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991.
Molecular cloning of bovine leukemia virus DNA integrated into the bovine tumor cell genome 总被引:3,自引:0,他引:3
The bovine leukemia virus (BLV) DNA harbored in the bovine tumor cell genome was cloned in lambda Charon 4A phage. Using either representative or 3' half-enriched BLV cDNA as a blot hybridization probe, clone lambda BLV-1 was shown to carry 9 kb of the BLV genome, flanked by cellular sequences at both ends. Restriction mapping with twelve endonucleases and hybridization of the DNA fragments to BLV cDNA representing a 3'-end portion of the viral genome revealed the presence and precise location of two long terminal repeats (LTRs) and virus-cell junctions. Thus, lambda BLV-1 appears to contain the complete BLV genome and flanking tumor cellular sequences. The restriction map of the cloned BLV proviral DNA closely resembles that previously reported for unintegrated linear proviral DNA, but differs significantly from that of the integrated provirus of another BLV isolate, the difference occurring preferentially in the putative gag and pol genes. 相似文献
992.
Abstract The antifungal antibiotic papulacandin β inhibited B(1,3)glucan-synthase activity, in vitro, from Schizosaccharomyces pombe . Levels of β(1,3)glucan-synthase from antibiotic-treated cultures were lower than the control cultures whereas mannan-synthase and β(1,3)glucanase activities were almost unaffected. The presence of an osmotic stabilizer reduced the inhibition of growth caused by the antibiotic. Addition of papulacandin β to a culture of S. pombe specifically inhibited incorporation of glucose into the β-glucan cell wall fraction. The fatty acids as well as the hydroxyl groups on the phenol residue of the papulacandin β molecule were essential for the inhibitory activity. 相似文献
993.
It is now generally accepted that the cell surface is involved in the interaction of the cells with the extracellular matrix. To identify and characterize cell-surface-associated components of corneal fibroblasts, several monoclonal antibodies were developed. Hybridomas were developed by fusing mouse myeloma cells SP2/OAg14 with spleen cells from mice immunized with membrane fractions of corneal fibroblasts grown in culture. Twenty-five hybridomas secreting monoclonal antibodies to cell-surface components were selected by an enzyme-linked immunosorbent assay using corneal fibroblasts grown in microtiter plates as the substrate. Immunohistochemical staining demonstrated that the antigenic determinants recognized by these antibodies were not present on corneal epithelial cells, but were present on skin fibroblasts. The antigenic determinants recognized by two of these antibodies, designated 10D2 and 716, were matrix components of the corneal stroma. Immunochemical characterization of the antigens was carried out by indirect precipitation of the radioactively labeled cellular proteins with the monoclonal antibodies and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the precipitates. Four antibodies were able to precipitate antigens from cell extract in detectable amounts. Antibodies designated 5E2, 9G2, and 10D2 recognized antigens consisting of polypeptides of approximate molecular weights 105K and 110K, while antibody 716 recognized an antigen of 100K molecular weight. However, based on the tissue distribution and cell-surface distribution, these antibodies reacted with different antigenic determinants. The antigen recognized by 716 was also secreted by cells in culture but consisted of 220K and 200K polypeptide chains. It was tentatively identified as cellular fibronectin, based on the reaction of this antigen with polyclonal antibodies to plasma fibronectin. 相似文献
994.
Expression of specific [125I]-prolactin-binding sites under culture conditions has been investigated for isolated mammary epithelial cells from virgin, pregnant, and lactating rabbits. Primary monolayer cultures were obtained by sequential enzymatic dispersion of mammary tissue followed by 48 hr incubation in a medium selective for epithelial cells. Scatchard analyses of binding data obtained from these cultures indicated a single class of receptor sites, the affinity constant of which (2.5 X 10(9) M-1) did not vary significantly during mammary development. The number of prolactin receptors, however, expressed by virgin and early pregnant epithelial cells was significantly increased over those from late pregnancy or lactation. Less differentiated cells also respond to growth in pregnant rabbit serum with an increase in specific [125I]-prolactin binding. The diminished receptor expression by cells obtained after 17 days of pregnancy coincides with the attainment of secretory capacity in the animal, and may reflect the influence of the low serum prolactin or high progesterone levels circulating during the last trimester in the rabbit, or be the cultural expression of secretory differentiation. 相似文献
995.
Rhodopsin in bovine photoreceptor disk membranes was subjected to limited proteolysis by thermolysin, removing twelve amino acids from rhodopsin's carboxyl terminus. (1) The rate of proteolysis is significantly faster with rhodopsin following exposure to light than with unbleached rhodopsin, provided that the incubation conditions (pH, temperature) favor the formation of metarhodopsin II. (2) If the disk membranes are illuminated under conditions in which metarhodopsin I is the predominant photoproduct (pH 8.5, 0°C), no increase in the rate of proteolysis is observed compared to unilluminated membranes. (3) The light-induced increase in the rate of proteolysis is transient: it slowly decays in the dark to the original rate found for unbleached rhodopsin. The enhanced susceptibility to proteolysis appears to measure a conformational change at rhodopsin's cytoplasmic surface which is first exhibited at the metarhodopsin II stage. This and possibly other light-dependent changes may allow rhodopsin to mediate its signal as a light-receptor protein by binding to and activating certain rod cell enzymes. 相似文献
996.
Pretreatment of Chang liver cells with (0.5 or 1 mM) stimulated Na+-independent uptake of leucine at low concentrations (?1 mM). The stimulatory effect of on the uptake of leucine measured in Na+-replete medium was completely blocked by the addition of (5 mM), which shows that the L system participates in the stimulation. The Na+-dependent uptake of glycine was depressed by pretreatment. The stimulation of the Na+-independent component of leucine uptake continued for at least 30 min after treatment, while the inhibition of glycine uptake was progressive with time and the Na+-dependent uptake of leucine became depressed later, after the treatment. It has been demonstrated that treatment of cells with is capable of increasing the Na+-independent influx of leucine and at the same time slightly decreasing the efflux of it. These results suggest that attacks the Na+-independent system of amino acid transport at the reactive SH groups(s) of relevant protein(s) in favor of specific activation of that system in this cell. 相似文献
997.
Binding, internalization and intracellular processing of 125I-epidermal growth factor purified by isoelectric focusing 总被引:1,自引:0,他引:1
B E Magun S R Planck L M Matrisian J S Finch 《Biochemical and biophysical research communications》1982,108(1):299-306
When epidermal growth factor (EGF) which had been extensively purified by HPLC was subjected to iodination with sodium 125iodide, 5 major species of differing isoelectric points were produced. Some of these species bound to rat fibroblasts with different affinities but were internalized with equal efficiency. Examination of the internalized 125I-labelled molecules revealed processing of all the 125I-EGF species to macromolecules with more acidic isoelectric points. The 125I-EGF species with a pI of 4.5 corresponded in electrofocusing behavior with intact non-iodinated EGF. Other EGF species probably represented molecules which were covalently modified as a result of the iodination procedure. 相似文献
998.
Thomas E. Gray David G. Thomassen Marc J. Mass J. Carl Barrett 《In vitro cellular & developmental biology. Plant》1983,19(7):559-570
Summary A cell culture system is described for the growth of rat tracheal epithelial (RTE) cells at clonal density. The system uses
normal, early passage RTE cells grown on feeder layers of lethally irradiated 3T3 cells. The RTE cells have a high colony
forming efficiency (5 to 10%) in culture, can be passaged up to 5 times, and are capable of more than 20 cumulative doublings
per colony forming cell. The epithelial nature of the cells was confirmed by cell and colony morphology, immunoperoxidase
staining of intracellular keratin, and cellular ultrastructural studies. The cytotoxic response of RTE cells to a variety
of carcinogens, including a direct acting chemical carcinogen, a physical carcinogen, and a series of polycyclic aromatic
hydrocarbons, was quantitated. A linear decrease in the logarithm of survival was observed with increasing doses ofN-methyl-N′-nitro-N-nitrosoguanidine (MNNG), γ-irradiation, 7,12-dimethylbenz(a)anthracene, and a diol-epoxide of benzo(a)pyrene. No toxicity was observed after treatment with benzo(a)pyrene or 3-methylcholanthrene over the concentration range examined. In contrast, phorbol ester tumor promoters stimulated
cell growth markedly. Based on these and other studies, the RTE cell culture system represents a model system that will be
useful for quantitative studies of epithelial cell growth, differentiation, and carcinogenesis. 相似文献
999.
Summary To optimize culture conditions and gain a more reliable culturing system for studies of metabolic properties of neuronal cells,
a simplified perfusion chamber was developed. It consists of two parts: a perfusion block and a standard plastic culture dish.
To confirm the suitability of this chamber for continuous culturing of anchorage-dependent cells, the growth and morphology
of the four neuronal cell lines glioma C6 and glioma 138MG, neuroblastoma C1300, clones N1E115 and N18 were followed for 4
d using both traditional and perfusion techniques. A marked increase in growth and a decrease in the degree of morphological
differentiation were obtained with the latter technique compared to the former.
This work was supported by grants from the National Swedish Board for Technical Development (Grant 81-5009), the Swedish Work
Environmental Foundation (Grant 76-53), and Ollie and Elof Ericssons Foundation for Scientific Research. 相似文献
1000.
Cultivation of mosquito cell lines in serum-free media and their effects on dengue virus replication
Goro Kuno 《In vitro cellular & developmental biology. Plant》1983,19(9):707-713
Summary Seven mosquito cell lines from five species (Aedes aegypti, Ae. albopictus, Ae. pseudoscutellaris, Culex tarsalis, andToxorhynchites amboinensis) were adapted to three kinds of serum-free media (SEM), which were composed of equal volumes of tryptose phosphate broth
and of either Leibovitz (L15) medium, Eagle’s minimum essential medium, or Medium 199 with Hanks’ salts. Population growth
rates of the cells cultivated in the SMFs were generally slower than those of original cell cultures maintained in conventional
media containing bovine sera. A karyological study showed a significant shift to heteroploidy in two of the four cell lines
examined. Four SMF-adapted sublines were compared with parental cultures for replication of dengue viruses.Ae. aegypti RML-12,Ae. albopictus C6/36,Ae. pseudoscutellaris AP-61, andTx. amboinensis TRA-171 demonstrated different levels of alteration in virus replication ranging from lower titers (as inAe. albopictus C6/36) to comparable or higher titers (as inAe. aegypti RML-12) when they were simultaneously inoculated with four dengue serotypes.
Use of trade names and commercial sources is for identification only and does not constitute endorsement by the Public Health
Service or by the U.S. Department of Health and Human Services. 相似文献