全文获取类型
收费全文 | 34086篇 |
免费 | 2856篇 |
国内免费 | 1014篇 |
出版年
2024年 | 88篇 |
2023年 | 480篇 |
2022年 | 702篇 |
2021年 | 1104篇 |
2020年 | 1318篇 |
2019年 | 1669篇 |
2018年 | 1420篇 |
2017年 | 955篇 |
2016年 | 938篇 |
2015年 | 1238篇 |
2014年 | 2021篇 |
2013年 | 2160篇 |
2012年 | 1237篇 |
2011年 | 1663篇 |
2010年 | 1172篇 |
2009年 | 1547篇 |
2008年 | 1654篇 |
2007年 | 1610篇 |
2006年 | 1579篇 |
2005年 | 1361篇 |
2004年 | 1184篇 |
2003年 | 992篇 |
2002年 | 862篇 |
2001年 | 636篇 |
2000年 | 592篇 |
1999年 | 444篇 |
1998年 | 491篇 |
1997年 | 476篇 |
1996年 | 510篇 |
1995年 | 498篇 |
1994年 | 478篇 |
1993年 | 427篇 |
1992年 | 443篇 |
1991年 | 379篇 |
1990年 | 371篇 |
1989年 | 325篇 |
1988年 | 282篇 |
1987年 | 279篇 |
1986年 | 227篇 |
1985年 | 277篇 |
1984年 | 270篇 |
1983年 | 140篇 |
1982年 | 239篇 |
1981年 | 195篇 |
1980年 | 178篇 |
1979年 | 175篇 |
1978年 | 115篇 |
1977年 | 117篇 |
1976年 | 108篇 |
1973年 | 81篇 |
排序方式: 共有10000条查询结果,搜索用时 31 毫秒
991.
The object of this study was to determine the kinetics of chromosome decondensation during the G1 period of the HeLa cell cycle. HeLa cells synchronized in the G1 period following the reversal of mitotic block were fused with Colcemid-arrested mitotic HeLa cells at 1.5, 3, 5, and 7 h
after the reversal of N2O block. The resulting prematurely condensed chromosomes (PCC) were classified into six categories depending on the degree
of their condensation. The frequency of occurrence of each category was plotted as a function of time after mitosis. The results
of this study indicate that the process of chromosome decondensation, initiated during the telophase of mitosis continues
throughout the G1 period without any interruption, thus the chromatin reaches an ultimate state of decondensation by the end of G1 period, when DNA synthesis is initiated. 相似文献
992.
S. A. Weiss T. L. Lester S. S. Kalter R. L. Heberling 《In vitro cellular & developmental biology. Plant》1980,16(7):616-628
Summary Chemically defined media SFRE-199-1 for the growth and SFRE-199-2 for the maintenance of primary baboon kidney (Bak) cell
cultures were formulated by supplementing medium M199 with insulin, sodium pyruvate, zinc sulfate, and increasing arginine-HCl,
cysteine, cystine,l-glutamine,l-glutamic acid, glycine, histidine, tyrosine, and glucose to maximally active nontoxic concentrations. For prolonged maintenance
of the cells, physiological pH control, and blocking of excessive lactic acid accumulation in the spent medium of the cell
cultures, it was necessary to supplement the medium containing Earle's balanced salts withd-(+) galactose.
The cells grew and were maintained equally well on glass or polystyrene surfaces. Selenium, when added to growth medium or
substituted for insulin and zinc sulfate, did not stimulate cell growth. Electron microscopy showed that numerous dense particles,
approximately 250 to 400 ? in diameter, with the appearance of glycogen, were found throughout the cytoplasm in the cells
grown in SFRE-199-1 and maintained in SFRE-199-2. Echovirus types 1 to 3, poliovirus types 1 to 3, coxsackievirus types B2,
B4, B5,Herpesvirus hominis type 1, simian herpesvirusH. simiae and SA8, and simian adenovirus SV34 when titrated in primary Bak cells and grown and maintained in SFRE-199-1 and 2, respectively,
developed titers comparable to those obtained in conventionally grown and maintained cells.
This study was supported in part by National Institute of Health Grant RR00361 and World Health Organization Grant V4/181/38.
This laboratory serves as the NIH/WHO Collaborating Center for Reference and Research in Simian Viruses. 相似文献
993.
Selective isolation and culture of a proliferating epithelial cell population from the hamster trachea 总被引:7,自引:0,他引:7
William E. Goldman Joel B. Baseman 《In vitro cellular & developmental biology. Plant》1980,16(4):313-319
Summary A reliable cell isolation technique was developed to allow the cultivation of cells from the hamster respiratory tract. Repeated
thermolysin treatments and gradient centrifugation yielded a cell culture completely free from contamination by fibroblasts.
Viable cells could be isolated from as little tissue as a single hamster trachea, but in vitro proliferation occurred only
if the hamster was less than 4 months of age. The cultured cells could be repeatedly passaged and subcultured for weeks by
employing normal tissue culture techniques. Morphologically, the monolayers appeared to be a homogeneous population of epithelial
cells, and successful cloning of freshly isolated single cells resulted in apparently identical cultures. The epithelial origin
of these cells was also suggested by continued growth in minimum essential medium withd-valine substituted forl-valine. The relative ease with which this cell type can be isolated, cultured, and manipulated in vitro should encourage
its application as a model of the respiratory epithelium.
This research was supported by Public Health Service Grant P50-HL 19171 and Research Career Development Award 1-K04-AI 00178
to J. B. B. 相似文献
994.
995.
Jason Yang Raphael Guzman James Richards S. Nandi 《In vitro cellular & developmental biology. Plant》1980,16(6):502-506
Summary Mammary tumor epithelial cells from BALB/cfC3H mice were dispersely embedded inside the collagen gels in Ham's F-12 medium
containing horse serum. A sustained cell growth leading to a 5- to 10-fold increase in cell number over initial level was
observed in less than 2 weeks. The extent of this growth was found to be dependent on serum concentration. However, addition
of various protein and steroid hormones, both singly and in combination, to low-serum-containing medium failed to achieve
a comparable level of growth to that promoted by higher serum concentration. Mammary tumor cells can now be consistently propagated
in primary culture.
This investigation was supported by Grants CA05388 and CA09041 awarded by the National Cancer Institute, Department of Health,
Education and Welfare, and by cancer research funds of the University of California. 相似文献
996.
Asynchronous 9L cells were separated into relatively homogeneously-sized populations using centrifugal elutriation with both
a conventional collection method and a long collection method. A substantial increase in the homogeneity of the volume distributions
and in the degree of synchrony of the separated fractions was obtained using the long collection method. Autoradiographic
data indicated that fractions containing ≥97% G1 cells, ≥80% S cells, and 70–75% G2 cells could be routinely recovered with this procedure. Recovery in these fractions varied from 5 to 8% of the total number
of cells elutriated. The colony forming efficiency (CFE) of cells from fractions representing each phase of the cell cycle
was a constant 60–70%, which was comparable to the 60–80% usually found for asynchronous 9L cells. The percentage of cells
in the G1, S, and G2 phases in the elutriated fractions was more accurately determined from the volume distribution than from computer fits of
the DNA histogram obtained from flow cytometry. In general, the degree of synchrony was related to the coefficient of variation
(CV) of the volume distributions of the elutriated fractions. The CV was about 14% for all elutriated fractions. When the
≥97% G1 population was allowed to progress to S and G2, the CVs were about 17 and 20.2%, respectively. Thus, the best nonperturbing method for obtaining synchronous 9L cells in
the S or G2 phases was direct elutriation with the long collection method. 相似文献
997.
Cytochrome P-448 from Saccharomyces cerevisiae in permeabilized whole cell, microsomal fraction and in a highly purified reconstituted benzopyrene-3-monooxygenase (EC 1.14.14.1) system have been immobilized on various supports. Calcium alginate was found to be especially useful and the kinetics of hydroxylation were close to that of the free enzyme system with all three forms of enzyme, even with permeabilized whole yeast cells (V max of 664 pmol 3-hydroxybenzo(a)pyrene produced per h per nmol cytochrome P-448 compared with 1000 for free highly purified reconstituted enzyme system). Only the highly purified reconstituted form was successfully immobilized by BrCN-activated Sepharose-4B or by acrylamide. Both of these supports stabilized the highly purified reconstituted cytochrome P-448 benzopyrene-3-monooxygenase activity in prolonged storage at 4°C. Applications for various immobilized enzymes and cells are assessed. 相似文献
998.
Bo Mattiasson Per-Olof Larsson Lennarth Lindahl Peter Sahlin 《Enzyme and microbial technology》1982,4(3):153-157
A vitamin B1 (thiamin)-sensitive electrode has been devised by combining an oxygen electrode with a yeast-containing membrane. The assembly was used for assaying thiamin at concentrations down to 10?11 gl?1. The analytical procedure developed should allow the measurement of 10–20 samples per hour. The performance of the yeast electrode was improved when alginate membranes reinforced with a nylon network were used. An apparatus for preparing such membranes is described together with a magnetic membrane holder facilitating handling of membranes in combination with electrodes. 相似文献
999.
Differentiation between the somatostatin inhibition and the post-somatostatin rebound observed on growth hormone secretion in vitro 总被引:1,自引:0,他引:1
A rebound in growth hormone secretion following somatostatin treatment has been shown in several systems where somatostatin suppresses secretion of the hormone. We have developed an in vitro system in which isolated and cultured pituitary cells were perfused after mild trypsinization. After washing, these cells retained their sensitivity and secreted growth hormone (GH) in response to physiological activators (norepinephrine, dopamine, serotonin) or inhibitors (somatostatin) as well as pharmacological activators (PGE2). The variation in GH secretion occurred within a minute after commencement of the infusion and was as rapidly reversible and repeatable minutes later. During somatostatin infusion the GH secretion was not totally suppressed (residual secretion (mean +/- S.D.) 34 +/- 7%). After the infusion a rapid rebound in GH secretion occurred, reaching levels in excess of the pretreatment value of 138 +/- 13%. This rebound effect occurred at doses higher than (10(-10)M) but not at lower doses, even when significant inhibition was observed. The inhibitory effect is of greater magnitude than the rebound effect (rebound = inhibition X 57 +/- 7% (mean +/- S.D.)). Furthermore, rebound was not enhanced by prolongation of somatostatin infusion. These latter results indicate that the rebound in secretion cannot be explained on the sole basis of storage of intracellular GH during somatostatin infusion and in fact suggest the involvement of a process of GH degradation and/or an inhibition of GH synthesis. 相似文献
1000.
External ATP causes passive permeability change in several transformed cells, but not in untransformed cells. We studied the effect of external ATP on the passive permeability of CHO-K1 cells, a transformed clone of Chinese hamster ovary cells. Treatment of the cells with external ATP alone did not produce a permeability change, and this was observed only when a mitochondrial inhibitor, such as rotenone or oligomycin, was present together with ATP. These inhibitors reduced the concentration of intracellular ATP and a permeability change by external ATP was observed when intracellular ATP was decreased more than 70%. This requirement for permeability change of CHO-K1 cells was quite unique, since passive permeability change of other transformed cells so far tested was induced by ATP alone. Treatment of CHO-K1 cells with cyclic AMP analogues increased their sensitivity to external ATP about 2-fold. The roles of external and intracellular ATP in controlling passive permeability are discussed. 相似文献