全文获取类型
收费全文 | 6940篇 |
免费 | 260篇 |
国内免费 | 378篇 |
专业分类
7578篇 |
出版年
2023年 | 59篇 |
2022年 | 95篇 |
2021年 | 116篇 |
2020年 | 150篇 |
2019年 | 215篇 |
2018年 | 181篇 |
2017年 | 134篇 |
2016年 | 186篇 |
2015年 | 191篇 |
2014年 | 389篇 |
2013年 | 564篇 |
2012年 | 208篇 |
2011年 | 380篇 |
2010年 | 295篇 |
2009年 | 385篇 |
2008年 | 423篇 |
2007年 | 400篇 |
2006年 | 361篇 |
2005年 | 331篇 |
2004年 | 248篇 |
2003年 | 254篇 |
2002年 | 260篇 |
2001年 | 132篇 |
2000年 | 116篇 |
1999年 | 122篇 |
1998年 | 105篇 |
1997年 | 97篇 |
1996年 | 76篇 |
1995年 | 93篇 |
1994年 | 101篇 |
1993年 | 93篇 |
1992年 | 82篇 |
1991年 | 60篇 |
1990年 | 51篇 |
1989年 | 61篇 |
1988年 | 43篇 |
1987年 | 46篇 |
1986年 | 34篇 |
1985年 | 69篇 |
1984年 | 112篇 |
1983年 | 78篇 |
1982年 | 42篇 |
1981年 | 46篇 |
1980年 | 30篇 |
1979年 | 12篇 |
1978年 | 15篇 |
1977年 | 8篇 |
1976年 | 12篇 |
1975年 | 6篇 |
1973年 | 4篇 |
排序方式: 共有7578条查询结果,搜索用时 0 毫秒
951.
Comparison of neoeriocitrin and naringin on proliferation and osteogenic differentiation in MC3T3-E1
Naringin is considered the main effective compound of Drynaria Rhizome, which is used commonly in the treatment of osteoporosis in traditional Chinese medicine. However, we found neoeriocitrin, a new compound isolated from Drynaria Rhizome, showed a better activity than naringin on proliferation and osteogenic differentiation in MC3T3-E1. Both neoeriocitrin and naringin exhibited the best effect on proliferation and osteogenic differentiation at concentration of 2 μg/ml. Neoeriocitrin more significantly improved proliferation and alkaline phosphatase (ALP) activity as well as up-regulated Runx2, COLI and OCN expression by 56%, 37% and 14% respectively than naringin. Furthermore, neoeriocitrin could rescue the inhibition effect of cell differentiation induced by PD98059 to some degree. Therefore, neoeriocitrin may be a new promising candidate drug for treatment of osteoporosis. 相似文献
952.
Tadamichi Sonoda Hiroyuki Osada Junji Magae Kiyoshi Isono 《Bioscience, biotechnology, and biochemistry》2013,77(5):1259-1263
To investigate the immunosuppressive effects of glutarimide antibiotics including a new antibiotic named epiderstatin, we tested these antibiotics for inhibition of the blastogenesis of mouse spleen cells induced by mitogen stimulation (concanavalin A or lipopolysaccharide). The inhibitory activity was measured by colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Among the glutarimide antibiotics tested, epiderstatin and acetoxycycloheximide especially strongly inhibited the blastogenesis of mouse spleen cells induced by concanavalin A and lipopolysaccharide, however, selectivity between T and B lymphocytes was not observed. 相似文献
953.
Jasmin Strotmeier Stefan Mahrhold Nadja Krez Constantin Janzen Jianlong Lou James D. Marks Thomas Binz Andreas Rummel 《FEBS letters》2014
Botulinum neurotoxins (BoNTs) inhibit neurotransmitter release by hydrolysing SNARE proteins. The most important serotype BoNT/A employs the synaptic vesicle glycoprotein 2 (SV2) isoforms A-C as neuronal receptors. Here, we identified their binding site by blocking SV2 interaction using monoclonal antibodies with characterised epitopes within the cell binding domain (HC). The site is located on the backside of the conserved ganglioside binding pocket at the interface of the HCC and HCN subdomains. The dimension of the binding pocket was characterised in detail by site directed mutagenesis allowing the development of potent inhibitors as well as modifying receptor binding properties. 相似文献
954.
955.
Jane C. Spetzler Vibeke Westphal Jakob R. Winther Morten Meldal 《Journal of peptide science》1998,4(2):128-137
Protein disulphide isomerase is an enzyme that catalyses disulphide redox reactions in proteins. In this paper, fluorogenic and interchain disulphide bond containing peptide libraries and suitable substrates, useful in the study of protein disulphide isomerase, are described. In order to establish the chemistry required for the generation of a split-synthesis library, two substrates containing an interchain disulphide bond, a fluoroescent probe and a quencher were synthesized. The library consists of a Cys residue flanked by randomized amino acid residues at both sides and the fluoroescent Abz group at the amino terminal. All the 20 natural amino acids except Cys were employed. The library was linked to PEGA‒beads via methionine so that the peptides could be selectively removed from the resin by cleavage with CNBr. A disulphide bridge was formed between the bead‒linked library and a peptide containing the quenching chromophore (Tyr(NO2)) and Cys(pNpys) activated for reaction with a second thiol. The formation and cleavage of the interchain disulphide bonds in the library were monitored under a fluoroescence microscope. Substrates to investigate the properties of protein disulphide isomerase in solution were also synthesized. © 1998 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
956.
Célia Parinot Quentin Rieu Jonathan Chatagnon Silvia C. Finnemann Emeline F. Nandrot 《Journal of visualized experiments : JoVE》2014,(94)
Analysis of one of the vital functions of retinal pigment epithelial (RPE) cells, the phagocytosis of spent aged distal fragments of photoreceptor outer segments (POS) can be performed in vitro. Photoreceptor outer segments with stacks of membranous discs containing the phototransduction machinery are continuously renewed in the retina. Spent POS are eliminated daily by RPE cells. Rodent, porcine/bovine and human RPE cells recognize POS from various species in a similar manner. To facilitate performing large series of experiments with little variability, a large stock of POS can be isolated from porcine eyes and stored frozen in aliquots. This protocol takes advantage of the characteristic of photopigments that display an orange color when kept in the dark. Under dim red light, retinae are collected in a buffer from opened eyecups cut in halves. The retinal cell suspension is homogenized, filtered and loaded onto a continuous sucrose gradient. After centrifugation, POS are located in a discrete band in the upper part of the gradient that has a characteristic orange color. POS are then collected, spun, resuspended sequentially in wash buffers, counted and aliquoted. POS obtained this way can be used for phagocytosis assays and analysis of protein activation, localization or interaction at various times after POS challenge. Alternatively, POS can be labeled with fluorophores, e.g., FITC, before aliquoting for subsequent fluorescence quantification of POS binding or engulfment. Other possible applications include the use of modified POS or POS challenge combined with stress conditions to study the effect of oxidative stress or aging on RPE cells. 相似文献
957.
958.
Transgenic Bacillus thuringiensis Berliner (Bt) crops receive particular attention because they carry genes encoding insecticidal proteins that might negatively affect non‐target arthropods. Here, laboratory experiments were conducted to evaluate the impact of Cry1Ab‐expressing transgenic maize [5422Bt1 (event Bt11) and 5422CBCL (MON810)] on the biological parameters of two non‐target arthropods, the aphid Rhopalosiphum maidis (Fitch) (Hemiptera: Aphididae) and its predator the ladybeetle Propylea japonica (Thunberg) (Coleoptera: Coccinellidae). In a long‐term assay (three generations), no significant differences were found between R. maidis fed Bt maize and those fed a near‐isogenic line (5422) when individual parameters were compared, including nymph development time, adult longevity, aphid spawning period, and fecundity. No negative effects were detected throughout the life cycle of P. japonica in aphids’ feeding amount, development (nymphs, pupae, adults, and progeny eggs), fecundity, or egg hatching when they preyed on Bt maize‐fed aphids compared with non‐Bt maize treatments. A tritrophic assay revealed that Cry1Ab was highly diluted through the food chain (Bt maize leaves, R. maidis, and P. japonica), as detected by an enzyme‐linked immunosorbent assay (ELISA). In conclusion, although Cry1Ab concentrations in maize leaves increased as the plants developed, Cry1Ab levels were significantly reduced in the aphid R. maidis, and no traces of Cry1Ab were detected in P. japonica preying on Bt maize‐fed aphids. The two hybrids of Bt maize expressing Cry1Ab had no negative effects on the measured biological parameters of the aphid R. maidis or its predator, the ladybeetle P. japonica. 相似文献
959.
Fry SR Li J de las Heras R McCourt JA Arel E Kachab EH Hazell SL Huang CY 《Biochemical and biophysical research communications》2008,372(4):542-546
Enzyme immunoassays (EIAs) are widely used in the clinical laboratory and research institutes for the detection of biologically relevant analytes. Almost all EIAs are heterogeneous in nature and require multiple steps of process. In contrast, homogeneous immunoassays (HA) offer a simplified one-step approach with a number of potential advantages over contemporary heterogeneous EIAs such as higher throughput and greater clinical utility. Utilizing TEM-1 β-lactamase as a reporter enzyme, we have developed HAs based on in vitro protein fragment complementation (PCA) for the detection of antibodies and potentially be used for antigens or other biomarkers. In this proof-of-principle study we demonstrate the successful in vitro differentiation of anti-herpes simplex virus (HSV) type-1 and type-2 Immunoglobulin G (IgG) in human serum with high sensitivity and specificity. 相似文献
960.
Beta-lactoglobulin (beta-LG), one of the most investigated proteins, is a major bovine milk protein with a predominantly beta structure. The structural function of the only alpha-helix with three turns at the C-terminus is unknown. Vitamin D(3) binds to the central calyx formed by the beta-strands. Whether there are two vitamin D binding-sites in each beta-LG molecule has been a subject of controversy. Here, we report a second vitamin D(3) binding site identified by synchrotron X-ray diffraction (at 2.4 A resolution). In the central calyx binding mode, the aliphatic tail of vitamin D(3) clearly inserts into the binding cavity, where the 3-OH group of vitamin D(3) binds externally. The electron density map suggests that the 3-OH group interacts with the carbonyl of Lys-60 forming a hydrogen bond (2.97 A). The second binding site, however, is near the surface at the C-terminus (residues 136-149) containing part of an alpha-helix and a beta-strand I with 17.91 A in length, while the span of vitamin D(3) is about 12.51 A. A remarkable feature of the second exosite is that it combines an amphipathic alpha-helix providing nonpolar residues (Phe-136, Ala-139, and Leu-140) and a beta-strand providing a nonpolar (Ile-147) and a buried polar residue (Arg-148). They are linked by a hydrophobic loop (Ala-142, Leu-143, Pro-144, and Met-145). Thus, the binding pocket furnishes strong hydrophobic force to stabilize vitamin D(3) binding. This finding provides a new insight into the interaction between vitamin D(3) and beta-LG, in which the exosite may provide another route for the transport of vitamin D(3) in vitamin D(3) fortified dairy products. Atomic coordinates for the crystal structure of beta-LG-vitamin D(3) complex described in this work have been deposited in the PDB (access code 2GJ5). 相似文献