全文获取类型
收费全文 | 1000篇 |
免费 | 116篇 |
国内免费 | 3篇 |
出版年
2022年 | 7篇 |
2021年 | 11篇 |
2020年 | 8篇 |
2019年 | 8篇 |
2018年 | 7篇 |
2017年 | 11篇 |
2016年 | 15篇 |
2015年 | 27篇 |
2014年 | 27篇 |
2013年 | 44篇 |
2012年 | 52篇 |
2011年 | 43篇 |
2010年 | 34篇 |
2009年 | 36篇 |
2008年 | 34篇 |
2007年 | 48篇 |
2006年 | 39篇 |
2005年 | 37篇 |
2004年 | 38篇 |
2003年 | 38篇 |
2002年 | 34篇 |
2001年 | 29篇 |
2000年 | 31篇 |
1999年 | 34篇 |
1998年 | 13篇 |
1997年 | 21篇 |
1996年 | 12篇 |
1994年 | 9篇 |
1993年 | 15篇 |
1992年 | 23篇 |
1991年 | 22篇 |
1990年 | 15篇 |
1989年 | 26篇 |
1988年 | 20篇 |
1987年 | 14篇 |
1986年 | 21篇 |
1985年 | 15篇 |
1984年 | 9篇 |
1983年 | 13篇 |
1982年 | 13篇 |
1981年 | 16篇 |
1980年 | 9篇 |
1979年 | 11篇 |
1978年 | 9篇 |
1976年 | 8篇 |
1975年 | 7篇 |
1974年 | 10篇 |
1973年 | 10篇 |
1970年 | 8篇 |
1968年 | 7篇 |
排序方式: 共有1119条查询结果,搜索用时 296 毫秒
71.
Nandhakumar Thayanidhi Jared R. Helm Deborah C. Nycz Marvin Bentley Yingjian Liang Jesse C. Hay 《Molecular biology of the cell》2010,21(11):1850-1863
Toxicity of human α-synuclein when expressed in simple organisms can be suppressed by overexpression of endoplasmic reticulum (ER)-to-Golgi transport machinery, suggesting that inhibition of constitutive secretion represents a fundamental cause of the toxicity. Whether similar inhibition in mammals represents a cause of familial Parkinson''s disease has not been established. We tested elements of this hypothesis by expressing human α-synuclein in mammalian kidney and neuroendocrine cells and assessing ER-to-Golgi transport. Overexpression of wild type or the familial disease-associated A53T mutant α-synuclein delayed transport by up to 50%; however, A53T inhibited more potently. The secretory delay occurred at low expression levels and was not accompanied by insoluble α-synuclein aggregates or mistargeting of transport machinery, suggesting a direct action of soluble α-synuclein on trafficking proteins. Co-overexpression of ER/Golgi arginine soluble N-ethylmaleimide-sensitive factor attachment protein receptors (R-SNAREs) specifically rescued transport, indicating that α-synuclein antagonizes SNARE function. Ykt6 reversed α-synuclein inhibition much more effectively than sec22b, suggesting a possible neuroprotective role for the enigmatic high expression of ykt6 in neurons. In in vitro reconstitutions, purified α-synuclein A53T protein specifically inhibited COPII vesicle docking and fusion at a pre-Golgi step. Finally, soluble α-synuclein A53T directly bound ER/Golgi SNAREs and inhibited SNARE complex assembly, providing a potential mechanism for toxic effects in the early secretory pathway. 相似文献
72.
Mao H Graziano JJ Chase TM Bentley CA Bazirgan OA Reddy NP Song BD Smider VV 《Nature biotechnology》2010,28(11):1195-1202
Antibody discovery typically uses hybridoma- or display-based selection approaches, which lack the advantages of directly screening spatially addressed compound libraries as in small-molecule discovery. Here we apply the latter strategy to antibody discovery, using a library of ~10,000 human germline antibody Fabs created by de novo DNA synthesis and automated protein expression and purification. In multiplexed screening assays, we obtained specific hits against seven of nine antigens. Using sequence-activity relationships and iterative mutagenesis, we optimized the binding affinities of two hits to the low nanomolar range. The matured Fabs showed full and partial antagonism activities in cell-based assays. Thus, protein drug leads can be discovered using surprisingly small libraries of proteins with known sequences, questioning the requirement for billions of members in an antibody discovery library. This methodology also provides sequence, expression and specificity information at the first step of the discovery process, and could enable novel antibody discovery in functional screens. 相似文献
73.
Zhenrong Xu Salvacion Cacatian Jing Yuan Robert D. Simpson Lanqi Jia Wei Zhao Colin M. Tice Patrick T. Flaherty Joan Guo Alexey Ishchenko Suresh B. Singh Zhongren Wu Brian M. McKeever Boyd B. Scott Yuri Bukhtiyarov Jennifer Berbaum Jennifer Mason Reshma Panemangalore Maria Grazia Cappiello Ross Bentley David A. Claremon 《Bioorganic & medicinal chemistry letters》2010,20(2):694-699
Structure-guided drug design led to new alkylamine renin inhibitors with improved in vitro and in vivo potency. Lead compound 21a, has an IC50 of 0.83 nM for the inhibition of human renin in plasma (PRA). Oral administration of 21a at 10 mg/kg resulted in >20 h reduction of blood pressure in a double transgenic rat model of hypertension. 相似文献
74.
75.
76.
Galkin O Bentley AA Gupta S Compton BA Mazumder B Kinzy TG Merrick WC Hatzoglou M Pestova TV Hellen CU Komar AA 《RNA (New York, N.Y.)》2007,13(12):2116-2128
Ribosomal protein (rp) S5 belongs to a family of ribosomal proteins that includes bacterial rpS7. rpS5 forms part of the exit (E) site on the 40S ribosomal subunit and is essential for yeast viability. Human rpS5 is 67% identical and 79% similar to Saccharomyces cerevisiae rpS5 but lacks a negatively charged (pI approximately 3.27) 21 amino acid long N-terminal extension that is present in fungi. Here we report that replacement of yeast rpS5 with its human homolog yielded a viable yeast strain with a 20%-25% decrease in growth rate. This replacement also resulted in a moderate increase in the heavy polyribosomal components in the mutant strain, suggesting either translation elongation or termination defects, and in a reduction in the polyribosomal association of the elongation factors eEF3 and eEF1A. In addition, the mutant strain was characterized by moderate increases in +1 and -1 programmed frameshifting and hyperaccurate recognition of the UAA stop codon. The activities of the cricket paralysis virus (CrPV) IRES and two mammalian cellular IRESs (CAT-1 and SNAT-2) were also increased in the mutant strain. Consistently, the rpS5 replacement led to enhanced direct interaction between the CrPV IRES and the mutant yeast ribosomes. Taken together, these data indicate that rpS5 plays an important role in maintaining the accuracy of translation in eukaryotes and suggest that the negatively charged N-terminal extension of yeast rpS5 might affect the ribosomal recruitment of specific mRNAs. 相似文献
77.
Yuan J Simpson RD Zhao W Tice CM Xu Z Cacatian S Jia L Flaherty PT Guo J Ishchenko A Wu Z McKeever BM Scott BB Bukhtiyarov Y Berbaum J Panemangalore R Bentley R Doe CP Harrison RK McGeehan GM Singh SB Dillard LW Baldwin JJ Claremon DA 《Bioorganic & medicinal chemistry letters》2011,21(16):4836-4843
Structure-based design led to the discovery of a novel class of renin inhibitors in which an unprecedented phenyl ring filling the S1 site is attached to the phenyl ring filling the S3 pocket. Optimization for several parameters including potency in the presence of human plasma, selectivity against CYP3A4 inhibition and improved rat oral bioavailability led to the identification of 8d which demonstrated antihypertensive efficacy in a transgenic rat model of human hypertension. 相似文献
78.
Viegas TX Bentley MD Harris JM Fang Z Yoon K Dizman B Weimer R Mero A Pasut G Veronese FM 《Bioconjugate chemistry》2011,22(5):976-986
Polyoxazoline polymers with methyl (PMOZ), ethyl (PEOZ), and propyl (PPOZ) side chains were prepared by the living cationic polymerization method and purified by ion-exchange chromatography. The following properties of polyoxazoline (POZ) were measured: apparent hydrodynamic radius by aqueous size-exclusion chromatography, relative lipophilicity by reverse-phase chromatography, and viscosity by cone-plate viscometry. The PEOZ polymers of different molecular weights were first functionalized and then conjugated to model biomolecules such as bovine serum albumin, catalase, ribonuclease, uricase, and insulin. The conjugates of catalase, uricase, and ribonuclease were tested for in vitro activity using substrate-specific reaction methods. The conjugates of insulin were tested for glucose lowering activity by injection to nai?ve Sprague-Dawley rats. The conjugates of BSA were injected into New Zealand white rabbits and serum samples were collected periodically and tested for antibodies to BSA. The safety of POZ was also determined by acute and chronic dosing to rats. The results showed that linear polymers of POZ with molecular weights of 1 to 40 kDa can easily be made with polydispersity values below 1.10. Chromatography results showed that PMOZ and PEOZ have a hydrodynamic volume slightly lower than PEG; PEOZ is more lipophilic than PMOZ and PEG; and PEOZ is significantly less viscous than PEG especially at the higher molecular weights. When PEOZ was attached to the enzymes catalase, ribonuclease, and uricase, the in vitro activity of the resultant bioconjugates depended on the extent of protein modification. POZ conjugates of insulin lowered blood glucose levels for a period of 8 h when compared to 2 h for insulin alone. PEOZ, like PEG, was also able to successfully attenuate the immunogenic properties of BSA. The POZ polymers (10 and 20 kDa) are safe when administered intravenously to rats, and the maximum tolerated dose (MTD) was greater than 2 g/kg. Blood counts, serum chemistry, organ weights, and the histopathology of key organs were normal. These results conclude that POZ has the desired drug delivery properties for a new biopolymer. 相似文献
79.
Hom JR Quintanilla RA Hoffman DL de Mesy Bentley KL Molkentin JD Sheu SS Porter GA 《Developmental cell》2011,21(3):469-478
Although mature myocytes rely on mitochondria as the primary source of energy, the role of mitochondria in the developing heart is not well known. Here, we find that closure of the mitochondrial permeability transition pore (mPTP) drives maturation of mitochondrial structure and function and myocyte differentiation. Cardiomyocytes at embryonic day (E) 9.5, when compared to E13.5, displayed fragmented mitochondria with few cristae, a less-polarized mitochondrial membrane potential, higher reactive oxygen species (ROS) levels, and an open mPTP. Pharmacologic and genetic closing of the mPTP yielded maturation of mitochondrial structure and function, lowered ROS, and increased myocyte differentiation (measured by counting Z bands). Furthermore, myocyte differentiation was inhibited and enhanced with oxidant and antioxidant treatment, respectively, suggesting that redox-signaling pathways lie downstream of mitochondria to regulate cardiac myocyte differentiation. 相似文献
80.
Lamarque M Besteiro S Papoin J Roques M Vulliez-Le Normand B Morlon-Guyot J Dubremetz JF Fauquenoy S Tomavo S Faber BW Kocken CH Thomas AW Boulanger MJ Bentley GA Lebrun M 《PLoS pathogens》2011,7(2):e1001276
Obligate intracellular Apicomplexa parasites share a unique invasion mechanism involving a tight interaction between the host cell and the parasite surfaces called the moving junction (MJ). The MJ, which is the anchoring structure for the invasion process, is formed by secretion of a macromolecular complex (RON2/4/5/8), derived from secretory organelles called rhoptries, into the host cell membrane. AMA1, a protein secreted from micronemes and associated with the parasite surface during invasion, has been shown in vitro to bind the MJ complex through a direct association with RON2. Here we show that RON2 is inserted as an integral membrane protein in the host cell and, using several interaction assays with native or recombinant proteins, we define the region that binds AMA1. Our studies were performed both in Toxoplasma gondii and Plasmodium falciparum and although AMA1 and RON2 proteins have diverged between Apicomplexa species, we show an intra-species conservation of their interaction. More importantly, invasion inhibition assays using recombinant proteins demonstrate that the RON2-AMA1 interaction is crucial for both T. gondii and P. falciparum entry into their host cells. This work provides the first evidence that AMA1 uses the rhoptry neck protein RON2 as a receptor to promote invasion by Apicomplexa parasites. 相似文献