全文获取类型
收费全文 | 4660篇 |
免费 | 419篇 |
国内免费 | 2篇 |
专业分类
5081篇 |
出版年
2022年 | 35篇 |
2021年 | 59篇 |
2020年 | 54篇 |
2019年 | 59篇 |
2018年 | 48篇 |
2017年 | 65篇 |
2016年 | 112篇 |
2015年 | 169篇 |
2014年 | 205篇 |
2013年 | 257篇 |
2012年 | 336篇 |
2011年 | 332篇 |
2010年 | 204篇 |
2009年 | 190篇 |
2008年 | 263篇 |
2007年 | 268篇 |
2006年 | 253篇 |
2005年 | 254篇 |
2004年 | 257篇 |
2003年 | 221篇 |
2002年 | 230篇 |
2001年 | 63篇 |
2000年 | 47篇 |
1999年 | 71篇 |
1998年 | 64篇 |
1997年 | 56篇 |
1996年 | 38篇 |
1995年 | 38篇 |
1994年 | 30篇 |
1993年 | 51篇 |
1992年 | 34篇 |
1991年 | 37篇 |
1990年 | 34篇 |
1989年 | 39篇 |
1988年 | 27篇 |
1987年 | 24篇 |
1986年 | 28篇 |
1985年 | 28篇 |
1984年 | 30篇 |
1983年 | 25篇 |
1982年 | 27篇 |
1981年 | 24篇 |
1980年 | 19篇 |
1979年 | 24篇 |
1978年 | 19篇 |
1977年 | 22篇 |
1976年 | 21篇 |
1974年 | 16篇 |
1973年 | 24篇 |
1968年 | 15篇 |
排序方式: 共有5081条查询结果,搜索用时 15 毫秒
151.
Dryden KA Tihova M Nowotny N Matsui SM Mendez E Yeager M 《Journal of molecular biology》2012,422(5):650-658
Human astroviruses (HAstVs) are a major cause of gastroenteritis. HAstV assembles from the structural protein VP90 and undergoes a cascade of proteolytic cleavages. Cleavage to VP70 is required for release of immature particles from cells, and subsequent cleavage by trypsin confers infectivity. We used electron cryomicroscopy and icosahedral image analysis to determine the first experimentally derived, three-dimensional structures of an immature VP70 virion and a fully proteolyzed, infectious virion. Both particles display T = 3 icosahedral symmetry and nearly identical solid capsid shells with diameters of ~ 350 Å. Globular spikes emanate from the capsid surface, yielding an overall diameter of ~ 440 Å. While the immature particles display 90 dimeric spikes, the mature capsid only displays 30 spikes, located on the icosahedral 2-fold axes. Loss of the 60 peripentonal spikes likely plays an important role in viral infectivity. In addition, immature HAstV bears a striking resemblance to the structure of hepatitis E virus (HEV)-like particles, as previously predicted from structural similarity of the crystal structure of the astrovirus spike domain with the HEV P-domain [Dong, J., Dong, L., Méndez, E. &; Tao, Y. (2011). Crystal structure of the human astrovirus capsid spike. Proc. Natl. Acad. Sci. USA 108, 12681–12686]. Similarities between their capsid shells and dimeric spikes and between the sequences of their capsid proteins suggest that these viral families are phylogenetically related and may share common assembly and activation mechanisms. 相似文献
152.
Kwon EM Holt SK Fu R Kolb S Williams G Stanford JL Ostrander EA 《Cancer epidemiology》2012,36(4):347-353
Background: Prostate cancer (PC) is the most frequently diagnosed solid tumor in U.S. men. Genome-wide association studies (GWAS) have identified over 40 risk-associated single nucleotide polymorphisms (SNPs), including variants in androgen pathway genes (e.g., KLK3 and AR). Androgens are important in PC and genes involved in this pathway are therefore candidates for conferring susceptibility to PC. Methods: In this hypothesis-testing study, we evaluated PC risk in association with SNPs in 22 candidate genes involved in androgen metabolism or interactions with the androgen receptor (AR). A total of 187 SNPs were genotyped in 1458 cases and 1351 age-matched controls from a population-based study. PC risk was estimated using adjusted unconditional logistic regression and multinomial regression models. Results: Single SNP analyses showed evidence (p < 0.05) for associations with 14 SNPs in 9 genes: NKX3.1, HSD17B3, AKR1C3, SULT2A1, CYP17A1, KLK3, JAK2, NCOA4 and STAT3. The most significant result was observed for rs2253502 in HSD17B3 (odds ratio, OR = 0.57, 95% CI: 0.39–0.84). In addition, five SNPs in four genes (CYP17A1, HSD17B4, NCOA4, and SULT2A1) were associated with more aggressive disease (p < 0.01). Conclusions: Our results replicate previously reported associations for SNPs in CYP17A1, HSD17B3, ARK1C3, NKX3.1, NCOA4 and KLK3. In addition, novel associations were observed for SNPs in JAK2, HSD17B4, and SULT2A1. These results will require replication in larger studies. 相似文献
153.
Competition for Light between Toxic and Nontoxic Strains of the Harmful Cyanobacterium Microcystis 总被引:1,自引:0,他引:1 下载免费PDF全文
W. Edwin A. Kardinaal Linda Tonk Ingmar Janse Suzanne Hol Pieter Slot Jef Huisman Petra M. Visser 《Applied microbiology》2007,73(9):2939-2946
The cyanobacterium Microcystis can produce microcystins, a family of toxins that are of major concern in water management. In several lakes, the average microcystin content per cell gradually declines from high levels at the onset of Microcystis blooms to low levels at the height of the bloom. Such seasonal dynamics might result from a succession of toxic to nontoxic strains. To investigate this hypothesis, we ran competition experiments with two toxic and two nontoxic Microcystis strains using light-limited chemostats. The population dynamics of these closely related strains were monitored by means of characteristic changes in light absorbance spectra and by PCR amplification of the rRNA internal transcribed spacer region in combination with denaturing gradient gel electrophoresis, which allowed identification and semiquantification of the competing strains. In all experiments, the toxic strains lost competition for light from nontoxic strains. As a consequence, the total microcystin concentrations in the competition experiments gradually declined. We did not find evidence for allelopathic interactions, as nontoxic strains became dominant even when toxic strains were given a major initial advantage. These findings show that, in our experiments, nontoxic strains of Microcystis were better competitors for light than toxic strains. The generality of this finding deserves further investigation with other Microcystis strains. The competitive replacement of toxic by nontoxic strains offers a plausible explanation for the gradual decrease in average toxicity per cell during the development of dense Microcystis blooms. 相似文献
154.
Emma?M.M. Burkitt?Wright Helen?L. Spencer Sarah?B. Daly Forbes?D.C. Manson Leo?A.H. Zeef Jill Urquhart Nicoletta Zoppi Richard Bonshek Ioannis Tosounidis Meyyammai Mohan Colm Madden Annabel Dodds Kate?E. Chandler Siddharth Banka Leon Au Jill Clayton-Smith Naz Khan Leslie?G. Biesecker Meredith Wilson Marianne Rohrbach Marina Colombi Cecilia Giunta Graeme?C.M. Black 《American journal of human genetics》2011,89(2):346
155.
Population density and infection with parasites often are important factors affecting the growth and development of individuals.
How these factors co-occur and interact in nature should have important consequences for individual fitness and higher-order
phenomena, such as population dynamics of hosts and their interactions with other species. However, few studies have examined
the joint effects of density and parasitism on host growth and development. We examined the co-influences of rearing density
and parasitism, by the trematode Echinostoma trivolvis, on the growth and development of larval frogs, Rana (=Lithobates) pipiens. We also examined the potential role of parasite-mediated intraspecific competition by observing how unparasitized individuals
performed when housed with other unparasitized tadpoles, versus housing with a combination of unparasitized and parasitized
hosts. Mean mass and mean developmental stage were reduced under high rearing densities. The presence of parasitized conspecifics
had no significant effect, but there was a significant interaction of density and parasitism presence on host mass, due to
the fact that parasitized conspecifics grew poorly at high densities. Unparasitized individuals reared with parasitized and
unparasitized conspecifics fared no better than unparasitized individuals reared only with one another. This result indicates
that infected hosts compete as much as uninfected hosts for resources, even though infected individuals have reduced mass
under high-density conditions. Resource acquisition and resource allocation are different processes, and parasitism, if it
only affects the latter, might not have a discernible impact on competitive interactions. 相似文献
156.
Small KM Brown KM Forbes SL Liggett SB 《The Journal of biological chemistry》2001,276(34):31596-31601
Depressed G-protein-coupled receptor (GPCR) signaling has been implicated as a component of the pathophysiology of a number of complex diseases including heart failure and asthma, and augmentation or restoration of signaling by various means has been shown to improve organ function. Because some properties of native GPCRs are disadvantageous for ectopic therapeutic expression, we utilized the beta(2)-adrenergic receptor (beta(2)AR) as a scaffold to construct a highly modified therapeutic receptor-effector complex (TREC) suitable for gene therapy. Altogether, 19 modifications were made to the receptor. The ligand-binding site was re-engineered in TM-3 so that a beta-hydroxylmethyl side chain acts as a proton donor for the binding of a novel ligand. In addition, sites critical for agonist-promoted down-regulation in the amino terminus and for phosphorylation by GPCR kinases, and protein kinases A and C, in the third intracellular loop and the carboxyl terminus of the receptor were altered. These modifications of the receptor resulted in depressed agonist-stimulated adenylyl cyclase activity (26.8 +/- 2.1 versus 41.4 +/- 8 pmol/min/mg for wild-type beta(2)AR). This was fully restored by fusing the carboxyl terminus of the modified receptor to G alpha(s) (43.3 +/- 2.7 pmol/min/mg). The fully modified fused receptor was not activated by beta-agonists but rather by a nonbiogenic amine agonist that itself failed to activate the wild-type beta(2)AR. This two-way selectivity thus provides targeted activation based on physiologic status. Furthermore, the TREC did not display tachyphylaxis to prolonged agonist exposure (desensitization was 1 +/- 5% versus 55 +/- 4% for wild-type beta(2)AR). Thus, despite extensive alterations in regions of conformational lability, the beta(2)AR can be tailored to have optimal signaling characteristics for gene therapy. As a general paradigm, TRECs for enhancement of other G-protein signaling appear to be feasible for modification of other pathologic states. 相似文献
157.
Four juvenile male wolves (Canis lupus) each received an oral dose of 1.6-1.7 x 10(12) colony-forming units of Brucella abortus biovar 1 isolated from a bison (Bison bison) in Wood Buffalo National Park (Canada), and two others served as negative controls. Infected wolves did not show clinical signs of disease but did develop high Brucella antibody titers. Small numbers of B. abortus were excreted sporadically in feces until day 50 postinoculation (PI). Very small numbers of the bacterium were isolated from urine of only one wolf late on the same day that it was infected, and very small numbers of colonies of B. abortus were obtained from buccal swabs of three wolves for up to 48 hr PI. Two infected wolves euthanized 6 mo after the start of the experiment had no lesions, and colonies of B. abortus were isolated from thymus and most major lymph nodes. The other two infected wolves euthanized 12 mo after the start of the experiment had no lesions, and smaller numbers of brucellae were recovered from fewer lymph nodes compared with the wolves killed 6 mo earlier. The sporadic excretion of very small numbers of brucellae by the wolves was insignificant when compared with the infective dose for cattle. Brucella abortus, brucellosis, Canis lupus, pathogenesis, serology, wolf. 相似文献
158.
Sonya B Dumanis Kelly A Chamberlain Yoo Jin Sohn Young Jin Lee Suzanne Y Guénette Toshiharu Suzuki Paul M Mathews Daniel TS Pak G William Rebeck Yoo-hun Suh Hee-Sae Park Hyang-Sook Hoe 《Molecular neurodegeneration》2012,7(1):1-15
Background
Loss-of-function mutations in PTEN-induced kinase 1 (PINK1) have been linked to familial Parkinson??s disease, but the underlying pathogenic mechanism remains unclear. We previously reported that loss of PINK1 impairs mitochondrial respiratory activity in mouse brains.Results
In this study, we investigate how loss of PINK1 impairs mitochondrial respiration using cultured primary fibroblasts and neurons. We found that intact mitochondria in PINK1?/? cells recapitulate the respiratory defect in isolated mitochondria from PINK1?/? mouse brains, suggesting that these PINK1?/? cells are a valid experimental system to study the underlying mechanisms. Enzymatic activities of the electron transport system complexes are normal in PINK1?/? cells, but mitochondrial transmembrane potential is reduced. Interestingly, the opening of the mitochondrial permeability transition pore (mPTP) is increased in PINK1?/? cells, and this genotypic difference between PINK1?/? and control cells is eliminated by agonists or inhibitors of the mPTP. Furthermore, inhibition of mPTP opening rescues the defects in transmembrane potential and respiration in PINK1?/? cells. Consistent with our earlier findings in mouse brains, mitochondrial morphology is similar between PINK1?/? and wild-type cells, indicating that the observed mitochondrial functional defects are not due to morphological changes. Following FCCP treatment, calcium increases in the cytosol are higher in PINK1?/? compared to wild-type cells, suggesting that intra-mitochondrial calcium concentration is higher in the absence of PINK1.Conclusions
Our findings show that loss of PINK1 causes selective increases in mPTP opening and mitochondrial calcium, and that the excessive mPTP opening may underlie the mitochondrial functional defects observed in PINK1?/? cells. 相似文献159.
Mette Nyegaard Nanna D. Rendtorff Morten S. Nielsen Thomas J. Corydon Ditte Demontis Anna Starnawska Anne Hedemand Annalisa Buniello Francesco Niola Michael T. Overgaard Suzanne M. Leal Wasim Ahmad Friedrik P. Wikman Kirsten B. Petersen Dorthe G. Crüger Jaap Oostrik Hannie Kremer Niels Tommerup Morten Fr?din Karen P. Steel Lisbeth Tranebj?rg Anders D. B?rglum 《PLoS genetics》2015,11(7)
160.
Paradis-Bleau C Markovski M Uehara T Lupoli TJ Walker S Kahne DE Bernhardt TG 《Cell》2010,143(7):1110-1120
Most bacteria surround themselves with a peptidoglycan (PG) exoskeleton synthesized by polysaccharide polymerases called penicillin-binding proteins (PBPs). Because they are the targets of penicillin and related antibiotics, the structure and biochemical functions of the PBPs have been extensively studied. Despite this, we still know surprisingly little about how these enzymes build the PG layer in?vivo. Here, we identify the Escherichia coli outer-membrane lipoproteins LpoA and LpoB as essential PBP cofactors. We show that LpoA and LpoB form specific trans-envelope complexes with their cognate PBP and are critical for PBP function in?vivo. We further show that LpoB promotes PG synthesis by its partner PBP in?vitro and that it likely does so by stimulating glycan chain polymerization. Overall, our results indicate that PBP accessory proteins play a central role in PG biogenesis, and like the PBPs they work with, these factors are attractive targets for antibiotic development. 相似文献