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131.
Jan Deussing Roth Wera Rommerskirch Winfried Wiederanders Bernd Figura von Kurt Christoph Peters 《Mammalian genome》1997,8(4):241-245
The mouse genes for the lysosomal cysteine proteinases cathepsin B, H, L, and S were mapped to Chromosomes (Chrs) 14, 9,
13, and 3, respectively. Two of the DNA probes used in this study detected an additional, independently segregating locus.
The cathepsin B-specific probe hybridized to a locus on Chr 2, and the cathepsin H probe to a locus on the X Chr. These loci
either correspond to pseudogenes or to cathepsin B- and cathepsin H-related genes. The four cysteine proteinases mapped in
this study lie within known regions of conserved synteny between mouse and human chromosomes, when compared with the corresponding
positions of their human homologs. Assuming that the genes of the cysteine proteinase gene family arose from a common ancestral
gene, our results suggest that these four cysteine proteinases had been dispersed over different chromosomes before separation
of mouse and human in evolution.
Received: 22 August 1996 / Accepted: 20 November 1996 相似文献
132.
133.
Maurizio Pellecchia Gerhard Wider Hideo Iwai Kurt Wüthrich 《Journal of biomolecular NMR》1997,10(2):193-197
A novel 2D NMR experiment, 2D HE(NE)HGHH, is presented for the assignment ofarginine side chain 1H and 15N resonances inuniformly 15N-labeled proteins. Correlations between1H, 1Hand 1H are established on the basis of3J(15N,1H) heteronuclear scalarcoupling constants, and sequence-specific assignments are obtained by overlapof these fragments with 1H chemical shiftsobtained by assignment procedures starting from the polypeptide backbone.Since guanidino protons exchange quite rapidly with the bulk water, the 2DHE(NE)HGHH pulse scheme has been optimized to avoid saturation and dephasingof the water magnetization during the course of the experiment. As anillustration, arginine side chain assignments are presented for two uniformly15N-labeled proteins of 7 and 23 kDa molecular weight. 相似文献
134.
In the giant-celled marine algae Valonia utricularis the turgor-sensing mechanism of the plasmalemma and the role of the tonoplast in turgor regulation is unknown because of
the lack of solid data about the individual electrical properties of the plasmalemma and the vacuolar membrane. For this reason,
a vacuolar perfusion technique was developed that allowed controlled manipulation of the vacuolar sap under turgescent conditions
(up to about 0.3 MPa). Charge-pulse relaxation studies on vacuolarly perfused cells at different turgor pressure values showed
that the area-specific resistance of the total membrane barrier (tonoplast and plasmalemma) exhibited a similar dependence
on turgor pressure as reported in the literature for nonperfused cells: the resistance assumed a minimum value at the physiological
turgor pressure of about 0.1 MPa. The agreement of the data suggested that the perfusion process did not alter the transport
properties of the membrane barrier.
Addition of 16 μm of the H+-carrier FCCP (carbonylcyanide p-trifluoromethoxyphenyhydrazone) to the perfusion solution resulted in a drop of the total membrane potential from +4 mV to
−22 mV and in an increase of the area-specific membrane resistance from 6.8 × 10−2 to 40.6 × 10−2Ωm2. The time constants of the two exponentials of the charge pulse relaxation spectrum increased significantly. These results
are inconsistent with the assumption of a high-conductance state of the tonoplast (R. Lainson and C.P. Field, J. Membrane Biol.
29:81–94, 1976).
Depending on the site of addition, the pore-forming antibiotics nystatin and amphotericin B affected either the time constant
of the fast or of the slow relaxation (provided that the composition of the perfusion solution and the artificial sea water
were replaced by a cytoplasma-analogous medium). When 50 μm of the antibiotics were added externally, the fast relaxation process disappeared. Contrastingly, the slow relaxation process
disappeared upon vacuolar addition. The antibiotics cannot penetrate biomembranes rapidly, and therefore, the findings suggested
that the fast and slow relaxations originated exclusively from the electrical properties of the plasmalemma and the tonoplast
respectively. This interpretation implies that the area-specific resistance of the tonoplast is significantly larger than
that of the plasmalemma (consistent with the FCCP data) and that the area-specific capacitance of the tonoplast is unusually
high (6.21 × 10−2 Fm−2 compared to 0.77 × 10−2 Fm−2 of the plasmalemma). Thus, we have to assume that the vacuolar membrane of V. utricularis is highly folded (by a factor of about 9 in relation to the geometric area) and/or contains a fairly high concentration of
mobile charges of an unknown electrogenic ion carrier system.
Received: 22 October 1996/Revised: 16 January 1997 相似文献
135.
Astrid Zimmermann Anke Haina Ute Gröschel-Stewart 《Development genes and evolution》1995,204(4):271-275
The development of the autonomic ganglia of Auerbach's plexus and gizzard smooth muscle was studied in chicken embryos. Nervous system and smooth-muscle-specific antibodies were employed in immunofluorescence stainings on tissue sections to investigate the temporal and spatial frame of neural and muscular differentiation in relation to each other. Subserosal clusters of neural cells were clearly demonstrable at embryonic day 5 (ED5), the earliest stage analysed, with the monoclonal antibody El (SGIII-1). Fine nerve fibres (ED6) and, later, large axon bundles projecting from subserosal neuron clusters towards the lumen were followed and found to reach the luminal border by ED11. Already in early development the area of the future laminar tendons on the ventral and dorsal surface of the gizzard was devoid of neuroblasts, and nerve fibres were not extending to the muscle-tendon borderline until ED16. Double stainings with antibodies to smooth muscle myosin (SMM) and El revealed that SMM expression, taken as an indicator for muscle differentiation, followed neural growth. It was first detectable in close apposition to the differentiating neuroblasts in the caudal and cranial portion of the gizzard at ED6. With further development, myosin expression proceeded inward towards the lumen in a wave which followed the ingrowth of E1-positive nerve fibres from the prospective Auerbach plexus. Neuromuscular differentiation deviated from this pattern in the lateral tendon area where nerve growth was delayed and myosin expression preceeded the arrival of E1-positive nerve fibres. The findings suggest that the gizzard could serve as a model system for the analysis of potential early nervous system imprints on smooth premuscle mesenchyme differentiation. 相似文献
136.
DISGEO is a new implementation of a distance geometry algorithm which has been specialized for the calculation of macromolecular conformation from distance measurements obtained by two-dimensional nuclear Overhauser enhancement spectroscopy. The improvements include (1) a decomposition of the complete embedding process into two successive, more tractable calculations by the use of “substructures”, (2) a compact data structure for storing incomplete distance information on a structure, (3) a more efficient shortest-path algorithm for computing the triangle inequality limits on all distances from this information, (4) a new algorithm for selecting random metric spaces from within these limits, (5) the use of chirality constraints to obtain good covalent geometry without the use ofad hoc weights or excessive optimization. The utility of the resultant program with nuclear magnetic resonance data is demonstrated by embedding complete spatial structures for the protein basic pancreatic trypsin inhibitor vs all 508 intramolecular, interresidue proton-proton contacts shorter than 4.0 Å that were present in its crystal structure. The crystal structure could be reproduced from this data set to within 1.3 Å minimum root mean square coordinate difference between the backbone atoms. We conclude that the information potentially available from nuclear magnetic resonance experiments in solution is sufficient to define the spatial structure of small proteins. 相似文献
137.
Lun He Kurt J. Isselbacher Jack R. Wands Howard M. Goodman Chiaho Shih Andrea Quaroni 《In vitro cellular & developmental biology. Plant》1984,20(6):493-504
Summary A human hepatocellular carcinoma cell line (FOCUS—Friendship of China and United States) was derived from a patient with primary
hepatocellular carcinoma. This cell line has been in continuous culture over an 18-mo period. The morphological and ultrastructural
features of FOCUS are consistent with its neoplastic hepatocellular orgin. FOCUS cells contain aspartate aminotransferase
and glucose-6-phosphatase activity. In addition, α1-antitrypsin, fibrinogen, alpha fetoprotein, and carcinoembryonic antigens were detectble in the cytoplasm of the cultured
cells by immunochemical staining techniques. The karyotype of the FOCUS cell is human in origin and it contains human DNA
sequences as detected by molecular hybridization analysis. The FOCUS cells do not show evidence of density-dependent inhibition
of growth under confluent conditions. Repeated growth curves over an 18-mo period were identical, revealing a doubling time
of 42 to 48 h. The malignant potential of FOCUS cells was further demonstrated by their ability to lead to gross tumor formation
after subcutaneous infection into nude mice. From one of the solid tumors grown in nude mice, recultured cell lines have been
established and found to have properties identical to the original FOCUS cell line. This FOCUS cell line represents an additional
model for further investigation of tumor specific antigens and the relationship between hepatitis B virus (HBV) and hepatocellular
carcinoma. Preliminary molecular characterization has indicated the existence of integrated HBV sequences within the FOCUS
genome. 相似文献
138.
J.M. Gennity Nestor R. Bottino Ralph A. Zingaro Andrew E. Wheeler Kurt J. Irgolic 《Biochemical and biophysical research communications》1984,118(1):176-182
Axenic cultures of the green algae and red algae were grown in the presence of sublethal quantities of selenite. All purified lipids from both algae were found to contain bound selenium, except for saturated hydrocarbons. Of the lipids which contain selenium, carotenoid pigments contain the greatest concentrations. Lipid-associated selenium is not metabolically incorporated. The selenium is probably non-covalently bound to the lipids. 相似文献
139.
Kurt A. Santarius 《Planta》1984,161(6):555-561
Freezing of isolated spinach thylakoids in the presence of NaCl uncoupled photophosphorylation from electron flow and increased the permeability of the membranes to protons. Addition of ATP prior to freezing diminished membrane inactivation. On a molar basis, ATP was at least 100 times more effective in protecting thylakoids from freezing damage than low-molecularweight carbohydrates such as sucrose and glucose. The cryoprotective effectiveness of ATP was increased by Mg2+. In the absence of carbohydrates, preservation of thylakoids during freezing in 100 mM NaCl was saturated at about 1–2 mM ATP, but under these conditions membranes were not fully protected. However, in the presence of small amounts of sugars which did not significantly prevent thylakoid inactivation during freezing, ATP concentrations considerably lower than 0.5 mM caused nearly complete membrane protection. Neither ADP nor AMP could substitute for ATP. These findings indicate that cryoprotection by ATP cannot be explained by a colligative mechanism. It is suggested that ATP acts on the chloroplast coupling factor, either by modifying its conformation or by preventing its release from the membranes. The results are discussed in regard to freezing injury and resistance in vivo.Abbreviations CF1
chloroplast coupling factor
- Hepes
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- PMS
phenazine methosulfate
- Tris
2-amino-2-(hydroxymethyl)-1,3-propandiol 相似文献
140.
The alpha 1-adrenergic receptor ligand, 3H-WB4101, and the alpha 2-adrenergic receptor ligand, 3H-para-aminoclonidine, were utilized at a 1.0 nM incubation concentration to determine relative alpha 1-and alpha 2-adrenergic receptor binding by cell membranes from selected tissues within the brain, ovary and oviduct of the domestic fowl. Significant specific alpha 1-adrenergic binding was observed in the hypothalamus, anterior pituitary, pineal, cerebrum and cerebellum but only the cerebrum had significant alpha 2-receptor binding. Significant levels of alpha 1-adrenergic binding were observed in the granulosa cells of the three largest ovarian follicles and in the postovulatory follicle. Significant specific alpha 2-adrenergic binding was measured in the infundibulum, magnum, isthmus and shell gland of the oviduct. The physiological implications of alpha-adrenergic receptors in these tissues are discussed. 相似文献