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71.
The DNase I sensitivity of three different chromatin regions in mouse testicular cells was analysed by in situ nick translation with biotin-dUTP combined with various counterstaining techniques. The regions were: (i) the constitutive centromeric heterochromatin, (ii) an interstitial C-band positive insertion on chromosome 1, Is(HSR1;C5)1Lub, and (iii) the chromatin containing rDNA (designated nucleolar chromatin herein). Incorporated biotin was detected either by the horseradish peroxidase reaction with diaminobenzidine (DAB) or the alkaline phosphatase reaction with fast red. The latter resulted in a water insoluble red precipitate, which was easily removable by any organic solution thus allowing the application of various counterstaining protocols. DNase I sensitivity of the three chromatin regions was screened in different cell types of the mouse testis. The interstitial Is(HSR) region was highly DNase I sensitive when it was recognizable by strong mithramycin fluorescence. The centromeric heterochromatin was DNase I resistant when it was compacted into microscopically visible chromosomal structures (mitosis, pachytene, metaphase I and II). In interphase nuclei from Sertoli cells and spermatogonia it became highly DNase I sensitive. In round spermatids it displayed medium DNase I sensitivity. Nucleolar chromatin was not labelled by in situ nick translation when silver staining demonstrated strong protein production. Sperm cells were highly DNase I sensitive from stages 11 to 15, but resistant as mature spermatozoa. 相似文献
72.
Dr. Laura Curatolo Christine Chaponnier Maria Benedetta Donati Luciano Morasca Giulio Gabbiani 《Cell and tissue research》1982,223(3):665-673
Summary It is known that human and animal fibroblasts are able to induce the retraction of a fibrin clot. In the present study the correlation between (i) fibrinclot retractile (FCR) activity, (ii) the number of actin stress-lines in mouse fibroblasts during growth in culture, and (iii) the sensitivity of actin stress-lines to a powerful actin-depolymerizing factor (ADF), present in plasma and serum of humans and laboratory animals was investigated. Fibroblasts at early passages (2–4) were tested for these parameters at various intervals after seeding (24, 96, and 168 h). The number of actin stress-lines was progressively higher, while the sensitivity to ADF action was progressively lower in cells cultured from 24 to 168 h; the FCR capacity was significantly decreased at 168 h. These data suggest that cells containing weakly polymerized and/or stabilized actin are more active than those containing highly polymerized and/or stabilized actin in triggering fibroblast contraction. 相似文献
73.
74.
Of the total adenylate-kinase activity in 10-d-old barley and wheat leaves, 40–50% is localised in the chloroplasts, while in mature spinach leaves 50–70% of the enzyme is chloroplastic. The extra-chloroplastic adenylate-kinase activity is associated with the mitochondria, very little, if any, is freely soluble in the cytoplasm. The adenylate pool of the cytoplasm could have access to adenylate-kinase activity in the intermitochondrial space because of the free permeation of adenylates across the outer mitochondrial membrane. Thus the adenylate pool of the cytoplasm could be subject to adenylate-kinase equilibrium. The mitochondrial adenylate kinase appeared to the localised exclusively in the intermembrane space. 相似文献
75.
Six of the eight transfer RNAs coded by bacteriophage T4 are synthesized via three dimeric precursor molecules. The sequences of two of these have been determined. Both of these precursors give rise to equimolar amounts of the cognate tRNA molecules in vivo. In contrast, even in wild-type infections, tRNAIle is present in ≤ 30% the amount of tRNAThr, with which it is processed from a common dimeric precursor.We have now determined the sequence of this dimer. In addition to the nucleotides present in tRNAThr and tRNAIle, it contains nine precursor-specific residues, located at the 5′ and 3′ termini and at the interstitial junction of the two tRNA sequences. While the three dimers share the majority of structural features in common, pre-tRNAThr + Ile is the only case in which an encoded tRNA 3′ -C-C-A terminus is present in the interstitial region.The processing of this dimer in various biosynthetic mutants has been analyzed in vivo and in vitro and shown to be anomalous in several respects. These results suggest that the apparent underproduction of tRNAIle can be explained by a novel processing pathway that generates a metabolically unstable tRNAIle product. Data from DNA sequence analysis of the T4 tRNA gene cluster (Fukada & Abelson, 1980) support the conclusion that the asymmetric maturation of this precursor is a consequence of the unique disposition of the -C-C-A sequence. These results argue that gene expression can be modulated at the level of RNA processing. The biological significance of this phenomenon is discussed in relation to evidence that tRNAIle has a unique physiological role. 相似文献
76.
77.
L. Barbieri Anna Gasperi-Campani M. Derenzini Christine M. Betts F. Stirpe 《Virchows Archiv. B, Cell pathology including molecular pathology》1979,30(1):15-24
Rats poisoned with abrin (2.5 micrograms/100 g body weight) died within 36 h with severe necrosis of acinar pancreatic cells. Incorporation in vivo of labelled amino acids into pancreatic protein was greatly impaired 6 h after poisoning. Microsomes isolated from the pancreas of poisoned rats at 6 h had a reduced capacity for protein synthesis in vitro. Incorporation in vivo of orotic acid into pancreatic RNA was decreased 12 h after poisoning. 相似文献
78.
79.
Shuttling RNAs are recognized as molecules that migrate against steep concentration gradients from one nucleus (through the cytoplasm) into another nucleus in the same cell. In previous work these molecules were identified through experiments involving the separation of two kinds of nuclei utilizing differences in the nuclei that may have produced misleading results. In the experiments reported here normal, randomly-chosen ameba (A. proteus) nuclei containing [32P]RNA were implanted into unlabeled normal, randomly-chosen cells and, after suitable incubation, the labeled RNAs present in each kind of nucleus were characterized by gel electrophoresis. The previously obtained results were confirmed: i.e. (a) the recipient cell nuclei acquired the same four small, distinct RNAs, which are recognized as shuttling ones because they migrate from one nucleus to the other; (b) the grafted nuclei possess, in addition to the four shuttling RNAs, three small, distinct RNAs, which are recognized as non-shuttling RNAs. New evidence also is presented to show that the acquisition by a nucleus of labeled RNAs in the above kind of experiment is not a result of new synthesis of RNAs from the labeled turnover products emanating from the transplanted nucleus. 相似文献
80.