全文获取类型
收费全文 | 4581篇 |
免费 | 450篇 |
国内免费 | 2篇 |
出版年
2022年 | 31篇 |
2021年 | 64篇 |
2020年 | 42篇 |
2019年 | 48篇 |
2018年 | 41篇 |
2017年 | 36篇 |
2016年 | 91篇 |
2015年 | 158篇 |
2014年 | 167篇 |
2013年 | 207篇 |
2012年 | 306篇 |
2011年 | 278篇 |
2010年 | 211篇 |
2009年 | 172篇 |
2008年 | 248篇 |
2007年 | 297篇 |
2006年 | 250篇 |
2005年 | 242篇 |
2004年 | 214篇 |
2003年 | 196篇 |
2002年 | 188篇 |
2001年 | 57篇 |
2000年 | 50篇 |
1999年 | 59篇 |
1998年 | 57篇 |
1997年 | 42篇 |
1996年 | 50篇 |
1995年 | 34篇 |
1994年 | 52篇 |
1993年 | 41篇 |
1992年 | 46篇 |
1991年 | 53篇 |
1990年 | 44篇 |
1989年 | 34篇 |
1988年 | 36篇 |
1986年 | 48篇 |
1985年 | 53篇 |
1984年 | 52篇 |
1983年 | 40篇 |
1982年 | 48篇 |
1981年 | 47篇 |
1980年 | 45篇 |
1979年 | 35篇 |
1978年 | 33篇 |
1977年 | 29篇 |
1976年 | 28篇 |
1975年 | 35篇 |
1974年 | 33篇 |
1973年 | 36篇 |
1971年 | 24篇 |
排序方式: 共有5033条查询结果,搜索用时 20 毫秒
31.
We have shown, in a preliminary report, that macrophages can induce strand breaks in the DNA of co-cultured tumor cells (Chong et al., 1988). The present study is designed to determine if oxygen-centered species generated by the cell-free enzyme-substrate combination of hypoxanthine and xanthine oxidase can induce similar lesions and to identify the specific mediator(s). We report that co-incubation of murine mammary tumor cell lines with hypoxanthine and xanthine oxidase leads to the induction of DNA-strand breaks as determined by fluorescence analysis of DNA unwinding (FADU) assay or alkaline elution techniques. This damage is preventable by catalase which removes hydrogen peroxide but no protection is provided by agents to remove or prevent the formation of superoxide anion (superoxide dismutase), or hydroxyl radical (mannitol or the iron chelator o-phenanthroline). Likewise, cyclooxygenase or lipoxygenase inhibitors of arachidonate metabolism (indomethacin, nordihydroguaiaretic acid, caffeic acid) or bromophenacyl bromide do not alter the degree of DNA scission. Treatment with higher doses of oxygen species leads to significant toxicity as determined by evaluation of cell growth potential or colony-forming ability. Again, toxicity is prevented only by the presence of catalase. Tumor cells are able to rejoin strand breaks at lower, less toxic doses. When comparing different tumor cell subpopulations at various stages of progression, i.e., metastatic vs. nonmetastatic, for sensitivity to hydrogen peroxide-induced strand breakage, we found that at lower concentrations (less than 5μM) metastatic populations are sensitive whereas nonmetastatic populations exhibit no significant breakage. At higher concentrations of hydrogen peroxide, all lines were sensitive, suggesting that a lower threshold of sensitivity may exist for more progressed tumour cell lines. 相似文献
32.
When young wheat (Triticum aestivum L.) or barley (Hordeum vulgare L.) plants were deprived of an external sulphate supply (-S plants), the capacity of their roots to absorb sulphate, but not phosphate or potassium, increased rapidly (derepression) so that after 3–5 d it was more than tenfold that of sulphate-sufficient plants (+S plants). This increased capacity was lost rapidly (repression) over a 24-h period when the sulphate supply was restored. There was little effect on the uptake of L-methionine during de-repression of the sulphate-transport system, but S input from methionine during a 24-h pretreatment repressed sulphate influx in both+S and-S plants.Sulphate influx of both+S and-S plants was inhibited by pretreating roots for 1 h with 4,4-diisothiocyanatostilbene-2,2-disulphonic acid (DIDS) at concentrations > 0.1 mol · m-3. This inhibition was substantially reversed by washing for 1 h in DIDS-free medium before measuring influx. Longer-term pretreatment of roots with 0.1 mol·m-3 DIDS delayed de-repression of the sulphatetransport system in-S plants but had no influence on+S plants in 3 d.The sulphydryl-binding reagent, n-ethylmaleimide, was a very potent inhibitor of sulphate influx in-S roots, but was much less inhibitory in +S roots. Its effects were essentially irreversible and were proportionately the same at all sulphate concentrations within the range of operation of the high-affinity sulphate-transport system. Inhibition of influx was 85–96% by 300 s pretreatment by 0.3 mol·m-3
n-ethylmaleimide. No protection of the transport system could be observed by including up to 50 mol·m-3 sulphate in the n-ethylmaleimide pre-treatment solution. A similar differential sensitivity of-S and+S plants was seen with p-chloromercuriphenyl sulphonic acid.The arginyl-binding reagent, phenylglyoxal, supplied to roots at 0.25 or 1 mol·m-3 strongly inhibited influx in-S wheat plants (by up to 95%) but reduced influx by only one-half in+S plants. The inhibition of sulphate influx in-S plants was much greater than that of phosphate influx and could not be prevented by relatively high (100 mol·m-3 sulphate concentrations accompanying phenylglyoxal treatment. Effects of phenylglyoxal pretreatment were unchanged for at least 30 min after its removal from the solution but thereafter the capacity for sulphate influx was restored. The amount of new carrier appearing in-S roots was far greater than in+S roots over a 24-h period.The results indicate that, in the de-repressed state, the sulphate transporter is more sensitive to reagents binding sulphydryl and arginyl residues. This suggests a number of strategies for identifying the proteins involved in sulphate transport.Abbreviations DIDS
4,4-diisothiocyanatostilbene-2,2-disulphonic acid
- NEM
n-ethylmaleimide
- PCMBS
p-chloromercuriphenyl sulphonic acid 相似文献
33.
34.
G R Newton P J Hansen F W Bazer M V Leslie D C Stephenson B G Low 《Biology of reproduction》1989,40(2):417-424
Allantoic and amniotic fluids were collected on Days 60 (n = 3), 100 (n = 4), and 140 (n = 3) of pregnancy. The presence of uterine milk proteins (UTM-proteins) in these samples was evaluated by Ouchterlony immunodiffusion and enzyme-linked immunoabsorbant assay (ELISA). Eight of ten samples of allantoic fluid and three of ten samples of amniotic fluid produced one or two immunoprecipitin bands against antiserum to UTM-proteins. Each band fused with immunoprecipitin bands from UTM-proteins purified from uterine fluid. Data from a semi-quantitative ELISA indicated that allantoic fluid from all ewes and amniotic fluid from six of ten ewes contained immunoreactive UTM-proteins. Concentrations of UTM-proteins in these fluids were not statistically affected by day of gestation (p greater than 0.10), but tended to decline as gestation advanced. Greater concentrations of UTM-proteins were detected in allantoic fluid than in amniotic fluid (p less than 0.05). The physical characteristics of the immunoreactive material in allantoic and amniotic fluids were examined by polyacrylamide gel electrophoresis and Western blotting. The immunoreactive material was found to possess pIs and molecular weights identical to UTM-proteins. These results indicate that fetal fluids contain material that reacts with antiserum to UTM-proteins and has physical properties similar to UTM-proteins. It is likely, therefore, that the UTM-proteins are transported across the placenta during gestation, perhaps to serve some function in the fetal compartment. 相似文献
35.
Molecular cloning of the human transmembrane secretory component (poly-Ig receptor) and its mRNA expression in human tissues 总被引:6,自引:0,他引:6
P Krajci R Solberg M Sandberg O Oyen T Jahnsen P Brandtzaeg 《Biochemical and biophysical research communications》1989,158(3):783-789
A 2.5 kilobase (kb) cDNA clone containing 92% of the coding region for human transmembrane secretory component (SC) or poly-Ig receptor, was isolated from a mammary gland cDNA library. The cDNA clone encoded a protein of 693 amino acids which showed 99% homology with the primary amino acid sequence of human free SC as reported by Eiffert et al. (1), and 54% homology with the deduced amino acid sequence of rabbit transmembrane SC for which cDNA was cloned by Mostov et al. (2). Northern blot analysis showed mRNA expression in various human exocrine tissues in good agreement with our previous immunohistochemical studies of SC. 相似文献
36.
Common expression of a tumor necrosis factor resistance mechanism among gynecological malignancies 总被引:2,自引:0,他引:2
C. Bethan Powell David G. Mutch L. Stewart Massad Ming-Shian Kao John Leslie Collins 《Cancer immunology, immunotherapy : CII》1990,32(2):131-136
Summary The efficacy of tumor necrosis factor (TNF) as an anticancer agent is limited. This limitation might be related to the expression of a protein-synthesis-dependent resistance mechanism that prevents the lysis of tumor cells by TNF. To test this possibility eight randomly selected human cell lines, three derived from ovarian carcinomas and five derived from cervical carcinomas, were tested for their in vitro sensitivity to TNF-mediated lysis. The results of this analysis showed that all eight cell lines are normally resistant to lysis by TNF. However, in the presence of inhibitors of protein synthesis, seven of them showed a significant increase in TNF-mediated lysis. Measurement of protein synthesis showed that there is a linear correlation between the level of inhibition of protein synthesis and the level of TNF-mediated lysis. The fact that seven of eight randomly selected cell lines are resistant to TNF because they express a protein-synthesis-dependent resistance mechanism suggests that this mechanism of resistance may be common among gynecological cancers. The results also suggest that a therapy involving TNF and inhibitors of protein synthesis might be useful for the treatment of gynecological malignancies. 相似文献
37.
Guinea-pig hepatocytes whose plasma membranes were rendered permeable by treatment with saponin, accumulated 45calcium in the presence of potassium oxalate and ATP. The uptake was linear with time for up to one hour when high-capacity EGTA buffers were used (5mM). In the presence of a supra-maximal concentration of inositol 1,4,5-trisphosphate, under conditions minimising metabolism of this calcium-mobilising messenger, 45calcium accumulation was inhibited by about 40% for a period of one hour. Electron microscopic examination of the cells, revealed the presence of electron dense precipitates. Electron microprobe analysis of the precipitates indicated that they constituted the majority of the oxalate-dependent calcium uptake. The precipitates were located throughout the non-nuclear regions of the cells. Cells treated with inositol 1,4,5-trisphosphate contained fewer precipitates, but high cell-to-cell variability prevented conclusions as to the precise location of the pool sensitive to inositol 1,4,5-trisphosphate. These results support the previous contention that a fraction of endoplasmic reticulum is completely emptied of calcium by maximal concentrations of inositol 1,4,5-trisphosphate, while another fraction is insensitive to this action. In addition, these findings indicate that the pool of intracellular calcium on which inositol 1,4,5-trisphosphate acts is oxalate-permeable, and that the calcium-releasing action of inositol 1,4,5-trisphosphate does not desensitise within one hour. 相似文献
38.
Judith R. Turnlund Leslie Wada Janet C. King William R. Keyes Lorra L. Acord 《Biological trace element research》1988,17(1):31-41
Copper absorption was measured at two levels of dietary zinc in six healthy young men who were confined to a metabolic unit
for a 75 d study of zinc utilization. A diet of conventional foods was fed, providing either 16.5 or 5.5 mg zinc and 1.3 mg
copper daily. Copper absorption was determined by feeding65Cu, a stable isotope of copper, once during the 16.5 mg Zn diet and near the beginning and end of the 5.5 mg Zn diet. Apparent
copper absorption averaged 48.1% when the 16.5 mg Zn diet was fed. This was significantly higher than the averages of 37.2
and 38.5% when the 5.5 mg Zn diet was fed. Absorption also differed significantly among subjects. Fecal copper did not differ
between diets or among subjects. All subjects were in positive copper balance at both levels of dietary zinc. These results
suggest that a dietary zinc intake slightly above the Recommended Dietary Allowance of 15 mg/d does not increase fecal copper
loss and does not interfere with copper absorption. 相似文献
39.
Leslie S. Indrasith Takuji Sasaki Toshinobu Yaginuma Okitsugu Yamashita 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1988,158(1):1-7
Summary Analysis of yolk proteins of the silkworm,Bombyx mori, by SDS-polyacrylamide gel electrophoresis and immunoblotting showed that there was a developmental change in subunit composition of egg-specific protein; egg-specific protein consisting of 72 kDa subunits alone (premature form) was found in vitellogenic follicles, whereas the protein in mature eggs was composed of 72 kDa and 64 kDa subunits (mature form). The premature form of egg-specific protein was purified from young ovaries to homogeneity using a high performance liquid chromatography system. The purified protein had an apparent molecular mass of 225 kDa which could not be distinguished from that of the mature form. By circular dichroism analysis, both egg-specific proteins were estimated to have about 30% -helix and 20% -sheet, but the mature form showed a relatively rigid conformation in the aromatic region. The premature egg-specific protein purified from vitellogenic ovaries, consisted of three 72 kDa subunits, whereas mature egg-specific protein was composed of two 72 kDa subunits and one 64 kDa subunit. All of these subunits showed the same immunoreactivity towards antiserum raised against the mature form. An identical NH2-terminal amino acid sequence was found in both 72 kDa polypeptides and 64 kDa polypeptide for the initial 10 amino acids.Abbreviations
SDS
sodium dodecyl sulfate
-
PMSF
phenylmethylsulfonyl fluoride
-
PAGE
polyacrylamide gel electrophoresis
-
HPLC
high performance liquid chromatography
-
ESP
egg-specific protein
-
Vtn
vitellin 相似文献
40.
Allison A. Welder Tina Machu Steven W. Leslie Richard E. Wilcox June Bradlaw Daniel Acosta 《In vitro cellular & developmental biology. Plant》1988,24(8):771-777
Summary Primary mycolardial cell cultures and freshly isolated cardiac cells in suspension resprensent two isolated, whole cell models
for investigating cellular transsarcolemmal45Ca++ exchange in response to a receptor-coupled stimulus. Studies were performed to characterize beta-adrenergic receptor binding,
beta-adrenergic receptor mediated cellular calcium (45Ca++) exchange, and viability in purified primary myocardial cell cultures and freshly isolated cardiac cells in suspension obtained
from 3-to 3-d-old Sprague-Dawley rats. In addition, beta-adrenergic receptor binding was characterized in whole-heart crude
membrane preparations. All three preparations had saturable beta-adrenergic binding sites with the antagonist [125I]iodopindolol ([125I]IPIN). The suspensions had a significantly lower B
max
(42±6 fmol/mg protein) than the membranes and cultures (77±8 and 95±10 fmol/mg protein, respectively). The K
D
of the cultures (218±2.0 pM) was significantly higher than that for the suspensions (107 ±1.3 pM) and membranes (93±1.3 pM). Viability was significantly lower in the suspensions (57%) when compared to 94% viability in myocardial cell cultures after
3 h of incubation in Kreb's Henseleit buffer. Incubation of the cultures with 5.0×10−7
M isoproterenol resulted in a significant increase in45Ca++ exchange as early as 15 s. In contrast,45Ca++ exchange into the suspensions was not increased. Although both primary cell cultures and cardiac cells in suspension possess
saturable beta-adrenergic receptors, only the monolayer cultures exhibited functional beta-adrenergic receptor-mediated45Ca++ exchange. Of the two intact cell models investigated, these data suggest that primary myocardial cell cultures are more suitable
than cell suspensions for investigating beta-adrenergic receptor binding and functions in the postnatal rat heart.
This research was supported by The University of Texas Research Institute, a grant from the Texas Advanced Research Technology
Program awarded to S. W. Leslie and R. E. Wilcox, and contract 223-86-2109 from the Food and Drug Administration. 相似文献