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991.
Katarina Jansson Gunnar Kratz Anders Haegerstrand 《In vitro cellular & developmental biology. Animal》1996,32(9):534-540
Summary Reepithelialization of artificial partial thickness wounds made in biopsies of human skin was determined after 3, 5, or 7
d of incubation, submerged or elevated to the air-liquid interface. The biopsies were reepithelialized within 5–7 d, with
a more complete epidermal healing in wounds exposed to air. Both types of wounds showed similar time-course in deposition
of basement membrane components, as detected by immunofluorescence labeling. Laminin and collagen type VII were deposited
underneath the migrating tips, whereas collagen type IV was detected after reepithelialization. Markers of terminal differentiation
showed a pattern close to normal in the air-liquid incubated wounds after reepithelialization. Involucrin was detected in
the suprabasal regions of the migrating epidermis and thereafter in the upper half of neo-epidermis in the air-liquid incubated
wound. Filaggrin could not be detected in the submerged wounds at any time during healing, whereas wounds exposed to air showed
a well-differentiated epidermis by Day 7. Tritiated thymidine-incorporation indicated proliferation of epidermal and dermal
cells during reepithelialization and a maintained viability, as shown by cultivation of endothelial- and fibroblast-like cells
obtained from the dermis 7 d after wounding.
Reepithelialization in this humanin vitro model is supported by a matrix close to normal with the possibility of extracellular influences and cell-cell interactions
and, in addition, the technique is simple and reproducible. Therefore, we suggest this model for studies of regeneration in
culture and as a complement toin vivo studies on epidermal healing. 相似文献
992.
Jeffery R. Cook Robert G. van Buskirk 《In vitro cellular & developmental biology. Animal》1996,32(5):300-306
Summary Laminin synthesis and deposition are concomitant with the development of a basal lamina between the human epidermis and the
underlying dermis. One of the challenges in tissue engineering of human epidermal models is to develop substrates and conditions
that encourage the development of a basement membrane. The purpose of this study was to determine if actin filaments and/or
microtubules are involved in the synthesis/secretion of laminin by normal human epidermal keratinocytes (NHEK)in vitro. NHEK synthesize and secrete laminin subunits B1, B2, and M but little, if any, of laminin subunit A. Data indicate that
disruption of microfilaments by the destabilizing agent, cytochalasin D, had no apparent effect on the relative synthesis
rates of most cytosolic proteins as, revealed by one-dimensional sodium dodecyl sulfate (SDS) gel electrophoresis. This drug,
however, increased laminin B2 synthesis several fold over untreated controls. This enhanced synthetic rate was independent
of the type of collagen, matrix on which the NHEK were grown. Similar increases in synthesis of the M and B1 laminin chains
were not observed. To determine if this increase in synthesis lead to increases in laminin B2 secretion, laminin B2 was immunoprecipitated
from both the apical and basal domains of NHEK cells grown on microporous membranes. While more laminin B1, B2, and M were
secreted basally than apically, an observation consistent with laminin’s role in basal lamina formation, cytochalasin D had
no apparent effect on either basal or apical laminin B2 secretion. Experiments with the microtubule destabilizer, nocodazole,
showed no similar effects on laminin synthesis and/or secretion. We conclude that (a) disruption of the actin network in NHEK
selectively increases the synthesis of laminin B2, (b) the secretion of laminin B2 from NHEK cells is not governed by either
the microfilamentous cytoskeleton or the amount of laminin synthesized by NHEK, and (c) disruption of the microtubular network
does not alter laminin synthesis or secretion. 相似文献
993.
During conjugation, the micronucleus of Tetrahymena thermophila undergoes five consecutive nuclear divisions: meiosis, third prezygotic division (pregamic mitosis) and two postzygotic mitoses of the synkaryon. The four products of the synkaryon differentiate into macronuclear anlagen and new micronuclei and the old macronucleus is resorbed. The protein synthesis inhibitor cycloheximide, applied during conjugation, induced several developmental blocks. Pairs shifted to the drug during early meiotic prophase (stages I–III) were arrested at prophase. Cycloheximide applied to cells at pachytene (stages IV-VI) to metaphase arrested the conjugants at the stage of modified prometaphase/metaphase with overcondensed, swollen bivalents. In contrast to other systems, in the presence of cycloheximide, separation of chromatids, decondensation of chromosomes and exit from metaphase I were inhibited in both diploid and haploid cells. Pairs shifted to the drug after metaphase I were arrested at postmeiotic interphase after completing one nuclear cycle. The same rule applied to the subsequent cycle; then cells were arrested at the stage of pronuclei, and those pairs with functional pronuclei and synkarya were arrested at the stage of two products of the first postzygotic division (pronuclei were not arrested in nuclear transfer and karyogamy). Only pairs with two products of the first postzygotic division were arrested at the same stage after the cycloheximide treatment. Pairs shifted to cycloheximide during the second postzygotic division were arrested in development of macronuclear anlagen and resorption of old macronuclei. The postmeiotic conjugants pulse-treated with cycloheximide (2 h) yielded heterokaryons retaining parental macronuclei (i.e. they exhibited macronuclear retention). 相似文献
994.
Cell signaling and heat shock protein expression 总被引:5,自引:0,他引:5
Exposure of cells and organs to heat shock is associated with numerous changes in various cellular metabolic parameters and overexpression of proteins collectively known as heat shock proteins (HSP). In this communication we review the cell-signaling events that are altered in response to heat shock as they relate to the subsequent induction of HSP 70 kd (HSP-70) expression. We also review the mechanisms by which HSP-70 is involved in conferring cytoprotective effects. The possibility of altering HSP expression through manipulations of the cell-signal process has clinical importance.The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or reflecting the views of the Department of the Army or Department of Defense. 相似文献
995.
Mark A. Brown Alan Carne Geoffrey K. Chambers 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1996,113(4):731-736
α2-Macroglobulin (α2-M), a large molecular mass proteinase-binding protein, was identified in plasma from tuatara (Sphenodon), a rare reptile endemic to New Zealand. In this genus, α2-M constitutes 11–13% of total plasma protein (∼2.2–3.9 mg/ml). Analysis of blood samples collected at approximately monthly intervals from individual tuatara indicated that the plasma level of α2-M remains fairly constant. The subunits of tuatara α2-M have an apparent molecular mass of ∼160 kDa as determined by SDS-polyacrylamide gel electrophoresis and the intact protein is an oligomer that contains inter-chain disulfide bonds. N-terminal sequence analyses of tuatara α2-M revealed a distinct similarity to α-macroglobulins of other vertebrates and that at least two types of α2-M subunits are present in plasma of tuatara. 相似文献
996.
Summary Examination was made of the structure and assembly of the cortical microtubule (MT) cytoskeleton in the coenocytic green algaBoodlea coacta (Dickie) Murray et De Toni by immunofluorescence microscopy. Cortical MTs inBoodlea protoplasts are arranged randomly but some show a meridional arrangement within 6 h after protoplast formation. At 6–9 h such MTs become highly concentrated and parallel to each other in certain areas. At 12 h the concentration is uniformly high throughout the cell, indicating the completion of high density meridional arrangement of cortical MTs. Cortical MTs exhibiting a high density, meridional arrangement show characteristic disassembly by treatment with 10 M amiprophos-methyl (APM) or cold treatment (0 °C). Disassembly occurs by each MT unit at positions skipping 30–40 m in the transverse direction, and neighboring MTs subsequently disassemble to form MT groups. Each group becomes slender and then disappears completely within the following 24 h. The meridional arrangement of cortical MTs is disrupted by N-ethylmaleimide (NEM) accompanied by a remarkable reduction in density. The remaining MTs form groups at 30–40 m intervals from each other, as also occurs with drug or cold treatment, but disruption and density return to normal levels following removal of NEM. It appears that there are meridionally oriented channels, anchor-rich and anchor-poor, in the plasma membrane. The channels could be distributed alternately and anchors could be deposited in a cross-linking manner with cortical MTs to form a stable cortical MT-cytoskeleton. MTs comprising the cortical MT cytoskeleton could be oriented by meridionally oriented channels of anchors which are distributed following establishment of cell polarity.Abbreviations APM
amiprophos-methyl
- MT
microtubule
- MTOC
microtubule organizing center
- NEM
N-ethylrnaleimide 相似文献
997.
J. C. Rodríguez-Aguilera F. Navarro A. Arroyo F. J. Alcaín J. M. Villalba P. Navas 《Protoplasma》1995,184(1-4):229-232
Summary Ascorbate is stabilized in the presence of HL-60 cells. Our results showed that cAMP derivatives and agents that increase cAMP stimulate the ability of HL-60 cells to stabilize ascorbate. On the other hand, tunicamycin, a glycosilation-interfering agent, inhibited this ability. The ascorbate stabilization in the presence of HL-60 cells has been questioned as a simple chemical effect. Further properties and controls about the enzymatic nature of this stabilization are described and discussed. This data, together with hormonal regulation, support the hypothesis that an enzymatic redox system located at the plasma membrane is responsible of the extracellular ascorbate stabilization by HL-60 cells.Abbreviations AFR
ascorbate free radicals
- FCS
fetal calf serum
- Sp-cAMPS
Sp-cyclic adenosine monophosphothionate
- Rp-cAMPS
Rp-cyclic adenosine monophosphothionate 相似文献
998.
P. Fleurat-Lessard S. Bouché-Pillon C. Leloup W. J. Lucas R. Serrano J. -L. Bonnemain 《Protoplasma》1995,188(3-4):180-185
Summary Immunocytochemical techniques were employed to study the spatial distribution of the plasma membrane H+-ATPase within various cell types of the young reactive primary pulvinus ofMimosa pudica L. These cells were interconnected by large numbers of plasmodesmata, being concentrated within pit-fields. Although we could routinely detect evidence of the H+-ATPase along the plasma membrane, immunolabelling was rarely, if ever, observed along the plasma membranes of the plasmodesmata. This finding is discussed with respect to the likely specialized supramolecular structure of the plasmodesma.Abbreviations SEL
size exclusion limit of plasmodesmata 相似文献
999.
Summary Plasma membranes of maize (Zea mays L., cv. Sil Anjou 18) roots were isolated by aqueous two-phase partitioning. Multi elemental analysis by proton induced X-ray emission (PIXE) was used for the investigation of elemental composition of plasma membranes. Fe, Cu, and Zn as well as P, S, and Ca were identified. We did not find significant amounts of V, Mn, Se, Mo, or W.Abbreviations EDTA
ethylenediaminetetraacetic acid
- HCF III
hexacyanoferrate III (ferricyanide, K3[FeCN6])
- Hepes
2-[4-(2-hydroxyethyl)-1-piperazine]-ethanesulfonic acid
- PIXE
proton induced X-ray emission (proton microprobe)
- STA
siliciotungstic acid
- Tris
tris (hydroxymethyl)aminomethane 相似文献
1000.
Binding-protein expression is subject to temporal,developmental and stress-induced regulation in terminally differentiated soybean organs 总被引:3,自引:0,他引:3
Andrew Kalinski Daniel L. Rowley Deborah S. Loer Carolyn Foley George Buta Eliot M. Herman 《Planta》1995,195(4):611-621
Binding protein (BiP) is a widely distributed and highly conserved endoplasmic-reticulum luminal protein that has been implicated in cotranslational folding of nascent polypeptides, and in the recognition and disposal of misfolded polypeptides. Analysis of cDNA sequences and genomic blots indicates that soybeans (Glycine max L. Merr.) possess a small gene family encoding BiP. The deduced sequence of BiP is very similar to that of other plant BiPs. We have examined the expression of BiP in several different terminally differentiated soybean organs including leaves, pods and seed cotyledons. Expression of BiP mRNA increases during leaf expansion while levels of BiP protein decrease. Leaf BiP mRNA is subject to temporal control, exhibiting a large difference in expression in a few hours between dusk and night. The expression of BiP mRNA varies in direct correlation with accumulation of seed storage proteins. The hybridization suggests that maturing-seed BiP is likely to be a different isoform from vegetative BiPs. Levels of BiP protein in maturing seeds vary with BiP mRNA. High levels of BiP mRNA are detected after 3 d of seedling growth. Little change in either BiP mRNA or protein levels was detected in maturing soybean pods, although BiP-protein levels decrease in fully mature pods. Persistent wounding of leaves by whiteflies induces massive overexpression of BiP mRNA while only slightly increasing BiP-protein levels. In contrast single-event puncture wounding only slightly induces additional BiP expression above the temporal variations. These observations indicate that BiP is not constitutively expressed in terminally differentiated plant organs. Expression of BiP is highest during the developmental stages of leaves, pods and seeds when their constituent cells are producing seed or vegetative storage proteins, and appears to be subject to complex regulation, including developmental, temporal and wounding.The mention of vendor or product does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over vendors of similar products not mentioned.Abbreviations BiP
binding protein
The sequences reported in this paper have been submitted to Gen-Bank and are identified with the accession numbers BiP-A (U08384), BiP-B (U08383), BiP-C (U08382) and -1,3 glucanase (U08405). 相似文献