全文获取类型
收费全文 | 21959篇 |
免费 | 1502篇 |
国内免费 | 9篇 |
专业分类
23470篇 |
出版年
2024年 | 19篇 |
2023年 | 126篇 |
2022年 | 311篇 |
2021年 | 580篇 |
2020年 | 352篇 |
2019年 | 470篇 |
2018年 | 594篇 |
2017年 | 478篇 |
2016年 | 811篇 |
2015年 | 1216篇 |
2014年 | 1346篇 |
2013年 | 1659篇 |
2012年 | 1949篇 |
2011年 | 1894篇 |
2010年 | 1178篇 |
2009年 | 1018篇 |
2008年 | 1376篇 |
2007年 | 1288篇 |
2006年 | 1181篇 |
2005年 | 1081篇 |
2004年 | 981篇 |
2003年 | 927篇 |
2002年 | 754篇 |
2001年 | 117篇 |
2000年 | 106篇 |
1999年 | 135篇 |
1998年 | 125篇 |
1997年 | 116篇 |
1996年 | 89篇 |
1995年 | 86篇 |
1994年 | 92篇 |
1993年 | 89篇 |
1992年 | 53篇 |
1991年 | 56篇 |
1990年 | 63篇 |
1989年 | 45篇 |
1988年 | 50篇 |
1987年 | 39篇 |
1986年 | 42篇 |
1985年 | 51篇 |
1984年 | 39篇 |
1983年 | 44篇 |
1982年 | 32篇 |
1981年 | 42篇 |
1980年 | 26篇 |
1979年 | 37篇 |
1978年 | 33篇 |
1977年 | 30篇 |
1976年 | 25篇 |
1975年 | 23篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
971.
Anna Rita Rossi Eleonora Perrone Luciana Sola 《Central European Journal of Biology》2006,1(4):636-647
The gilthead seabream, Sparus aurata, represents an important economic resource for Mediterranean aquaculture. In spite of its wide geographic distribution and
economic importance, only recently studies have been carried out on the genetic composition of natural populations, which
have revealed a picture of a heterogeneous degree of genetic differentiation among S. aurata populations. In this study an allozyme analysis of samples from six different collecting sites along the Italian and Croatian
coasts was carried out, covering an area in the Central Mediterranean sea that has yet to be investigated through gene-enzyme
systems. Data on 26 gene loci, 10 of which are polymorphic, indicate a slight but significant genetic structure (FST = 0.0167) of the species. A hierarchical analysis of population subdivision made it possible to identify three different
assemblages found in the Adriatic Sea, Tyrrhenian Sea and Sardinian Channel, though an isolation by distance model can be
rejected. The results are discussed in the light of previous literature and taking conservation into consideration. 相似文献
972.
Kilanczyk E Filipek S Filipek A 《Biochemical and biophysical research communications》2011,404(1):179-183
Recently, we have reported that the CacyBP/SIP protein binds ERK1/2 (Kilanczyk et al., BBRC, 2009). In this work we show that CacyBP/SIP exhibits a phosphatase activity toward ERK1/2 kinases while its E217K mutant does not. The Km and Vmax values established for a standard phosphatase substrate, p-NPP, are 16.9 ± 3.6 mM and 4.3 ± 0.4 μmol/min, respectively. The CacyBP/SIP phosphatase activity is decreased by okadaic acid (IC50 = 45 nM). Our experimental results are supported by a theoretical analysis which revealed important sequence similarities between CacyBP/SIP and the phosphatase-like proteins as well as certain MAP kinase phosphatases. 相似文献
973.
Wrenger C Müller IB Schifferdecker AJ Jain R Jordanova R Groves MR 《Journal of molecular biology》2011,405(4):956-971
Aspartate aminotransferases (AspATs; EC 2.6.1.1) catalyze the conversion of aspartate and α-ketoglutarate into oxaloacetate and glutamate and are key enzymes in the nitrogen metabolism of all organisms. Recent findings suggest that the plasmodial enzyme [Plasmodium falciparum aspartate aminotransferase (PfAspAT)] may also play a pivotal role in energy metabolism and in the de novo biosynthesis of pyrimidines. However, while PfAspAT is a potential drug target, the high homology between the active sites of currently available AspAT structures hinders the development of specific inhibitors of these enzymes. In this article, we report the X-ray structure of the PfAspAT homodimer at a resolution of 2.8 Å. While the overall fold is similar to the currently available structures of other AspATs, the structure presented shows a significant divergence in the conformation of the N-terminal residues. Deletion of these divergent PfAspAT N-terminal residues results in a loss of activity for the recombinant protein, and addition of a peptide containing these 13 N-terminal residues results in inhibition both in vitro and in a lysate isolated from cultured parasites, while the activity of human cytosolic AspAT is unaffected. The finding that the divergent N-terminal amino acids of PfAspAT play a role in catalytic activity indicates that specific inhibition of the enzyme may provide a lead for the development of novel compounds in the treatment of malaria. We also report on the localization of PfAspAT to the parasite cytosol and discuss the implications of the role of PfAspAT in the supply of malate to the parasite mitochondria. 相似文献
974.
Bernardetta Maresca Luisa Cigliano Maria Stefania Spagnuolo Fabrizio Dal Piaz Maria M. Corsaro Nicola Balato Massimiliano Nino Anna Balato Fabio Ayala Paolo Abrescia 《PloS one》2012,7(12)
Improved diagnosis of psoriasis, by new biomarkers, is required for evaluating the progression rate of the disease and the response to treatment. Haptoglobin (Hpt), a glycoprotein secreted by hepatocytes and other types of cells including keratinocytes, was found with glycan changes in psoriasis and other diseases. We previously reported that Hpt isolated from plasma of psoriatic patients is more fucosylated than Hpt of healthy subjects. The aim of this study was to compare the glycosylation pattern of Hpt isolated from skin scales or plasma of patients with psoriasis with that of Hpt from cornified epidermal layer or plasma of healthy subjects. High performance liquid chromatography analysis of the glycans isolated from the protein backbone revealed that glycan patterns from skin and plasma of patients were similar, and mostly displayed quantitative rather than qualitative differences from normal pattern. Biotin-labeled lectins were used to evaluate quantitative differences in the glycoforms of Hpt from plasma and psoriatic skin scales. Hpt from skin and plasma of patients showed more fucosylated and branched glycans than Hpt from plasma of healthy subjects. Tryptic glycopeptides of Hpt were also analyzed by mass spectrometry, and a decreased amount of sialylated glycan chains was found in glycopeptides of skin Hpt, as compared with Hpt from plasma. High levels of glycans with fucosylated and tetra-antennary chains were detected on the peptide NLFLNHSENATAK from Hpt of psoriatic patients. Our data demonstrate that specific changes in glycan structures of Hpt, such as enhanced glycan branching and fucose content, are associated with psoriasis, and that differences between circulating and skin Hpt do exist. A lower extent of glycan fucosylation and branching was found in Hpt from plasma of patients in disease remission. Altered glycoforms might reflect changes of Hpt function in the skin, and could be used as markers of the disease. 相似文献
975.
Matthias Oelze Swenja Kr?ller-Sch?n Philipp Welschof Thomas Jansen Michael Hausding Yuliya Mikhed Paul Stamm Michael Mader Elena Zin?ius Saule Agdauletova Anna Gottschlich Sebastian Steven Eberhard Schulz Serge P. Bottari Eric Mayoux Thomas Münzel Andreas Daiber 《PloS one》2014,9(11)
Objective
In diabetes, vascular dysfunction is characterized by impaired endothelial function due to increased oxidative stress. Empagliflozin, as a selective sodium-glucose co-transporter 2 inhibitor (SGLT2i), offers a novel approach for the treatment of type 2 diabetes by enhancing urinary glucose excretion. The aim of the present study was to test whether treatment with empagliflozin improves endothelial dysfunction in type I diabetic rats via reduction of glucotoxicity and associated vascular oxidative stress.Methods
Type I diabetes in Wistar rats was induced by an intravenous injection of streptozotocin (60 mg/kg). One week after injection empagliflozin (10 and 30 mg/kg/d) was administered via drinking water for 7 weeks. Vascular function was assessed by isometric tension recording, oxidative stress parameters by chemiluminescence and fluorescence techniques, protein expression by Western blot, mRNA expression by RT-PCR, and islet function by insulin ELISA in serum and immunohistochemical staining of pancreatic tissue. Advanced glycation end products (AGE) signaling was assessed by dot blot analysis and mRNA expression of the AGE-receptor (RAGE).Results
Treatment with empagliflozin reduced blood glucose levels, normalized endothelial function (aortic rings) and reduced oxidative stress in aortic vessels (dihydroethidium staining) and in blood (phorbol ester/zymosan A-stimulated chemiluminescence) of diabetic rats. Additionally, the pro-inflammatory phenotype and glucotoxicity (AGE/RAGE signaling) in diabetic animals was reversed by SGLT2i therapy.Conclusions
Empagliflozin improves hyperglycemia and prevents the development of endothelial dysfunction, reduces oxidative stress and improves the metabolic situation in type 1 diabetic rats. These preclinical observations illustrate the therapeutic potential of this new class of antidiabetic drugs. 相似文献976.
Z. Dave Sharp Anna Meriwether Jerry Ware M. Donald Cave 《Biochemical genetics》1986,24(11-12):911-923
An analysis of a 29-kilobase nontranscribed spacer fragment in the ribosomal DNA (rDNA) of the house cricket, Acheta domesticus, revealed a highly repetitious structure. A total of eight EcoRI repeats of three different size classes measuring 259, 420, and 508 base pairs (bp) was mapped to a region 2 kilobases (kb) from the 18 S coding region. The repeats were oriented in a nonrandom manner and had sequences homologous to DNA located immediately adjacent to the repetitive array. DNA sequence analysis showed that the repetitive region was composed of smaller direct repeats 66, 67, and 383 bp in length. There was minor length heterogeneity of the chromosomal restriction fragments containing the entire array, indicating that a variable number of EcoRI repeats is a minor contributor to the total repeat-unit length heterogeneity. Immediately upstream from the EcoRI array there is a 17-kb region composed of 50 to 60 subrepeat elements recognized by a variety of restriction endonucleases. A subcloned SmaI repeat from the array was not homologous to any other part of the rDNA repeat unit or other chromosomal DNA. There was little length heterogeneity in restriction fragments containing the chromosomal 17-kb repetitions region. Immediately upstream from the 17-Kb region there is a 4.1-kb segment with sequences homologous to the EcoRI repeats. 相似文献
977.
A standard type II polyketide synthase (PKS) gene cluster was isolated while attempting to clone the biosynthetic gene for lipstatin from Streptomyces toxytricini NRRL 15,443. This result was observed using a Southern blot of a PstI-digested S. toxytricini chromosomal DNA library with a 444 bp amplified probe of a ketosynthase (KS) gene fragment. Four open reading frames [thioesterase (TE), beta-ketoacyl systhase (KAS), chain length factor (CLF), and acyl carrier protein (ACP)], were identified through the nucleotide sequence determination and analysis of a 4.5 kb cloned DNA fragment. In order to confirm the involvement of a cloned gene in lipstatin biosynthesis, a gene disruption experiment for the KS gene was performed. However, the resulting gene disruptant did not show any significant difference in lipstatin production when compared to wild-type S. toxytricini. This result suggests that lipstatin may not be synthesized by a type II PKS. 相似文献
978.
979.
Vojtch Vejvoda Ondej Kaplan Karel Bezouka Petr Pompach Miroslav ulc Maria Cantarella Oldich Benada Bronislava Uhnkov Anna Ringelov Sabine Lutz-Wahl Lutz Fischer Vladimír Ken Ludmila Martínkov 《Journal of Molecular Catalysis .B, Enzymatic》2008,50(2-4):99-106
An intracellular nitrilase was purified from a Fusarium solani O1 culture, in which the enzyme (up to 3000 U L−1) was induced by 2-cyanopyridine. SDS-PAGE revealed one major band corresponding to a molecular weight of approximately 40 kDa. Peptide mass fingerprinting suggested a high similarity of the protein with the putative nitrilase from Gibberella moniliformis. Electron microscopy revealed that the enzyme molecules associated into extended rods. The enzyme showed high specific activities towards benzonitrile (156 U mg−1) and 4-cyanopyridine (203 U mg−1). Other aromatic nitriles (3-chlorobenzonitrile, 3-hydroxybenzonitrile) also served as good substrates for the enzyme. The rates of hydrolysis of aliphatic nitriles (methacrylonitrile, propionitrile, butyronitrile, valeronitrile) were 14–26% of that of benzonitrile. The nitrilase was active within pH 5–10 and at up to 50 °C with optima at pH 8.0 and 40–45 °C. Its activity was strongly inhibited by Hg2+ and Ag+ ions. More than half of the enzyme activity was preserved at up to 50% of n-hexane or n-heptane or at up to 15% of xylene or ethanol. Operational stability of the enzyme was examined by the conversion of 45 mM 4-cyanopyridine in a continuous and stirred ultrafiltration-membrane reactor. The nitrilase half-life was 277 and 10.5 h at 35 and 45 °C, respectively. 相似文献
980.
Winton MJ Van Deerlin VM Kwong LK Yuan W Wood EM Yu CE Schellenberg GD Rademakers R Caselli R Karydas A Trojanowski JQ Miller BL Lee VM 《FEBS letters》2008,582(15):2252-2256
TAR DNA-binding protein-43 (TDP-43) is a highly conserved, ubiquitously expressed nuclear protein that was recently identified as the disease protein in frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS). Pathogenic TDP-43 gene (TARDBP) mutations have been identified in familial ALS kindreds, and here we report a TARDBP variant (A90V) in a FTLD/ALS patient with a family history of dementia. Significantly, A90V is located between the bipartite nuclear localization signal sequence of TDP-43 and the in vitro expression of TDP-43-A90V led to its sequestration with endogenous TDP-43 as insoluble cytoplasmic aggregates. Thus, A90V may be a genetic risk factor for FTLD/ALS because it predisposes nuclear TDP-43 to redistribute to the cytoplasm and form pathological aggregates. 相似文献