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991.
John J. Malinowski Bruce L. Grasberger Gary Trakshel Edward E. Huston Tracey M. Banks Patricia G. Brake Richard B. Ciccarelli Barry N. Jones James A. Koehn Diane Kratz Nicole Lundberg Panayiotis E. Stevis Carla T. Helaszek Mark A. Ator Angela M. Small Wood Travis Stams Byron Rubin Richard S. Alexander 《Protein science : a publication of the Protein Society》1995,4(10):2149-2155
992.
Androgen can directly modulate the induction of steroidogenic enzymes by FSH (follicle stimulating hormone) in ovary granulosa cells. In studies of its mechanism of action, we examined the androgen effect on granulosa cell interaction with lipoproteins, the physiologic source of cholesterol. After granulosa cells were cultured for 48 hours with and without androgen and/or FSH, the cells were incubated for 24 hours with 125I-lipoproteins [human high density lipoprotein (HDL), rat HDL, or human low density lipoprotein (LDL)]. The media were then analyzed for lipoprotein protein coat degradation products (mainly 125I-monoiodotyrosine) and progestin [mainly 20α-dihydroprogesterone (20α-DHP)]. In the absence of FSH and androgen, 2 × 105 granulosa cells degraded basal levels of all three lipoproteins, but produced no measurable 20α-DHP. The addition of 10?7 M androstenedione (A), testosterone (T), or 5α-dihydrotestosterone (DHT) had no effect on lipoprotein protein degradation or 20α-DHP production. FSH alone stimulated lipoprotein protein degradation by 50 to 300% while the addition of androgen synergistically augmented the FSH-stimulated 20α-DHP production as well as protein coat degradation of all three lipoproteins. DHT and T were both effective, indicating that androgens themselves, and not estrogen products, were responsible for the effect on lipoprotein protein degradation and 20α-DHP production. The addition of a 10-fold excess cyproterone acetate (an anti-androgen) inhibited the effect of T, suggesting that the action of T was mediated by the granulosa cell androgen receptor. Androgen and FSH also synergistically stimulated the production of 3H-progestin when the granulosa cells were incubated with either 3H-cholesterol ester core labeled human HDL or similarly labeled human LDL. This report demonstrates that androgen, in combination with FSH, augments the steroidogenic pathway of the granulosa cell from the degradation of lipoprotein and utilization of the cholesterol ester core, to the production of progestin product. 相似文献
993.
Applicability of the fluorescein diacetate method of detecting active bacteria in freshwater 总被引:6,自引:0,他引:6
Thomas H. Chrzanowski Rhonda D. Crotty James G. Hubbard Robert P. Welch 《Microbial ecology》1984,10(2):179-185
Fluorescein diacetate (FDA) hydrolysis was evaluated as a means to detect actively metabolizing bacteria in freshwater. Fluorescein diacetate, a nonfluorescent derivative of fluorescein, can be transported across cell membranes and deacetylated by nonspecific esterases. Resultant fluorescein accumulates within cells and allows direct visualization by epifluorescent microscopy. Application of FDA to a variety of freshwater habitats yielded estimates of active cells ranging from 6–24% of the total population. These estimates were 49–61% lower than estimates of active cells obtained from measures of electron transport activity. The difference was attributed to low permeability of the fluorogen through the outer membrane of heterotrophic gram-negative cells. Data suggest that FDA hydrolysis as a means of detecting active bacteria may be limited to environments rich in eucaryotes and gram-positive cells. 相似文献
994.
John M. Beale Jr James M. Hewitt John P. Rosazza 《Enzyme and microbial technology》1984,6(12):543-548
One hundred microorganisms have been screened for their abilities to selectively modify the structure of the sesquiterpene lactone known as quadrone. The only products obtained were those formed when the 4-ketone functional group was reduced to the stereoisometric 4-quadronols. Quadrone alcohol isomers of (S) or (R) absolute configurations were identified by proton and carbon n.m.r., and high performance liquid chromatography (h.p.l.c.) was used to separate and quantitate these compounds in extracts of fermentations. Microorganisms were categorized according to their abilities to achieve Re- or Si-face carbonyl reduction to yield (S)- or (R)-alcohol isomers by h.p.l.c. Three groups of microorganisms were identified: those yielding only the (R)-alcohol isomer; those yielding only the (S)-alcohol isomer; and those providing mixtures of the two alcohol isomers. With quadrone as substrate, Mucor and Curvularia spp. may contain either Re- or Si-face reductases. The selection of microorganisms for their abilities to achieve enantiospecific reductions of ketones to alcohol products is discussed. 相似文献
995.
Protoplasts were separately stained with the fluorescent dyes fluorescein iso-thiocyanate (FITC) and tetramethylrhodamine isothiocyanate (TRITC). Following fusion, doubly stained heterokaryons were identified under fluorescence microscopy by using the Zeiss filter set 48 77 05 (excitation filter 450-490 nm, dichroic reflector 510 nm, and barrier filter 520 nm) which allowed simultaneous fluorochrome emissions. Previously, either emisson spectrum, but not both, was possible for any single filter set. 相似文献
996.
R. Van Potter Theresa Ruh Evanson Debra P. Gayda James A. Gurr 《In vitro cellular & developmental biology. Plant》1984,20(9):723-731
Summary The induction and decay of ornithine decarboxylase (ODC) by insulin and asparagine in cultures of H4-II-EC3 (H35) hepatoma
cells was studied in a modified Waymouth medium in the presence of fetal bovine serum (FBS) and in serum-free media. The insulin
response was enhanced by the presence of asparagine although the effect of asparagine was not so much on the initial increase
as it was on a slowing of the decline after the maximum was reached at 6 to 8 h after the supplements were added together
with fresh medium. In all cases the initial ODC activity was zero at zero time for addition of media and supplements, and,
after reaching the maximum, activity declined to near zero by 24 h. Fetal bovine serum gave induction that followed a similar
time course but was inferior to the combintion, of insulin plus asparagine and, in fact, FBS inhibited the latter response.
Putrescine (the product formed from ornithine by ODC), at 10−5
M, markedly inhibited the induction of ODC by insulin or FBS, but the inhibition was less when asparagine was present.
This work was supported in part by Grants CA-07175, CA-22484, and CA-17334 from the National Cancer Institute. D. P. G. is
a Predoctoral Fellow at the Food Research Institute, supported by a fellowship from the Monsanto Fund and by NIH Grant R01-AI
15693 to Prof. Michael W. Pariza, Food Research Institute, University of Wisconsin, Madison. 相似文献
997.
Roger A. Williamson James K. Koehler W. Dianne Smith Laurence E. Karp 《Molecular reproduction and development》1984,10(3):319-325
We studied six men whose spermatozoa were immotile and possessed a variety of sperm tail structural abnormalities by electron microscopy. The semen of all six subjects had a normal percentage of oval forms and sperm undergoing capacitation and acrosome reaction. Despite the absence of motility, when incubated sperm from these subjects was added to a microdrop of medium containing zona pellucida-free hamster ova, sperm penetration or entry into the cytoplasm of from 1–9% of the eggs was evident with phase contrast microscopy. This latter finding suggests that, at least in this system, oocytes actively facilitate sperm incorporation. Penetration was absent when sperm of fertile men were rendered immotile, though still viable, by heat treatment. 相似文献
998.
This study was conducted to determine the optimal concentration of sperm to use for the insemination of females to detect differences among strains of mice in the percentage of eggs fertilized. Female ICR mice were inseminated with sperm of concentrations ranging from 0.25 to 8 × 106/50 μl from males of either DBA/2N, CF1, or C57BL/6N strains. Differences among strains were detected only when approximately 50% of the eggs were fertilized but not when each of the strains fertilized either a high or low percentage of eggs. The optimal concentration of sperm therefore was the concentration that gave approximately 50% fertilized eggs. 相似文献
999.
Edith Wallace Harold I. Calvin Miklos P. Salgo James E. Dennis Karin Ploetz 《Molecular reproduction and development》1984,9(4):375-386
Zinc is required for spermatogenesis in mammals and is concentrated in the dense outer fibers of the sperm tail, where it is associated with cysteine-rich protein. To investigate the effects of marginal zinc deficiency upon dense fiber formation and upon sperm quality in general, weanling Sprague-Dawley rats were administered a commercial low-zinc diet, supplemented with phytate, for approximately 60 days, and were compared with controls fed the same diet plus 50 ppm zinc in their drinking water. The following characteristics of the zinc-deficient rats were significantly lower than in the controls: body weight, testis weight, epididymis weight, seminal vesicle weight, sperm content of the cauda epididy-midis, sperm motility, testis zinc, and hair zinc. By contrast, the levels of sperm zinc and sperm sulfhydryls were the same in the zinc-deficient and control rats. The zinc-deficient rats displayed a highly variable spectrum of sperm defects, which included decapitation, disorganized and redundant tail elements, and superfluous cytoplasm. However, abortive dense fiber development was only rarely observed. Apparently, even when availability of zinc is limited and reduced sperm production ensues, elaboration of dense fibers rich in zinc and sulfhydryls continues to be obligatory for the completion of spermiogenesis. 相似文献
1000.
James S. Clegg 《Cell biochemistry and biophysics》1984,6(3):153-169
Cysts of the crustaceanArtemia are a useful model for studies on intracellular water because they are capable of essentially complete and reversible desiccation.
We have used a variety of techniques on this system, the present work being an attempt to estimate the density of intracellular
water (ρw). The density of individual cysts was evaluated from sedimentation velocity. Heptane displacement methods were used to determine
the volume of a known mass of cysts, from which the density was calculated. The two methods produce comparable results. It
was shown that the densities and water contents of large masses of cysts accurately reflect those of individual cysts. Cyst
densities (ρc) were determined over the entire range of water content from 0 to 0.63 weight fraction of water (W
f), and temperature dependence was measured for 0.61W
f over 2–41°C. The following refer to 25°C. No marked change was detected in ρc until the water content exceeded 0.15W
f, at which ρc decreased as a linear function of Wf to maximum water content. However, the cyst does not behave ideally in the sense that
the densities of the nonaqueous components and added water are not additive as a function ofW
f. The partial specific volume of water in cysts at maximum hydration was estimated to be 3% larger than that of pure water.
These observations are compared with density measurements on other systems, and with previous findings on the physical properties
of water in this system. 相似文献