全文获取类型
收费全文 | 16502篇 |
免费 | 1124篇 |
国内免费 | 1篇 |
专业分类
17627篇 |
出版年
2023年 | 116篇 |
2022年 | 244篇 |
2021年 | 481篇 |
2020年 | 323篇 |
2019年 | 381篇 |
2018年 | 484篇 |
2017年 | 442篇 |
2016年 | 681篇 |
2015年 | 1014篇 |
2014年 | 1093篇 |
2013年 | 1344篇 |
2012年 | 1551篇 |
2011年 | 1434篇 |
2010年 | 900篇 |
2009年 | 763篇 |
2008年 | 950篇 |
2007年 | 914篇 |
2006年 | 870篇 |
2005年 | 704篇 |
2004年 | 669篇 |
2003年 | 590篇 |
2002年 | 483篇 |
2001年 | 128篇 |
2000年 | 101篇 |
1999年 | 123篇 |
1998年 | 92篇 |
1997年 | 95篇 |
1996年 | 79篇 |
1995年 | 60篇 |
1994年 | 62篇 |
1993年 | 45篇 |
1992年 | 52篇 |
1991年 | 37篇 |
1990年 | 31篇 |
1989年 | 27篇 |
1988年 | 24篇 |
1987年 | 27篇 |
1986年 | 14篇 |
1985年 | 21篇 |
1984年 | 25篇 |
1983年 | 20篇 |
1982年 | 15篇 |
1981年 | 16篇 |
1980年 | 17篇 |
1979年 | 9篇 |
1978年 | 12篇 |
1977年 | 10篇 |
1975年 | 6篇 |
1974年 | 7篇 |
1972年 | 10篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
41.
Crystallization and preliminary X-ray diffraction studies of aSFP, a bovine seminal plasma protein with a single CUB domain architecture. 下载免费PDF全文
J. M. Dias A. L. Carvalho I. Klln J. J. Calvete E. Tpfer-Petersen P. F. Varela A. Romero C. Urbanke M. J. Romo 《Protein science : a publication of the Protein Society》1997,6(3):725-727
Bovine acidic seminal fluid protein (aSFP) is a 1.29 kDa polypeptide of the spermadhesin family built by a single CUB domain architecture. The CUB domain is an extracellular module present in 16 functionally diverse proteins. To determine the three-dimensional structure of aSFP, the protein was crystallized at 21 degrees C by vapor diffusion in hanging drops, using ammonium sulfate, pH 4.7, and polyethyleneglycol 4,000 as precipitants, containing 10% dioxane to avoid the formation of clustered crystals. Elongated prismatic crystals with maximal size of 0.6 x 0.3 x 0.2 mm3 diffract to beyond 1.9 A resolution and belong to space group P2(1)2(1)2(1), with cell parameters a = 52.4 A, b = 41.5 A, c = 48.2 A. There is one aSFP molecule per asymmetric unit, which corresponds to a crystal volume per unit molecular mass of 2.04 A3/Da, and analytical ultracentrifugation analysis show that aSFP is a monomeric protein. 相似文献
42.
Several reports have claimed that the mitochondrial chaperonin cpn60, or a close homolog, is also present in some other subcellular compartments of the eukaryotic cell. Immunoelectron microscopy studies, using a polyclonal serum against cpn60, revealed that the protein is exclusively localized within the mitochondria of rat liver and embryonic Drosophila cells (SL2). Furthermore, no cpn60 immunoreactive material could be found within the nucleus of SL2 cells subjected to a 1 h 37°C heat-shock treatment. In contrast to these findings, immunoelectron microscopy studies, using a cpn60 monoclonal antibody, revealed mitochondrial and extramitochondrial (plasma membrane, nucleus) immunoreactive material in rat liver cells. Surprisingly, the monoclonal antibody also reacted with fixed proteins of the mature red blood cell. The monoclonal antibody, as well as cpn60 polyclonal sera, only recognize mitochondrial cpn60 in Western blots of liver proteins. Furthermore, none of the cpn60 antibodies used in this study recognized blotted proteins from rat red blood cells. Therefore, we suggest that the reported extramitochondrial localization of cpn60 in metazoan cells may be due to cross-reactivity of some of cpn60 antibodies with conformational epitopes also present in distantly related cpn60 protein homologs that are preserved during fixation procedures of the cells. © 1995 Wiley-Liss, Inc. 相似文献
43.
The distribution of aphidicolin-induced chromosomal lesions was analyzed to determine the relative breakage susceptibility of euchromatin and heterochromatin in the cactus mouse, Peromyscus eremicus. The observed breakage was tested against expected distributions corresponding to the karyotypic proportions of autosomal euchromatin, autosomal heterochromatin, X-chromosomal euchromatin, and X-chromosomal heterochromatin. The distribution of induced breakage was independent of sex but dependent on the individual. In all individuals, there was a highly significant (P0.0001) deficiency in the number of breaks observed as compared to expected in autosomal heterochromatin. Sparse observations in the X chromosome and the absence of breaks in the Y chromosome precluded valid statistical tests of the sex-chromosomal distribution of induced breakage. These data indicate that the autosomal heterochromatin of Peromyscus is resistant to aphidicolin-induced chromosomal breakage and argue against a simple relationship between late replication and a general mechanism for chromosomal fragility. 相似文献
44.
In mixed culture of Lactobacillus hilgardii X1B and Leuconostoc oenos X2L, isolated from Argentinian wines, an amensalistic growth response was observed: Leuconostoc oenos did not grow, and after 24 h of incubation at 30°C no viable cells were detected. In pure and mixed cultures, Lactobacillus hilgardii produced hydrogen peroxide early in the growth cycle, reaching the maximum at 24 h. The values of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for the action of hydrogen peroxide on the growth of Leuconostoc oenos were: 4.08g ml-1 and 17.00 g ml-1 respectively. 相似文献
45.
The structure of the grasses megagametophyte is considered to be characteristic enough as to deserve a particular place in the megagametophyte typology. Furthermore, it is compared with those of other Monocotyledonous families to point out embryological affinities.Both are members of the Carrera del Investigador (Conicet, Argentina). 相似文献
46.
Dissatisfaction with the hemodynamic characteristics of available porcine valves prompted a clinical trial of the Ionescu-Shiley percardial xenograft (ISPX) valve. Three hundred fifty-six ISPX valves were implanted consecutively in 326 patients. Operative mortality was 2.6% (2/75) for aortic valve replacement alone and 7.7% (12/155) for aortic valve replacements that included reoperations and combined procedures such as mitral commissurotomy, annuloplasty, and coronary artery bypass. Operative mortality for all patients who underwent mitral valve replacement was 9.5% (14/147). The mean peak systolic gradient pressure in the aortic position was 5.4 mm Hg overall and 4.27 mm Hg with the size 19 mm valve. There were no embolic episodes in patients who received the ISPX valve in the aortic position. The available data indicate that the rate of peripheral embolism with the ISPX valve compares favorably with that of porcine valves. Considering its hemodynamic advantage, if the longterm durability of the full-orifice Ionescu-Shiley pericardial xenograft valve continues to be confirmed by follow-up studies, it is our opinion that it is the biologic valve of choice. 相似文献
47.
Mauricio Goihman-Yahr Leo Pine Maria Cecilia Albornoz Luis Yarzabal Maria Helena De Gomez Blanca San Martin Ana Ocanto Tulio Molina Jacinto Convit 《Mycopathologia》1980,71(2):73-83
Mild sonication was used to obtain single cell suspensions of Paracoccidioides brasiliensis. These cells were intact by microscopic criteria. Direct cell counts in a given inoculum and colony formation on various media were used to determine plating efficiency. Sonicated and nonsonicated cell suspensions were used to study plating efficiency and to estimate viability by means of vital dyes. Methylene blue, Erythrosin B, and Janus green were unreliable when used with P. brasiliensis, but vital dyes were accurate when tested with Candida albicans.Acridine orange gave more meaningful results of viability. Estimates of viability, however, changed significantly as a result of relatively minor alterations in the composition of the suspending medium.In initial experiments, the plating efficiency of P. brasiliensis was dismally low. It descended abruptly with increasing dilution of inoculum. Efficiency was much improved if horse serum was added to brain heart infusion plates or if glucose glycine yeast extract (GGY) plates were incubated at room temperature and mycelial colonies were counted. With the technique we report, current plating efficiency of sonicated suspensions is of the order of 25 %. Our results and procedures have an important bearing upon those studies concerned with in vitro killing of P. brasiliensis in suspensions or with isolating this fungus from clinical or environmental specimens. 相似文献
48.
Ana Maria Andia-Waltenbaugh Charlotte A. Tate Naomi Kraus Friedmann 《Molecular and cellular biochemistry》1981,36(3):177-184
Summary Previous work by this and other laboratories has shown that glucagon administration stimulates calcium uptake by subsequently
isolated hepatic mitochondria. This stimulation of hepatic mitochondrial Ca2+ uptake byin vivo administration of glucagon was further characterized in the present report. Maximal stimulation of mitochondrial Ca2+ accumulation was achieved between 6–10 min after the intravenous injection of glucagon into intact rats. Under control conditions,
Ca2+ uptake was inhibited by the presence of Mg2+ in the incubation medium. Glucagon treatment, however, appeared to obliterate the observed inhibition by Mg2+ of mitochondrial Ca2+ uptake. Kinetic experiments revealed the usual sigmoidicity associated with initial velocity curves for mitochondrial calcium
uptake. Glucagon treatment did not alter this sigmoidal relationship. Glucagon treatment significantly increased the Vmax for Ca2+ uptake from 292±22 to 377±34 nmoles Ca2+ /min per mg protein (n=8) but did not affect the K0.5, (6.5–8.6 μM). Since the major kinetic change in mitochondrial Ca2+ uptake evoked by glucagon is an increase in Vmax, the enhancement mechanism is likely to be an increase either in the number of active transport sites available to Ca2+ or in the rate of Ca2+ carrier movement across the mitochondrial membranes. 相似文献
49.
Lettuce ferredoxin has been purified to homogeneity, with a yield of 18 mg/kg of denerved leaves. It crystallizes in magnificent needles, often clustered in broom-like sheaves. The absorption spectrum showed maxima at 460, 422, 330 and 274 nm,with a ratio A422/A274, of 0.46. The mM absorption coefficient was 9.74 at 422 nm, and 21.62 at 274 nm. This ferredoxin showed a pI = 4.7 and an E′0 = ?425 mV (at pH = 7.7). MWs of 12 400, 11480 and 13000 were obtained by sucrose gradient centrifugation, and on the basis of the amino acid composition and the iron content, respectively, with an average of 12 300. The amino acid analysis showed the existence of one methionine residue per mole, with 105 amino acid residues. There are two iron atoms and two labile sulfide groups per mole; 4 half-cystine residues were found by performic acid oxidation, and 5 cysteine groups when determined by titration with pHMB. The native protein is not fixed on thiol-Sepharose 4B, but it is quantitatively retained after incubation with 8 M urea. Lettuce ferredoxin showed a 62, 58 and 78% effectiveness with the spinach ferredoxin-NADP reductase, nitrite reductase and fructose-1,6-diphosphatase (FDPase), respectively, when compared with the spinach ferredoxin. This different behaviour of both ferredoxins is joined to genetic-structural relationships, and suggests that the role of ferredoxin in FDPase activation is more sophisticated than that of a mere nonspecific reductant. 相似文献
50.