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121.
122.
By coupling Anisakis larval hemoglobin with cyanogen bromide-activated Sepharose, a stable and sensitive antigen for the indirect fluorescent antibody test for anisakiasis was prepared. Using this antigen, cross-reaction with other helminth infections was minimized and the test was greatly simplified.Furthermore, the results obtained indicate that this technique may be useful not only in the sero-diagnosis of anisakiasis but may also be applicable to the diagnosis of other helminthic diseases. 相似文献
123.
Grazyna Adamus Z. Suzanne Zam Scotts Emerson Paul A. Hargrave 《In vitro cellular & developmental biology. Plant》1989,25(12):1141-1146
Summary We present a practical method for the rescue of previosly stable hybridoma clones which increases the proportion of desired
cells in the population before cloning by limiting dilution. When the antibody activity of a culture supernatant was lower
than that previously obtained, a precloning distribution at a density of 10 cells per microtiter well greatly improved the
chances of obtaining a single active clone by subsequent limiting dilution. The Poisson distribution model was used to evaluate
the method. Probabilities calculated clearly demonstrate the advantage of this precloning distribution step when attempting
to isolate a hydridoma cell line that is relatively rare in a population.
This work was supported in part by grants EY 06225 and EY 06226 from the National Eye Institute of the National Institutes
of Health, Bethesda, MD and by an unrestricted departmental award from Research to Prevent Blindness, Inc. 相似文献
124.
Carol L. Williams Vanda A. Lennon Mark R. Pittelkow 《In vitro cellular & developmental biology. Plant》1989,25(5):397-401
Summary A time-dependent redistribution of microfilaments was observed in cultured human keratinocytes using a human monoclonal autoantibody
specific for myosin. Immunofluorescent staining revealed that 5 days after plating keratinocytes in either 0.1 mM or 2.0 mM
Ca++, myosin was distributed uniformly throughout the cytoplasm. At day 6, parallel arrays of myosin-containing microfilaments
were prominent in the cell peripheries. At day 7 the microfilaments formed circumferential rings. The distribution of the
microfilaments was disrupted by cytochalasin but not by colchicine, indicating that this novel distribution of myosin was
not dependent on colchicine-sensitive vimentin intermediate filaments. The time-dependent redistribution of myosin was not
influenced by cell population density, cell shape or cell cycle phase, except for mitotic cells in which myosin was distributed
diffusely through the cytoplasm. If, as suggested by Kolega (9), microfilaments align parallel to the direction of applied
tension, the redistribution of myosin-containing microfilaments in cultured keratinocytes may reflect the increased tension
between cells resulting from increasing strength of cell-cell junctions over time. In sectioned human skin, myosin was localized
in the peripheral cytoplasm of stratified epidermal cells. Tensions arising from the numerous desmosomal junctions between
cellsin vivo could account for this distribution of myosin.
Supported by grant NS-23537 (V. A. L.) from the National Institutes of Health, Bethesda, MD, and by the Mayo Foundation. C.
L. W. is recipient of the Kermit E. Osserman and Blanche McClure Fellowship, 1987, National Myasthenia Gravis Foundation. 相似文献
125.
Alston-Mills Brenda Li Qi Chang Ottinger Mary Ann 《In vitro cellular & developmental biology. Plant》1989,25(10):934-938
Summary Production of antibodies against peptides or poorly antigenic proteins by conventional methods often requires either large
quantities of the native immunogen or some chemical modification to increase their antigenicity. In this study an in vivo
and in vitro immunization protocol has been used to generate monoclonal antibodies against the decapeptide luteinizing hormone-releasing
hormone (LHRH). Two injections of 100 μg of avian LHRH-I into BALB/c mice were given 7 d apart. Dissociated splenocytes were
collected under sterile conditions. They were incubated with 100 μg of the immunogen in 75-cm2 tissue culture flasks in thymocyte-conditioned media. After 5 to 8 d exposure to the antigen, splenocytes were fused with
SP2/O myeloma cells by polyethylene glycol. The cells were plated into 24 wells and then incubated in hypoxanthine aminopterin
and thymidine selective media. After 14 d an initial screening was done by enzyme immunoassay. The positive wells (6/24) were
expanded into 96-well plates and rescreened. Selected lines were cloned out 3 times by limiting dilution and the most positive
expanded for ascites production. The antibody was affinity purified in a protein A column. The antibody cross-reacted with
LHRH-I and II but preferentially to LHRH-I, as shown by competitive assay. A hypothalamic extract from a mature chick showed
a higher response than preparations from whole brain explants of 1- to 3-d posthatched chicks, mature quail, and mature mouse.
This work was funded by the Maryland Agricultural Experiment Station artical no. A4975, contribution no. 8019. 相似文献
126.
八倍体小黑麦×普通小麦杂种后代群体中的染色体易位 总被引:3,自引:0,他引:3
用改良的Giemsa C-带技术以单株为基础分析了八倍体小黑麦×普通小麦的杂种BC_1,F_(?)和F_(?)代植株的核型。在鉴定了C-带核型的1098株杂种后代植株中,发现了78条小麦-黑麦和277条黑麦-黑麦易位染色体。在不同的世代和株系中,小麦-黑麦染色体易位率变化在4.35—14.07%之间,平均7.10%;黑麦-黑麦染色体易位率在0.48—52.78%之间,平均25.23%。鉴定的小麦-黑麦易位染色体涉及了黑麦的14条不同的染色体臂和小麦的A、B和D组染色体。易位的48.57%发生在小麦和黑麦的部分同源染色体之间,51.43%发生在非部分同源染色体之间。不同的黑麦染色体臂参与易位的频率不同。小麦-黑麦染色体易位主要发生在杂种的早期世代,使用适当的选择技术在F_3获得了纯合的易位植株。文中讨论了快速选育易位系的技术和它们在小麦育种中的应用问题。 相似文献
127.
128.
以NT方法为基础比较了ELISA和FIA方法,共检测84份猴B病毒相关抗体的敏感性,结果ELISA和EIA阳性各50份(59.5%),NT阳性45份(53.6%)。三种方法相符者71份,符合率84.5%。ELISA和FIA均较NT敏感,而且快速,简便,经济,可应用于大批标本的检查。 相似文献
129.
Andrea Streit reas Faissner Bernd Gehrig Melitta Schachner 《Journal of neurochemistry》1990,55(5):1494-1506
The monoclonal L5 antibody reacts with an N-glycosidically linked carbohydrate structure which is present on the neural cell adhesion molecule L1, neural chondroitin sulfate proteoglycans, and other not yet identified glycosylated proteins. Using this antibody, we isolated and characterized proteoglycans from adult mouse brain and cultured astrocytes biosynthetically labeled with Na2 35SO4 and a 3H-amino acid mixture. Our data suggest that the L5 proteoglycans of both sources are identical in their biochemical properties. The apparent molecular mass of the L5 proteoglycan is approximately 500 kDa. Digestion of the iodinated L5 proteoglycan from mouse brain and of the [35S]methionine-labeled L5 proteoglycan from cultured astrocytes with proteinase-free chondroitinases ABC and AC revealed three major core proteins with apparent molecular masses of approximately 380, 360, and 260 kDa. These represent molecularly distinct protein cores. 相似文献
130.
Summary A cloned gene with an insertion, which was made by introducing cat, was ligated to the cloning site of the phage gt11. P1 phage grown on cells lysogenized with the recombinant phage could transduce the mutant gene into the original site on the Escherichia coli chromosome. 相似文献