, suggesting that ASR1 might be linking ABA and ethylene-dependent ripening. These results revealed that ARF2A interconnects signals of ethylene and additional hormones to co-ordinate the capacity of fruit tissue to initiate the complex ripening process.     

黄土丘陵区不同土地利用下土壤释放N2O潜力的影响因素

采用室内培养试验,研究了不同水热条件对黄土丘陵区林地、草地和果园土壤释放N2O的影响,同时测定了土壤中不同氮素形态的变化,旨在探讨影响土壤释放N2O潜力的因素。结果表明：土样中N2O通量与温度显著相关（r=0.1599, P<0.05）,均随温度的升高不断增大,35℃时达到最大。N2O通量与土壤水分含量极显著相关（r=0.2499,P<0.0001）,在土壤水分含量较低时,各土样中N2O通量与土壤水分含量呈正相关,土壤水分接近田间持水量时N2O通量最大,超过田间持水量时N2O通量急剧下降。土壤水分和温度对N2O通量的影响可用拟合方程F=a bT cT^2 dT^3 eT^4 fW来描述。在培养条件下,土壤中N2O的释放总量大小依次为：果园土>林地土>草地土,果园土释放N2O的总量分别比林地土和草地土的释放总量多30%,14%。土壤氮素形态与N2O的释放量有一定关系,但规律不明显。     

单克隆人胰腺干细胞分化特性的研究

研究1例来源于4月龄男性流产胎儿胰腺组织的单克隆人胰腺干细胞（monoclonal human pancreatic stem cell,mhPSC）系的体内外分化特性。将mhPSCs接种在铺有0．1％明胶的培养皿内,扩增培养3d后,加高糖DMEM诱导液诱导培养25d。相差显微镜下．观察细胞生长状况。采用双硫腙染色法、RT—PCR及葡萄糖刺激释放胰岛素和C肽实验．对体外定向诱导mhPSCs分化为功能性胰岛进行检测。将mhPSCs悬液注射在成年雄性裸鼠腹股沟皮下．注射30d时,取出移植物,采用SP法进行免疫组织化学反应,以检测mhPSCs的体内自然分化潜能。体外扩增培养,mhPSCs贴壁生长,呈多角形上皮样。生长至单层．呈“铺路石”状。体外定向诱导,细胞逐渐由多角形变成圆形,并聚集成类胰岛。诱导培养15d时．形成的类胰岛中少数细胞分化为B细胞,双硫腙染色阳性。诱导培养25d时,多数细胞分化为8细胞,双硫腙染色阳性,转录表达胰岛素的mRNA。用不同浓度葡萄糖刺激．诱导胰岛不仅释放胰岛素和C肽,而且其释放量随糖刺激浓度升高显著增加（0．01〈P〈0．05）。体内分化实验显示,mhPSCs在裸鼠背部形成类畸胎瘤。类畸胎瘤易与裸鼠分离,色白,血管丰富。显著表达pdx1、胰岛素、胰高血糖素、CK、MBP及NF蛋白。该研究结果证实单克隆人胰腺干细胞系体外定向诱导分化为包含大量β细胞的功能性类胰岛,在体内自然分化为胰岛、上皮及神经组织细胞。     

具有抗HIV活性的天花粉蛋白在大肠杆菌中的表达及纯化

目的:天花粉蛋白(TCS)有较强的抗HIV活性。利用基因工程技术在大肠杆菌中表达TCS并进行纯化。方法:从新鲜栝楼叶片中获取TCS基因组DNA,利用PCR技术扩增其全长基因,经BamHⅠ和EcoRⅠ双酶切后与原核表达载体pRSET-A连接,转化感受态E.coliDH5α,提取质粒进行酶切鉴定及测序;将所获阳性重组质粒转化感受态E.coliBL21(DE3)得到工程菌,经IPTG诱导表达后,对表达产物进行SDS-PAGE及Western印迹鉴定;用Ni-NTA柱对所获目的蛋白进行纯化。结果:获得了目的蛋白的可溶性高效表达,并通过了Western印迹鉴定。经Ni-NTA柱纯化后,得到大量均一的6His-TCS融合蛋白。结论:TCS在大肠杆菌中的表达与纯化,为通过基因工程方法研制具有抗HIV活性的药物奠定了基础。