全文获取类型
收费全文 | 4548篇 |
免费 | 467篇 |
国内免费 | 434篇 |
出版年
2023年 | 100篇 |
2022年 | 142篇 |
2021年 | 116篇 |
2020年 | 146篇 |
2019年 | 182篇 |
2018年 | 232篇 |
2017年 | 166篇 |
2016年 | 150篇 |
2015年 | 155篇 |
2014年 | 327篇 |
2013年 | 403篇 |
2012年 | 247篇 |
2011年 | 270篇 |
2010年 | 234篇 |
2009年 | 251篇 |
2008年 | 251篇 |
2007年 | 294篇 |
2006年 | 201篇 |
2005年 | 163篇 |
2004年 | 128篇 |
2003年 | 108篇 |
2002年 | 84篇 |
2001年 | 60篇 |
2000年 | 54篇 |
1999年 | 70篇 |
1998年 | 46篇 |
1997年 | 52篇 |
1996年 | 57篇 |
1995年 | 35篇 |
1994年 | 40篇 |
1993年 | 52篇 |
1992年 | 41篇 |
1991年 | 34篇 |
1990年 | 25篇 |
1989年 | 26篇 |
1988年 | 24篇 |
1987年 | 21篇 |
1986年 | 19篇 |
1985年 | 31篇 |
1984年 | 55篇 |
1983年 | 36篇 |
1982年 | 45篇 |
1981年 | 34篇 |
1980年 | 43篇 |
1979年 | 40篇 |
1978年 | 23篇 |
1977年 | 28篇 |
1976年 | 22篇 |
1974年 | 24篇 |
1973年 | 23篇 |
排序方式: 共有5449条查询结果,搜索用时 46 毫秒
61.
Leda Guzman Rodrigo Bustos Ricardo B. Maccioni 《Molecular and cellular biochemistry》1994,131(2):105-113
The changes in the levels of microtubule-associated proteins (MAPs) during advanced embryonic stages, neonatal and adult organisms reflect the importance of these cytoskeletal proteins in relation to the morphogenesis of the central nervous system. MAP-1B is found in prenatal brains and it appears to have the highests levels in neonatal rat brains, being a developmentally-regulated protein. In this research, a fast procedure to isolate MAP-1B, as well as MAP-2 and MAP-3 from neonatal rat brains was designed, based on the differential capacity of poly L-aspartic acid to release MAPs during temperature-dependent cycles of microtubule assembly in the absence of taxol. The high molecular weight MAP-1B was recovered in the warm supernatants after microtubular protein polymerization in the presence of low concentrations of polyaspartic acid. Instead, MAP-2 and a 180 kDa protein with characteristics of MAP-3 remained associated to the polymer after the assembly. Further purification of MAP-1B was attained after phosphocellulose chromatography. Isolation of MAP-2 isoforms together with MAP-3 was achieved on the basis of their selective interactions with calmodulin-agarose affinity columns. In addition, MAP-2 and MAP-3 were also purified on the basis of their capacities to interact with the tubulin peptide -II (422–434) derivatized on an Affigel matrix. However, MAP-1B did not interact with the -II tubulin fragment, but it showed interaction with the Affigel-conjugated -I (431–444) tubulin peptide. The different MAPs componentes were characterized by western blots using specific monoclonal antibodies. A salient feature of neonatal rat brain MAP-3 was its interactions with site-directed antibodies that recognize binding epitopes on the repetitive sequences of tau and MAP-2. However, these site-specific antibodies did not interact with MAP-1B from the neonatal rat brain tissue.Abbreviations PAA
poly (L-aspartic acid)
- HMW-MAPs
high molecular weight microtubule associated proteins 相似文献
62.
Chronic Administration of Lithium Chloride Increases Immunodetectable Glial Fibrillary Acidic Protein in the Rat Hippocampus 总被引:3,自引:0,他引:3
Abstract: We studied the effect of treating rats with lithium salts on the content and in vitro phosphorylation rate of the astrocyte cell marker, glial fibrillary acidic protein (GFAP), in brain slices. Rats were fed a diet incorporating lithium chloride until the concentration of Li+ in serum reached 0.6–1.2 m M , a range similar to that achieved in clinical practice. Hippocampal tissue was analyzed for immunoreactive GFAP by a dot assay, and slices of hippocampus and caudate nucleus were labeled with [32 P]-phosphate to determine the in vitro rate of phosphorylation of GFAP. Compared with controls, the level of immunoreactive GFAP in the hippocampus from lithium-treated rats was increased 34%, and GFAP in hippocampal slices incorporated 39% more 32 P. This effect of lithium was apparently not confined to the hippocampus because the in vitro rate of phosphorylation of GFAP in caudate slices was also increased in the treated rats. 相似文献
63.
Mondher Jaziri Kayo Yoshimatsu Jacques Homès Koichiro Shimomura 《Plant Cell, Tissue and Organ Culture》1994,38(2-3):257-262
Hairy root cultures of Atropa belladonna L. were established by infection either with Agrobacterium rhizogenes ATCC 15834 or MAFF 03-01724, and transgenic plants were obtained from both hairy root cultures. Doubly transformed roots were induced by re-infection of the leaf segments of transgenic Atropa belladonna plants (A. rhizogenes 15834) with MAFF 03-01724. Shoots and viviparous leaves were regenerated from the doubly transformed roots. The genetic transformation was determined by the opine assay (agropine, mannopine and/or mikimopine) and polymerase chain reaction. Physiological changes and tropane alkaloid biosynthesis in the hairy roots (singly and doubly transformed) were investigated. The alkaloid content in the doubly transformed root strain was intermediate as compared to the root strains which were singly transformed. On the other hand endogenous IAA levels in doubly transformed roots were significantly decreased compared to both singly transformed roots.Abbreviations BA
benzyladenine
- IAA
indoleacetic acid
- NAA
naphthaleneacetic acid
- PCR
polymerase chain reaction
-
t-ZR
trans-zeatin 相似文献
64.
Soraya C. M. Leal David J. Bertioli Brenda V. Ball Tariq M. Butt 《Biocontrol Science and Technology》1994,4(1):89-94
Nucleic acids from 41 strains of Metarhizium anisopliae, obtained from different parts of the world were extracted and examined by electrophoresis. Strong bands of double‐stranded RNA (dsRNA) were detected in two isolates from Brazil, V215 and V291, which had, respectively, 13 and 9 distinct bands ranging in size from ca. 0.75 to 3.5 kb. Icosahedral virus‐like particles (VLPs) (ca. 33 nm in diameter) were observed by transmission electron microscopy in extracts of these isolates. The VLPs and dsRNA were both absent from a clone of the isolate V291 which had been subcultured successively on solid medium. Bioassays against the aphid Myzus persicae showed no detectable difference in virulence between the clone of V291 which contained dsRNA and the clone that did not. 相似文献
65.
本文就萤叶甲亚科中柱萤叶甲属鞘翅具黑色刻点的种类进行研究,共记述4种,我国已记录3种,其中1种为新种。 相似文献
66.
几种濒危植物及其近缘类群总DNA的提取与鉴定 总被引:122,自引:0,他引:122
用低pH 介质,高盐沉淀蛋白质方法成功地从银杉(Cathaya argyrophylla Chun etKuang)、矮牡丹(Paeonia suffruticosa var. spontanea)、南川升麻(Cim icifuga nanchuanensisHsiao)、裂叶沙参(Adenophora lobophylla)的同属种泡沙参(A. potaninii)等植物中提取和部分纯化了细胞总DNA,并对其产率、质量和纯度作了鉴定。此方法的关键是用了一个低pH提取介质,它能有效防止组织破碎及沉淀大量材料时的电离化作用及酚化合物的进一步氧化。所得DNA 不需经氯化铯梯度离心或柱层析,直接可用于限制性片断长度多态性(RFLP)及随机扩增的DNA多态性(RAPD)等分子水平的遗传标记。为检测濒危植物的遗传多样性提供了一套迅速、简便和可靠的技术方案 相似文献
67.
用共振拉曼和紫外-可见吸收光谱研究了水溶金属卟啉4-N-乙酸乙酯基-吡啶基铜卟啉和镍卟啉[简称Cu(NEAE)和Ni(NEAE)]及4-N-乙腈基-吡啶基铜卟啉[简称Cu(NACN)]与小牛胸腺DNA的相互作用。分析表明Cu(NEAE)、Ni(NEAE)和Cu(NACN)分别以外部键联、部分插入和沟槽方式与DNA作用;卟啉插入DNA时吡啶基团向卟啉环平面转动但不可能转成与之共面;而以非插入方式作用时吡啶基团会向垂直于或者平行于卟啉环平面转动。吡啶取代基的大小和空间位阻是影响相互作用方式的关键因素之一。 相似文献
68.
Junichiro J. Kazama Takashi Aikata Masaaki Arakawa Hidehiro Ozawa 《Biotechnic & histochemistry》1994,69(6):324-328
We describe a new technique for immunohistochemical and enzyme-histochemi-cal double staining using confocal laser scanning microscopy in the reflection mode. As an example, we investigated the immunoreactivity for Spot 35-calbindin-D28K, a vitamin D-dependent calcium binding protein, and the enzyme activity for ma+-ATPase in the rat kidney. The lead precipitation method for Ca2+-ATPase was initially used to process kidney slices. Each specimen was then dehydrated and embedded in a water soluble resin. Thin sections were cut from the resin block, and an indirect immunocolloidal gold method with silver enhancement for Spot 35-calbindin-D28K antigen was carried out on the glass slides. Results were then observed by confocal laser scanning microscopy in the reflection mode. The three-dimensional distribution of the reaction products was detected by serial optic slice images. Lead phosphate particles, which represented the location of Ca2+-ATPase, were distributed deep in the section. The most intense signals from the silver partkles were detected from the surface slice of the section. A stereoscopic image generated from the serial optic slices clearly showed the differences in their distribution. 相似文献
69.
Corinne Beaudouin Gisèle Haurat Jean Alex Laffitte Bernard Renaud 《Journal of neurochemistry》1993,61(3):928-935
Abstract: (+)-S-Adenosyl- l -methionine [(+)-SAM] was isolated from rat brain and was quantified by HPLC followed by UV spectrophotometric measurements and by 1 H-NMR. Its estimated ratio in brain is 3% of total SAM. Because of its commercial unavailability, (+)-SAM was also prepared from chemically synthesized SAM by separation of the two diastereoisomers on a preparative reverse-phase Nucleosil C8 column. The (+) diastereoisomer thus obtained was then assayed in vitro both as an inhibitor and a substrate of phenylethanolamine N -methyltransferase. Enzymatic activity was measured by HPLC analysis. It was shown that (+)-SAM has no effect on phenylethanolamine N -methyltransferase activity; therefore, it is unlikely that (+)-SAM plays any possible role in regulation of adrenaline synthesis in the brain. 相似文献
70.
The behaviour of a highly purified -l-fucosidase (E.C. 3.2.1.51) extracted from octopus hepatopancreas was studied with phospholipid vesicles composed of phosphatidylcholine (PC) and phosphatidylserine (PS) containing the fucosylated ganglioside FucGM1, a potential natural substrate of the enzyme. The substrate recognition and hydrolysis take place only with PS/FucGM1 mixtures via an association process of the enzyme with the vesicles at acidic pH; the enzyme rapidly and stably binds to PS vesicles but not to PC vesicles. The data suggest that only the PS-associated enzyme is able to hydrolyse FucGM1 embedded in the same bilayer. The enzyme association with FucGM1/PS vesicles is a prerequisite for ganglioside hydrolysis but is followed by irreversible enzyme inactivation. 相似文献