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951.
A. Palamidessi I. Testa E. Frittoli S. Barozzi M. Garrè D. Mazza P. P. Di Fiore A. Diaspro G. Scita Mario Faretta 《European biophysics journal : EBJ》2010,39(6):947-957
The dissection of the molecular circuitries at the base of cell life and the identification of their abnormal transformation
during carcinogenesis rely on the characterization of biological phenotypes generated by targeted overexpression or deletion
of gene products through genetic manipulation. Fluorescence microscopy provides a wide variety of tools to monitor cell life
with minimal perturbations. The observation of living cells requires the selection of a correct balance between temporal,
spatial and “statistical” resolution according to the process to be analyzed. In the following paper ad hoc developed optical
tools for dynamical tracking from cellular to molecular resolution will be presented. Particular emphasis will be devoted
to discuss how to exploit light–matter interaction to selectively target specific molecular species, understanding the relationships
between their intracellular compartmentalization and function. 相似文献
952.
Yeasts isolated from olive mill wastewaters from southern Italy: technological characterization and potential use for phenol removal 总被引:1,自引:0,他引:1
Milena Sinigaglia Nilde Di Benedetto Antonio Bevilacqua Maria Rosaria Corbo Angela Capece Patrizia Romano 《Applied microbiology and biotechnology》2010,87(6):2345-2354
Olive mill wastewater (OMW) samples from two traditional varieties (Peranzana and Ogliarola Garganica) of Apulian region (southern
Italy) and produced through continuous and traditional methods were microbiologically and chemically examined; thus, 104 yeasts
were isolated and selected for further analyses. The strains were identified as Candida boidinii, Pichia holstii, Pichia membranifaciens, and Saccharomyces cerevisiae and analyzed to assess their suitability to metabolize phenols. Based on phenol metabolism, 27 strains were selected and
inoculated into OMW aliquots to determine their ability to reduce phenols in vivo; then, five strains (identified with the
codes 682—C. boidinii and 625, 642, 647, and 941—P. holstii) were used as a cocktail in wastewaters for a final validation step. In this last experiment, the effects of the temperature
(10–30°C) and (NH4)2SO4 (0.0–6.0 g l−1) were studied through a central composite design approach, and the results highlighted that the cocktail was able to reduce
phenols by 40% at 10°C with 6.0 g l−1 of (NH4)2SO4 added. 相似文献
953.
954.
955.
Zhang D Kato Y Zhang L Fujimoto M Tsutsumi N Sodmergen Sakamoto W 《The Plant cell》2010,22(11):3710-3725
FtsH is an ATP-dependent metalloprotease present as a hexameric heterocomplex in thylakoid membranes. Encoded in the Arabidopsis thaliana YELLOW VARIEGATED2 (VAR2) locus, FtsH2 is one isoform among major Type A (FtsH1/5) and Type B (FtsH2/8) isomers. Mutants lacking FtsH2 (var2) and FtsH5 (var1) are characterized by a typical leaf-variegated phenotype. The functional importance of the catalytic center (comprised by the zinc binding domain) in FtsH2 was assessed in this study by generating transgenic plants that ectopically expressed FtsH2(488), a proteolytically inactive version of FtsH2. The resulting amino acid substitution inhibited FtsH protease activity in vivo when introduced into Escherichia coli FtsH. By contrast, expression of FtsH2(488) rescued not only leaf variegation in var2 but also seedling lethality in var2 ftsh8, suggesting that the protease activity of Type B isomers is completely dispensable, which implies that the chloroplastic FtsH complex has protease sites in excess and that they act redundantly rather than coordinately. However, expression of FtsH2(488) did not fully rescue leaf variegation in var1 var2 because the overall FtsH levels were reduced under this background. Applying an inducible promoter to our complementation analysis revealed that rescue of leaf variegation indeed depends on the overall amount of FtsH. Our results elucidate protein activity and its amount as important factors for the function of FtsH heterocomplexes that are composed of multiple isoforms in the thylakoid membrane. 相似文献
956.
Guoyong Hu Jiaqin Shen Li Cheng Di Xiang Zhonghui Zhang Miao He Huili Lu Shunying Zhu Mingyuan Wu Yan Yu Xingpeng Wang Wei Han 《Protein expression and purification》2010,69(2):186-190
Regenerating gene (Reg) IV is a newly discovered member of the regenerating gene family belonging to the calcium (C-type) dependent lectin superfamily. Reg IV is highly expressed in the gastrointestinal tract and markedly up-regulated in colon adenocarcinoma, pancreatic cancer, gastric adenocarcinoma, and inflammatory bowel disease. However, the physiological and pathological functions of Reg IV are largely unknown, partly due to the limited access of the bioactive protein. We report here the first expression and purification of Reg IV proteins using a prokaryotic system. Human Reg IV was expressed in Escherichia coli as an insoluble protein which was identified in the fraction of inclusion body after ultrasonication of the bacteria. After the protein aggregate was solubilized by guanidine–HCl, it was refolded by sucrose and arginine-assisted procedures and purified using cation-exchange chromatography. The protein identity and purity of the final preparation were confirmed by analysis of the protein mass and immune specificity in SDS–PAGE, Western blotting, and HPLC assay. The biological activity of the protein was determined by the HCT116 and HT29 cell proliferation assays. The highly purified bioactive human Reg IV should aid in further characterization of its physiological and pathological functions. 相似文献
957.
Di Xiang Jing Zhang Yizhe Chen Yiping Guo Adrian Schalow Zhonghui Zhang Xiaojia Hu Hongjing Yu Mei Zhao Shunying Zhu Huili Lu Mingyuan Wu Yan Yu Anja Moldenhauer Wei Han 《Protein expression and purification》2010,69(2):153-158
Chemerin is a novel chemokine that binds to the G protein-coupled receptor (GPCR) ChemR23, also known as chemokine-like receptor 1 (CMKLR1). It is secreted as a precursor and executes pro-inflammatory functions when the last six amino acids are removed from its C-terminus by serine proteases. After maturation, Chemerin attracts dendritic cells and macrophages through binding to ChemR23. We report a new method for expression and purification of mature recombinant human Chemerin (rhChemerin) using a prokaryotic system. After being expressed in bacteria, rhChemerin in inclusion bodies was denatured using 6 M guanidine chloride. Soluble rhChemerin was prepared by the protein-specific renaturation solution under defined conditions. It was subsequently purified using ion-exchange columns to more than 95% purity with endotoxin level <1.0 EU/μg. We further demonstrated its biological activities for attracting migration of human dendritic cells and murine macrophages in vitro using established chemotaxis assays. 相似文献
958.
Collagen and mature elastic fibre organisation as a function of depth in the human cornea and limbus
Christina S. Kamma-Lorger Craig Boote Sally Hayes Julian Moger Manfred Burghammer Carlo Knupp Andrew J. Quantock Thomas Sorensen Emanuela Di Cola Nick White Robert D. Young Keith M. Meek 《Journal of structural biology》2010,169(3):424-430
A network of circumferentially oriented collagen fibrils exists in the periphery of the human cornea, and is thought to be pivotal in maintaining corneal biomechanical stability and curvature. However, it is unknown whether or not this key structural arrangement predominates throughout the entire corneal thickness or exists as a discrete feature at a particular tissue depth; or if it incorporates any elastic fibres and how, with respect to tissue depth, the circumcorneal annulus integrates with the orthogonally arranged collagen of the central cornea. To address these issues we performed a three-dimensional investigation of fibrous collagen and elastin architecture in the peripheral and central human cornea using synchrotron X-ray scattering and non-linear microscopy. This showed that the network of collagen fibrils circumscribing the human cornea is located in the posterior one-third of the tissue and is interlaced with significant numbers of mature elastic fibres which mirror the alignment of the collagen. The orthogonal arrangement of collagen in the central cornea is also mainly restricted to the posterior stromal layers. This information will aid the development of corneal biomechanical models aimed at explaining how normal corneal curvature is sustained and further predicting the outcome of surgical procedures. 相似文献
959.
960.
Interactions between quercetin and Warfarin for albumin binding: A new eye on food/drug interference
The interaction between quercetin, a popular antioxidant flavonoid, and human serum albumin (HSA) is investigated and characterized by means of induced circular dichroism and saturation transfer difference NMR. These techiques demonstrate the reversible binding of quercetin to the carrier protein, which is responsible for its dissolution in aqueous medium. Competition experiments with two classical probes for HSA binding sites, namely Ibuprofen and Warfarin (a common anticoagulant coumarin), demonstrate that quercetin has a primary binding site located in the subdomain IIA, where coumarins are hosted. The affinity for this site is large and we found that quercetin may effectively displace warfarin from HSA. This may have relevant consequences in rationalizing the interferences of common dietary compounds and food supplements to anticoagulant treatments. Chirality, 2010. © 2009 Wiley‐Liss, Inc. 相似文献