全文获取类型
收费全文 | 1674篇 |
免费 | 70篇 |
国内免费 | 4篇 |
出版年
2021年 | 11篇 |
2020年 | 7篇 |
2019年 | 7篇 |
2018年 | 16篇 |
2017年 | 7篇 |
2016年 | 37篇 |
2015年 | 41篇 |
2014年 | 54篇 |
2013年 | 116篇 |
2012年 | 94篇 |
2011年 | 119篇 |
2010年 | 65篇 |
2009年 | 53篇 |
2008年 | 95篇 |
2007年 | 92篇 |
2006年 | 127篇 |
2005年 | 97篇 |
2004年 | 104篇 |
2003年 | 108篇 |
2002年 | 97篇 |
2001年 | 28篇 |
2000年 | 22篇 |
1999年 | 16篇 |
1998年 | 19篇 |
1997年 | 26篇 |
1996年 | 20篇 |
1995年 | 21篇 |
1994年 | 21篇 |
1993年 | 18篇 |
1992年 | 34篇 |
1991年 | 25篇 |
1990年 | 12篇 |
1989年 | 16篇 |
1988年 | 10篇 |
1987年 | 11篇 |
1986年 | 6篇 |
1985年 | 6篇 |
1984年 | 15篇 |
1983年 | 9篇 |
1982年 | 7篇 |
1981年 | 14篇 |
1980年 | 6篇 |
1979年 | 6篇 |
1978年 | 4篇 |
1977年 | 4篇 |
1975年 | 5篇 |
1974年 | 3篇 |
1972年 | 3篇 |
1963年 | 3篇 |
1961年 | 2篇 |
排序方式: 共有1748条查询结果,搜索用时 15 毫秒
41.
Kazuyuki Nakajima Masato Shimojo Makoto Hamanoue Shoichi Ishiura Hideo Sugita Shinichi Kohsaka 《Journal of neurochemistry》1992,58(4):1401-1408
In the course of studying the secretory products of microglia, we detected protease activity in the conditioned medium. Various proteins (casein, histone, myelin basic protein, and extracellular matrix) were digested. The protease activity was characterized by using purified myelin basic protein as a substrate. Maximal activity was observed at neutral pH levels (7-8), which was different from the optimum pH level of proteolytic activity observed in the cell homogenate. The activity was inhibited approximately 60 and 50% by 1 mM phenylmethylsulfonyl fluoride and 40 microM elastatinal, respectively. In gel filtration, the major activity, which was inhibited in the presence of N-methoxysuccinyl-Ala-Ala-Pro-Val-methyl chloride, eluted at a position corresponding to a molecular mass of approximately 25 kDa. These results suggest that the major protease present in microglial conditioned medium is elastase or an elastase-like protease. This suggestion was confirmed by the finding that the 25-kDa protein band was stained with anti-elastase antiserum by western blotting. De novo synthesis of elastase in microglia was supported by [35S]methionine incorporation. In the presence of lipopolysaccharide, the secretory elastase decreased. These results demonstrate that microglia secrete proteases, one of which was identified as elastase. The significance of this enzyme production in physiological and pathological conditions is discussed. 相似文献
42.
Takashi Tobe Shigeo Nakajo Atsutaka Tanaka Akira Mitoya Kumiko Omata Kazuyasu Nakaya Motowo Tomita Yasuharu Nakamura 《Journal of neurochemistry》1992,59(5):1624-1629
A new acidic protein with a molecular weight of 14,000 was purified from rat brain, in which it was specifically expressed, and partially sequenced by protein sequencing. On the basis of results obtained from the amino acid sequences, mixed oligonucleotides were synthesized and used as probes to clone a cDNA from a rat brain cDNA library. The cloned cDNA provided the full-length sequence of the 14-kDa protein. Northern blot hybridization using total RNA from several tissues of the rat provided evidence that the 14-kDa protein was expressed specifically in rat brain. Transfection of this cDNA into mammalian cells resulted in expression of the 14-kDa protein. The amino acid sequence predicted from the cDNA of the rat brain 14-kDa protein contained 137 amino acid residues. A hydropathy profile revealed a hydrophobic domain (amino acids 60-80) flanked by highly hydrophilic stretches on both sides. Whereas the N-terminal region of the 14-kDa protein contained four repeating motifs, EKTKEGV, the C-terminal domain was rich in glutamic acid and proline. A computer search of the amino acid sequence of the 14-kDa protein indicated no homology to any other protein reported so far. 相似文献
43.
M Hashiramoto T Kadowaki A E Clark A Muraoka K Momomura H Sakura K Tobe Y Akanuma Y Yazaki G D Holman 《The Journal of biological chemistry》1992,267(25):17502-17507
The structure-function relationship of the HepG2/erythrocyte-type glucose transporter (GLUT1) has been studied by in vitro site-directed mutagenesis. Chinese hamster ovary clones in which glucose transporters were transfected were shown by Western blotting with a GLUT1 anti-COOH-terminal peptide antibody to have expression levels of Gln282----Leu, Asn288----Ile, and Asn317----Ile mutations that were comparable with the wild type. All three mutant GLUT1 clones had high 2-deoxy-D-glucose transport activity compared with a nontransfected clone, suggesting that these residues are not absolutely required for the transport function. We have examined the possibility that the inner and outer portions of the transport pathway are structurally separate by measuring the interaction of the mutant transporters with the inside site-specific ligand cytochalasin B and the outside site-specific ligand 2-N-4-(1-azi-2,2,2-trifluoroethyl)benzoyl-1,3-bis(D-mannos-4 -yloxy)-2- propylamine (ATB-BMPA). All three mutant GLUT1 clones showed high levels of cytochalasin B labeling, and the N288I and N317I mutants showed high levels of ATB-BMPA labeling. In contrast to the transport and cytochalasin B labeling results, the transmembrane helix 7 Gln282----Leu mutant was labeled by ATB-BMPA to a level that was only 5% of the level observed in the wild type. We have confirmed that this mutant was defective in the outer site by comparing the inhibition of wild-type and mutant 2-deoxy-D-glucose transport by the outside site-specific ligand 4,6-O-ethylidene-D-glucose. 4,6-O-Ethylidene-D-glucose inhibited wild-type transport with a Ki of approximately 12 mM, but this was increased to greater than 120 mM in the Gln282----Leu mutant. Thus, of the 3 residues mutated in this study, only glutamine 282 substitution causes a major perturbation in function, and this is a specific and striking reduction in the affinity for the outside site-specific ligands ATB-BMPA and 4,6-O-ethylidene-D-glucose. 相似文献
44.
Structural and Kinetic Properties of NADP-Malic Enzyme from the Inducible Crassulacean Acid Metabolism Plant Mesembryanthemum crystallinum L. 总被引:1,自引:0,他引:1
Saitou Kazuyuki; Agata Waichi; Asakura Masae; Kubota Fumitake 《Plant & cell physiology》1992,33(5):595-600
NADP-malic enzyme (EC 1.1.1.40
[EC]
), which is involved in Crassulaceanacid metabolism (CAM), was purified to electrophoretic homogeneityfrom the leaves of the inducible CAM plant Mesembryanthemumcrystallinum. The NADP-malic enzyme, which was purified 1,146-fold,has a specific activity of 68.8 µmol (mg protein)1min1. The molecular weight of the subunits of the enzymewas 64 kDa. The native molecular weight of the enzyme was determinedby gel-filtration to be 390 kDa, indicating that the purifiedNADP-malic enzyme is a hexamer of identical subunits. The optimalpH for activity of the enzyme was around 7.2. Double-reciprocalplots of the enzymatic activity as a function of the concentrationof L-malate yielded straight lines both at pH 7.2 and at pH7.8 and did not reveal any evidence for cooperativity of bindingof L-malate. The Km value for L-malate was 0.35 mM. Hill plotsof the activity as a function of the concentration of NADP+indicated positive cooperativity in the binding of NADP+ tothe enzyme with a Hill coefficient (nH) of 2.0. An S0.5 value(the concentration giving half-maximal activity) of 9.9 µMfor NADP+ was obtained. Oxaloacetate inhibited the activityof the NADP-malic enzyme. Effects of succinate and NaHCO3 onthe activity of NADP-malic enzyme were small. (Received October 30, 1991; Accepted May 1, 1992) 相似文献
45.
46.
Photoaffinity labeling of allatostatin receptor proteins in the corpora allata of the cockroach, Diploptera punctata. 总被引:2,自引:0,他引:2
M Cusson G D Prestwich B Stay S S Tobe 《Biochemical and biophysical research communications》1991,181(2):736-742
Membranes of corpora allata, obtained from Diploptera punctata virgin females, were incubated for 45 min with a bioactive, radioiodinated azidosalicylamide photoaffinity analogue of allatostatin-1. Irradiation of the incubation mixture at 254 nm, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography, revealed the presence of two bands (59 and 39 kDa) whose specific labeling was demonstrated by addition of excess allatostatin. In brain preparations, clear competition by excess allatostatin was shown for a 41 kDa protein. The abdominal fat body also contained proteins (38, 41, 60 kDa) which bound specifically to the photoaffinity analogue. The proteins identified in corpora allata are likely involved in allatostatin recognition, but the functions of those identified in the brain and fat body are as yet undetermined. 相似文献
47.
Y Nakano Y Sugita Y Ishikawa N H Choi T Tobe M Tomita 《Biochimica et biophysica acta》1991,1074(2):326-330
Decay-accelerating factor (DAF) was purified from human pooled urine by conventional techniques. The urine DAF was separated into two peaks, pool I and pool II, by gel chromatography. DAF-U1 was isolated from pool I by hydrophobic chromatography, and DAF-U2 from pool II by anti-DAF IgG column. The specific activities of DAF-U1 and DAF-U2 to decay membrane-phase C5 convertase were about 3% and 70% of membrane form DAF, respectively. However, both urine DAFs revealed a similar activity to each other and slightly higher activity than that of membrane form DAF in decay-accelerating fluid-phase C3 convertase of the alternative pathway. 相似文献
48.
Summary The kinetic process of folding of bovine pancreatic ribonuclease A in a2H2O environment at pH 1.2 was examined by a recently developed temperature-jump NMR method (Akasaka et al., (1990) Rev. Sci. Instrum.61, 66–68). Upon temperature-jump down from 45°C to 29°C, which was attained within 6 s, the proton NMR spectral changes were followed consecutively in time intervals of seconds. There was a rapid spectral change, which was finished within the jump period, followed by a much slower process which lasted for a minute or longer. Rates of the slower process were measured at different positions of the polypeptide chain as intensity changes of individual His and Tyr proton signals of the folded conformer and as intensity changes of aliphatic and His protons of the unfolded conformer. Most of these rates coincided with each other within experimental error with an average value of 2.8×10–2s–1. The result gave clear experimental evidence that the slow folding of RNase A at low pH is a cooperative process involving most regions of the molecule, not only thermodynamically, but kinetically as well. 相似文献
49.
50.