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991.
J. COSSON 《Cell biology international》1996,20(2):83-94
Beating of cilia and flagellae allows movement of the fluid surrounding isolated cells (for example: protists) or epithelia (bronchial tissue) but is also responsible for the movement of unicellular organisms in this medium (such as spermatozoa or protists). This paper aims to describe: (1) the biochemical and structural elements of the ‘9 +2’ structure called the axoneme; (2) the mechanisms of wave generation and propagation along the axoneme of cilia and flagellae are then described, stating that in most models of wave propagation, a clear distinction is made between the dynein-dependent microtubule sliding which represents the oscillatory motor and the bending mechanism which regulates wave propagation. In current models, the bending propagation is supported by a bind /relax cyclic mechanism which propagates in register, but frame-shifted, with the powering action of the dynein motor along the axoneme. While a large amount of knowledge was accumulated about the motor, little is known about the resisting elements regulating the bending. (3) The present study also puts forward ideas as to how these organelles have been highly conserved throughout eucaryotic evolution, and concludes with suggestions for further fields of investigation into this unique mechanical device used for cell movement. 相似文献
992.
Shur-Jen Wang Peter Greer Robert Auerbach 《In vitro cellular & developmental biology. Animal》1996,32(5):292-299
Summary We report on the isolation and propagation of endothelial cells from the mouse embryonic yolk sac, the earliest site of blood
vessel development, and on the advantages of a hypervascular transgenic mouse source of these cells. These transgenic mice
express multiple copies of an activated allele of the humanfps/fes proto-oncogene and display hypervascularity progressing to multifocal hemangiomas. This phenotype suggested a role of thefps/fes proto-oncogene in vasculogenesis and angiogenesis and led us to investigate the growth characteristics of yolk-sac-derived
endothelial cells from transgenicfps/fes embryos. We have established eight independent cell clones from a mixture of transgenic and control yolk sacs from Day 12
embryos. Southern blot hybridization analysis showed all eight clones to be derived from transgenic cells suggesting a growth
advantage of cells carrying the activatedfps/fes gene. A cell line, Clone 166 (C166), established from one of these clones, was more fully characterized. C166 exhibits normal
endothelial characteristics, such as rearrangement into tubelike structures when placed on Matrigel, expression of angiotensin
converting enzyme, retention of cobblestone morphology at confluence, and the presence of cell surface receptors for acetylated
low density lipoprotein. The cells constitutively express murine endothelial cell adhesion molecule VCAM-1 and the vascular
addressin identified by antibody MECA-99. As expected, the cell line expresses high levels of the cytoplasmic protein-tyrosine
kinase encoded by thefps/fes proto-oncogene. The clone we have described as well as other endothelial cell lines that we have established from the mouse
embryonic yolk sac should prove useful for the study of endothelial cell differentiation and for the determination of the
mechanisms underlying the establishment of organ-specific endothelial cell heterogeneity. 相似文献
993.
Summary Techniques for the isolation of ahhighly pure population of viable osteoclasts are limited. For this reason, we developed
an isolation procedure that results in a high yield of osteoclast-like cells, up to 92% pure, from 3-wk-old chicken tibias.
The unique feature of the method is the migration of cells from marrow-free endosteal surfaces to vitronectin-coated plates.
The cells retain the osteoclast phenotype and remain viable in culture for a minimum of 1 wk. The cells were characterized
and compared to two populations of authentic avian osteoclasts, which were isolated on the basis of association with fibronectin-coated
plates. The cells contained substantial amounts of tartrate-resistant acid phosphatase. Alkaline phosphatase levels were negligible,
suggesting little contamination by osteoblasts. Response to parathyroid hormone, dibutyryl cyclic adenosine monophosphate,
calcitonin, acetazolamide, 17β-estradiol, and prostaglandin E2 was evident, as detected by measuring acid production. The vitronectin-associating cells contained numerous mitochondria,
had the ability to resorb bone in anin vitro bone slice assay, and specifically bound biotinylated vitronectin. At 5 d of culture, the cells demonstrated marginal multinuclearity,
having two to three nuclei. A large number (∼1×106 cells/tibia) of viable cells that exhibit characteristics of authentic osteoclasts can be obtained by the method described.
Potentially, this method could be applied to other species. 相似文献
994.
Human and animal mesenchymal progenitor cells from bone marrow: Identification of serum for optimal selection and proliferation 总被引:14,自引:0,他引:14
Donald P. Lennon Stephen E. Haynesworth Scott P. Bruder Neelam Jaiswal Arnold I. Caplan 《In vitro cellular & developmental biology. Animal》1996,32(10):602-611
Summary An undifferentiated subset of cells within the stromal cell population of bone marrow in postnatal mammals retains the capacity
to differentiate along osteogenic, adipogenic, fibroblastic, and chondrogenic lines. These cells, which are referred to as
mesenchymal stem cells (MSCs), can be maintainedin vitro and expanded in number through a process of subculturing. MSCs are maintained in culture in medium supplemented with 10%
fetal bovine serum (FBS). It is believed that certain, as yet unidentified, serum components play critical roles in the attachment
and proliferation of MSCs. Commercially available FBS is poorly characterized and may vary in composition and quality from
lot to lot. This study describes a method for the selection of lots of FBS that best support maintenance of the undifferentiated
state, mitotic expansion of MSCsin vitro, and retention of multilineage developmental potential in response to appropriate cues. 相似文献
995.
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
996.
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
997.
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
998.
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
999.
Joji Sekine Kazuo Sano Masataka Uehara Tsugio Inokuchi 《Biotechnic & histochemistry》1996,71(3):152-156
A technique Is described for rapid detection of S-pha?e cells of tumor tissues in smear specimens using bromodeoxyuridine (BrdU) immunostaining. Mouse NR-S1 tumors and human tumor specimens were prepared for smear cytology after incubation in RPMI 1640 culture medium containing 200 μM BrdU at 37 °C under 3 atm for 1 hr. Samples were fixed in 70% ethanol for 30 min and used immediately or air dried for 30 min. Samples were then denatured in either 4 N HC1 or 0.07 N NaOH to prepare partially single-stranded DMA. Fixation with air drying for 30 min followed by 30 min in 70% ethanol and 1 min denaturation with 0.07 N NaOH resulted in satisfactory staining quality. Cultured tumor specimens were processed for routine paraffin sections after smears were made for cytology. The labeling indices of the smear specimens and of the paraffin sections gave similar results. This technique should be useful in evaluating the cell proliferative potential of tumor tissue in smear cytology without processing paraffin sections. 相似文献
1000.
JÜRGEN ROHWEDEL ULRICH SEHLMEYER JIN SHAN ARMIN MEISTER ANNA M. WOBUS 《Cell biology international》1996,20(8):579-587
Embryonic germ (EG) cells of line EG-1 derived from mouse primordial germ cells were investigated for theirin vitrodifferentiation capacity. By cultivation as embryo-like aggregates EG-1 cells differentiated into cardiac, skeletal muscle and neuronal cells accompanied by the expression of tissue-specific genes and proteins as shown by RT-PCR analysis and indirect immunofluorescence. In comparison to embryonic stem (ES) cells of line D3 the efficiency of differentiation into cardiac and muscle cells was comparatively low, whereas spontaneous neuronal differentiation was more efficient than in D3 cells. Furthermore, the distribution of cell cycle phases as a parameter for the differentiation state was analysed in undifferentiated EG cells and ES cells and compared to data obtained for embryonic carcinoma (EC) cells of line P19 and differentiated, epithelioid EPI-7 cells. Flow cytometric analysis revealed similar cell cycle phase distributions in EG, EC and ES cells. In contrast, the somatic differentiated EPI-7 cells showed a longer G1-phase and shorter S- and G2/M-phases. Together, our results demonstrate that the differentiation state and capacity of EG cellsin vitroresemble that of totipotent ES cells. 相似文献