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951.
Intra‐trophic isotopic discrimination of 15N/14N for amino acids in autotrophs: Implications for nitrogen dynamics in ecological studies 下载免费PDF全文
Yuko Takizawa Prarthana S. Dharampal Shawn A. Steffan Yoshinori Takano Naohiko Ohkouchi Yoshito Chikaraishi 《Ecology and evolution》2017,7(9):2916-2924
The differential discrimination of nitrogen isotopes (15N/14N) within amino acids in consumers and their diets has been routinely used to estimate organismal tropic position (TP). Analogous isotopic discrimination can occur within plants, particularly in organs lacking chloroplasts. Such discrimination likely arises from the catabolic deamination of amino acids, resulting in a numerical elevation of estimated TP, within newly synthesized biomass. To investigate this phenomenon, we examined the 15N/14N of amino acids (δ15NAA) in spring leaves and flowers from eight deciduous and two annual plants. These plants were classified on the basis of their time of bloom, plants that bloomed when their leaves were absent (Type I) versus plants that bloomed while leaves were already present (Type II). Based on the δ15NAA values from leaves, both plant types occupied comparable and ecologically realistic mean TPs (=1.0 ± 0.1, mean ± 1σ). However, the estimated TPs of flowers varied significantly (Type I: 2.2 ± 0.2; Type II: 1.0 ± 0.1). We hypothesize that these results can be interpreted by the following sequence of events: (1) Type I floral biomass is synthesized in absence of active photosynthesis; (2) the catabolic deamination of amino acids in particular, leaves behind 15N in the residual pool of amino acids; and (3) the incorporation of these 15N‐enriched amino acids within the biomass of Type I flowers results in the numerical elevation of the TPs. In contrast, the actively photosynthesizing Type II leaves energetically sustain the synthesis of Type II flower biomass, precluding any reliance on catabolic deamination of amino acids. Amino acids within Type II flowers are therefore isotopically comparable to the Type II leaves. These findings demonstrate the idiosyncratic nature of the δ15NAA values within autotrophic organs and have implications for interpreting trophic hierarchies using primary producers and their consumers. 相似文献
952.
Tomoyuki Bando Setsuko Fujita Naoko Nagano Soichiro Yoshikawa Yoshinori Yamanishi Masashi Minami Hajime Karasuyama 《Biochemistry and Biophysics Reports》2017
Basophils have been erroneously considered as minor relatives of mast cells, due to some phenotypic similarity between them. While recent studies have revealed non-redundant roles for basophils in various immune responses, basophil-derived effector molecules, including lipid mediators, remain poorly characterized, compared to mast cell-derived ones. Here we analyzed and compared eicosanoids produced by mouse basophils and mast cells when stimulated with IgE plus allergens. The production of 5-LOX metabolites such as LTB4 and 5-HETE was detected as early as 0.5 h post-stimulation in both cell types, even though their amounts were much smaller in basophils than in mast cells. In contrast, basophils and mast cells showed distinct time course in the production of COX metabolites, including PGD2, PGE2 and 11-HETE. Their production by mast cells was detected at both 0.5 and 6 h post-stimulation while that by basophils was detectable only at 6 h. Of note, mast cells showed 8–9 times higher levels of COX-1 than did basophils at the resting status. In contrast to unaltered COX-1 expression with or without stimulation, COX-2 expression was up-regulated in both cell types upon activation. Importantly, when activated, basophils expressed 4–5 times higher levels of COX-2 than did mast cells. In accordance with these findings, the late-phase production of the COX metabolites by basophils was completely ablated by COX-2 inhibitor whereas the early-phase production by mast cells was blocked by COX-1 but not COX-2 inhibitor. Thus, the production of COX metabolites is differentially regulated by COX-1 and COX-2 in basophils and mast cells. 相似文献
953.
Shiro Okumura Tetsuyuki Akao Satoko Yamashita Tokio Ichimatsu Kuniyo Inouye 《Cytotechnology》2005,47(1-3):59-67
Proteins of plasma membrane could be an index of purification of the plasma membrane of animal cells. A convenient method
is proposed for determining the plasma membrane proteins by a surface plasmon resonance (SPR) biosensor. Biotinylated proteins
were observed only in the peripheral areas of MOLT-4 cells which were treated by 5-[5-(N-succinimidyloxycarbonyl) pentylamido] hexyl-d-biotinamide. The proteins on HeLa cells were also biotinylated. And then the membrane samples of the HeLa cells were injected
onto the avidin-immobilized SPR-surface, and components bound non-specifically on the surface were removed by a washout solution.
The amount of biotinylated protein (BP) was determined directly from the absolute resonance unit (RU) after injection of the
washout solution. In the method a reference surface was not needed. The amount of BP bound to the surface was gradually attenuated
with the repeated injection, and a method for calibrating the RU value was introduced by considering the ratio of attenuation
by every injection. The correlation between the BP titer calculated by the calibration and the theoretically-estimated one
was greatly improved. Three cycles of the BP determination on a sensor surface was performed successfully. During the purification
process of membrane fractions, the degree of purification as judged by the BP titer was in good agreement with the degree
of increase in aminopeptidase N activity in the membrane fraction. Thus, the BP titer could be used as an index for purification
of plasma membrane. 相似文献
954.
Takenori Yamamoto Satsuki Terauchi Aiko Tachikawa Kikuji Yamashita Masatoshi Kataoka Hiroshi Terada Yasuo Shinohara 《Journal of bioenergetics and biomembranes》2005,37(5):299-307
N,N′-dicyclohexylcarbodiimide (DCCD) was earlier reported to have stimulatory effects on mitochondrial respiration and to induce
mitochondrial swelling, when it was added to mitochondrial suspensions. These data seem to imply that DCCD caused the mitochondrial
permeability transition (PT), but this possibility had never been investigated. In the present study, effects of DCCD on the
mitochondrial structure and function were studied in detail. DCCD was found to induce mitochondrial PT in a cyclosporine A-insensitive
manner. Electron microscopic analysis also supported the induction of the mitochondrial PT by DCCD. However, different from
many other PT inducers, DCCD failed to cause massive release of mitochondrial cytochrome c. To understand the relationship between the induction of mitochondrial PT and the release of mitochondrial cytochrome c, we compared the actions of DCCD on mitochondrial structure and function with those of Ca2+, known as an ordinary PT inducer. As a result, two parameters considered to be critical for controlling the release of mitochondrial
cytochrome c on the induction of PT were mitochondrial volume and the velocity of mitochondrial oxygen consumption. 相似文献
955.
Kiichiro Teruya Yoshihito Daimon Xiao-Yan Dong Yoshinori Katakura Takumi Miura Akira Ichikawa Tsukasa Fujiki Makiko Yamashita Tetsuya Mori Hideya Ohashi Sanetaka Shirahata 《Cytotechnology》2005,47(1-3):29-36
The cell line D29, which was easily and rapidly established by the promoter-activated production and glutamine synthetase
hybrid system, secreted recombinant human interleukin-6 (hIL-6) at a productivity rate of 39.5 μg 10−6 cells day−1, one of the highest reported levels worldwide. The productivity rate was about 130-fold higher than that of the cell line
A7, which was established without both promoter activation and gene amplification. Although D29 cells had a high copy number
and high mRNA level of the hIL-6 gene as well as a high secretion rate of hIL-6, large amounts of intracellular hIL-6 protein
accumulated in D29 cells compared to A7 cells. Northern blotting analysis showed no change in the GRP78/BiP expression level
in D29 cells. In contrast, an electrophoresis mobility shift assay revealed strong activation of NF-κB in D29 cells. These
results suggest that large amounts of hIL-6 translated from large amounts of hIL-6 mRNA cause excess accumulation of intact
hIL-6 in the endoplasmic reticulum (ER), and that subsequent negative feedback signals via the ER overload response inhibit
hIL-6 protein secretion. To enhance the hIL-6 productivity rate of D29 cells by releasing the negative feedback signals, the
effect of pyrrolidinedithiocarbamate, an inhibitor of NF-κB activation, was examined. Suppression of NF-κB activation in D29
cells produced a 25% augmentation of the hIL-6 productivity rate. Therefore, in highly productive cells like D29 cells, the
release of negative feedback signals could increase the total amount of recombinant protein secretion. 相似文献
956.
Application of heat-induced antigen retrieval to aldehyde-fixed fresh frozen sections. 总被引:1,自引:0,他引:1
Shuji Yamashita Yasunori Okada 《The journal of histochemistry and cytochemistry》2005,53(11):1421-1432
We applied the heat-induced antigen retrieval (HIAR) to aldehyde-fixed fresh frozen sections based on a new approach (i.e., a rapid and complete immobilization of antigen followed by heating). Frozen sections were fixed with 10% formalin in 0.1 M cacodylate buffer (pH 7.4) containing 25 mM CaCl(2) for 30 min, or with 0.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 1 min at room temperature, and then autoclaved in 20 mM Tris-HCl buffer (pH 9.0) for 10 min at 120 C. Both fixatives yielded good tissue structure after autoclaving. In the sections fixed with formalin containing CaCl(2), 20 of 22 antigens located in the nucleus, cytoplasm, membranes, and extracellular matrix greatly recovered their antigenicity after autoclaving; only two antigens exhibited stronger immunoreaction in acetone-fixed fresh frozen sections than these sections. Heating also retrieved the immunoreactivity of at least 14 antigens in the sections fixed with glutaraldehyde. We used the similar procedures to localize ligand-free estrogen receptor alpha (ERalpha) and glucocorticoid receptors (GR). Mouse uterine cells exhibited almost the same nuclear ERalpha immunostaining regardless of the hormonal status in glutaraldehyde-fixed fresh frozen sections and unliganded GR was localized mainly in the nucleus of mouse hepatocytes in fresh frozen sections fixed with 20% formalin containing 50 or 75 mM CaCl(2) at 40 C, after autoclaving. These results demonstrate that HIAR is useful for the immunohistochemistry of many antigens in aldehyde-fixed fresh frozen sections. 相似文献
957.
958.
959.
Pitavastatin up-regulates the induction of iNOS through enhanced stabilization of its mRNA in pro-inflammatory cytokine-stimulated hepatocytes. 总被引:1,自引:0,他引:1
960.
In many physiological and disease processes, TGF-beta usurps branches of MAP kinase pathways in conjunction with Smads to induce apoptosis and epithelial-to-mesenchymal transition, but the detailed mechanism of how a MAP kinase cascade is activated by TGF-beta receptors is not clear. We report here that TRAF6 is specifically required for the Smad-independent activation of JNK and p38, and its carboxyl TRAF homology domain physically interacts with TGF-beta receptors. TGF-beta induces K63-linked ubiquitination of TRAF6 and promotes association between TRAF6 and TAK1. Our results indicate that TGF-beta activates JNK and p38 through a mechanism similar to that operating in the interleukin-1beta/Toll-like receptor pathway. 相似文献