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101.
Jean F. Emly Wendy A. Ratcliffe Elaine Green Sarah J. Bowden David A. Heath Ann Blight Susan Hughes John G. Ratcliffe 《生物化学与生物物理学报:疾病的分子基础》1992,1180(1):58-64
The molecular forms of parathyroid hormone-related protein (PTHRP) in conditioned media from the BEN human lung cancer cell line, rat parathyroid cells (PT-r) and human keratinocytes were studied by gel-filtraton chromatography with assay of PTHRP by immunoassays and bioassay. Immunoreactivity (1–86 and 1–34) and bioactivity (1–34) in conditioned media eluted as a coincident major peak (approx. molecular mass 19–22 kDa) and there was evidence of amino-terminal species in the molecular mass range 10–16 kDa in BEN and keratinocyte media. Western blotting of PTHRP affinity purified by monoclonal antibodies directed at regions 1–34 or 37–67, identified a major species in all cell cytosols and media with an apparent molecular mass of 24–25 kDa, consistently slighty larger than recombinant PTHRP(1–141) (mobility of 21 kDa) which may represent an intact or native form of PTHRP. Additional amino-terminal species were identified in medium from keratinocytes (16 and 7 kDa), BEN cells (18 and 14 kDa) and PT-R cells (17 kDa), suggesting that processing occurs at the C-terminus and within the mid-region to form a range of amino-terminal fragments. 相似文献
102.
Thérèse E. Malliavin Marc A. Delsuc Jean Y. Lallemand 《Journal of biomolecular NMR》1992,2(4):349-360
Summary The structural determination of biological molecules in solution by NMR relies on the determination of a set of interatomic distances obtained by measurement of intramolecular nuclear Overhauser effects (NOE). It is shown in this paper that it is possible to obtain the accurate relaxation rate (and hence the interatomic distance) from the direct measurement of a single NOE signal. The precise analysis of a NOESY peak evolution with respect to the mixing time allows the evaluation of the relaxation parameters for the pair of spins under consideration. This is done without any assumption on the relaxation of unmeasured spins, or on the movement of the molecule. The theoretical basis of this method is presented. In order to evaluate the proposed method, a simulated case on the protein BPTI is studied, which shows that the method performs very well even in the case of noisy data sets. 相似文献
103.
Claude Chachaty 《Molecular Engineering》1992,2(1):65-94
The order parameters as well as the rates of overall and internal motions of aggregated surfactants can be obtained from deuteron and carbon-13 nuclear relaxation experiments. The main contribution to the relaxation is generally the quadrupolar coupling (2H) or the short range dipolar interaction with protons (13C). In some cases it is convenient to derive the same information from the13C relaxation induced by long range dipolar interactions with a paramagnetic probe exchanging rapidly among the polar heads of surfactant molecules. This paper outlines the methods of interpretation of relaxation data by means of a rotational jump model of internal motions, taking into account most of the accessible conformers. The conformational and dynamical parameters are obtained from the magnetic field dependence of the longitudinal relaxation rates (micelles) or from the simultaneous fit of these rates and of the dipolar or quadrupolar splittings (liquid crystals). Some examples of application of these methods are given from recent works on single and double detailed surfactants. 相似文献
104.
105.
A 126 bp fragment of a plant histone gene promoter confers preferential expression in meristems of transgenic Arabidopsis 总被引:3,自引:0,他引:3
Rossitza Atanassova Nicole Chaubet Claude Gigot 《The Plant journal : for cell and molecular biology》1992,2(3):291-300
The tissue-specific pattern of expression directed by the H4A748 Arabidopsis histone promoter was investigated by analysis of beta-glucuronidase (GUS) activity in transgenic Arabidopsis containing H4A748-GUS gene fusions. As determined by fluorimetric and histochemical tests, the H4A748 promoter directs preferential expression in meristems of young seedlings and adult plants. The low activity found in nonproliferating tissues may relate to basal constitutive expression of the histone promoter and/or to endoreduplication occurring in some tissues. The endogenous histone mRNA levels parallel the GUS activity found in different tissues. Analysis of the regulatory properties of 5' deleted promoters showed that multiple positive elements exist between -900 and -219 and that the proximal region of the promoter to -219 is sufficient to establish the full tissue-specific pattern of expression. Further deletion to -93 nearly abolished the promoter activity thus suggesting that the 126 bp fragment located between -219 and -93 contains the elements responsible for the specific expression pattern. The presence of several remarkable sequences within this fragment is discussed. 相似文献
106.
The serum of the channel catfish (Ictalurus punctatus) was examined for heme- and hemoglobin-binding proteins. Electrophoretic mobility retardation assays failed to detect a hemoglobin-binding material similar to mammalian haptoglobin; however, a heme-binding component (not previously described) was identified in catfish seru. The heme-binding component was purified by gel filtration chromatography; electrophoretic analyses suggested it to be composed of two polypeptide subunits of molecular masses about 115 and 98 kDa. This composition is inconsistent with hemopexin, the known heme-binding serum protein of mammals. Although it was not fully saturated with heme, the catfish component contained detectable heme in normal sera. When complexed by the binding material, heme was used as an iron source by isolates of the bacterial Gram-negative genusAeromonas; the capacity of other bacteria to use the complex was not tested. The physiological function of the catfish heme-binding serum protein is presently not clear. 相似文献
107.
Valérie Maxime Jean-Pierre Pennec Claude Peyraud 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(6):557-568
Summary The effects of increased ambient salinity (35 mg · ml-1) were studied at 1, 6, and 24 h after direct transfer of rainbow trout from freshwater to seawater. Two series of experiments were carried out successively. The first series was designed to simultaneously study all the respiratory (except Hb affinity for O2), circulatory, and acid-base variables in each fish. In this series, fish were fitted with catheters chronically inserted into the cardiac bulbus, the dorsal aorta, and the opercular and buccal cavities. In the second series, designed to study haemoglobin O2 affinity, fish were fitted with only a dorsal aorta catheter. The ventilatory flow (
) was markedly increased just after transfer (by 55% at 1 h), then more moderately (by 20% at 6 h and 32% at 24 h). The initial hyperventilation peak was associated with frequent couphing motions. These ventilatory changes resulted essentially from increase in ventilatory amplitude. Initially, standard oxygen consumption (MM}O2) decreased slightly, the moderately increased (by 12% at 24 h), so that the oxygen convection requirement (
) increased substantially. In spite of an increased ventilation, the partial pressure of oxygen in arterial blood (P
aO2) decreased slightly at 1 h, prior to returning to control levels, while partial pressure of carbon dioxide in arterial blood (P
aCO2) was not significantly decreased. Gill oxygen transfer factor decreased substantially at 1 h (by 35%) then more moderately (by 7% at 1 h and 12% at 24 h). These results suggest a decrease in gas diffusing capacity of the gills. As P
aCO2 remained approximatively unchanged, the gradual decrease in arterial pH (pHa) from 7.94 to 7.67 at 24 h must therefore be regarded as a metabolic acidosis. The strong ion difference decreased markedly because the concentration of plasma chloride increased more than that of sodium. Arterial O2 content (C
aO2) gradually decreased (by 38% at 24 h) simultaneously with the decrease in pHa, while the ratio P
aO2/C
aO2 increased. In parallel, seawater exposure induced a marked decrease in affinity of haemoglobin for O2, so that at 24 h, P50 was increased by 26% above the value obtained in freshwater-adapted trout. The increase in
could be ascribed initially (at 1 h) to the decrease of P
aO2 and later to a stimulation of respiratory neurons resulting from the lowered medullary interstitial pH. The decrease in C
aO2 could be interpreted mainly as a consequence of a decreased affinity of haemoglobin for O2, likely to be due to the blood acidosis and a predictable increase in chloride concentration within erythrocytes. Cardiac output (
) slightly decreased at 1 h, then progressively increased by 30% at 24 h. Branchial vascular resistance increased at 1 h by 28%, then decreased by 18% of the control value at 24 h. Systemic vascular resistance decreased markedly by 40% at 24 h. As heart rate (HR) remained significantly unchanged, the cardiac stroke volume initially decreased then increased in relation to the changes in
. The increase of
, allowing compensation for the effect of decreased C
aO2 in tissue O2 supply, was interpreted as a passive consequence of the decrease in total vascular resistance occurring during seawater exposure.Abbreviations a.u.
arbitrary units
-
C
aO2
arterial oxygen content
- pH50
arterial pH at P50
-
C
vO2
venous oxygen content
- Hb
haemoglobin
- HR
heart rate
- Hct
hematocrit
- nHill
Hill coefficient
- O2
standard oxygen consumption
-
P
aCO2
arterial partial pressure of carbon dioxide
-
P
aO2
arterial partial pressure of oxygen
-
P
vO2
oxygen partial pressure in mixed venous blood
- P50
oxygen tension at half saturation of haemoglobin
-
P
VA, P
DA
blood pressure in ventral and dorsal aorta
- pHa
arterial pH
-
PIO2, PEO2
oxygen partial pressure of inspired and expired water
-
PO2
oxygen partial pressure
-
cardiac output
- SEM
standard error of mean
- S.I.D.
strong ion difference
- SV
cardiac stroke volume
- TO2
gill oxygen transfer factor
- U
oxygen extraction coefficient
- VA
ventilatory amplitude
- VF
ventilatory frequency
- VRG, VRS
branchial and systemic vascular resistances
-
ventilatory flow
-
ventilatory oxygen convection requirement 相似文献
108.
C Jean-Faucher C Guilbaud M Manin C Jean 《The International journal of developmental biology》1991,35(2):77-82
This paper is concerned with hormonal regulation of the developmental pattern of major proteins of the mouse vas deferens (mouse vas deferens protein: MVDP, 34.5 kD) and seminal vesicle (15.5, 120 and 140 kD) whose expression is regulated by testosterone at adulthood. The ontogeny of these proteins, studied by SDS-polyacrylamide gel electrophoresis, appeared to be uncoordinated. MVDP was not accumulated until animals were 20 days old and its concentration increased sharply from 20 to 30 days of age. In seminal vesicle, the 15.5 kD protein did not accumulate before day 30 whereas 120 and 140 kD proteins appeared and accumulated between 30 and 40 days. In 30-day-old mice castrated at birth or treated with cyproterone acetate over 29 days, MVDP levels were not abolished and were similar to those measured in 20-day-old males. Testosterone administration, from 1 to 10 days of age, did not induce precocious expression of MVDP. These results suggest that the neonatal expression of MVDP is independent of androgens. In seminal vesicle, the first expression of the 3 proteins studied was dependent upon testicular androgens as shown by neonatal castration and injection experiments. The marked increase in the levels of the 4 proteins studied, during sexual maturation, was not associated with quantitative or qualitative changes in tissular androgen concentrations, suggesting that other factors may be necessary for protein expression. Whereas thyroxine may induce a precocious accumulation of MVDP, prolactin had no stimulatory effect on the accumulation of proteins from vas deferens and seminal vesicle. The results suggest that during sexual maturation gene activation by androgens was progressive. 相似文献
109.
Henri Debray Danuta Dus Jean-Michel Wieruszeski Gérard Strecker Jean Montreuil 《Glycoconjugate journal》1991,8(1):29-37
Four bi-antennary glycan fractions of theN-acetyllactosamine-type, derived from a Lewis lung carcinoma (LL2) cell subline resistant to theAleuria aurantia agglutinin were studied by 400 MHz1H-NMR spectroscopy. By this method, their antennae were found to be terminated either by (2-3 or 6)-linkedN-acetylneuraminic acid or (1-3)-linked galactose residues. The primary structure of glycans of these four glycopeptide or derived oligosaccharide-alditols has been determined in full detail.Abbreviations NAc
N-acetyl group
- NGc
N-glycolyl group
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- NeuGc
N-glycolylneuraminic acid
- Man
mannose
- Gal
galactose
- Fuc
fucose
- Con A
concanavalin A
- LCA
Lens culinaris agglutinin
- AAA
Aleuria aurantia agglutinin
- WGA
Wheat germ agglutinin
- RCA II
Ricinus communis agglutinin II
- PBS
phosphate buffered saline, 0.01m Na2HPO4/0.14m NaCl, pH 7.2
- HPLC
high performance liquid chromatography
- EMEM
Eagle's Minimal Essential Medium
- LecR
lectin resistant
- MG
-methylglycoside 相似文献
110.
Claude Sauvage Jean-François Rumigny Michel Maitre 《Molecular and cellular biochemistry》1991,107(1):65-77
Summary Three G proteins from human brain membranes were purified to near homogeneity by conventional techniques including preparative
electrophoresis. These G proteins were characterized by their ability to bind GTP, GDP and GTP analogs. Two of these proteins
have molecular weights of 50,000 (G50) and 36,000 (G36), as determined on SDS-gels. G36 was ADP-ribosylated by pertussis toxin. Thus, G50 could represent a Gsα subunit, whereas G36 could be Giα or Goα. G50 was phosphorylated by cAMP dependent protein kinase and protein kinase C. G36 was phosphorylated by a protein kinase independent of calcium and phospholipid, a proteolytic product of protein kinase C,
analogous to protein kinase M. Phosphorylation of G36 by this protein kinase induced a dramatic decrease in its GTPase activity. The third G protein, of molecular weight 22,000
probably belongs to the group of monomeric G proteins possessing functional similarities withras gene products. The regulation of G proteins involving calcium-dependent and independent pathways is delineated. 相似文献