全文获取类型
收费全文 | 43916篇 |
免费 | 3284篇 |
国内免费 | 10篇 |
专业分类
47210篇 |
出版年
2023年 | 228篇 |
2022年 | 385篇 |
2021年 | 742篇 |
2020年 | 527篇 |
2019年 | 581篇 |
2018年 | 1350篇 |
2017年 | 1130篇 |
2016年 | 1373篇 |
2015年 | 2030篇 |
2014年 | 2028篇 |
2013年 | 2729篇 |
2012年 | 3183篇 |
2011年 | 3042篇 |
2010年 | 1914篇 |
2009年 | 1638篇 |
2008年 | 2442篇 |
2007年 | 2382篇 |
2006年 | 2322篇 |
2005年 | 2042篇 |
2004年 | 2005篇 |
2003年 | 1801篇 |
2002年 | 1657篇 |
2001年 | 865篇 |
2000年 | 796篇 |
1999年 | 729篇 |
1998年 | 438篇 |
1997年 | 334篇 |
1996年 | 313篇 |
1995年 | 318篇 |
1994年 | 259篇 |
1993年 | 244篇 |
1992年 | 423篇 |
1991年 | 424篇 |
1990年 | 351篇 |
1989年 | 319篇 |
1988年 | 303篇 |
1987年 | 264篇 |
1986年 | 250篇 |
1985年 | 257篇 |
1984年 | 203篇 |
1983年 | 162篇 |
1982年 | 148篇 |
1981年 | 175篇 |
1980年 | 142篇 |
1979年 | 190篇 |
1978年 | 165篇 |
1977年 | 138篇 |
1976年 | 146篇 |
1975年 | 132篇 |
1974年 | 134篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
1.
G. Larivière-Gauthier A. Thibodeau É. Yergeau P. Fravalo 《Letters in applied microbiology》2021,72(2):113-120
Recent studies have shown that Salmonella shedding status affects sows’ microbiota during gestation and that these modifications are reflected in the faecal microbiota of their piglets at weaning. The aims of this study were: (a) to evaluate the persistence, up to the fattening period, of the previously measured link between the microbiota of piglets and their mothers’ Salmonella shedding status; and (b) measure the impact of the measured microbiota variations on their Salmonella excretion at this stage. To achieve this, 76 piglets born from 19 sows for which the faecal microbiota was previously documented, were selected in a multisite production system. The faecal matter of these swine was sampled after 4 weeks, at the fattening stage. The Salmonella shedding status and faecal microbiota of these animals were described using bacteriological and 16S rRNA gene amplicon sequencing respectively. The piglet digestive microbiota association with the Salmonella shedding status of their sows did not persist after weaning and did not affect the risk of Salmonella excretion during fattening, while the birth mother still affected the microbiota of the swine at fattening. This supports the interest in sows as a target for potentially transferrable microbiota modifications. 相似文献
2.
E Estébanez-Perpi?á A Bayés J Vendrell M A Jongsma D P Bown J A Gatehouse R Huber W Bode F X Avilés D Reverter 《Journal of molecular biology》2001,313(3):629-638
The cotton bollworm Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) is one of the most serious insect pests in Australia, India and China. The larva causes substantial economical losses to legume, fibre, cereal oilseed and vegetable crops. This pest has proven to be difficult to control by conventional means, mainly due to the development of pesticide resistance. We present here the 2.5 A crystal structure from the novel procarboxypeptidase (PCPAHa) found in the gut extracts from H. armigera larvae, the first one reported for an insect. This metalloprotease is synthesized as a zymogen of 46.6 kDa which, upon in vitro activation with Lys-C endoproteinase, yields a pro-segment of 91 residues and an active carboxypeptidase moiety of 318 residues. Both regions show a three-dimensional structure quite similar to the corresponding structures in mammalian digestive carboxypeptidases, the most relevant structural differences being located in the loops between conserved secondary structure elements, including the primary activation site. This activation site contains the motif (Ala)(5)Lys at the C terminus of the helix connecting the pro- and the carboxypeptidase domains. A remarkable feature of PCPAHa is the occurrence of the same (Ala)(6)Lys near the C terminus of the active enzyme. The presence of Ser255 in PCPAHa instead of Ile and Asp found in the pancreatic A and B forms, respectively, enlarges the S1' specificity pocket and influences the substrate preferences of the enzyme. The C-terminal tail of the leech carboxypeptidase inhibitor has been modelled into the PCPAHa active site to explore the substrate preferences and the enzymatic mechanism of this enzyme. 相似文献
3.
4.
Sebastián Moran Miguel Vizoso Anna Martinez-Cardús Antonio Gomez Xavier Matías-Guiu Sebastián M Chiavenna Andrés G Fernandez Manel Esteller 《Epigenetics》2014,9(6):829-833
A formalin-fixed paraffin-embedded (FFPE) sample usually yields highly degraded DNA, which limits the use of techniques requiring high-quality DNA, such as Infinium Methylation microarrays. To overcome this restriction, we have applied an FFPE restoration procedure consisting of DNA repair and ligation processes in a set of paired fresh-frozen (FF) and FFPE samples. We validated the FFPE results in comparison with matched FF samples, enabling us to use FFPE samples on the Infinium HumanMethylation450 Methylation array. 相似文献
5.
6.
E. Molina Grima J. A. Sánchez Pérez F. García Camacho F. G. Acién Fernández J. M. Fernández Sevilla F. Valdés Sanz 《Biotechnology letters》1994,16(10):1035-1040
Summary Eicosapentaenoic acid (EPA) volumetric productivity from an outdoor chemostat culture ofPhaeodactylum tricornutum UTEX 640 in a 50-l tubular photobioreactor varies with dilution rate, reaching a maximum of 47.8 mg l–1 d–1 at D=0.36 d–1. Continuous culture at high dilution rates' is proposed as the most adequate operating mode to maximize polyunsaturated fatty acid production. 相似文献
7.
Carmen de Sena-Tomás Mónica Navarro-González Ursula Kües José Pérez-Martín 《Genetics》2013,195(1):47-57
The fungal fruiting body or mushroom is a multicellular structure essential for sexual reproduction. It is composed of dikaryotic cells that contain one haploid nucleus from each mating partner sharing the same cytoplasm without undergoing nuclear fusion. In the mushroom, the pileus bears the hymenium, a layer of cells that includes the specialized basidia in which nuclear fusion, meiosis, and sporulation occur. Coprinopsis cinerea is a well-known model fungus used to study developmental processes associated with the formation of the fruiting body. Here we describe that knocking down the expression of Atr1 and Chk1, two kinases shown to be involved in the response to DNA damage in a number of eukaryotic organisms, dramatically impairs the ability to develop fruiting bodies in C. cinerea, as well as other developmental decisions such as sclerotia formation. These developmental defects correlated with the impairment in silenced strains to sustain an appropriated dikaryotic cell cycle. Dikaryotic cells in which chk1 or atr1 genes were silenced displayed a higher level of asynchronous mitosis and as a consequence aberrant cells carrying an unbalanced dose of nuclei. Since fruiting body initiation is dependent on the balanced mating-type regulator doses present in the dikaryon, we believe that the observed developmental defects were a consequence of the impaired cell cycle in the dikaryon. Our results suggest a connection between the DNA damage response cascade, cell cycle regulation, and developmental processes in this fungus. 相似文献
8.
Eugénie Bassères Giuseppe Coppotelli Thorsten Pfirrmann Jens B. Andersen Maria Masucci Teresa Frisan 《Cellular microbiology》2010,12(11):1622-1633
Invasion of eukaryotic target cells by pathogenic bacteria requires extensive remodelling of the membrane and actin cytoskeleton. Here we show that the remodelling process is regulated by the ubiquitin C‐terminal hydrolase UCH‐L1 that promotes the invasion of epithelial cells by Listeria monocytogenes and Salmonella enterica. Knockdown of UCH‐L1 reduced the uptake of both bacteria, while expression of the catalytically active enzyme promoted efficient internalization in the UCH‐L1‐negative HeLa cell line. The entry of L. monocytogenes involves binding to the receptor tyrosine kinase Met, which leads to receptor phosphorylation and ubiquitination. UCH‐L1 controls the early membrane‐associated events of this triggering cascade since knockdown was associated with altered phosphorylation of the c‐cbl docking site on Tyr1003, reduced ubiquitination of the receptor and altered activation of downstream ERK1/2‐ and AKT‐dependent signalling in response to the natural ligand Hepatocyte Growth Factor (HGF). The regulation of cytoskeleton dynamics was further confirmed by the induction of actin stress fibres in HeLa expressing the active enzyme but not the catalytic mutant UCH‐L1C90S. These findings highlight a previously unrecognized involvement of the ubiquitin cycle in bacterial entry. UCH‐L1 is highly expressed in malignant cells that may therefore be particularly susceptible to invasion by bacteria‐based drug delivery systems. 相似文献
9.
10.
A new purification method for cytosolic malate dehydrogenases from several sources has been developed. The procedure, employing chromatographies on 5'AMP-Sepharose, DEAE-Sephacel and Blue-Sepharose, allows for a rapid isolation of the enzyme (approximately 40 hours), in large quantities, with good yields (45-54%). The specific activity of final preparations were around 1300 I.U./mg and were judged homogeneous by polyacrylamide gradient gel and sodium dodecyl sulfate polyacrylamide gel electrophoresis, high performance size exclusion chromatography and isoelectric focusing. 相似文献