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201.
Microculture of single protoplasts of Brassica napus 总被引:1,自引:0,他引:1
Protoplasts of Brassica napus L. were cultured individually in a microdroplet system using a synthetic medium with survival rates of more than 70% and division frequencies of up to 65%. Microcallus formation occurred at frequencies of up to 50%. Factors affecting the survival and division of individually cultured protoplasts, such as composition and volume of culture medium, pH, buffering system, osmolarity and genotype, were analyzed. 相似文献
202.
I. Cacciari M. Del Gallo S. Ippoliti D. Lippi T. Pietrosanti W. Pietrosanti 《Plant and Soil》1986,90(1-3):107-116
Summary
Azospirillum brasilense andArthrobacter giacomelloi were grown in single and mixed succinate-limited continuous cultures at a partial oxygen pressure of 0.01atm. Growth, viability and survival during nutrient starvation were examined at various dilution rates. At D=0.05 h–1, Ks values for succinate consumed were calculated.Arthrobacter giacomelloi viability was inversely related to dilution rate whereasAzo. brasilense was directly related. Slightly lower values of viability were obtained in mixed culture, but the ratio between the microorganisms was constant. The survival ofArth. giacomelloi in single culture decreased with increasing growth rate while survival ofAzo. brasilense was directly related to dilution rate. Acetylene reduction activity was generally very low in both single and mixed cultures. Respiration rate was also determined and the mixed culture showed an oxygen uptake rate higher than that of single cultures.Research work supported by CNR, Italy. Special grant I.P.R.A. Sub-project 1. Paper N. 317. 相似文献
203.
204.
H. O. Jauregui P. N. McMillan J. Driscoll S. Naik 《In vitro cellular & developmental biology. Plant》1986,22(1):13-22
Summary Long-term monolayer cultures of adult rat hepatocytes were tested for their ability to glucuronize phenol red and to maintain
initial levels of cell proteins, glucose consumption, and lactic acid production. Lactate dehydrogenase leakage served as
an index of culture status because a high value indicates cell death. Three tissue culture (TC) media formulations were the
main variables introduced to determine ideal conditions for cell survival in vitro. Investigations of long-term cultures were
preceded by studies of hepatocyte attachment to polystyrene surfaces. This attachment was influenced by the amount of substrate
deposited and the number of cells seeded, but not by the uniformity of the substrate coating. A statistical analysis of our
data revealed that in the absence of fetal bovine serum (FBS), air dried collagen (ADC) and Biomatrix (BMX) were superior
to saline precipitated collagen and fibronectin as attachment substrates. In the presence of 10% FBS, all of the substrates
performed equally.
Chee's Medium (CEM) proved to be the best for preserving cell proteins over a time course of 28 d and Williams' E medium also
performed adequately up to 14 d. The glucuronization of phenol red was at 50% of initial values at Day 7 in CEM-ADC hepatocytes
in contrast to 30% for cells in Williams' E medium and 5% for cells grown in Waymouth's. At 14 d glucuronization was still
present at 40% of original values in CEM-ADC cells but had ceased in the other two media. When BMX was used, none of the TC
media supported glucuronization levels comparable to ADC cells.
This research was supported in part by grant 1R01-AM-26520 from the National Institute of Arthritis, Diabetes and Digestive
Kidney Diseases, NIH, Bethesda, Maryland. 相似文献
205.
Michael M. Lipsky Talia R. Sheridan Richard O. Bennett Eric B. May 《In vitro cellular & developmental biology. Plant》1986,22(6):360-362
Summary Comparisons were made of attachment and viability of rainbow trout (Salmo gairdneri) hepatocytes in short-term (2 days), primary culture on plastic, collagen-coated or extracellular matrix (ECM) coated dishes.
Hepatocyte isolation routinely yielded cells with good viability (96%). Cells plated on ECM attached with high efficiency
(93%) in contrast to cells cultured on plastic or collagen (∼20%). The cells plated on ECM flattened out and formed monolayers,
while the cells on plastic and collagen rounded up and formed multi-cell aggregates in suspension. Viability of cells in all
substrates remained high over the 2 day culture period. ECM is the first substrate to support trout-hepatocyte attachment
in primary culture. Differentiated liver function was maintained in cells cultured on ECM as evidence by the induction of
tyrosine aminotransferase by hydrocortisone (200%).
This work was supported in part by research grant R809599010 from the U. S. Environmental Protection Agency.
Editor's Statement This paper reports improved methods for culture of trout liver-derived cells that make in vitro investigations
of fish metabolism, carcinogenesis and chemical toxicity more feasible than previously applied techniques. Recent interest
in fish as models for study and indicators of effects of envionmental and food-related toxins make this work timely, poarticularly
since many of the compounds of interest are primarily metabolized by hepatocytes or act on liver as a major target. David
W. Barnes 相似文献
206.
William D. Meek Walter L. Davis 《In vitro cellular & developmental biology. Plant》1986,22(12):725-737
Summary The potent fungal metabolite cytochalasin D (CD) and cationized ferritin (CF) are used in combination to test for negative
charge distribution on blebs (knobs). Two established human epithelial cell lines, WISH and HeLa, that display blebs in various
phases of the cell cycle or under certain culture conditions (37,46) are investigated. CD alone, applied at a low concentration
(1.0 μg/ml) and for a short time period (3 min), causes blebs to appear as the prevalent surface feature. These are filled
mainly with free ribosomes. Additionally, feltlike mats, presumed to be disorganized, compacted microfilaments, are formed
directly beneath the cell membrane. These are especially evident in the cortical cytoplasm below the blebs or bleb clusters.
CF (0.345 mg/ml), applied for a 5-min period after CD administration (1.0 μg/ml) for 3 min, appears along the surface of microvilli,
at the base of blebs, and in vesicles beneath the bleb clusters. In some cases, microfilaments (6 nm in diameter) are closely
related to the vesicles. CF does not preferentially bind to the apical cell membrane of blebs. Above areas of the subplasmalemmal
microfilaments, CF membrane binding is apparent, even under circumstances where the filaments are disorganized by cytochalasin
treatment. These results seem to show the following: (a) bleb membranes are different from the remainder of the cell and do
exhibit a loss of negative charge and (b) surface charge may be dependent on the presence or structural integrity of membrane-related
6-nm microfilaments.
The support of this research by a grant from the Baylor College of Dentistry and The Oklahoma College of Osteopathic Medicine
and Surgery is gratefully acknowledged. The assistance of Dr. J. H. Martin, Department of Pathology, Baylor University Medical
Center, is also greatly appreciated. 相似文献
207.
Summary Cultures generated from tissues consisting of multiple types of cells are often heterogeneous. Unless the cell type of interest
has or can be given some selective growth advantage it may be overgrown by other cells. While developing techniques for the
tissue culture of microvascular endothelial cells we evaluated an electrosurgical generator (diathermy) to selectively kill
nonendothelail cells. Primary cell cultures were observed at ×100 magnification under phase contrast microscopy and a needle
electrode apposed to the cell to be destroyed. A return electrode was constructed by placing a sterile clip in contact with
the culture medium. The diathermy power setting controlled the area of lysis. Use of this technique allowed weeding of unwanted
cells without damage to endothelial cells, which were able to grow to confluence in pure culture.
Dr. Marks receives a Medical Postgraduate Research Scholarship from the National Health and Medical Research Council of Australia.
Financial support was received from the Leo Leukaemia and Cancer Research Trust and the Scleroderma Association of New South
Wales. 相似文献
208.
棒头草幼穗在含2,4-D的MS培养基上诱导出了胚性、非胚性和中间型愈伤组织。根据形态、淀粉粒等指标可将组成这些愈伤组织的细胞分为三类。改变培养基中2,4-D的浓度,能诱导三类愈伤组织相互转变。从胚性愈伤组织中诱导形成了大量体细胞胚;体细胞胚是从单个原胚细胞直接发育而来,它们能正常萌发、再生小植株。这种再生能力现已保持了34个月。小植株移植在土壤中可以正常生长、分蘖、开花和结实。 相似文献
209.
Albert E. Smith 《Physiologia plantarum》1985,65(2):124-128
The differential response of white clover ( Trifolium repens L. cv. Regal Ladino) and berseem clover ( Trifolium alexandrinum L. cv. Mississippi ecotype) was investigated by treating greenhouse cultured plants with 4-(2,4-dichlorophenoxy)butyric acid (2,4-DB). Berseem clover plants were significantly injured by a treatment concentration of 0.6 kg ha-1 of 2,4-DB, whereas white clover plants were not injured by treatment levels below 2.4 kg ha-1 . The metabolism of 2,4-DB in cell suspension cultures of white clover and berseem clover was investigated using [ring-14 C]-2,4-DB and non-labeled 2,4-DB. White clover cell cultures metabolized ca 4-fold more 2,4-DB than berseem cultures over a 44-h treatment period. The decrease in berseem cell population was 4-fold greater than the decrease in white clover cell population in response to the 8 μ M 2,4-DB treatment. The herbicide and its [ring-14 C]-labeled metabolites were isolated from treated cells and medium after 44 h by partition and thin-layer chromatography. White clover cells metabolized 90% of the [14 C]-2,4-DB and berseem clover cells metabolized 22% of the herbicide. The major portion of the radiolabel was in the glycoside fractions from extracts of both species. The differential response of Trifolium species to 2,4-DB is implied to be due to the differential rate of 2,4-DB metabolism to a glycoside by the clover plants. 相似文献
210.
The action of light in the initiation of floral buds in vitro has been studied using monochromatic light qualities on root explants of a long day plant, Cichorium intybus L. cv. Witloof. Red light (660 nm, 0.30 W m-2 ) promotes flowering, while far-red (730 nm, 0.31 W m-2 ) and irradiation with combined red + far-red (0.20 + 0.41 W m-2 ) have no effect. In short day conditions floral response can be obtained in two ways: 1) by interrupting the dark period with 5 brief irradiations of red light (0.45 W m-2 , 12 min) at regular intervals, although these are counteracted by far-red irradiations of equal intensity and duration; 2) by interrupting the long night with 5 h red light applied during the second third of the night, while at the beginning or at the end it is ineffective. Red light efficiency appears to depend on the photosynthetic activity of the tissues, so that flowering increases with increasing intensity of white light and is suppressed if no white light is supplied. The reproductive development is determined by the coordination of proper irradiation conditions with sufficient sensitivity of the perceiving meristematic cells. The period of highest sensitivity to environmental light conditions in the life cycle of a Cichorium root explant occurs between the 8th and the 16th day after the start of the culture. The data strongly suggest that phytochrome is involved in flower induction of Cichorium in vitro. 相似文献