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991.
F. Fierro E. Montenegro S. Gutiérrez J. F. Martín 《Applied microbiology and biotechnology》1996,44(5):597-604
The organization of the genes of the penicillin cluster has been studied in three different mutants of P. chrysogenum impaired in penicillin biosynthesis. The three blocked mutants (derived from the parental strain P. chrysogenum Bb-1) lacked the genes pcbAB, pcbC and penDE of the penicillin biosynthetic pathway and were unable to form isopenicillin N synthase and isopenicillin N acyltransferase. All strains were identified as P. chrysogenum derivatives by fingerprinting analysis with (GTG)n as a probe. The borders of the deleted region were cloned and sequenced, showing the same junction point in the three mutants. The deleted DNA region was found to be identical to that described in P. chrysogenum npe10. The frequent deletion of the pen gene cluster at this point may indicate that this cluster is located in an unstable genetic region, flanked by hot spots of recombination, that is easily lost by mutagen-induced recombination. 相似文献
992.
Living yeast cells can be selectively stained with the lipophilic cationic cyanine dye DiOC6(3) in a mitochondrial membrane potential-dependent manner. Our study extends the use of flow cytometric analysis and sorting to DiOC6(3)-stained yeast cells. Experimental conditions were developed that prevented the toxic side effect of the probe and gave a quantitative correlation between fluorescence and mitochondrial membrane potential, without any staining of other membranes. The localization of the fluorochrome was checked by confocal microscopy and image cytometry. The mitochondrial membrane alterations were also tested through cardiolipin staining with nonyl acridine orange. Differences in light scattering and in fluorescence were detected in mutants (rho-, rho degrees, mit-, or pet-) and wild-type (rho+mit+) populations of yeast. The dye uptake of respiratory-deficient yeast strains was significantly reduced as compared to that of the wild-type. Application of an uncoupler (mCICCP), which collapsed the mitochondrial membrane potential (alphapsi(m)), led to a drastic reduction of the dye uptake. It was observed that a decrease in deltapsi(m), was usually correlated with a decrease in cardiolipin stainability by nonyl acridine orange (NAO). Quantitative flow cytometry is a fast and reproducible technique for rapid screening of yeast strains that might be suspected of respiratory dysfunction and/or mitochondrial structural changes. We give evidence that it is an adequate method to characterize and isolate respiratory mutants through sorting procedure, with selective enrichment of the population studied in respiring or non-respiring yeast cells. Confocal microscopy and image cytometry corroborate the flow cytometry results. 相似文献
993.
Settler agriculture and the dynamics of resource allocation in frontier environments 总被引:1,自引:0,他引:1
Francisco J. Pichón 《Human ecology: an interdisciplinary journal》1996,24(3):341-371
This article attempts to conceptualize the dynamics of resource allocation by colonist farmers under the unique conditions of land abundance and labor scarcity which characterize frontier environments, such as the smallholder agricultural settlement areas in the Amazon basin. In contrast, most previous theoretical literature on household agricultural decision making and land-use change in rural areas considers conditions of high population density and land scarcity, and is not, therefore, adequate for understanding critical land-use changes which may be occurring in frontier regions. This article first discusses the appropriateness and inadequacies of the analytical frameworks commonly used to explain the expansion of settler agriculture into remote forest regions and the unsustainable land-use practices observed in these areas. This review serves as the basis for characterizing resource allocation under the particular conditions of frontier environments. A conceptual advance in the analysis is its consideration of the way institutional/policy factors and farm-level characteristics can interact to produce land-use outcomes. This knowledge is essential to understand not only the social and economic factors affecting present land use and choice of technology, but also those factors influencing farmers' demand for more optimal systems of land use which are consistent with varying agro-ecological potentials, demographic situations, and their own management capacity. 相似文献
994.
The majora2–6 sialoglycoproteins in detergent-extracts of Kurloff cells were purified by anion-exchange andSambucus nigra agglutinin-affinity chromatographies. The similar ultrastructural localisations of (1)S. nigra agglutinin-gold conjugates and (2) acid phosphatase activities on the Kurloff body and particularly on its myelin figures indicated that the majora2-6 sialoglycoproteins of the Kurloff cell had acid phosphatase activity. Two-dimensional electrophoresis showed that these tartrate-sensitive phosphatases corresponded to 2 acidic (pI 3.4–3.7) polypeptides of 36 and 34 kDa. Hydrolysis with peptide-N-glycosidases F gave a 33 kDa apoprotein rich in alanine, glutamic acid, tyrosine and lysin. A lectin-affinity study demonstrated that they contained hybrid type bisected and fucosylatedN-linked oligosaccharides. Cytotoxic properties were previously attributed to Kurloff cells and other studies suggested that not only acid phosphatases but alsoa2-6-linked sialic acid residues themselves may participate in natural killer activity. 相似文献
995.
Neural control of the expression of a Ca2+-activated K+ channel involved in the induction of myotonic-like characteristics 总被引:2,自引:0,他引:2
Beatriz U. Ramírez Maria Isabel Behrens Cecilia Vergara 《Cellular and molecular neurobiology》1996,16(1):39-49
Summary 1. Expression of the apamin-sensitive K+ channel (SK+) in rat skeletal muscle is neurally regulated. The regulatory effect of the nerve over the expression of some muscle ion
channels has been attributed to the electrical activity triggered by the nerve and/or to a trophic effect of some molecules
transported from the soma to the axonal endings.
2. SK+ channels apparently are involved in myotonic dystrophy (MD), therefore understanding the factors that regulate their expression
may ultimately have important clinical relevance.
3. To establish if axoplasmic transport is involved in this process, we used two experimental approaches in adult rats: (a)
Both sciatic nerves were severed, leaving a short or a long nerve stump attached to the anterior tibialis (AT). (b) Colchicine
or vinblastine (VBL), two axonal transport blockers of different potencies, was applied on one leg to the sciatic nerve. To
determine whether electrical activity affects the expression of SK+ channels, denervated AT were directly stimulated. The corresponding contralateral muscles were used as controls.
4. With these experimental conditions we measured (a) apamin binding to muscle membranes, (b) muscle contractile characteristics,
and (c) electromyographic activity.
5. In the short- and long-nerve stump experiments, 5 days after denervation125I-apamin binding to AT membranes was 2.0 times higher in the short-stump side. This difference disappeared at longer times.
The delayed expression of SK+ channels in the muscle left with a longer nerve stump can be attributed to the extra axoplasm contained in the longer stump,
which maintains a normally repressive signal for a longer period of time. Ten to 15 days after application of axonal transport
blockers we found that the muscle half-relaxation time increased in the drug-treated side and apamin partially reverted the
prolonged relaxation. Myotonic-like discharges specifically blockable by apamin were always present in the drug-treated leg.125I-Apamin binding, which is undetectable in a microsomal preparation from hind leg control muscles, was increased in the drug-treated
preparations. Apamin binding to denervated and stimulated AT muscles was lower than in the contralateral unstimulated muscles
[3.3±1.0 vs 6.8±0.8 (n=4) fmol/mg protein].
6. Our results demonstrate that electrical activity and axoplasmic transport are involved in the control of expression of
SK+ in rat skeletal muscle. However, the increased expression of this channel induces myotonic-like characteristics that are
reversed by apamin. This myotonic activity could be a model for MD. 相似文献
996.
Signalling by protein kinase C isoforms in the heart 总被引:11,自引:0,他引:11
Understanding transmembrane signalling process is one of the major challenge of the decade. In most tissues, since Fisher and Krebs's discovery in the 1950's, protein phosphorylation has been widely recognized as a key event of this cellular function. Indeed, binding of hormones or neurotransmitters to specific membrane receptors leads to the generation of cytosoluble second messengers which in turn activate a specific protein kinase. Numerous protein kinases have been so far identified and roughly classified into two groups, namely serine/threonine and tyrosine kinases on the basis of the target amino acid although some more recently discovered kinases like MEK (or MAP kinase kinase) phosphorylate both serine and tyrosine residues.Protein kinase C is a serine/threonine kinase that was first described by Takai et al. [1] as a Ca- and phospholipid-dependent protein kinase. Later on, Kuo et al. [2] found that PKC was expressed in most tissues including the heart. The field of investigation became more complicated when it was found that the kinase is not a single molecular entity and that several isoforms exist. At present, 12 PKC isoforms and other PKC-related kinases [3] were identified in mammalian tissues. These are classified into three groups. (1) the Ca-activated -, -,and -PKCs which display a Ca-binding site (C2); (2) the Ca-insensitive -, -, -, -, and -PKCs. The kinases that belong to both of these groups display two cystein-rich domains (C1) which bind phorbol esters (for recent review on PKC structure, see [4]). (3) The third group was named atypical PKCs and include , , and -PKCs that lack both the C2 and one cystein-rich domain. Consequently, these isoforms are Ca-insensitive and cannot be activated by phorbol esters [5]. In the heart. evidence that multiple PKC isoforms exist was first provided by Kosaka et al. [6] who identified by chromatography at least two PKC-related isoenzymes. Numerous studies were thus devoted to the biochemical characterization of these isoenzymes (see [7] for review on cardiac PKCs) as well as to the identification of their substrates.This overview aims at updating the present knowledge on the expression, activation and functions of PKC isoforms in cardiac cells. (Mol Cell Biochem 157: 65–72, 1996) 相似文献
997.
Applications of intrinsic fluorescence measurements in the study of Ca2+-transport ATPases are reviewed. Since the initial reports showing that the fluorescence emission was sensitive to Ca2+ binding, a substantial amount of work has focused on the use of both steady-state and time-resolved fluorescence spectroscopy to investigate structure-function relationships in sarcoplasmic reticulum and plasma membrane Ca2+-ATPases. These studies have revealed ligand-induced conformational changes, as well as provided information on protein-protein, protein-solvent and/or protein-lipid interactions in different functional states of these proteins. The main results of these studies, as well as possible future prospects are discussed. 相似文献
998.
V. Alvarez E. Coto S. González-Roces C. López-Larrea F. Setién C. López-Larrea 《Immunogenetics》1996,44(6):446-452
N-formyl peptides (FMLP) and complement fragment C5a are neutrophil chemoattractants. In humans, a single-copy gene was identified
for the C5a receptor, and the receptor for FMLP (FPR1) is encoded by a single gene that shows 53% amino acid similarity to
the C5aR. Two other humanFPR1 homologues,FPR-like 1 (FPR2/FPRL1) andFPR-like 2 (FPRL2) have been cloned. The human C5aR, FPR1, FPRL1, and FPRL2 are physically linked. By direct sequencing or by sequencing plasmid
clones we studied theC5aR andFPR genes from four non-human primates (chimpanzee, gorilla, orangutan, and macaque). The sequences showed 95%–99% similarity
to the human homologues, with the major divergences observed in macaque. In these genes, the transmembrane and the cytoplasmic
domains are highly conserved, while the highest divergence corresponded to the extracellular loops involved in ligand binding.
Additionally, we constructed a physical map of these genes in non-human primates. In all species the four genes were physically
linked and we defined the relative orientation of the four genes in primates:C5aR>FPR1>FPR2 (FPRL1)>FPRL2.
The nucleotide sequence data reported in this paper have been submitted to the EMBL/GenBank nucleotide sequence databases
and have assigned the accession numbers X97730 (PTC5aR), X97731 (MMC5aR), X97732 (PPC5aR), X97730 (GGC5aR), X97734 (MMFPR1), X97735 (PPFPR1), X97736 (GGFPR1), X97737 (MMFPRL1), X97738 (GGFPRL1), X97739 (PTFPRL1), X97740 (MMFPRL2), X97741 (PPFPRL2), X97742 (GGFPRL2), X97743 (PTFPRL2), X97744 (PPFPRL1), and X97745 (PTFPR1) 相似文献
999.
1000.
Summary Fe (III) and Cu (II) each at 50 M in four commercial strains of Saccharomyces cerevisiae induced an increase of NAD(P)+ reduction in one strain (Turkish), but two others (Chilean and Brazilian), the presence of Fe(III) and/or Cu(II) diminished NAD(P)+ reduction presumably due to free radicais formation inside these living cells. Suprisingly, in the American strain, Fe(III) induced a decrease and Cu (II) an increase of NAD(P)+ reduction. 相似文献