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21.
Anthony B. Thompson Edward H. Allison Ben P. Ngatunga 《Environmental Biology of Fishes》1996,47(3):235-254
Synopsis Lake Malawi/Niassa is the second largest rift valley lake in Africa, with an area of 28 800 km2, and an average and maximum depth of 292 m and>700 m, respectively. The lake is well known for the great diversity of fish occurring in the inshore zone. However, the offshore fish community is poorly documented. To rectify this, regular sampling was undertaken over two years, using trawl and gillnets at six offshore locations. This paper reports on the species composition, spatial distribution and breeding biology of the dominant cichlids species from the offshore pelagic zone. Cichlids formed approximately 88% of the offshore fish biomass. Most abundant were two species of zooplanktivores in the genus Diplotaxodon that made up 71% of the offshore fish biomass. An undescribed species, given the cheironym D. bigeye, was mainly found at a depth of 220 m during the day, but moved into near surface waters at night when the moon was full. This species was absent from the shallow regions of the lake. The most abundant offshore species was D. limnothrissa, which was distributed evenly throughout the lake to depths of 220 m. A less common offshore zooplanktivore was Copadichromis quadrimaculatus that formed 5% of the biomass and was confined to the upper 100 m of the water column. The main piscivores were in the genus Rhamphochromis and formed approximately 10% of the offshore fish biomass. The two dominant taxa were R. longiceps and the large Rhamphochromis group, and both were more common in the southern half of the lake. The former occurred mainly in the upper 100 m of the water column and the latter mainly at depths of 100–150 m. The length at maturity and fecundity for the dominant offshore species were estimated and seasonal breeding cycles determined from gonad activity and gonado-somatic indices. 相似文献
22.
Anthony P. Fordham-Skelton F. Safadi M. Golovkin A. S. N. Reddy 《Plant Molecular Biology Reporter》1994,12(4):358-366
Calmodulin labeled with125I or34S has been used to screen expression libraries to isolate cDNAs encoding calmodulin-binding proteins (CBPs) from several eukaryotic
systems. The use of radiolabeled calmodulin has, however, several disadvantages. We have developed a nonradiactive method
to isolate cDNAs for CBPs using biotinylated calmodulin. Screening of a cDNA library in an expression vector with biotinylated
calmodulin resulted in the isolation of cDNAs encoding CBPs. Avidin and biotin blocking steps, prior to incubation of the
filters with biotinylated calmodulin, are found to be essential to eliminate the cDNAs that code for biotin-containing polypeptides.
The cDNA clones isolated using this nonradioactive method bound calmodulin in a calcium-dependent manner. The binding of biotinylated
calmodulin to these clones was completely abolished by ethylene glycolbis(\-aminoethylether)-N,N′-tetraacetic acid (EGTA),
a calcium chelator. Furthermore, the isolated cDNAs were confirmed by probing the clones with35S-labeled calmodulin. All the isolated clones bound to radiolabeled calmodulin in the presence of calcium but not in the presence
of EGTA. The method described here is simple, fast, and does not involve preparation and handing of radiolabeled calmodulin.
All the materials used in this method are commercially available; hence, this procedure should be widely applicable to isolate
cDNAs encoding CBPs from any eukaryotic organism. 相似文献
23.
Metallothionein (MT) is a ubiquitous mammalian protein comprising 61 or 62 nonaromatic amino acids of which 20 are cysteine residues. The high sulfhydryl content imparts to this protein a unique and remarkable ability to bind multiple metal ions in structurally significant metal–thiolate clusters. MT can bind seven divalent metal ions per protein molecule in two domains with exclusive tetrahedral metal coordination. The domain stoichiometries for the M7S20 structure are M4(Scys)11 (α domain) and M3(Scys)9 (β domain). Up to 12 Cu(I) ions can displace the 7 Zn2+ ions bound per molecule in Zn7–MT. The incoming Cu(I) ions adopt a trigonal planar geometry with domain stoichiometries for the Cu12S20 structure of Cu6(Scys)11 and Cu6(Scys)9 for the α and β domains, respectively. The circular dichroism (CD) spectra recorded as Cu+ is added to Zn7–MT to form Cu12–MT directly report structural changes that take place in the metal binding region. The spectrum arises under charge transfer transitions between the cysteine S and the Cu(I); because the Cu(I)–thiolate cluster units are located within the chiral binding site, intensities in the CD spectrum are directly related to changes in the binding site. The CD technique clearly indicates stoichiometries of several Cu(I)–MT species. Model Cu(I)–thiolate complexes, using the tripeptide glutathione as the sulfhydryl source, were examined by CD spectroscopy to obtain transition energies and the Cu(I)–thiolate coordination geometries which correspond to these bands. Possible structures for the Cu(I)–thiolate clusters in the α and β domains of Cu12–MT are proposed. © 1994 Wiley-Liss, Inc. 相似文献
24.
Anthony T.W. Cheung Richard B. Moss Geoffrey Kurland Albin B. Leong William J. Novick 《Journal of medical primatology》1993,22(4):257-262
We have recently established a rhesus monkey model of chronic Pseudomonas aeruginosa (PA) endobronchitis by bronchoscopic instillation of PA-embedded agar beads. All experimental animals developed chronic neutrophilic endobronchitis similar to chronic PA endobronchitis in cystic fibrosis (CF). Histopathologic studies further confirmed similarities to chronic PA endobronchitis in CF, including marked peribronchial inflammation, epithelial damage, presence of degraded cilia and ciliary abnormalities, appearance of PA bacterial clusters, mucosal hyperplasia, goblet cell hypertrophy/hypersecretion, airway obstruction, alveolar abnormalities, bronchiectasis, and fibrosis. 相似文献
25.
Stuart B. Moss Brenda L. Burnham Anthony R. Bellv 《Molecular reproduction and development》1993,34(2):164-174
The presence of lamin proteins in mouse spermatogenic cells has been examined by using an anti-lamin AC and an anti-lamin B antisera which recognize somatic lamins A and C, and somatic lamin B, respectively. Anti-lamin B binds to the nuclear periphery of all cell types examined, including Sertoli cells, primitive type A spermatogonia, preleptotene, leptotene, zygotene and pachytene spermatocytes, and round spermatids. In sperm nuclei, the antigenic determinants are localized to a narrow domain of the nucleus. However, after removing the perinuclear theca, anti-lamin B localizes to the entire nuclear periphery in a punctate pattern, suggesting that it is binding to determinants previously covered by the theca constituents. On immunoblots anti-lamin B reacts with a ~ 68 kD polypeptide in all germ cells and, to a lesser extent, with four additional polypeptides present only in meiotic and post-meiotic nuclear matrices. Anti-lamin AC also reacts with the perinuclear region of the somatic cells in the testes, in particular, those of the interstitium and also the Sertoli cells of the seminiferous epithelium. In contrast to anti-lamin B, anti-lamin AC does not bind to the germ cells at any stage of spermatogenesis. In addition, nuclear matrix proteins from isolated spermatogenic cells do not bind anti-lamin AC on immunoblots, suggesting the lack of reactivity is not due to the masking of any antigenic sites. These data demonstrate that germ cells contain lamin B throughout spermatogenesis, even during meiosis and spermiogenesis when the nuclear periphery lacks a distinct fibrous lamina. © 1993 Wiley-Liss, Inc. 相似文献
26.
27.
Barry K Derham J Clive Ellory Anthony J Bron John J Harding 《European journal of biochemistry》2003,270(12):2605-2611
Alpha-crystallin, a molecular chaperone and lens structural protein protects soluble enzymes against heat-induced aggregation and inactivation by a variety of molecules. In this study we investigated the chaperone function of alpha-crystallin in a more physiological system in which alpha-crystallin was incorporated into red cell 'ghosts'. Its ability to protect the intrinsic membrane protein Na/K-ATPase from external stresses was studied. Red cell ghosts were created by lysing the red cells and removing cytoplasmic contents by size-exclusion chromatography. The resulting ghost cells retain Na/K-ATPase activity. alpha-Crystallin was incorporated in the cells on resealing and the activity of Na/K-ATPase assessed by ouabain-sensitive 86Rb uptake. Incubation with fructose, hydrogen peroxide and methylglyoxal (compounds that have been implicated in diabetes and cataract formation) were used to test inactivation of the Na/K pump. Intracellular alpha-crystallin protected against the decrease in ouabain sensitive 86Rb uptake, and therefore against inactivation induced by all external modifiers, in a dose-dependent manner. 相似文献
28.
29.
30.
José A. A. Sant''Ana Pereira Arnold De Haan Andy Wessels Antoon F. M. Moorman Anthony J. Sargeant 《The Histochemical journal》1995,27(9):715-722
Summary In the present study we report a novel histochemical method which, by sequential pre-incubations in alkaline and acidic media,
selectively differentiates muscle fibres expressing myosin heavy chain IIX, on the basis of a specific profile for myofibrillar
actomyosin ATPase (mATPase) activity. The enzyme reactions were tested for specificity by means of anti-myosin heavy chain
monoclonal antibodies, which were characterized on Western blots of muscle homogenates. Enzyme histochemical reactions with
the traditional pH buffers were compared to those of the new method and, in conjunction with the immunoreactions, used to
confirm the relationship between MyHC expression and the distinct profiles for mATPase. Imrnunohistochemical reactions demonstrated
that the new method only differentiates those fibres expressing myosin heavy chain IIX. The method revealed a continuum in
which the intermediate staining intensities corresponded to hybrid fibres expressing myosin heavy chain IIX in combination
with either the IIA or IIB forms. Quantitative histochemistry and immunohistochemistry (by image analysis), used to examine
the relationship between staining intensities for mATPase and amounts of myosin heavy chain IIX expression, revealed that
the new method discriminates well between hybrid fibres expressing variable amounts of the IIX isoform (r2 = 0.93). 相似文献