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991.
F. Sima Sariaslani Michael K. Trower Daniel A. Kunz 《Biocatalysis and Biotransformation》1989,2(2):151-160
Streptomyces griseus oxidizes the insecticide precocene II to its cis- and trans-dihydrodiols and 3-chromenol after growth on an enriched medium containing soybean flour. Oxidation of precocene II is dependent on the level of cytochrome P-450 in this organism. Extracts of cells grown on media lacking soybean flour were devoid of cytochrome P-450 and could not oxidize precocene II. In an in vitro reconstituted system containing NADPH, spinach ferredoxin reductase, spinach ferredoxin and ammonium sulfate fractions enriched in cytochrome P-450, precocene II was oxidized to its dihydrodiols. An aerial mycelium-negative variant of S. griseus (AMY mutant), that was unable to elicit cytochrome P-450 when grown on soybean flour-enriched medium, failed to oxidize precocene II. 相似文献
992.
The influence of the source of pregnant mares' serum gonadotropin (PMSG) on the num ber, quality, and in vitro development of mouse embryos before and after freezing was evaluated among three genotypes: N:NIH(S), C57BL/6N, and C3H/HeN-MTV?. Immature females were given PMSG from one of five commercial sources. Following col lection ( 116 hr later), embryos were evaluated for stage of development, and four-to eight-cell embryos were pooled within genotype and assigned to standardized fresh or freeze-thaw culture trials. Different PMSG sources stimulated the production of different num bers of total embryos (P < 0.05) but not necessarily more embryos suitable for freezing. Differences in embryo production among genotypes indicated that absolute embryo num bers using a single mouse genotype may not accurately reflect the potency of a specific gonadotropin source. The PMSG source also affected the ability of an embryo to survive in culture either immediately after collection or after frozen storage. The effect, however, was genotype specific, with some mouse strains being relatively insensitive to PMSG source, whereas gonadotropin source played a major role in determining in vitro viability in others. Development rates for freshly collected embryos differed, often inconsistently, from those of thawed embryos regardless of the PMSG source used, demonstrating that fresh embryo development cannot be used to estimate expected post-thaw survival. In vitro development of thawed embryos is influenced not only by genotype, but also the source of the gonadotropin used to promote follicular development and oocyte maturation. These findings may explain, in part, the wide variation in embryo viability and culture rates reported among laboratories and intraspecies animal populations. 相似文献
993.
Characterization of two soybean repetitive proline-rich proteins and a cognate cDNA from germinated axes 总被引:18,自引:7,他引:11
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We have resolved and analyzed two proline-rich proteins isolated from the walls of soybean cells in culture. The proteins are similar in amino acid content, containing 20% proline, 20% hydroxyproline, 20% lysine, 16% valine, 10% tyrosine, and 10% glutamate. The proteins undergo a rearrangement or a limited cleavage in dilute NaOH, but are otherwise remarkably stable to a high concentration of alkali. We have cloned and sequenced a cDNA from soybean axes germinated for 31 hours (1A10-2) coding for a protein that closely corresponds in its amino acid content to that of the proline-rich proteins. The cDNA sequence predicts a decameric repeat of Pro-Pro-Val-Tyr-Lys-Pro-Pro-Val-Glu-Lys. Consequently, this class of proteins is referred to as repetitive proline-rich proteins, i.e., RPRP2 and RPRP3. We have also analyzed RNA gel blots with probes that discriminate between the new cDNA clone and a related cDNA previously reported [SbPRP1; Hong, Nagao, and Key (1987). J. Biol. Chem. 262, 8367-8376]. Messenger RNAs from young seedlings and from soybean suspension cultures correspond primarily to the new RPRP clone (1A10-2), whereas the predominant mRNA accumulating later in the roots corresponds to SbPRP1. 相似文献
994.
Adrian S. Dobs Christiane Broussolle M. Daniel Lane 《In vitro cellular & developmental biology. Plant》1989,25(2):112-114
Summary The insulin-producing cell line RINm5F, has been used in short-term experiments to evaluate insulin secretion. We sought to
maintain the responsiveness of these cells to stimuli for up to 2 days. We examined the course of new insulin synthesis over
this period by measuring at intervals immunoreactive insulin (IRI) in two parts: IRI in the medium (M) and IRI extracted from
the cells (C). Control cells were incubated in RPMI 1640/2.8 mM glucose/10% fetal bovine serum/200 μg/ml bacitracin (to prevent
insulin degradation). The addition of dibutyryl cAMP 10 mM to the experimental dishes significantly increased total (M+C)
IRI at 48 hr to 37% above the insulin content of the control dishes (p<0.01). Theophylline 10 mM increased total (M+C) IRI
by 24% over control (p<0.05) after 24 hrs. Glucose, glyceraldehyde, leucine, arginine, glucagon and tolbutamide, other stimulants
of insulin production, had no effect. Under the experimental conditions reported here, including the use of bacitracin, IRI
synthesis can be studied for up to 48 hr.
Portions of this study have been published in abstract form for the 47th Annual Meeting of the American Diabetes Association,
Indianapolis, Indiana, 1987.
Supported in part by the American Diabetic Association, Maryland Affiliate. 相似文献
995.
Jila H. Boal Scott F. Deamond Daniel E. Callahan Sarah A. Bruce Paul O. P. Ts'o L. L. Kan 《Cell biochemistry and biophysics》1989,14(3):245-256
The nuclear magnetic resonance (NMR) parameters, spin-lattice (T1), and spin-spin (T2) relaxation time, are usually longer for neoplastic cells than for normal cells of the same cell type. This has generally
been true at low NMR frequencies (≤100 MHz) when comparisons have been made between normal and neoplastic cells that have
both spent a short time in culture. We have previously demonstrated that although the T1 values of paired normal and neoplastic Syrian hamster (SH) fibroblastic cells in culture are not significantly different
when measured at 300 MHz, the 300 MHz T2 values for the neoplastic cells are smaller than those of the normal cells. (Xin et al. (1986),Cell Biophysics
8, 213.) Since treatment of normal diploid cells with polypeptide growth factors or tumor promoters frequently results in reversible
expression of neoplasia-associated phenotypes, T1 and T2 were obtained at 300 MHz for treated and untreated SH cells to see if these compounds could also produce smaller 300 MHz
T2 values. Secondary culture SH fetal fibroblast cells were treated with epidermal growth factor (EGF), fibroblast growth factor
(FGF), phorbol-12,13-didecanoate (PDD) and 4-α-phorbol-12,13-didecanoate (4αPDD). Treatment with either growth factor resulted
in smaller T2 values, but a statistically significant decrease was not observed for PDD or 4αPDD. The observed reductions in T2 values were correlated with the morphological and growth-stimulatory effects of these compounds on the cells. 相似文献
996.
Elizabeth B. Gargus Douglas H. Robinson James K. Bubien Lawrence B. Bugaisky Dale J. Benos 《In vitro cellular & developmental biology. Plant》1989,25(5):435-441
Summary Six- and seven-day post-coitus (p.c.) rabbit embryos have been cultured in an attempt to establish a trophectodermal cell
line. Results indicate that cells with epithelial characteristics (i.e. positive staining for cytokeratin) will survive in
culture until Passage 3. At that time a fibroblastlike cell becomes predominant. In addition, we have found that the presence
of the inner cell mass is required for embryo explants often results in the development of cells that spontaneously contract.
These cells stain positively for myosin, which indicates that they may be precardiac cells. Maximum diastolic potential was
−59±1.2 mV and the threshold potential was −53±2.3 mV. Spontaneously contracting cells did not respond to atropine, acetylcholine,
epinephrine, isoproterenol, or propranolol. Action potential seems to be a result of an inward calcium current, because the
beating rate is decreased in a dose-related manner with the calcium channel blocker verapamil, whereas the voltage-sensitive
sodium channel blocker tetrodotoxin was without effect.
This work was supported by grants HD21302, HD07069, DK31091, and HL37320 from the National Institutes of Health, Bethesda,
MD, with additional support from a University of Alabama at Birmingham Cardviovascular Research and Training Center Award. 相似文献
997.
James C. Beck Howard L. Hosick Bruce A. Watkins 《In vitro cellular & developmental biology. Plant》1989,25(5):409-418
Summary We investigated the effects of conditioned media derived from mouse mammary fat pads on the proliferation of CL-S1 cells,
an epithelial cell line originally isolated from a preneoplastic mammary outgrowth line. Cell proliferation in vitro in serum-free
defined medium was compared to that in this medium conditioned using intact mammary fat pad pieces or isolated fat pad adipocytes.
Culture medium was conditioned by incubating the conditioning material in defined culture medium for 24 h at 37°C. Conditioned
medium induced CL-S1 proliferation as much as 10- to 20-fold above the minimal levels of growth in control cultures after
13 d of culture. The growth-stimulatory factor(s) had an apparent molecular weight of greater than 10 kDa. This growth-stimulatory
activity was both heat and trypsin stable. Because the role of adipose tissue is to store and release lipids, we next tested
whether lipids are released during medium conditioning. The lipid composition of the fat pad conditioned medium was characterized
using both thin layer and gas liquid chromatography. These lipid analyses indicated that the fat pad pieces released significant
amounts of fatty acids and phospholipids into the medium during the conditioning period. The free fatty acid composition included
both saturated and unsaturated molecules, and about 80% of the total fatty acids consisted of palmitate, stearate, oleate,
and linoleate. These same fatty acids were a structural component of the majority of phospholipid found in the medium. The
addition of palmitate or stearate to defined medium had no effect or was inhibitory for CL-S1 proliferation, depending on
the concentration used. Defined medium supplemented with oleate, arachidonate, or linoleate induced CL-S1 proliferation, and
the inhibitory effects of palmitate and stearate were overcome by addition of oleate and linoleate. These data indicate that
both unsaturated and saturated fatty acids are released from intact adipose cells of the mouse mammary fat pad and that fatty
acids can influence the growth of prenoplastic mouse mammary epithelium. Thus, unsaturated fatty acids, perhaps in conjunction
with other substances released simultaneously, are candidate molecules for the substances that mediate the effect of adipose
tissue on growth of epithelium.
This work was supported in part by a grant from the American Institute for Cancer Research; grant CA 46885 from the National
Institutes of Health, Bethesda, MD; and by State of Washington initiative 171. 相似文献
998.
Theneu oncogene is frequently found in certain types of human carcinomas and has been shown to be activated in animal models by nitrosourea-induced mutation. The activating mutation in theneu oncogene results in the substitution of a glutamic acid for a valine at position 664 in the transmembrane domain of the encoded protein product of 185 kda (designated p185), which, on the basis of homology studies, is presumed to be a receptor for an as yet unidentified growth factor. It has been proposed that activating amino acid substitutions in this region of p185 lead to a conformational change in the protein which causes signal transduction via an increase in tyrosine kinase activity in the absence of any external signal. Using conformational energy analysis, we have determined the preferred three-dimensional structures for the transmembrane decapeptide (residues 658–667) of the p185 protein with valine and glutamic acid at the critical position 664. The results indicate that the global minimum energy conformation of the decapeptide from the normal protein with Val at position 664 is an α-helix with a sharp bend (CD* conformation at residues 664 and 665) in this region, whereas the global minimum conformation for the decapeptide from the mutant transforming protein with Glu at position 664 assumes an all α-helical configuration. Furthermore, the second highest energy conformation for the decapeptide from the normal protein is identical to the global minimum energy conformation for the decapeptide from the transforming protein, providing a possible explanation why overexpression of the normal protein also has a transforming effect. These results suggest there may be a normal and a transforming conformation for theneu-encoded p185 proteins which may explain their differences in transforming activity. 相似文献
999.
Brain Cell Biology - The effects of the venom of the spider Latrodectus mactans hasselti on the superior cervical ganglion were studied in the guinea pig. Under anaesthesia the ganglion was bathed... 相似文献