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981.
Sekido C Nishimura N Takai M Hasumi K 《Bioscience, biotechnology, and biochemistry》2010,74(11):2320-2322
Plasma hyaluronan-binding protein (PHBP) is a serine protease implicated in proteolysis under inflammatory conditions. We identified laccaic acid, a widely used food coloring from scale insects, as a potent inhibitor of the protease in terms of both autoactivation of the PHBP proenzyme (IC(50) = 0.35-0.55 μg/ml) and the catalytic activity of the active enzyme (IC(50) = 1.1 μg/ml). 相似文献
982.
Naoko Kida Yousuke Katsuda Yuko Yoshikawa Seiji Komeda Takaji Sato Yoshihiro Saito Masahiko Chikuma Mari Suzuki Tadayuki Imanaka Kenichi Yoshikawa 《Journal of biological inorganic chemistry》2010,15(5):701-707
It is known that a 1,2,3-triazolato-bridged dinuclear platinum(II) complex, [{cis-Pt(NH3)2}2(μ-OH)(μ-1,2,3-ta-N
1,N
2)](NO3)2 (AMTA), shows high in vitro cytotoxicity against several human tumor cell lines and circumvents cross-resistance to cisplatin.
In the present study, we examined a dose- and time-dependent effect of AMTA on the higher-order structure of a large DNA,
T4 phage DNA (166 kbp), by adapting single-molecule observation with fluorescence microscopy. It was found that AMTA induces
the shrinking of DNA into a compact state with a much higher potency than cisplatin. From a quantitative analysis of the Brownian
motion of individual DNA molecules in solution, it became clear that the density of a DNA segment in the compact state is
about 2,000 times greater than that in the absence of AMTA. Circular dichroism spectra suggested that AMTA causes a transition
from the B to the C form in the secondary structure of DNA, which is characterized by fast and slow processes. Electrophoretic
measurements indicated that the binding of AMTA to supercoiled DNA induces unwinding of the double helix. Our results indicate
that AMTA acts on DNA through both electrostatic interaction and coordination binding; the former causes a fast change in
the secondary structure from the B to the C form, whereas the latter promotes shrinking in the higher-order structure as a
relatively slow kinetic process. The shrinking effect of AMTA on DNA is attributable to the possible increase in the number
of bridges along a DNA molecule. It is concluded that AMTA interacts with DNA in a manner markedly different from that of
cisplatin. 相似文献
983.
Mori S Iwase K Iwanami N Tanaka Y Kagechika H Hirano T 《Bioorganic & medicinal chemistry》2010,18(23):8158-8166
Histone modification, for example, by histone deacetylase (HDAC) and histone lysine methyltransferase (HMT), plays an important role in regulating gene expression. To obtain novel inhibitors as tools for investigating the physiological function of members of the HMT family, we designed and synthesized novel inhibitors, which are amine analogues of adenosylmethionine (AdoMet; the cofactor utilized in the methylation reaction) bearing various alkylamino groups coupled via an ethylene linker. The inhibitory activities of these compounds towards SET7/9, an HMT, were evaluated. It was found that introduction of an alkylamino group increased the inhibitory activity. 相似文献
984.
985.
Yosuke Masubuchi Yuko Nakagawa Jinhui Ma Tsutomu Sasaki Tadahiro Kitamura Yoritsuna Yamamoto Hitoshi Kurose Itaru Kojima Hiroshi Shibata 《PloS one》2013,8(1)
Background
Sweet taste receptor is expressed not only in taste buds but also in nongustatory organs such as enteroendocrine cells and pancreatic beta-cells, and may play more extensive physiological roles in energy metabolism. Here we examined the expression and function of the sweet taste receptor in 3T3-L1 cells.Methodology/Principal Findings
In undifferentiated preadipocytes, both T1R2 and T1R3 were expressed very weakly, whereas the expression of T1R3 but not T1R2 was markedly up-regulated upon induction of differentiation (by 83.0 and 3.8-fold, respectively at Day 6). The α subunits of Gs (Gαs) and G14 (Gα14) but not gustducin were expressed throughout the differentiation process. The addition of sucralose or saccharin during the first 48 hours of differentiation considerably reduced the expression of peroxisome proliferator activated receptor γ (PPARγ and CCAAT/enhancer-binding protein α (C/EBPα at Day 2, the expression of aP2 at Day 4 and triglyceride accumulation at Day 6. These anti-adipogenic effects were attenuated by short hairpin RNA-mediated gene-silencing of T1R3. In addition, overexpression of the dominant-negative mutant of Gαs but not YM-254890, an inhibitor of Gα14, impeded the effects of sweeteners, suggesting a possible coupling of Gs with the putative sweet taste-sensing receptor. In agreement, sucralose and saccharin increased the cyclic AMP concentration in differentiating 3T3-L1 cells and also in HEK293 cells heterologously expressing T1R3. Furthermore, the anti-adipogenic effects of sweeteners were mimicked by Gs activation with cholera toxin but not by adenylate cyclase activation with forskolin, whereas small interfering RNA-mediated knockdown of Gαs had the opposite effects.Conclusions
3T3-L1 cells express a functional sweet taste-sensing receptor presumably as a T1R3 homomer, which mediates the anti-adipogenic signal by a Gs-dependent but cAMP-independent mechanism. 相似文献986.
Masahiro Ikeda Yuichiro Habu Naoko Miyano-Kurosaki Hiroshi Takaku 《Nucleosides, nucleotides & nucleic acids》2013,32(4-6):427-437
We examined the combinatorial action of RNase P and tRNase ZL-mediated specific inhibition of HIV-1 in cultured cells. We designed two short extra guide sequences (sEGS) that specifically recognize the tat and vif regions of HIV-1 mRNA and mediate the subsequent cleavage of hybridized mRNA by the RNase P and tRNase ZL components. We constructed an RNase P and tRNase ZL-associated vif and tat sEGS expression vector, which used the RNA-polymerase III dependent U6 promoter, as an expression cassette for EGS. Together, the RNase P and tRNase ZL-associated sEGS molecules allow more efficient suppression of HIV-1 mRNA production when separately applied. The possibilities offered by the vector to encode sEGS will provide a powerful tool for gene therapy. 相似文献
987.
Hitoshi Suzuki Hiroyasu Kaneko Nobushige Tamai Kunitada Shimotohno Naoko Miyano-Kurosaki Hiroshi Takaku 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):815-820
Short hairpin RNAs (shRNAs) efficiently inhibit gene expression by RNA interference. Here, we report the efficient inhibition by DNA-based vector-derived shRNAs of core protein expression in Huh-7 cells. The shRNAs were designed to target the core region of the hepatitis C virus (HCV) genome. The core region is the most conserved region in the HCV genome, making it an ideal target for shRNAs. We identified an effective site on the core region for suppression of the HCV core protein. The HCV core protein in core protein-expressing Huh-7 cells was downregulated by core protein-shRNA expression vectors (core-shRNA-452, 479, and 503). Our results support the feasibility of using shRNA-based gene therapy to inhibit HCV core protein production. 相似文献
988.
989.
Akiko Kusunoki Naoko Miyano-kurosaki Tohru Kimura Kazuyuki Takai Naoki Yamamoto Hiroshi Gushima 《Nucleosides, nucleotides & nucleic acids》2013,32(10-12):1709-1719
Abstract The CXC chemokine receptor CXCR4 is used as a major co-receptor for fusion and entry by syncytia-inducing T-tropic (X4) isolates of HIV-1. In the present study, we report the effects of an antisense oligodeoxyribonucleotide on the inhibition of CXCR4 gene expression in X4 HIV-1 infected HeLa-CD4 cells, to find more efficacious therapeutic possibilities for Human Immunodeficiency Virus type 1 (HIV-1) infection. Antisense phosphorothioate oligodeoxyribonucleotides (anti-S-ODNs) corresponding to the sequence of bases 69 to 88 of the human CXCR4 mRNA gene were synthesized. When the naked anti-S-ODN was incubated with HeLa-CD4 cells, the surface levels of this chemokine receptor were reduced up to 50%, indicating sequence-specific inhibition. We also examined the concomitant use of a basic peptide transfection reagent, nucleosomal histone proteins (RNP), for delivery of anti-S-ODNs. The anti-S-ODN encapsulated with RNP had higher inhibitory effects on p24 products than the naked anti-S-ODN. 相似文献
990.
Yuichiro Habu Takashi Nagawa Norihiko Matsumoto Hiroaki Takeuchi Naoko Miyano-Kurosaki Hiroshi Takaku 《Nucleosides, nucleotides & nucleic acids》2013,32(10-12):1907-1917
We previously demonstrated the function of an HIV-1-dependent ribozyme expression vector, with which the site-specific excision of loxP sequences can be achieved by using the Cre-loxP system (ON/OFF) as a molecular switch in an acute HIV-1 infection. However, this expression system also revealed the lower, non-specific expression of the anti-HIV-1 ribozyme in the absence of tat. To circumvent this problem, we used the more efficient HIV-1-dependent Cre recombinase gene expression vector, encoding the LTR-gag-p17 (extending from the 5′-LTR to the middle of the gag gene (pLTR-gag-p17-Cre)). Comparatively, the pLTR-gag-p17-Cre induces a higher Cre-protein expression level in an HIV-1 infection-dependent manner than the minimal pLTR-Cre. Furthermore, we constructed the ploxP-Rz-U5 and pLTR-gag-p17-Cre plasmids and also combined them into a single vector, pLTR-gag-p17-Cre/loxP-Rz-U5, for a comparison of their anti-HIV-1 activities. The resultant simultaneous expression of the Cre protein and the homologous recombination of the two loxP sequences induced a high level of HIV-1 replication inhibition (95%). Significantly, a high steady-state of ribozyme expression was observed in the RT-PCR analysis. These data imply that targeting the HIV-1 genes with the pLTR-gag-p17-Cre/loxP-Rz-U5 vector, which mediates HIV-1-dependent ribozyme expression, would be a useful tool for HIV-1 gene therapy applications. 相似文献