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131.
Phagocytosis of Tetrahymena is inhibited by prednisolone-sodium-succinate and deoxy-corticosterone-glucoside, and stimulated by dexamethasone and prednisolone. Dexamethasone and estradiol enter the cells and are localized first in food vacuoles, and later on in the cytosol. They were never found in the nucleus. The demonstration by biochemical methods of a specific glucocorticoid binding protein failed in all three subcellular fractions examined. 相似文献
132.
M. Dürr K. Urech Th. Boller A. Wiemken J. Schwencke M. Nagy 《Archives of microbiology》1979,122(2):169-175
The conditions for synthesis, purification, and properties of tryptophanase by a marine organism (Vibrio K-7) were studied. Tryptophanase was induced by tryptophan and its analogs, and partially repressed by 0.5% glucose or glycerol. NaCl (0.4M) was required for optimal growth and tryptophanase activity in whole cells. The enzyme was purified to 92% homogeneity by heat treatment, hydroxyapatite chromatography and fractionation with ammonium sulfate. This tryptophanase has been found to have kinetic properties similar to the tryptophanase from other microorganisms. It carries out both , -elimination reactions (using tryptophan, serine, cysteine and S-methyl-cysteine as substrates) and -replacement reactions (forming tryptophan from indole and serine, cysteine or S-methyl-cysteine). The enzyme has a sedimentation coefficient of 9.2S and requires pyridoxal 5-phosphate as a cofactor. The optimal pH for the tryptophanase reaction is pH 8.0.Nonstandard Abbreviations PLP
pyridoxal 5-phosphate
- TPase
tryptophanase
- TSase
tryptophan synthase
- DHase
dehydratase
- TCA
tricarboxylic acid
- BSA
bovine serum albumin
Preliminary reports of this work have been presented (M. J. Klug and R. D. DeMoss, Bacteriol. Proc. 1971, p. 132; D. D. Whitt and R. D. DeMoss, Abstr. Annu. Meet. Am. Soc. Microbiol. 1973, p. 148) 相似文献
133.
The role of estrogens in the regulation of lactate dehydrogenase activity and its submolecular organization in rat anterior pituitary 总被引:1,自引:0,他引:1
Estrogens increase LDH activity and decrease H/M subunit ratio in rat anterior pituitary in both the experimental circumstances and the physiological conditions. The cellular messengers mediating estrogenic effect are structure- and stereospecific. The activity increasing and subunit ratio decreasing potency of the three tested estrogens was of following decreasing order: 17 beta-estradiol, estrone, and estriol. 17 alpha-estradiol did not affect activity parameters and submolecular organisation of the enzyme. The estrogen induced activity increase is consequence of the enhanced de novo enzyme protein synthesis which could be inhibited by Actinomycin-D. The lack of adrenocorticoids did not involve the alteration of LDH activity and H/M ratio in female rat anterior pituitary. Accordingly, these steroids do not mediate estrogenic action. 17 beta-estradiol had a substantial increasing effect on LDH activity in the subrenal implanted pituitary homografts and decreased H/M subunit ratio. Pituitary LDH activity in androgenized female rats decreased only after the removal of the polycystic ovaries. The two latter observations suggest that hypothalamic hormones are not involved in the regulation of pituitary LDH activity and its submolecular organization. De novo synthesis of LDH enzyme protein and the regulation its submolecular organization is induced by the direct action on anterior pituitary cells of the estrogens. 相似文献
134.
135.
Vladimir S. Gaitskhoki Felix I. Yershov Oleg I. Kisselev Olga V. Zaitseva Victor M. Zhdanov Solomon A. Neifakh 《Molecular and cellular biochemistry》1976,10(1):17-26
Summary The synthesis of virus-specific macromolecules was studied in the reconstituted system containing inner membrane-matrix fraction from rat liver mitochondria and infectious RNA of Venezuelian equine encephalomyelitis (VEE) virus. In a series of preliminary experiments it was shown that isolated submitochondrial fraction was completely free of interfering cytoplasmic contaminations and particularly, of cytoplasmic 80S ribosomes. VEE RNA when added to submitochondrial system caused significant stimulation of RNA and protein synthesis. These processes were resistant to actinomycin D which inhibited profoundly the synthesis of proper mitochondrial macromolecules. The stimulating effect of VEE RNA in experiments with submitochondrial system was about three times higher than that with intact mitochondria. The stimulation of14C-amino acid incorporation increased as a function of incubation time; a certain lag-period being observed. The newly formed virus-specific RNA's and ribonucleoproteins were identified with the aid of sedimentation analysis. In particular, radioactive RNA's with sedimentation coefficients 40S and 26-18S were isolated from the incubated system. These RNA's are similar respectively to VEE genome RNA and doublestranded VEE replicative RNA. In double labelling experiments with3H-uridine and14Camino acids it was shown that VEE RNA induced synthesis of ribonucleoproteins containing newly formed RNA and protein. These RNP possessed sedimentation coefficients 60-80S, 140S and 300S in sucrose gradient and buoyant densities 1.32 and 1.50 g/cm3 in cesium chloride gradients. These properties of ribonucleoproteins synthesized de novo in submitochondrial system are close to those of RNP intermediates of VEE virus reproduction in the infected cells. We concluded that viral RNA could program virus-specific synthesis in the submitochondrial system under conditions that eliminated the contribution of cytoplasmic ribosomes. 相似文献
136.
Walter D. Wosilait Carlos Soler-Argilaga Paul Nagy 《Biochemical and biophysical research communications》1976,71(2):419-426
The distribution of palmitate between the form bound by human serum albumin and the free form in plasma was calculated by use of 12 stepwise equilibrium constants and a computer program. Computations were carried out for molar ratios of palmitate to serum albumin of 0.5, 1,2,3, and 4. At most 0.0003% of the palmitate would be in the unbound form, and the remainder distributed among different complexes with albumin. At low molar ratios, the complexes with 1 to 2 moles of palmitate/albumin would predominate while at the highest ratio the complexes of 3 and 4 moles of palmitate/albumin would be most abundant. In the delivery of palmitate to tissues the relative contribution of the different complexes would change, as the molar ratio of fatty acid to albumin changed. 相似文献
137.
In this pilot study, the relationships between glucose and lactate concentrations of plasma, cerebrospinal fluid (CSF), cerebral cortex and subjacent white matter were investigated in one hyperglycaemic and three normoglycaemic anaesthetized rabbits. After a 90 min stabilization period, CSF was sampled and the brain frozen in situ. Triplet samples (n = 3 X 21) were obtained from the outer and inner halves of cortex and from the white substance and analysed for their water content as well as for glucose and lactate by enzymatic fluorescence methods. Preservation of the ATP content was demonstrated in brain slices by a bioluminescence method. The glucose and lactate levels of CSF seemed to reflect those of the outer half of the cortex. In the normoglycaemic animals, the tissue glucose and tissue lactate levels correlated inversely (r = 0.477: p less than 0.01). While the glucose concentrations were nearly identical in the inner cortex and white substance, there was a concentration difference of 0.54 mmol/kg tissue water between the outer half of the cortex and the white matter (p much less than 0.02). This might correspond to a steep intra-cortical glucose gradient starting from the CSF-facing surface and approximating the general cerebral glucose level in a depth of about 4-500 microns. The possible significance of this gradient in regulating CSF glucose is discussed. 相似文献
138.
The combination of vertical, one-dimensional isoelectric focusing and immunoblotting works very well for the evaluation of the phosphorylation state of the α-subunit of eIF2 using reticulocyte lysate or purified eIF2. However, the method is more difficult to apply to the analysis of eIF2α phosphorylation in cultured cells. In part this reflects the fact that the protein content of cultured cell extracts is rarely as high as that found in extracts produced from reticulocytes, and in part this reflects the fact that some component(s) of cell extracts interferes with the entry of eIF2α into the isoelectric focusing gel. To overcome these difficulties, we have modified the earlier method to include immunoprecipitation of eIF2 from cell extracts prior to isoelectric focusing, as well as a low sodium dodecyl sulfate concentration in the isoelectric focusing sample buffer. Since the PKR activation state and therefore the eIF2α phosphorylation state change with cell density and nutritional status, we routinely set up consistent feeding schedules and recommend the collection of data over a range of cell densities. 相似文献
139.
Joran Martijn Anders E. Lind Max E. Schön Ian Spiertz Lina Juzokaite Ignas Bunikis Olga V. Pettersson Thijs J. G. Ettema 《Environmental microbiology》2019,21(7):2485-2498
Amplicon sequencing of the 16S rRNA gene is the predominant method to quantify microbial compositions and to discover novel lineages. However, traditional short amplicons often do not contain enough information to confidently resolve their phylogeny. Here we present a cost-effective protocol that amplifies a large part of the rRNA operon and sequences the amplicons with PacBio technology. We tested our method on a mock community and developed a read-curation pipeline that reduces the overall read error rate to 0.18%. Applying our method on four environmental samples, we captured near full-length rRNA operon amplicons from a large diversity of prokaryotes. The method operated at moderately high-throughput (22286–37,850 raw ccs reads) and generated a large amount of putative novel archaeal 23S rRNA gene sequences compared to the archaeal SILVA database. These long amplicons allowed for higher resolution during taxonomic classification by means of long (∼1000 bp) 16S rRNA gene fragments and for substantially more confident phylogenies by means of combined near full-length 16S and 23S rRNA gene sequences, compared to shorter traditional amplicons (250 bp of the 16S rRNA gene). We recommend our method to those who wish to cost-effectively and confidently estimate the phylogenetic diversity of prokaryotes in environmental samples at high throughput. 相似文献
140.