全文获取类型
收费全文 | 2820篇 |
免费 | 236篇 |
国内免费 | 5篇 |
专业分类
3061篇 |
出版年
2024年 | 2篇 |
2023年 | 9篇 |
2022年 | 37篇 |
2021年 | 86篇 |
2020年 | 45篇 |
2019年 | 66篇 |
2018年 | 76篇 |
2017年 | 88篇 |
2016年 | 89篇 |
2015年 | 156篇 |
2014年 | 157篇 |
2013年 | 220篇 |
2012年 | 201篇 |
2011年 | 214篇 |
2010年 | 132篇 |
2009年 | 135篇 |
2008年 | 183篇 |
2007年 | 224篇 |
2006年 | 192篇 |
2005年 | 165篇 |
2004年 | 157篇 |
2003年 | 119篇 |
2002年 | 132篇 |
2001年 | 18篇 |
2000年 | 11篇 |
1999年 | 13篇 |
1998年 | 24篇 |
1997年 | 18篇 |
1996年 | 11篇 |
1995年 | 6篇 |
1994年 | 9篇 |
1993年 | 13篇 |
1992年 | 9篇 |
1990年 | 4篇 |
1989年 | 5篇 |
1987年 | 3篇 |
1986年 | 3篇 |
1983年 | 4篇 |
1982年 | 1篇 |
1981年 | 2篇 |
1980年 | 2篇 |
1979年 | 3篇 |
1978年 | 3篇 |
1977年 | 5篇 |
1975年 | 1篇 |
1972年 | 1篇 |
1971年 | 1篇 |
1969年 | 1篇 |
1967年 | 1篇 |
1964年 | 1篇 |
排序方式: 共有3061条查询结果,搜索用时 15 毫秒
81.
Alekseev S Luijsterburg MS Pines A Geverts B Mari PO Giglia-Mari G Lans H Houtsmuller AB Mullenders LH Hoeijmakers JH Vermeulen W 《Molecular and cellular biology》2008,28(24):7402-7413
Nucleotide excision repair (NER) is the principal pathway for counteracting cytotoxic and mutagenic effects of UV irradiation. To provide insight into the in vivo regulation of the DNA damage recognition step of global genome NER (GG-NER), we constructed cell lines expressing fluorescently tagged damaged DNA binding protein 1 (DDB1). DDB1 is a core subunit of a number of cullin 4-RING ubiquitin ligase complexes. UV-activated DDB1-DDB2-CUL4A-ROC1 ubiquitin ligase participates in the initiation of GG-NER and triggers the UV-dependent degradation of its subunit DDB2. We found that DDB1 rapidly accumulates on DNA damage sites. However, its binding to damaged DNA is not static, since DDB1 constantly dissociates from and binds to DNA lesions. DDB2, but not CUL4A, was indispensable for binding of DDB1 to DNA damage sites. The residence time of DDB1 on the damage site is independent of the main damage-recognizing protein of GG-NER, XPC, as well as of UV-induced proteolysis of DDB2. The amount of DDB1 that is temporally immobilized on damaged DNA critically depends on DDB2 levels in the cell. We propose a model in which UV-dependent degradation of DDB2 is important for the release of DDB1 from continuous association to unrepaired DNA and makes DDB1 available for its other DNA damage response functions. 相似文献
82.
Electron paramagnetic resonance study of radiation damage in photosynthetic reaction center crystals
Electron paramagnetic resonance (EPR) was used to simultaneously study radiation-induced cofactor reduction and damaging radical formation in single crystals of the bacterial reaction center (RC). Crystals of Fe-removed/Zn-replaced RC protein from Rhodobacter ( R.) sphaeroides R26 were irradiated with varied radiation doses at cryogenic temperature and analyzed for radiation-induced free radical formation and alteration of light-induced photosynthetic electron transfer activity using high-field (HF) D-band (130 GHz) and X-band (9.5 GHz) EPR spectroscopies. These analyses show that the formation of radiation-induced free radicals saturated at doses 1 order of magnitude smaller than the amount of radiation at which protein crystals lose their diffraction quality, while light-induced RC activity was found to be lost at radiation doses at least 1 order of magnitude lower than the dose at which radiation-induced radicals exhibited saturation. HF D-band EPR spectra provide direct evidence for radiation-induced reduction of the quinones and possibly other cofactors. These results demonstrate that substantial radiation damage is likely to have occurred during X-ray diffraction data collection used for photosynthetic RC structure determination. Thus, both radiation-induced loss of photochemical activity in RC crystals and reduction of the quinones are important factors that must be considered when correlating spectroscopic and crystallographic measurements of quinone site structures. 相似文献
83.
Kuznetsov VY Ivanov YD Bykov VA Saunin SA Fedorov IA Lemeshko SV Hoa HB Archakov AI 《Proteomics》2002,2(12):1699-1705
The application of atomic force microscopy (AFM) technique in proteomic research, identification and visualization of individual molecules and molecular complexes within the P450cam containing monooxygenase system was demonstrated. The method distinguishes between the binary protein complexes and appropriate monomeric proteins and, also, between the binary and ternary complexes. The AFM images of the components of a cytochrome P450cam containing monooxygenase system - cytochrome P450cam (P450cam), putidaredoxin (Pd) and putidaredoxin reductase (PdR) - were obtained on a mica support. The molecules of P450cam, Pd and PdR were found to have typical heights of 2.6 +/- 0.3 nm, 2.0 +/- 0.3 and 2.8 +/- 0.3 nm, respectively. The measured heights of the binary Pd/PdR and P450cam/PdR complexes were 4.9 +/- 0.3 nm and 5.1 +/- 0.3 nm, respectively. The binary P450cam/Pd complexes were found to have a typical height of about (3.9 / 5.7 nm) and the ternary PdR/Pd/P450cam complexes, a typical height of about 9.1 +/- 0.3 nm. 相似文献
84.
Andrei A. Rodionov Ekaterina V. Efimtseva Sergey N. Mikhailov 《Nucleosides, nucleotides & nucleic acids》2013,32(4-5):623-624
Abstract The first synthesis of O-β-D-ribofuranosyl-(1″-2′)-adenosine-5″-O-phosphate starting from protected 2′-O-β-D-ribofuranosyladenosine has been performed. 相似文献
85.
Evgenia V. Dolgova Yaroslav R. Efremov Konstantin E. Orishchenko Oleg M. Andrushkevich Ekaterina A. Alyamkina Anastasia S. Proskurina Sergey I. Bayborodin Valeriy P. Nikolin Nelly A. Popova Elena R. Chernykh Alexandr A. Ostanin Oleg S. Taranov Vladimir V. Omigov Alexandra M. Minkevich Vladimir A. Rogachev Sergey S. Bogachev Mikhail A. Shurdov 《Gene》2013
We previously reported that fragments of exogenous double-stranded DNA can be internalized by mouse bone marrow cells without any transfection. Our present analysis shows that only 2% of bone marrow cells take up the fragments of extracellular exogenous DNA. Of these, ~ 45% of the cells correspond to CD34 + hematopoietic stem cells. Taking into account that CD34 + stem cells constituted 2.5% of the total cell population in the bone marrow samples analyzed, these data indicate that as much as 40% of CD34 + cells readily internalize fragments of extracellular exogenous DNA. This suggests that internalization of fragmented dsDNA is a general feature of poorly differentiated cells, in particular CD34 + bone marrow cells. 相似文献
86.
VEGF-induced mobilization of caveolae and increase in permeability of endothelial cells 总被引:10,自引:0,他引:10
Chen J Braet F Brodsky S Weinstein T Romanov V Noiri E Goligorsky MS 《American journal of physiology. Cell physiology》2002,282(5):C1053-C1063
Glomerular epithelial cells (GEC) are aknown site of vascular endothelial growth factor (VEGF) production. Weestablished immortalized rat GEC, which retained the ability to produceVEGF. The isoforms expressed by GEC were defined as VEGF-205, -188, -120, and -164. The electrical resistance of endothelial cells culturedon GEC-conditioned matrix, an indicator of the permeability ofmonolayers to solutes, was significantly increased by the treatment with the neutralizing polyclonal antibodies to VEGF and decreased byVEGF-165. Transfection of endothelial cells with green fluorescence protein-caveolin construct and intravital confocal microscopy showedthat VEGF results in a rapid appearance of transcellular elongatedstructures decorated with caveolin. Transmission electron microscopy ofendothelial cells showed that caveolae undergo rapid internalizationand fusion 30 min after application of VEGF-165. Later (36 h),endothelial cells pretreated with VEGF developed fenestrae and showed adecrease in electrical resistance. Immunoelectron microscopy ofglomeruli confirmed VEGF localization to podocytes and in the basementmembrane. In summary, immortalized GEC retain the ability to synthesizeVEGF. Matrix-deposited and soluble VEGF leads to the enhancement ofcaveolae expression, their fission and fusion, formation of elongatedcaveolin-decorated structures, and eventual formation of fenestrae,both responsible for the increase in endothelial permeability. 相似文献
87.
Sergey M Zuev Stephen F Kingsmore Damian DG Gessler 《Theoretical biology & medical modelling》2006,3(1):8-15
Background
Sepsis (bloodstream infection) is the leading cause of death in non-surgical intensive care units. It is diagnosed in 750,000 US patients per annum, and has high mortality. Current understanding of sepsis is predominately observational and correlational, with only a partial and incomplete understanding of the physiological dynamics underlying the syndrome. There exists a need for dynamical models of sepsis progression, based upon basic physiologic principles, which could eventually guide hourly treatment decisions. 相似文献88.
Multidrug resistance driven by ABC membrane transporters is one of the major reasons for treatment failure in human malignancy. Some limited evidence has previously been reported on the cell cycle dependence of ABC transporter expression. However, it has never been demonstrated that the functional activity of these transporters correlates with the cell cycle position. Here, we studied the rate of intrinsic ABC transport in different phases of the cell cycle in cultured MCF-7 breast cancer cells. The rate was characterized in terms of the efflux kinetics from cells loaded with an ABC transporter substrate. As averaging the kinetics over a cell population could lead to errors, we studied kinetics of ABC transport at the single-cell level. We found that the rate of ABC transport in MCF-7 cells could be described by Michaelis-Menten kinetics with two classical parameters, V(max) and K(M). Each of these parameters showed similar unimodal distributions with different positions of maxima for cell subpopulations in the 2c and 4c states. Compared to the 2c cells, the 4c cells exhibited greater V(max) values, indicating a higher activity of transport. They also exhibited a greater V(max)/K(M) ratio, indicating a higher efficiency of transport. Our findings suggest that cell cycle-related modulation of MDR may need to be taken into account when designing chemotherapy regimens which include cytostatic agents. 相似文献
89.
Ogawa J Yamanaka H Mano J Doi Y Horinouchi N Kodera T Nio N Smirnov SV Samsonova NN Kozlov YI Shimizu S 《Bioscience, biotechnology, and biochemistry》2007,71(7):1607-1615
Arthrobacter simplex AKU 626 was found to synthesize 4-hydroxyisoleucine from acetaldehyde, alpha-ketobutyrate, and L-glutamate in the presence of Escherichia coli harboring the branched chain amino acid transaminase gene (ilvE) from E. coli K12 substrain MG1655. By using resting cells of A. simplex AKU 626 and E. coli BL21(DE3)/pET-15b-ilvE, 3.2 mM 4-hydroxyisoleucine was produced from 250 mM acetaldehyde, 75 mM alpha-ketobutyrate, and 100 mM L-glutamate with a molar yield to alpha-ketobutyrate of 4.3% in 50 mM Tris-HCl buffer (pH 7.5) containing 2 mM MnCl(2) x 4H(2)O at 28 degrees C for 2 h. An aldolase that catalyzes the aldol condensation of acetaldehyde and alpha-ketobutyrate was purified from A. simplex AKU 626. Mn(2+) and pyridoxal 5'-monophosphate were effective in stabilizing the enzyme. The native and subunit molecular masses of the purified aldolase were about 180 and 32 kDa respectively. The N-terminal amino acid sequence of the purified enzyme showed no significant homology to known aldolases. 相似文献
90.
Tetin SY Ruan Q Saldana SC Pope MR Chen Y Wu H Pinkus MS Jiang J Richardson PL 《Biochemistry》2006,45(47):14155-14165
Structure-function studies of antibody-antigen systems include the identification of amino acid residues in the antigen that interact with an antibody and elucidation of their individual contributions to binding affinity. We used fluorescence correlation spectroscopy (FCS) and alanine-scanning mutagenesis to characterize the interactions of brain natriuretic peptide (BNP) with two monoclonal antibodies. Human BNP is a 32 amino acid residue long cyclic polypeptide with the ring structure confined between cysteines in positions 10 and 26. It is an important cardiovascular hormone and a valuable diagnostic cardiac marker. We compare the binding strength of the N-terminus Alexa488-labeled BNP, native cyclic BNP, BNP alanine-substituted mutants, linear BNP, and its short fragments to determine the individual contributions of amino acid residues included in the continuous antigenic epitopes that are recognized by two different monoclonal antibodies raised toward BNP. Implementation of FCS for these studies offers all of the advantages of solution phase measurements, including high sensitivity, simplicity of manipulation with reagents, and elimination of solid phase interferences or separation steps. Significant differences in the molecular masses of the free and antibody bound BNP results in a substantial ( approximately 2.5-times) increase in the diffusion rates. Determination of the binding constants and inhibition effects by measuring the diffusion rates of the ligand at the single molecule level introduces the ultimate opportunity for researching systems where the fluorescence intensity and/or fluorescence anisotropy do not change upon interaction of the ligand with the protein. Monoclonal antibodies 106.3 and BC203 demonstrate high affinities to BNP and bind two distant epitopes forming robust antibody sandwiches. Both antibodies are used in Abbott diagnostic assays on AxSYM, IMx, and Architect platforms. 相似文献