Mechanisms of regulating the Raf kinase family

The MAP Kinase pathway is a key signalling mechanism that regulates many cellular functions such as cell growth, transformation and apoptosis. One of the essential components of this pathway is the serine/threonine kinase, Raf. Raf (MAPKK kinase, MAPKKK) relays the extracellular signal from the receptor/Ras complex to a cascade of cytosolic kinases by phosphorylating and activating MAPK/ERK kinase (MEK; MAPK kinase, MAPKK) that phosphorylates and activates extracellular signal regulated kinase (ERK; mitogen-activated protein kinase, MAPK), which phosphorylates various cytoplasmic and nuclear proteins. Regulation of both Ras and Raf is crucial in the proper maintenance of cell growth as oncogenic mutations in these genes lead to high transforming activity. Ras is mutated in 30% of all human cancers and B-Raf is mutated in 60% of malignant melanomas. The mechanisms that regulate the small GTPase Ras as well as the downstream kinases MEK and extracellular signal regulated kinase (ERK) are well understood. However, the regulation of Raf is complex and involves the integration of other signalling pathways as well as intramolecular interactions, phosphorylation, dephosphorylation and protein-protein interactions. From studies using mammalian isoforms of Raf, as well as C. elegans lin45-Raf, common patterns and unique differences of regulation have emerged. This review will summarize recent findings on the regulation of Raf kinase.     

金鱼精子质膜和核膜的区域特异性

本文结合采用扫描和透射电子显微镜(包括冷冻断裂-蚀刻、超薄切片以及细胞化学染色法),研究了金鱼精子的超微结构特征,结果表明金鱼精子的质膜和核膜都具有区域特异性:1)精子质膜内大部分区域含有许多蛋白颗粒,但在特定区域内,蛋白颗粒呈有序排列,构成晶格状结构。2)精子头颈部和尾部均有液泡密集之处,凡是覆盖着液泡区的质膜内,几乎都不含有蛋白颗粒。3)液泡区能被细胞化学方法染成致密色,表明内含糖蛋白。4)核膜孔只集中存在于靠近颈部的核膜上,而其他部分则没有。本文对上述诸点进行了讨论。     

Expression of branching enzyme I of maize endosperm in Escherichia coli.

The gene encoding for mature branching enzyme (BE) I (BEI) of maize (Zea mays L.) endosperm has been expressed in Escherichia coli using the T7 promoter. The expressed BEI was purified to near homogeneity so that amylolytic activity and bacterial BE could be completely eliminated from the BE preparation. The recombinant enzyme showed properties very similar to those of BEI purified from developing maize endosperm with respect to branching amylose and amylopectin. This result confirmed our earlier report that maize endosperm BEI had a higher rate of branching amylose and a much lower rate (less than 10% of that of branching amylose) of branching amylopectin. This study also showed a great advantage in purifying BEI from the bacterial expression system rather than from developing maize endosperm. Most important, this study has established the system with which to study the structure-function relationships of the maize BEI using site-directed mutagenesis.     

白豆杉的核型和性染色体的研究

白豆杉pseudotaxus chienii(Cheng)Cheng是我国裸子植物特有属之一,雌雄异常,根尖 细胞染色体分析表明：雌株有一对异形性染色体,异配性别,属ZW型;雄株是同配性别,属ZZ型,雌株的型为2n=2x=24=22m(2SAT ZW) 2T,雄株的核型为2n=2x=24=22m(2SAT ZZ) 2T。Giemsa C-带,显示,Z染色体长短臂均具端带,W染色体不显带。     

卵巢激素对肺泡巨噬细胞趋化活性的影响

为探讨卵巢激素对非性器官肺脏的防御功能有无影响,本研究以肺泡巨噬细胞（ＡＭ）趋化活性为指标,观察了卵巢激素对成年雄性大鼠离体ＡＭ的趋化活性的作用。结果显示：不同浓度的酵母多糖激活血清与ＡＭ在体外培养３．５ｈ,对ＡＭ趋化性有良好的线性关系。雌二醇能抑制ＡＭ的趋化活性,量效关系显著（ｒ＝－０．９２８０,Ｐ＜０．０１）;而孕酮则促进ＡＭ的趋化活性,亦具有剂量依从性（ｒ＝０．９９７５,Ｐ＜０．０１）。提示：卵巢激素除参与性器官功能的调节外,对于非性器官肺脏的防御功能亦具有一定的调控作用。     

白烨种群空间分布格局的研究

根据在白桦(Betula platyphylla)种群分布区内所选择的四个代表性地区的定位观测资料,本文运用数学分析方法,采用电子计算机进行运算,模拟,研究白桦种群的空间分布格局。结果表明:随着生境温度的逐渐降低或温度的逐渐升高,白桦种群由聚集分布过渡为随机分布,随着种群年龄的增加,种群空间分布格局表现出相同的变化规律。     

Inhibition of Nitrate Transporter 1.1-Controlled Nitrate Uptake Reduces Cadmium Uptake in Arabidopsis

Identification of mechanisms that decrease cadmium accumulation in plants is a prerequisite for minimizing dietary uptake of cadmium from contaminated crops. Here, we show that cadmium inhibits nitrate transporter 1.1 (NRT1.1)-mediated nitrate (NO₃⁻) uptake in Arabidopsis (Arabidopsis thaliana) and impairs NO₃⁻ homeostasis in roots. In NO₃⁻-containing medium, loss of NRT1.1 function in nrt1.1 mutants leads to decreased levels of cadmium and several other metals in both roots and shoots and results in better biomass production in the presence of cadmium, whereas in NO₃⁻-free medium, no difference is seen between nrt1.1 mutants and wild-type plants. These results suggest that inhibition of NRT1.1 activity reduces cadmium uptake, thus enhancing cadmium tolerance in an NO₃⁻ uptake-dependent manner. Furthermore, using a treatment rotation system allowing synchronous uptake of NO₃⁻ and nutrient cations and asynchronous uptake of cadmium, the nrt1.1 mutants had similar cadmium levels to wild-type plants but lower levels of nutrient metals, whereas the opposite effect was seen using treatment rotation allowing synchronous uptake of NO₃⁻ and cadmium and asynchronous uptake of nutrient cations. We conclude that, although inhibition of NRT1.1-mediated NO₃⁻ uptake by cadmium might have negative effects on nitrogen nutrition in plants, it has a positive effect on cadmium detoxification by reducing cadmium entry into roots. NRT1.1 may regulate the uptake of cadmium and other cations by a common mechanism.Cadmium is highly toxic to humans (Nicholson et al., 1983), and its primary route of entry into the body is through crops grown in cadmium-contaminated soil (Clemens et al., 2013). A recent survey indicated that vegetables and rice (Oryza sativa) account for approximately 40% and 38%, respectively, of total cadmium exposure in residents of Shanghai, China’s largest city (He et al., 2013). However, cadmium contamination of agricultural soils as a result of rapid industrial development and release of agrochemicals into the environment is an increasingly serious problem. Many strategies have been proposed for remediating cadmium-contaminated soil to prevent cadmium uptake by crops. These strategies include the dig-and-dump method or encapsulation of the contaminated soil, chemical immobilization or extraction of cadmium, and phytoremediation by cadmium-hyperaccumulating plants (Pulford and Watson, 2003). However, the dig-and-dump and chemical methods are expensive, whereas phytoremediation requires several growing seasons to be effective, making it impractical in regions where farmland is limited and food supply insufficient.The shortfalls of these strategies have prompted researchers to develop alternative techniques that are cost-effective and interfere less with crop production. Use of nitrogen fertilizers is one of the most important agronomic practices and it has been suggested that their appropriate use might provide a relatively inexpensive, time-saving, and effective strategy for reducing cadmium entry into, and accumulation in, crops because NO₃⁻ facilitates cadmium uptake in hydroponically grown plants (Sarwar et al., 2010; Luo et al., 2012). However, in a preliminary study, we found that, in plants grown in soil, the effect of the nitrogen form on cadmium accumulation was strongly associated with the pH-buffering capacity of the soil. In soil with a lower pH-buffering capacity, application of ammonium (NH₄⁺) resulted in higher cadmium levels in plants than application of NO₃⁻, probably as a result of soil acidification by NH₄⁺, and the opposite effect was seen when plants were grown in soil with higher pH-buffering capacity (S.K. Fan, S.T. Du, and C.W. Jin, unpublished data). This suggests that management of the use of nitrogen fertilizers to prevent cadmium entry into crops might be difficult because of the wide variation in soil pH-buffering capacity.Because NO₃⁻ facilitates cadmium uptake in hydroponically grown plants as described above, modification of NO₃⁻ uptake pathways in plants might also affect cadmium uptake, in which case modifying these pathways to reduce cadmium entry into crops could circumvent the risks and the difficulties involved in nitrogen fertilizer management. Exposure to cadmium has been shown to reduce NO₃⁻ uptake by roots (Hernández et al., 1997; Gouia et al., 2000; Rizzardo et al., 2012), but this has been assumed to be deleterious to plant growth (Finkemeier et al., 2003; Rizzardo et al., 2012). The process by which NO₃⁻ is taken up across the root plasma membrane is complex, and several nitrate transporters (NRTs) involved in NO₃⁻ uptake from the growth medium have been characterized. In Arabidopsis (Arabidopsis thaliana), NRT1.1 is a dual-affinity transporter involved in both high- and low-affinity uptake, NRT1.2 is involved only in low-affinity NO₃⁻ uptake, whereas NRT2.1, NRT2.2, and NRT2.4 are only involved in high-affinity NO₃⁻ uptake (Wang et al., 2012; Léran et al., 2014). However, the transporter responsible for the cadmium-induced decrease in NO₃⁻ uptake remains unknown. Given the presumed association between NO₃⁻ uptake and cadmium uptake, it is important to identify the molecular mechanism involved in this process, and it is particularly important to determine whether the modulation of relevant NO₃⁻ transporters affects cadmium entry into plants.In this study, we investigated the relationship between NO₃⁻ uptake and cadmium uptake in Arabidopsis roots. To our knowledge, our results reveal a new mechanism for resisting cadmium toxicity: Cadmium reduces NO₃⁻ uptake by inhibiting NRT1.1 activity, which in turn reduces cadmium entry into root cells. As a result, cadmium levels in plants are lower and plant growth is improved. Our findings may provide a strategy for minimizing cadmium accumulation in crops grown in contaminated soil using biotechnological pathways to decrease NO₃⁻ uptake.     

西天目山金钱松树轮中五种金属元素的含量分析

本文报道西天目山金钱松(Pseudolarix kaempferi Cord.)木质部中铝、铁、钙、锰、锌5种元素含量和累积量自1711年以来的变化情况。1711～1900年期间,钙的含量(μg/g)明显增大,铁和锰稍有上升,而铝和锌则略有下降;最近25年来,铝、铁、锰元素的含量升高,而锌含量下降。1711～1900年期间,5种元素的累积量(μg/Yr)都轻微下降,但最近20～25年来,铁、铝、锰的累积量又明显上升,锌元素变化不大。笔者认为铝、铁、钙、锰、锌元素在1711～1900年出现下降,可能是由于植被遭破坏的缘故,而在近25年中,铝、铁、锰元素的急速上升,则可能是工业污染的结果。     

糖皮质激素对肺泡巨噬细胞趋化活性的影响

采用测定肺泡巨噬细胞(AM)趋化性和AM产生中性粒细胞趋化因子的实验技术。观察到糖皮质激素体外直接作用于大鼠AM悬液或大鼠皮下注射7d都可抑制AM的趋化性,并且肺灌洗液中的AM数量明显减少;但注射3d组无上述效应。以糖皮质激素处理AM 16h可使AM产生的中性粒细胞趋化因子强度明显降低。结果表明糖皮质激素可抑制AM的趋化性和抑制AM产生中性粒细胞趋化因子。提示这可能是较长期应用糖皮质激素抑制免疫和炎症反应、降低肺部防御能力的部分机理。