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Lucie Parent Stéphane Supplisson Donald D. F. Loo Ernest M. Wright 《The Journal of membrane biology》1992,125(1):49-62
Summary Cystic fibrosis (CF) is characterized by abnormal epithelial Cl– conductance (GCl). In vitro studies that have shown that cAMP regulation is an intrinsic property of the CF-affected GCl(CF-GCl) have been carried out previously on cultured secretory cells and on nonepithelial cells. Even though GCl in absorption is defective in CF, a clear demonstration of cAMP regulation of CF-GCl in a purely absorptive tissue is lacking. We studied the cAMP regulation of CF-GCl in the microperfused intact human reabsorptive sweat duct. About 40% of the ducts responded to cAMP (responsive) while the remainder of the ducts did not. In responsive ducts, cAMP-elevating agents: -adrenergic agonist isoproterenol (IPR), CPT-cAMP, forskolin, theophylline or IBMX increased G
tby about 2.3-fold (n = no. of ducts = 8). Removal of media Cl–, but not amiloride pretreatment (in the lumen), abolished the cAMP response, indicating exclusive activation of GCl. cAMP activated both apical and basolateral GCl. cAMP hyperpolarized gluconate: Cl– (lumen: bath) transepithelial bionic potentials (V
t=–20.3±5.2 mV, mean ±se, n=9) and transepithelial 3 1 luminal NaCl dilution diffusion potentials (V
t=–8.8±2.9 mV, n=5). cAMP activated basolateral GCl as indicated by increased bi-ionic (gluconate: Cl–, bath: lumen) diffusion potentials (by about 12 mV). The voltage divider ratio in symmetric NaCl solutions increased by 60%. Compared to responsive ducts, nonresponsive ducts were characterized by smaller spontaneous transepithelial potentials in symmetrical Ringer's solution (V
t=–6.9±0.8 mV, n=24, nonresponsive vs. –19.4±1.8 mV, n=22, responsive ducts) but larger bi-ionic potentials (–94±6 mV, n=35, nonresponsive vs. –65±5 mV, n=17, responsive ducts) and dilution diffusion potentials (–40±5 mV, n=11, nonresponsive vs. –29±3 mV, n=7, responsive ducts). These results are consistent with an inherently (prestimulus) maximal activation of GCl in nonresponsive ducts and submaximal activation of GCl in responsive ducts. We conclude that cAMP activates CF-G
Cl which is expressed and abnormal in both apical and basal membranes of this absorptive epithelium in CF.Abbreviations CF
cystic fibrosis
-
G
t
transepithelial conductance
-
V
b
electrical potential across the basolateral membrane
-
V
a
electrical potential across the apical membrane
-
V
t
transepithelial potential
- V
b
transepithelial currentinduced voltage deflections across the basolateral membrane
- V
a
transepithelial current-induced voltage deflections across the apical membrane
- V
t
transepithelial current-induced voltage deflection across the epithelium
- VDR
voltage divider ratio
- GCl
transepithelial Cl– conductance
- CF-GCl
cystic fibrosis-affected Cl– conductance
- EMF
electromotive force
- IPR
isoproterenol
- IBMX
3-isobutyl-1-methylxanthine
- CPT-cAMP
chlorophenylthio-adenosine 3-5 cyclic monophosphate
- PGE2
prostaglandin E2 相似文献
24.
Patricia Maurer Corinne Royer Bernard Mauchamp Patrick Porcheron Danile Debieu Guy Riba 《Archives of insect biochemistry and physiology》1991,16(1):1-9
The major ecdysteroids in large worker pupae of the leaf-cutting ant Acromyrmex octospinosus were characterized at the peak ecdysteroid concentration by using high-performance liquid chromatography, enzyme immunoassay, and mass spectrometry. In decreasing amounts, they were determined to be makisterone A, an unidentified C28 ecdysteroid bearing a molecular weight of 494, 20-hydroxyecdysone (ratio of 1 to 6 as compared to makisterone A), and putative but negligible ecdysone. The presence of both C28 and C27 ecdysteroids is discussed in relation to the content of 4-desmethylsterols determined by gas chromatography and mass spectrometry to be ergosta-5,7,24 (28)-trien-3β-ol, ergosterol, ergosta-5,7-dien-3β-ol and ergosta-7,24(28)-dien-3β-ol for the main sterols, and with a small amount of cholesterol. 相似文献
25.
The 2-μm DNA plasmids from Saccharomyces cerevisiae strain H1 and strain HQ/5C were analyzed by electron microscopy for the presence of Escherichia coli RNA polymerase binding sites. On native 2-μm DNA isolated from strain HQ/5C five RNA polymerase binding sites were detected. One further site was mapped on cloned 2-μm DNA type 23 from S. cerevisiae strain H1. This additional site is located at a distance of 2.15 kilobases from EcoRI site B inside one of the inverted duplication (id) sequences. No such binding site could be detected in the other id sequence of the type 23 molecule, thus indicating that the two id sequences of strain H1 differ in at least one short region. The location of the id sequence carrying the RNA polymerase binding site was analyzed in native 2-μm DNA isolated from strain H1 and found to be present on HindIII fragment 2 and absent from HindIII fragment 5. This indicates that at least a part of the id sequences has a fixed position with respect to the unique S segment and further suggests a site specific recombination mechanism for the inversion of one of the unique segments. As a control for the specificity of RNA polymerase binding, we have mapped binding sites on vectors pBR313 and pBR322. The location of the E. coli RNA polymerase binding sites on 2-μm DNA is discussed in relation to the DNA regions expressed in E. coli minicells. 相似文献
26.
The minicell-producing Escherichia coli strain P 678-54 was transformed with a series of defined PTY chimeric plasmids consisting of yeast 2-μm DNA and E. coli plasmid pCR1. In minicells the integrated 2-μm DNA from yeast directed specifically the synthesis of six polypeptides with apparent molecular weights of 15 000, 17 500, 20 000, 22 000, 37 000, and 48 000. The specificity of five other polypeptides, which cover a molecular weight range of 19 000 to 28 000, has not yet been established with certainty. Neither the orientation of the integrated DNA, nor the inversion which distinguishes the two structural forms of 2-μm DNA affected the polypeptides synthesized. However, integration at a given EcoRI site appeared to be correlated with the absence of one particular polypeptide band; this suggests that at least one of these sites is located in an expressed region of the DNA. 相似文献
27.
28.
Ouimette Andrew P. Ollinger Scott V. Lepine Lucie C. Stephens Ryan B. Rowe Rebecca J. Vadeboncoeur Matthew A. Tumber-Davila Shersingh J. Hobbie Erik A. 《Ecosystems》2020,23(4):715-729
Ecosystems - Carbon (C) fluxes among different components of plant growth are important to forest ecosystem C cycling and are strongly influenced by species composition and resource availability.... 相似文献
29.
30.
Ondřej Korábek Lucie Juřičková Adam Petrusek 《Journal of Zoological Systematics and Evolutionary Research》2020,58(4):944-956
Exact locations of glacial refugia are relevant for the study of contemporary biodiversity, not only as places less disturbed during the climatic changes but also as sources of rapid expansion of the biota after the Last Glacial cycle. If continuously inhabited over several of the Quaternary glacial cycles, the refugia are readily identifiable by the accumulated genetic diversity. However, the sources of the Holocene range expansion, particularly important for the emergence of present-day bio- and phylogeographic patterns and for realistic estimation of species’ expansion rates, might have been located at the fringes of the glacial species ranges and lack unique lineages. This problem is pertinent when the variation is explored at slowly evolving genetic markers. We suggest that the location of such source refugia may be approximated by reconstructing the geographic location as a continuous trait evolving along the branches of a phylogenetic tree. We applied this approach, using the BEAST software, on two large southeast European land snail species: Caucasotachea vindobonensis and Helix thessalica. We found evidence for C. vindobonensis refugia in the western Balkans; notable is an apparently old refugium in Bosnia and Herzegovina. The plausible sources of the species’ Holocene range expansion, however, were located around the south-western end of the Carpathians. Although the source areas were likely similar in H. thessalica, some expansion sources suggested by the analyses (e.g., Podolia, Ukraine) appeared implausible and driven by sampling clustered in that area. The applied approach allows for additional exploitation of the mitochondrial data gathered during the past two decades of animal phylogeography studies. 相似文献