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991.
Rui Yan Mark Ottenbreit Bharati Hukku Michael Mally Sharong Chou Joseph Kaplan 《In vitro cellular & developmental biology. Animal》1996,32(10):656-662
Summary Methods for monitoring cell line identification and authentication include species-specific immunofluorescence, isoenzyme
phenotyping, chromosome analysis, and DNA fingerprinting. Most previous studies of DNA fingerprinting of cell lines have used
restriction fragment length polymorphism analysis. In this study, we examined the utility of an alternative and simpler method
of cell line DNA fingerprinting—polymerase chain reaction (PCR) amplification of fragment length polymorphisms. Fourteen human
cell lines previously found by other methods to be either related or disparate were subjected to DNA fingerprinting by PCR
amplification of selected fragment length polymorphism loci. Cell identification patterns by this method were concordant with
those obtained by isoenzyme phenotyping and restriction fragment length polymorphism-DNA fingerprinting, and were reproducible
within and between assays on different DNA extracts of the same cell line. High precision was achieved with electrophoretic
separation of amplified DNA products on high resolution agarose or polyacrylamide gels, and with fragment length polymorphism
(FLP) loci-specific “allelic ladders” to identify individual FLP alleles. Determination of the composite fingerprint of a
cell line at six appropriately chosen fragment length polymorphism loci should achieve a minimum discrimination power of 0.999.
The ability of PCR-based fragment length polymorphism DNA fingerprinting to precisely and accurately identify the alleles
of different human cell lines at multiple polymorphic fragment length polymorphism loci demonstrates the feasibility of developing
a cell line DNA fingerprint reference database as a powerful additional tool for future cell line identification and authentication. 相似文献
992.
Human papilloma virus E6/E7 genes can expand the lifespan of human corneal fibroblasts 总被引:2,自引:0,他引:2
Donna M. Peters Nathan Dowd Curtis Brandt Teresa Compton 《In vitro cellular & developmental biology. Animal》1996,32(5):279-284
Summary Human corneal fibroblasts were infected with a retroviral delivery vector containing the E6 and E7 genes from human Papilloma
virus type 16 in order to produce cell lines that have an expanded lifespan in culture. Morphologically, some of the trasfected
corneal fibroblast lines appeared to have the normal spindle-shape morphology of diploid fibroblasts, whereas other lines
appeared to have a more elongated morphology. All the cell lines were anchorage-dependent. Cells that had a normal morphology
grew at a rate similar to normal diploid human corneal fibroblasts and had a population doubling time of 48 h. All E6/E7 expressing
cell lines, regardless of morphology, produce types I, III, and V collagen, at levels similar to those observed in the parent
corneal diploid fibroblast. These corneal fibroblast lines will be a usefulin vitro system to study collagen expression and fibril formation, as well as normal stroma development. These results also demonstrate
that the use of E6/E7 genes to expand a cell’s lifespan can be a powerful tool because it does not appear to alter either
the growth rate of the cell or collagen expression. 相似文献
993.
Effect of anoxia and reoxygenation on antioxidant enzyme activities in immortalized brain endothelial cells 总被引:1,自引:0,他引:1
Olivier Rabin Michèle Piciotti Katy Drieu Jean-Marie Bourre Françoise Roux 《In vitro cellular & developmental biology. Animal》1996,32(4):221-224
Summary The effects of anoxia and reoxygenation on major antioxidant enzyme activities were investigatedin vitro in immortalized rat brain endothelial cells (RBE4 cells). A sublethal anoxic period of 12 h was assessed for RBE4 cells using the neutral red uptake test. Anoxia markedly influenced the specific activity of catalase and superoxide dismutase,
with no major effect on glutathione peroxidase or glutathione reductase. After 24 h postanoxia, the superoxide dismutase activity
modulated by the presence or absence of oxygen returned to control value.
Damage and recovery of RBE4 immortalized rat brain endothelial cells in culture after exposure to free radicals and other oxygen-derived species provides
a usefulin vitro model to study anoxia-reoxygenation trauma at the cellular level. 相似文献
994.
Yamato Kikkawa Kotaro Akaogi Hiroto Mizushima Naoki Yamanaka Makoto Umeda Kaoru Miyazaki 《In vitro cellular & developmental biology. Animal》1996,32(1):46-52
Summary Ladsin is a laminin-like cell-adhesive scatter factor with potent cell motility-stimulating ability and was purified from
serum-free conditioned medium of a malignant human gastric adenocarcinoma cell line STKM-1. To test its possible role in tumor
angiogenesis, we investigated its effect on primary culture of endothelial cells (human umbilical vein endothelial cells)
and endothelial cell line ECV304 in this study. Cell adhesion and motility effects of ladsin were observed in both types of
endothelial cells. In cell-attachment assay, ladsin interacted with integrin α3β1 that was expressed on the endothelial cell
surface. In Boyden chambers, ladsin stimulated both directed and random migration of ECV304 cells. Ladsin induced repair of
artificial wounds generated in ECV304 cell monolayers by stimulating cell migration. Ladsin did not affect the growth rate
of ECV304 cells at a low cell density but significantly increased the saturation cell density. These results suggest that
ladsin may be involved in the adhesion and migration of endothelial cells under some physiological and pathological conditions. 相似文献
995.
KAZUYUKI MIKAMI 《The Journal of eukaryotic microbiology》1996,43(1):43-48
ABSTRACT. The germinal micronucleus divides six times during conjugation of Paramecium caudatum : this includes two meiotic divisions and one mitosis of haploid nuclei during mating, and three mitoses of a fertilization nucleus (synkaryon). Microsurgical removal of the macronucleus showed that micronuclei were able to divide repeatedly in the absence of the macronucleus, after metaphase of meiosis I of the micronucleus and also after synkaryon formation. When the macronucleus was removed after the first division of synkaryon, in an extreme case the synkaryon divided five times and produced 32 nuclei, compared to three divisions and eight nuclei produced in the presence of the macronucleus. Treatment with actinomycin D (100 μ /ml) inhibited the morphological changes of the macronucleus during conjugation and induced a multimicronucleate state in exconjugants. However, in other cells, it induced production of a few giant micronuclei. We conclude that the micronucleus is able to undergo repeated divisions at any stage of conjugation in the absence of the macronucleus once the factor(s) for induction of the micronuclear division has been produced by the macronucleus. The macronucleus may also produce a regulatory factor required to stop micronucler division. 相似文献
996.
玉米浸渍过程中乳酸杆菌作用的研究 总被引:1,自引:0,他引:1
为了加强乳酸杆菌在玉米浸渍中的促进作用,我们对自己选育的一株乳酸杆菌HW—106进行了增殖培养。在浸渍开始时,把该茵液以10%量接种于玉米浸渍水中,浸渍液中SO2的浓度为0.10%;浸渍温度为50±1℃。在此条件下,玉米的浸渍时间由传统的68h,缩短到32h。 相似文献
997.
Payne DA Chan Ts Ostermeyer Schoaib B Patten BM Tyring SK 《Journal of biomedical science》1996,3(5):319-322
When monoclonal gammopathies arise in persons without evidence of plasma cell malignancy or lymphoproliferative disease, the term monoclonal gammopathy of unknown significance (MGUS) can be used. MGUS is believed to be the preneoplastic phase of lymphoproliferative diseases because many of these patients eventually develop malignant disease, mainly multiple myeloma. We have previously identified human papillomavirus (HPV) in a chronic benign plasma cell tumor of the cervix and in the bone marrow of multiple-myeloma patients. In the following study, we expanded upon our initial observation by analyzing 14 patients with MGUS. Bone marrow biopsies of the patients were analyzed for HPV sequences using polymerase chain reaction (PCR) and in situ hybridization. Normal controls included 26 bone marrow specimens, 24 analyzed by PCR and two by in situ hybridization. A significant association was found to exist between HPV and MGUS (p=0.001). Among 14 patients iwth MGUS, HPV sequences have been identified in 10 of the bone marrow biopsies. These results suggest that HPV can reside in the bone marrow of a premalignant lymphoproliferative disease. 相似文献
998.
The Nucleotide Sequence of Human Acylamino Acid-Releasing Enzyme 总被引:3,自引:0,他引:3
Mitta Masanori; Ohnogi Hiroshi; Mizutani Shigetoshi; Sakiyama Fumio; Kato Ikunoshin; Tsunasawa Susumu 《DNA research》1996,3(1):31-35
The nucleotide sequence of a cDNA coding for the human acylaminoacid-releasing enzyme (AARE, also known as acylpeptide hydrolase)[EC 3.4.19.1] subunit has been determined. The amino acid sequenceof human AARE subunit deduced from its cDNA nucleotide sequenceshowed a high degree of identity (91.5%) with both the correspondingproteins from the pig and the rat. The AARE cDNA shows 99.2%identity with a 3.3 kb cDNA transcribed from a locus (DNF15S2)on the short arm of human chromosome 3, whose deletion is associatedwith small cell lung cancer, taking into consideration thatthe sequence of the 3.3-kb cDNA previously reported was causedby misreading. 相似文献
999.
远交系小鼠胚胎干细胞系的建立及嵌合鼠的获得 总被引:2,自引:0,他引:2
ES细胞(EmbryonicStemCells)是来源于小鼠早期胚胎的多潜能干细胞,它可以在体外大量培养。并以单细胞的形式注射到早期胚胎里,发育为嵌合体。到目前为止,通常使用的129小鼠品系是来源于近交系(inbred)小鼠的胚胎.与之相比,远交系小鼠应当具有较强的生命力和抗病能力。曾有人报道过建成了远交系小鼠胚胎干细胞系,但是尚没有见到获得嵌合鼠的报道。有人甚至认为:由于不同品系小鼠所具有的遗传背景不同,有的小鼠不能建成ES细胞系。最近,本实验室在这方面做了有益的探索,成功地建成了远交系小鼠胚胎干细胞系,并在这里报导首例用远交系小鼠胚胎干细胞系培育成功嵌合体小鼠。采用源于Swiss小鼠远交群的昆明(KM)品系小鼠囊胚建成了三个小鼠胚胎干细胞系(KE1.KE2.KE5)。核型正常率均达到70%以上。自第八代起分批冻存,复苏后,培养至第12代,消化成单细胞,通过囊胚显微注射,将其注射到615品系小鼠胚胎。在幸存的幼鼠中获得了一只来源于KE1细胞的嵌合鼠(Table1).其毛色表现为受体鼠(615)的白色中嵌合有供体鼠(KM)的黑褐色(PlateI-A).嵌合鼠与受体鼠的杂交后代鼠中仍然出现了受体鼠的毛色类型( 相似文献
1000.
Petra R. Moog Tom A. W. van der Kooij Wolfgang Brüggemann John W. Schiefelbein Pieter J. C. Kuiper 《Planta》1995,195(4):505-513
Arabidopsis thaliana (L.) Heynh. Columbia wild type and a root hair-less mutant RM57 were grown on iron-containing and iron-deficient nutrient solutions. In both genotypes, ferric chelate reductase (FCR) of intact roots was induced upon iron deficiency and followed a Michaelis-Menten kinetic with a K
m of 45 and 54 M FeIII-EDTA and a V
max of 42 and 33 nmol Fe2+·(g FW)–1·min–1 for the wild type and the mutant, respectively. The pH optimum for the reaction was around pH 5.5. The approximately four fold stimulation of FCR activity was independent of formation of root hairs and/or transfer cells induced by iron deficiency. Iron-deficiency-induced chlorosis and the development of a rigid root habit disappeared when ferric chelate was applied to the leaves, while FCR activity remained unchanged. The time course of the responses to iron deficiency showed that morphological and physiological responses were controlled separately.Abbreviations FCR
ferric chelate reductase
- FW
fresh weight
Thanks are due to Klaas Sjollema (Department of Electronmicroscopy, University of Groningen, The Netherlands) for help with the electron microscopy sample preparation and especially to Dr. Uwe Santore (Heinrich-Heine-University for electron microscopy. This work was supported by the SCIENCE programm of the European community; P.R.M.) and a Personal Research Grant by the Ministerium für Wissenschaft und Forschung of Nordrhein-Westfalen (P.R.M.) and last, not least by the productive discussions in ECOTRANS B.V. 相似文献