首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   4502篇
  免费   563篇
  国内免费   687篇
  2024年   9篇
  2023年   48篇
  2022年   116篇
  2021年   227篇
  2020年   211篇
  2019年   234篇
  2018年   217篇
  2017年   203篇
  2016年   213篇
  2015年   290篇
  2014年   363篇
  2013年   336篇
  2012年   417篇
  2011年   349篇
  2010年   221篇
  2009年   200篇
  2008年   246篇
  2007年   226篇
  2006年   170篇
  2005年   144篇
  2004年   203篇
  2003年   211篇
  2002年   255篇
  2001年   194篇
  2000年   118篇
  1999年   90篇
  1998年   57篇
  1997年   29篇
  1996年   30篇
  1995年   19篇
  1994年   25篇
  1993年   15篇
  1992年   11篇
  1991年   14篇
  1990年   12篇
  1989年   11篇
  1988年   1篇
  1987年   1篇
  1985年   5篇
  1984年   2篇
  1983年   2篇
  1982年   3篇
  1981年   3篇
  1980年   1篇
排序方式: 共有5752条查询结果,搜索用时 193 毫秒
231.
The Hengduan Mountains comprise one of the world's most important hot spots of biodiversity. Tibetia (Ali) H.P. Tsui (Fabaceae), which has four or five species in two sections, is one of the genera endemic to the region. This paper describes for the first time the karyotype of three of those species. The chromosome counts of all three are 2 n = 16. The karyotypes of the species examined contain chromosomes of variable karyotypic symmetry with centromeres at median and submedian positions that correlate with the morphological characteristics of the species. Karyotypic variation at the diploid level appears to be the predominant feature of chromosome evolution in the genus and may provide a clue to the study of evolutionary patterns of plants in this region.  相似文献   
232.
Recently, glycine has been shown to prevent liver injury after endotoxin treatment in vivo. We demonstrated that ethanol and endotoxin stimulated Kupffer cells to release PGE(2), which elevated oxygen consumption in parenchymal cells. Because glycine has been reported to protect renal tubular cells, isolated hepatocytes, and perfused livers against hypoxic injury, the purpose of this study was to determine whether glycine prevents increases in intracellular free Ca(2+) concentration ([Ca(2+)](i)) in hepatic parenchymal cells by agonists released during stress, such as with PGE(2) and adrenergic hormones. Liver parenchymal cells isolated from female Sprague-Dawley rats were cultured for 4 h in DMEM/F12 medium, and [Ca(2+)](i) in individual cells was assessed fluorometrically using the fluorescent calcium indicator fura 2. PGE(2) caused a dose-dependent increase in [Ca(2+)](i) from basal values of 130 +/- 10 to maximal levels of 434 +/- 55 nM. EGTA partially prevented this increase, indicating that either extracellular calcium or agonist binding is Ca(2+) dependent. 8-(Diethylamino)octyl 3,4,5-trimethoxybenzoate (TMB-8), an agent that prevents the release of Ca(2+) from intracellular stores, also partially blocked the increase in [Ca(2+)](i) caused by PGE(2), suggesting that intracellular Ca(2+) pools are involved. Together, these results are consistent with the hypothesis that both the intracellular and extracellular Ca(2+) pools are involved in the increase in [Ca(2+)](i) caused by PGE(2). Interestingly, glycine, which activates anion (i.e., chloride) channels, blocked the increase in [Ca(2+)](i) due to PGE(2) in a dose-dependent manner. Low-dose strychnine, an antagonist of glycine-gated chloride channel in the central nervous system, partially reversed the inhibition by glycine. When extracellular Cl(-) was omitted, glycine was much less effective in preventing the increase in [Ca(2+)](i) due to PGE(2). Phenylephrine, an alpha(1)-type adrenergic receptor agonist, also increased [Ca(2+)](i), as expected, from 159 +/- 20 to 432 +/- 43 nM. Glycine also blocked the increase in [Ca(2+)](i) due to phenylephrine, and the effect was also reversed by low-dose strychnine. Together, these data indicate that glycine rapidly blocks the increase in [Ca(2+)](i) in hepatic parenchymal cells due to agonists released during stress, most likely by actions on a glycine-sensitive anion channel and that this may be a major aspect of glycine-induced hepatoprotection.  相似文献   
233.
Basic fibroblast growth factor (bFGF) is a potent mitogen of many cell types and plays an important role in angiogenesis. To help identify proteins that bind to bFGF and mediate its intracellular transport and signaling, we overexpressed and purified a bFGF fusion protein in Escherichia coli. The fusion protein consists of bFGF fused to the C-terminus of glutathione S-transferase (GST). The GST-bFGF fusion protein was purified using SP-Sepharose and glutathione-Sepharose affinity chromatography. The ability of the purified GST-bFGF to stimulate the growth of human umbilical vein endothelial cells (HUVECs) was equivalent to that of purified recombinant 18 kDa bFGF.  相似文献   
234.
极度濒危植物五针白皮松的保护遗传学研究   总被引:10,自引:0,他引:10  
用随机扩增多态(RAPD)方法对极度濒危植物五针白皮松(Pinus squamata X.W.Li)的遗传多样性和居群遗传结构进行了研究。14个随机引物共获得93条RAPD谱带,其中6条为多态带,多态位点百分率仅为6.45%,遗传多样性极低。Shannon指数I和Nei指数h在种内也只有0.020和0.030,两个亚居群间(半阴坡亚居群与半阳坡亚居群)遗传分化程度不高,遗传分化系数Gst只有0.110,与大多数松科植物近似,居群每代迁移数为4.032。五针白皮松极低的遗传多样性可能是由于它在演化过程中遭受过严重的灾害,造成严重的瓶颈效应,丧失其大部分遗传变异。在随后的演化过程中由于遗传漂变、自交衰退等小种群现象,导致遗传多样性的进一步丧失。另外,阔叶树种对其生存的挤压和人类的干扰也是导致五针白皮松遗传多样性降低的因素之一。本文最后对该结果的保护生物学意义进行了评价。  相似文献   
235.
Bones of the postcranial skeleton of higher vertebrates originate from either somitic mesoderm or somatopleural layer of the lateral plate mesoderm. Controversy surrounds the origin of the scapula, a major component of the shoulder girdle, with both somitic and lateral plate origins being proposed. Abnormal scapular development has been described in the naturally occurring undulated series of mouse mutants, which has implicated Pax1 in the formation of this bone. Here we addressed the development of the scapula, firstly, by analysing the relationship between Pax1 expression and chondrogenesis and, secondly, by determining the developmental origin of the scapula using chick quail chimeric analysis. We show the following. (1) The scapula develops in a rostral-to-caudal direction and overt chondrification is preceded by an accumulation of Pax1-expressing cells. (2) The scapular head and neck are of lateral plate mesodermal origin. (3) In contrast, the scapular blade is composed of somitic cells. (4) Unlike the Pax1-positive cells of the vertebral column, which are of sclerotomal origin, the Pax1-positive cells of the scapular blade originate from the dermomyotome. (5) Finally, we show that cells of the scapular blade are organised into spatially restricted domains along its rostrocaudal axis in the same order as the somites from which they originated. Our results imply that the scapular blade is an ossifying muscular insertion rather than an original skeletal element, and that the scapular head and neck are homologous to the 'true coracoid' of higher vertebrates.  相似文献   
236.
This article presents an analytical method for the determination of the relative concentrations of trace elements in plasma protein by gel chromatography combined with SXRF (synchrotron radiation X-ray fluorescence). The fraction of plasma protein of male Kunming mice (body weight 24.2±0.3 g), treated with a cisplatin ip injection at a dose of 10 mg/kg, was obtained by the separation of a Sephadex G-50 column (buffered with ammonium acetate, pH 5.7). The SXRF experiments were performed at the Beijing Electron Positron Collider synchrotron radiation facility. The elements (Pt, S, Ca, Fe, Ni, Cu, Zn, Se, Br, and Sr) in the fraction of the plasma proteins (>22 kDa) were assayed using highly sensitive SXRF. The relative concentrations of elements were calculated by a normalization of Compton scattering intensity around 22 keV, after the normalization for the collection time of the X-ray spectrum and the counting of the ion chamber, and subtracting the contribution of the polycarbonate film for the supporting sample. The determination could prove that the element Pt in plasma was bound with macromolecular protein. Cu and S were present in the fraction of the protein in mice treated with cisplatin increase, and their ratios of treated/control were 1.66±0.06 and 1.78±0.33, respectively; Zn decreased to a ratio of 0.78±0.09. Our results are in agreement with others that cisplatin exposure leads to a marked loss of kidney copper and a moderate rise in kidney zinc. However, this article primarily describes one of the analytical methods used; it does not emphasize the results of the effect of cisplatin on trace elements in plasma protein.  相似文献   
237.
脑型一氧化氮合成酶的钙调蛋白结合区的表达及活性鉴定   总被引:1,自引:0,他引:1  
用PCR法克隆出nNOS的CaM结合区基因(nNOS 2455~2988bp),并在大肠杆菌中进行了高效表达。经金属离子螯合亲和层析得到纯度为90%以上的重组蛋白.分子量为22kDa,CaM Oveday assay证实该蛋白具有CaM的结合活性。由于所表达的重组蛋白既具有序列特异性又具有CaM的结合活性.因此。可将它作为筛选nNOS特异性抑制肽的靶蛋白,亦可用于特异性抗体的制备。  相似文献   
238.
观察不同盐浓度培养液中生长的杜氏盐藻可溶性蛋白SDS-PAG图谱,发现高盐与低盐相比,其51kD和63kD蛋白含量高,而23kD的蛋白含量则低。高渗骤变后,51kD蛋白的含量减少,而23kD蛋白的含量增加两倍或两倍以上,骤变23h后含量增加已非常明显;低渗骤变后24h,51kD和23kD蛋白的含量变化不明显。另外,有一分子量约为26kD的蛋白在高盐下易降解。分析了这些蛋白与社氏盐藻渗透调节的可能关系。  相似文献   
239.
DNA、RNA和PRO的合成、积累及相互关系是调控细胞周期动力学最主要的三个参数。同时检测这些组成部分能够更精确细致地评判细胞的周期动力学特征。本文探索了人正常骨髓CD34~ 造血细胞周期动力学相关大分子DNA、RNA和PRO的含量,以便认识CD34~ 造血细胞周期动力学的特征。用新型CIMS-100免疫磁性分离系统高效富集人骨髓CD34~ 造血细胞,经FCM及APAAP鉴定,富集的CD34~ 造血细胞的纯度达90~95%。随之采用碘化丙啶(PI)、派若宁Y(PY)及异硫氰荧光素(FITC)分别进行标记DNA、RNA和PRO并在FCM上检测。结果表明,DNA、RNA和PRO在CD34~ 造血细胞中的含量明显低于单个核细胞,分别仅占后者的34±3%、48±21%及62±14%。结合我们以往的结果,我们认为CD34~ 造血细胞的确是一独特的体细胞群,不仅表现在重建造血与免疫学功能上,而且表现在细胞周期动力学上。据我们所知,这是目前国际上首次有关人CD34~ 造血细胞周期动力学相关大分子的系统分析报道,提供了大多数CD34~ 造血细胞处于静止期的直接证据。  相似文献   
240.
Most existing human mobility literature focuses on exterior characteristics of movements but neglects activities, the driving force that underlies human movements. In this research, we combine activity-based analysis with a movement-based approach to model the intra-urban human mobility observed from about 15 million check-in records during a yearlong period in Shanghai, China. The proposed model is activity-based and includes two parts: the transition of travel demands during a specific time period and the movement between locations. For the first part, we find the transition probability between activities varies over time, and then we construct a temporal transition probability matrix to represent the transition probability of travel demands during a time interval. For the second part, we suggest that the travel demands can be divided into two classes, locationally mandatory activity (LMA) and locationally stochastic activity (LSA), according to whether the demand is associated with fixed location or not. By judging the combination of predecessor activity type and successor activity type we determine three trip patterns, each associated with a different decay parameter. To validate the model, we adopt the mechanism of an agent-based model and compare the simulated results with the observed pattern from the displacement distance distribution, the spatio-temporal distribution of activities, and the temporal distribution of travel demand transitions. The results show that the simulated patterns fit the observed data well, indicating that these findings open new directions for combining activity-based analysis with a movement-based approach using social media check-in data.  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号