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91.
A steady-state kinetic analysis of the activation of bovine Factor X, by bovine Factor Xa, was undertaken. The activation was found to be dependent on the presence of divalent cations; Ca2+ showing the greatest stimulatory effect and Mn2+ exhibiting a lower degree of activity for this reaction. Although Sr2+ and Mg2+ were ineffective when present alone, each contributed synergistically to the activation rate at suboptimal levels of Ca2+. The effect of phospholipid (phosphatidylcholine:phosphatidylserine, 4:1, w:w) on the rate of activation and on the activation pathway was investigated. Phospholipid (PL) concentrations of up to 40 μm had no effect on the activation rate; whereas, concentrations of 40–180 μm were slightly inhibitory. In the absence of PL, the major product of the activation was Factor α-Xa, while in the presence of PL, lower-molecular-weight forms of Factor X (Factor β-X) and Factor Xa (Factor β-Xa were produced. At saturating levels of Ca2+, the Km app for the activation, at pH 7.4 and 37 °C, in the absence of PL, was found to be 0.6 ± 0.1 μm and the V was 1.7 ± 0.3 mol Factor X cleaved min?1 mol?1 Factor Xa. The corresponding values, in the presence of 90 μm PL, were 1.4 ± 0.2 μm and 2.2 ± 0.2 mol Factor X cleaved min?1 mol?1 Factor Xa. 相似文献
92.
93.
Michael Solursh Karen L. Jensen Carl T. Singley Thomas F. Linsenmayer Rebecca S. Reiter 《Developmental biology》1982,94(2):311-325
The effect of developmental stage on chondrogenic capacity in high-density cell cultures of chick embryonic wing bud mesenchyme is examined. Mesenchyme from stage 19 embryos forms aggregates of closely associated cells which do not form cartilage matrix, nor contain significant levels of type II collagen that are detectable by immunofluorescence, unless they are treated with dibutyryl cyclic AMP. Mesenchyme from stage 24 embryonic wing buds in high-density cell cultures will spontaneously form cartilage, as defined by electron microscopy and immunofluorescence with antibody to type II collagen. Cultures prepared from stage 26 wings form numerous aggregates which fail to accumulate an Alcian blue-staining matrix and which resemble mesenchyme cells morphologically. However, because these cells show considerable intracellular immunofluorescence for type II collagen, they are actually unexpressed cartilage cells. Several treatments, including exposure to dibutyryl cyclic AMP, ascorbic acid and an atmosphere of 5% oxygen, or mixture with small numbers of stage 24 wing mesenchyme cells, stimulate expression, as determined by the accumulation of an Alcian blue-staining matrix and an ultrastructurally recognizable cartilage matrix. Since the addition of similar numbers of differentiated cartilage cells does not stimulate expression of stage 26 cells, it is proposed that initial cartilage expression is dependent on a mesenchyme-specific influence which might be removed by cell dissociation. These studies demonstrate that there are at least two distinct transitions in cartilage differentiation: one involves the conversion of mesenchyme to unexpressed chondrocytes and the second involves mesenchyme-dependent expression of chondrogenic differentiation. 相似文献
94.
While the apical ectodermal ridge (AER) is well known for its required role in the development of distal parts of the limb and for its ability to stimulate limb duplications, the mechanism of its action is unknown. In this study we use a culture system previously developed by M. Globus and S. Vethamany-Globus (1976, Differentiation6, 91–96) in which an AER grafted onto a high-density cell culture of limb mesenchyme stimulates the formation of an outgrowth. Time-lapse movies taken during the outgrowth period demonstrated no cellular activities other than cell division. Both the mitotic index and labeling index in the mesenchyme were significantly elevated under the AER as compared to that without AER, indicating that the AER provides a growth-promoting stimulus which increases the proportion of dividing cells. On the other hand, nonridge ectoderm had no detectable effect on the mitotic index. Treatment of cultures with cytosine arabinoside both inhibited DNA synthesis and prevented AER-induced outgrowth. These results demonstrate a mitogenic capacity of AER tissue and suggest that mesenchymal outgrowth requires this activity. The mitogenic property of the AER is considered in relation to limb outgrowth in situ. 相似文献
95.
Rebecca W. Dolan 《Brittonia》1991,43(1):54-56
The Friesner Herbarium (BUT) of Butler University is a collection of over 100,000 specimens built from the personal herbarium
of Ray C. Friesner. He and other botanists at Butler amassed one of the largest and most complete collections of Indiana plants.
Active exchange from the 1920’s through the 1940’s increased the holdings of plants from other states. Although the collection
does not contain many type specimens, it is rich in vouchers from floristic and ecological studies conducted in the first
half of the 20th century and published in the scientific journal,Butler University Botanical Studies. 相似文献
96.
A review of the salt sensitivity of the Australian freshwater biota 总被引:13,自引:7,他引:6
Barry T. Hart Paul Bailey Rick Edwards Kent Hortle Kim James Andrew McMahon Charles Meredith Kerrie Swadling 《Hydrobiologia》1991,210(1-2):105-144
In Victoria, Australia, both dryland salinity and salinity in irrigation regions are serious agricultural problems. One option
to control the latter is to pump groundwater to maintain it below the surface. However, this leaves a saline wastewater for
disposal, probably into local streams or wetlands. This review of the salt sensitivity of the biota of Australian streams
and wetlands gives information of interest to those responsible for developing controls on these discharges. The review addresses
the lethal and sub-lethal effects of salinity on microbes (mainly bacteria), macrophytes and micro-algae, riparian vegetation,
invertebrates, fish, amphibians, reptiles, mammals, and birds. Data suggest that direct adverse biological effects are likely
to occur in Australian river, stream and wetland ecosystems if salinity is increased to around 1 000 mg L−1. The review highlights a general lack of data on the sensitivity of freshwater plants and animals to salinity increases. 相似文献
97.
Endopeptidase-24.11 is a 90-kDa surface glycoprotein with the ability to hydrolyze a variety of biologically active peptides. Interest in this enzyme is based on the consensus that it may play a role in the termination of peptide signals in the central nervous system. In the present study, we have investigated the distribution of endopeptidase-24.11 in two nerves of the peripheral nervous system of newborn pigs: the sciatic, composed of a mixture of myelinated and nonmyelinated axons, and cervical sympathetic trunk in which greater than 99% of the axons are nonmyelinated. The endopeptidase was monitored enzymatically, as well as by immunoblotting and immunocytochemistry using mono- and polyclonal anti-endopeptidase antibodies. Endopeptidase-24.11 was detected in both the sciatic nerve and the cervical sympathetic trunk. Membrane preparations from sciatic nerve hydrolyzed 125I-insulin B-chain, and more than 50% of the activity was inhibited by phosphoramidon with an IC50 concentration of 3.2 nM. Moreover, a 90-kDa polypeptide was detected by immunoblotting of sciatic nerve membranes. The type of cells expressing the endopeptidase was determined by immunohistochemistry. In teased nerve preparations, these cells were identified morphologically as myelin- and non-myelin-forming Schwann cells. Endopeptidase-24.11 was also expressed by cultured Schwann cells from sciatic nerve and cervical sympathetic trunk maintained for 3 h in vitro. The presence of endopeptidase-24.11 on the Schwann cell surface raises the possibility of a potential role for the enzyme in nerve development and/or regeneration. 相似文献
98.
Niels C. Krejci Lynne Smith Rebecca Rudd Robert Langdon Joseph McGuire 《In vitro cellular & developmental biology. Animal》1991,27(12):933-938
Summary To investigate the regulation of epithelial differentiation, normal human epidermal keratinocytes were cultured floating on
the surface of culture medium without attachment to a solid substrate. Keratinocytes spread out on the surface of the medium,
proliferated and differentiated either into several flat lacy sheets 1 to 3 cells thick (on medium containing 0.15 mM calcium) or formed one single aggregate of cells from 5 to 15 cells in thickness on medium containing 1.15 mM calcium. The cell aggregates demonstrated a pattern of ordered epithelial differentiation. Levels of progressive differentiation
resembling the structure of normal human epidermis were identified by light microscopy, immunohistochemistry, and electron
microscopy. Differentiation proceeded from cells at the air side toward cells at the medium side with basal appearing cells
on the air side and keratinocytes expressing filaggrin and involucrin on the side toward the medium. These results demonstrate
that organized epithelial differentiation can occur in the absence of extracellular matrix. In contrast with other air-liquid
interface cultures, epithelial differentiation in the absence of extracellular matrix progresses from air towards medium. 相似文献
99.
The structure of the vacuolar ATPase from mesophyll tonoplasts of Mesembryanthemum crystallinum has been studied by electron microscopy using negatively stained specimens of membrane-bound and detergent-solubilized ATPase molecules. We observed a high density of particles on the surface of tonoplast vesicles and “head and stalk” structures on the edge of the membrane, similar to the F0F1-ATPases of mitochondrial and chloroplast membranes. The staining conditions, which are often critical for such small objects, were improved by using methylamine tungstate as negative stain for the membrane-bound ATPase. Compared to other staining solutions generally applied, dissociation of the F1-like enzyme complex from the membrane was best prevented and structural damage of the vesicles was least observed with methylamine tungstate. In freeze-fracture electron microscopy of tonoplast vesicles, where dissociation never occurs since no detergent is used, we also observed “head and stalk” structures on the edge of the membranes, beside many particles on the fracture faces. The detergent-solubilized ATPase forms string-like structures, caused by the aggregation of the hydrophobic membrane-embedded F0-like part of the enzyme. After negative staining the F1-like enzyme complex is arranged alternately along both sides of the string and connected by a narrow stalk. 相似文献
100.
A genetic linkage map of human chromosome 5 with 60 RFLP loci. 总被引:6,自引:0,他引:6
B Weiffenbach K Falls A Bricker L Hall J McMahon J Wasmuth V Funanage H Donis-Keller 《Genomics》1991,10(1):173-185
A genetic map of human chromosome 5 that contains 60 restriction fragment length polymorphism (RFLP) loci in one linkage group has been constructed. Segregation data using these markers and 40 large multigenerational families supplied by the Centre d'Etude du Polymorphisme Humain have been collected. Linkage analyses were performed with the program package CRI-MAP; using odds greater than 1000:1, 30 RFLP loci could be placed on the map. This genetic map spans 289 cM sex-equal, 353 cM in females, and 244 cM in males. While the relative rate of recombination for female meioses is nearly twice that of males over much of the chromosome, several instances of statistically significant excess male recombination were observed. The order of probes on the genetic map has been confirmed by their physical order as determined by somatic cell hybrid lines containing deletions of normal chromosome 5. There is concordance between the physical positions of markers and their genetic positions. Our most distal probes on the genetic map are cytologically localized to the most distal portions of the chromosome. This suggests that our genetic map spans most of chromosome 5. 相似文献