全文获取类型
收费全文 | 30797篇 |
免费 | 2449篇 |
国内免费 | 853篇 |
专业分类
34099篇 |
出版年
2024年 | 41篇 |
2023年 | 281篇 |
2022年 | 520篇 |
2021年 | 890篇 |
2020年 | 622篇 |
2019年 | 802篇 |
2018年 | 879篇 |
2017年 | 835篇 |
2016年 | 1164篇 |
2015年 | 1778篇 |
2014年 | 1901篇 |
2013年 | 2293篇 |
2012年 | 2736篇 |
2011年 | 2571篇 |
2010年 | 1631篇 |
2009年 | 1524篇 |
2008年 | 1847篇 |
2007年 | 1724篇 |
2006年 | 1658篇 |
2005年 | 1418篇 |
2004年 | 1304篇 |
2003年 | 1215篇 |
2002年 | 1105篇 |
2001年 | 271篇 |
2000年 | 187篇 |
1999年 | 237篇 |
1998年 | 283篇 |
1997年 | 258篇 |
1996年 | 207篇 |
1995年 | 176篇 |
1994年 | 162篇 |
1993年 | 165篇 |
1992年 | 130篇 |
1991年 | 115篇 |
1990年 | 98篇 |
1989年 | 86篇 |
1988年 | 69篇 |
1987年 | 60篇 |
1986年 | 55篇 |
1985年 | 93篇 |
1984年 | 83篇 |
1983年 | 62篇 |
1982年 | 68篇 |
1981年 | 75篇 |
1980年 | 58篇 |
1979年 | 48篇 |
1978年 | 45篇 |
1977年 | 39篇 |
1975年 | 31篇 |
1973年 | 40篇 |
排序方式: 共有10000条查询结果,搜索用时 7 毫秒
121.
Ferreira-Gomes MS González-Lebrero RM de la Fuente MC Strehler EE Rossi RC Rossi JP 《The Journal of biological chemistry》2011,286(37):32018-32025
In this work, we set out to identify and characterize the calcium occluded intermediate(s) of the plasma membrane Ca(2+)-ATPase (PMCA) to study the mechanism of calcium transport. To this end, we developed a procedure for measuring the occlusion of Ca(2+) in microsomes containing PMCA. This involves a system for overexpression of the PMCA and the use of a rapid mixing device combined with a filtration chamber, allowing the isolation of the enzyme and quantification of retained calcium. Measurements of retained calcium as a function of the Ca(2+) concentration in steady state showed a hyperbolic dependence with an apparent dissociation constant of 12 ± 2.2 μM, which agrees with the value found through measurements of PMCA activity in the absence of calmodulin. When enzyme phosphorylation and the retained calcium were studied as a function of time in the presence of La(III) (inducing accumulation of phosphoenzyme in the E(1)P state), we obtained apparent rate constants not significantly different from each other. Quantification of EP and retained calcium in steady state yield a stoichiometry of one mole of occluded calcium per mole of phosphoenzyme. These results demonstrate for the first time that one calcium ion becomes occluded in the E(1)P-phosphorylated intermediate of the PMCA. 相似文献
122.
Carvalho WA Ianella P Arnoldi FG Caetano AR Maruyama SR Ferreira BR Conti LH da Silva MR Paula JO Maia AA Santos IK 《Immunogenetics》2011,63(5):319-324
Bovines present contrasting, heritable phenotypes of infestations with the cattle tick, Rhipicephalus (Boophilus) microplus. Tick salivary glands produce IgG-binding proteins (IGBPs) as a mechanism for escaping from host antibodies that these ectoparasites ingest during blood meals. Allotypes that occur in the constant region of IgG may differ in their capacity to bind with tick IGBPs; this may be reflected by the distribution of distinct allotypes according to phenotypes of tick infestations. In order to test this hypothesis, we investigated the frequency of haplotypes of bovine IgG2 among tick-resistant and tick-susceptible breeds of bovines. Sequencing of the gene coding for the heavy chain of IgG2 from 114 tick-resistant (Bos taurus indicus, Nelore breed) and tick-susceptible (B. t. taurus, Holstein breed) bovines revealed SNPs that generated 13 different haplotypes, of which 11 were novel and 5 were exclusive of Holstein and 3 of Nelore breeds. Alignment and modeling of coded haplotypes for hinge regions of the bovine IgG2 showed that they differ in the distribution of polar and hydrophobic amino acids and in shape according to the distribution of these amino acids. We also found that there was an association between genotypes of the constant region of the IgG2 heavy chain with phenotypes of tick infestations. These findings open the possibility of investigating if certain IgG allotypes hinder the function of tick IGBPs. If so, they may be markers for breeding for resistance against tick infestations. 相似文献
123.
目的本试验旨在研究不同蛋白水平对实验藏酋猴生长性能和血液生化指标的影响并确定幼年实验藏酋猴的蛋白营养需要量。方法选用15只幼年实验藏酋猴随机分为5个处理组,每个处理3个重复,分别饲喂蛋白水平为11.1%、16.1%、20.5%、24.5%和29.8%的日粮,试验期90d。结果随着日粮中蛋白水平的提高,实验藏酋猴的增重、血清白蛋白、球蛋白、天门冬氨酸氨基转移酶、丙氨酸氨基转移酶和尿素氮含量显著增加(P〈0.05),而血清总蛋白、甘油三酯、γ-谷氨酰胺转移酶和碱性磷酸酶无显著差异。应用折线法确定幼年实验藏酋猴的蛋白营养需要量为23.69%。结论日粮蛋白水平对实验藏酋猴生长性能有显著影响,从生物学和经济学角度考虑,初步认为在本试验条件下,未成年实验藏酋猴日粮蛋白水平以23.69%最合适。 相似文献
124.
García-Rodríguez FM Barrientos-Durán A Díaz-Prado V Fernández-López M Toro N 《Applied and environmental microbiology》2011,77(3):854-861
The group IIA intron Ll.LtrB from Lactococcus lactis and the group IIB intron EcI5 from Escherichia coli have intron-encoded proteins (IEP) with a DNA-binding domain (D) and an endonuclease domain (En). Both have been successfully retargeted to invade target DNAs other than their wild-type target sites. RmInt1, a subclass IIB3/D intron with an IEP lacking D and En domains, is highly active in retrohoming in its host, Sinorhizobium meliloti. We found that RmInt1 was also mobile in E. coli and that retrohoming in this heterologous host depended on temperature, being more efficient at 28°C than at 37°C. Furthermore, we programmed RmInt1 to recognize target sites other than its wild-type site. These retargeted introns efficiently and specifically retrohome into a recipient plasmid target site or a target site present as a single copy in the chromosome, generating a mutation in the targeted gene. Our results extend the range of group II introns available for gene targeting. 相似文献
125.
Summary
1H-NMR spectra of bovine pancreatic trypsin inhibitor (BPTI) both native and oxidized by chloramine T, are reported. The spectrum of the oxidized form is characterized by the appearance of two singlets for methyl group shifted 0.60 and 0.46 ppm downfield with respect to the native form. 相似文献
126.
127.
Pedro Del Bianco Benedeti Lorrayny Galoro da Silva Eduardo Marostegan de Paula Teshome Shenkoru Marcos Inácio Marcondes Hugo Fernando Monteiro Brad Amorati Yenling Yeh Simon Roger Poulson Antonio Pinheiro Faciola 《PloS one》2015,10(11)
The objective of this study was to evaluate the effects of partially replacing dry ground corn with glycerin on ruminal fermentation using a dual-flow continuous culture system. Six fermenters (1,223 ± 21 ml) were used in a replicated 3x3 Latin square arrangement with three periods of 10 d each, with 7 d for diet adaptation and 3 d for sample collections. All diets contained 75% concentrate and three dietary glycerin levels (0, 15, and 30% on dry matter basis), totaling six replicates per treatment. Fermenters were fed 72 g of dry matter/d equally divided in two meals/d, at 0800 and 2000 h. Solid and liquid dilution rates were adjusted daily to 5.5 and 11%/h, respectively. On d 8, 9, and 10, samples of 500 ml of solid and liquid digesta effluent were mixed, homogenized, and stored at -20°C. Subsamples of 10 ml were collected and preserved with 0.2 mL of a 50% H2SO4 solution for later determination of NH3-N and volatile fatty acids. Microbial biomass was isolated from fermenters for chemical analysis at the end of each experimental period. Data were analyzed using the MIXED procedure in SAS with α = 0.05. Glycerin levels did not affect apparent digestibility of DM (P
Lin. = 0.13; P
Quad. = 0.40), OM (P
Lin. = 0.72; P
Quad. = 0.15), NDF (P
Lin. = 0.38; P
Quad. = 0.50) and ADF (P
Lin. = 0.91; P
Quad. = 0.18). Also, glycerin inclusion did not affect true digestibility of DM (P
Lin. = 0.35; P
Quad. = 0.48), and OM (P
Lin. = 0.08; P
Quad. = 0.19). Concentrations of propionate (P < 0.01) and total volatile fatty acids (P < 0.01) increased linearly and concentrations of acetate (P < 0.01), butyrate (P = 0.01), iso-valerate (P < 0.01), and total branched-chain volatile fatty acids, as well as the acetate: propionate ratio (P < 0.01) decreased with glycerin inclusion. Linear increases on NH3-N concentration in digesta effluent (P < 0.01) and on NH3-N flow (P < 0.01) were observed due to glycerin inclusion in the diets. Crude protein digestibility (P = 0.04) and microbial N flow (P = 0.04) were greater in the control treatment compared with the other treatments and responded quadratically with glycerin inclusion. Furthermore, the inclusion of glycerin linearly decreased (P = 0.02) non-ammonia N flow. Glycerin levels did not affect the flows of total N (P
Lin. = 0.79; P
Quad. = 0.35), and dietary N (P
Lin. = 0.99; P
Quad. = 0.07), as well as microbial efficiency (P
Lin. = 0.09; P
Quad. = 0.07). These results suggest that partially replacing dry ground corn with glycerin may change ruminal fermentation, by increasing total volatile fatty acids, and propionate concentration without affecting microbial efficiency, which may improve glucogenic potential of beef cattle diets. 相似文献
128.
129.
Annalisa Cossu Anna Maria Posadino Roberta Giordo Costanza Emanueli Anna Maria Sanguinetti Amalia Piscopo Marco Poiana Giampiero Capobianco Antonio Piga Gianfranco Pintus 《PloS one》2012,7(11)
The cardiovascular benefits associated with diets rich in fruit and vegetables are thought to be due to phytochemicals contained in fresh plant material. However, whether processed plant foods provide the same benefits as unprocessed ones is an open question. Melanoidins from heat-processed apricots were isolated and their presence confirmed by colorimetric analysis and browning index. Oxidative injury of endothelial cells (ECs) is the key step for the onset and progression of cardiovascular diseases (CVD), therefore the potential protective effect of apricot melanoidins on hydrogen peroxide-induced oxidative mitochondrial damage and cell death was explored in human ECs. The redox state of cytoplasmic and mitochondrial compartments was detected by using the redox-sensitive, fluorescent protein (roGFP), while the mitochondrial membrane potential (MMP) was assessed with the fluorescent dye, JC-1. ECs exposure to hydrogen peroxide, dose-dependently induced mitochondrial and cytoplasmic oxidation. Additionally detected hydrogen peroxide-induced phenomena were MMP dissipation and ECs death. Pretreatment of ECs with apricot melanoidins, significantly counteracted and ultimately abolished hydrogen peroxide-induced intracellular oxidation, mitochondrial depolarization and cell death. In this regard, our current results clearly indicate that melanoidins derived from heat-processed apricots, protect human ECs against oxidative stress. 相似文献
130.