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991.
992.
Liu Q Zhan J Chen X Zheng S 《Biochemical and biophysical research communications》2006,343(3):857-863
Ricin is a potent ribosome inactivating protein and now has been widely used for synthesis of immunotoxins. To target ribosome in the mammalian cytosol, ricin must firstly retrograde transport from the endomembrane system to reach the endoplasmic reticulum (ER) where the ricin A chain (RTA) is recognized by ER components that facilitate its membrane translocation to the cytosol. In the study, the fusion gene of enhanced green fluorescent protein (EGFP)-RTA was expressed with the pET-28a (+) system in Escherichia coli under the control of a T7 promoter. The fusion protein showed a green fluorescence. The recombinant protein can be purified by metal chelated affinity chromatography on a column of NTA. The rabbit anti-GFP antibody can recognize the fusion protein of EGFP-RTA just like the EGFP protein. The cytotoxicity of EGFP-RTA and RTA was evaluated by the MTT assay in HeLa and HEP-G2 cells following fluid-phase endocytosis. The fusion protein had a similar cytotoxicity of RTA. After endocytosis, the subcellular location of the fusion protein can be observed with the laser scanning confocal microscopy and the immuno-gold labeling Electro Microscopy. This study provided important evidence by a visualized way to prove that RTA does reach the endoplasmic reticulum. 相似文献
993.
Sheng-Bin Kou Gang Xu Xiao-Dan Jiang Ru-Xiang Xu Yan-Ping Tang Gang Xu Ying-Qian Cai Mou-Xuan Du Zhi-Cheng Xiao 《Cellular and molecular neurobiology》2010,30(2):275-282
Myelin-derived proteins, such as tenascin-R (TN-R), myelin associate glycoprotein (MAG), oligodendrocyte-myelin glycoprotein
(OMgp), and Nogo-A, inhibit the central nervous system regeneration. In this study, the DNA vaccine encoding for oligodendrocyte
and myelin-related antigens was employed to attenuate the axonal growth inhibitory properties of myelin in the setting of
spinal cord injury. Using a rat spinal cord dorsal hemisection model, the vaccine directed against the inhibitory epitopes
of Nogo-A, MAG, OMgp, and TN-R was administered intramuscularly once a week following spinal cord injury, supplemented with
local application of specific anti-sera against the four antigens. Anterograde labeling of dorsal column fibers showed active
axonal regeneration through the lesion site at the eighth week following the treatment in experimental group but not in control
groups. Light microscopic and ultrastructural analysis revealed that vaccination with these myelin-related antigens did not
lead to demyelinating disease. OMgp and TN-R levels were down-regulated at the lesion site together with a parallel increase
in growth-associated protein 43 levels in the treatment groups. This study reveals the effective approach of a DNA vaccine
strategy by attaining the special antibody to direct neutralization of the myelin inhibitors during spinal cord injury. 相似文献
994.
This proof-of-concept study explores the feasibility of using a drug-loaded hydrophilic polymeric layer sandwiched between two hydrophobic layers for improving film drug load while achieving sustained release of poorly water-soluble drug. Such films having total thickness in range ~?146–250 μm were prepared by slurry-based casting using hydrophilic hydroxypropyl methylcellulose (HPMC) as matrix layer containing fenofibrate (FNB) as the model drug, encased between two very thin rate-limiting layers of 10 μm each of hydrophobic poly-?-caprolactone (PCL). Film precursor slurry consisted of HPMC with plasticizer and water along with micronized FNB powders, which were dry-coated with hydrophilic silica. Characterization techniques demonstrated the presence of homogeneously dispersed crystalline FNB in films. The films are very thin and hence two-dimensional; hence, average drug load per unit area in range ~?5 to ~?9 mg/cm2 could be achieved by altering the thickness of the drug matrix layer. Drug amount and drug content uniformity were measured through assay of ten circular samples ~?0.712 cm2 in area punched out using a circular-shaped punch tool. Drug release rate was investigated using USP IV flow-through cell and surface dissolution imaging system. Thinner films followed Fickian diffusion, and thicker films followed non-Fickian anomalous diffusion. Overall, the application of middle layer thickness could be used as a tool to manipulate drug load without the need for altering its formulation or precursor preparation by changing its thickness, hence achieving relatively high drug loading yet having sustained release of drug. 相似文献
995.
DNA methylation changes in photoperiod-thermo-sensitive male sterile rice PA64S under two different conditions 总被引:1,自引:0,他引:1
Epigenetic modification can occur at a high frequency in crop plants and might generate phenotypic variation without changes in DNA sequences. DNA methylation is an important epigenetic modification that may contribute to environmentally-induced phenotypic variations by regulating gene expression. Rice Photoperiod-Thermo-Sensitive Genic Male Sterile (PTGMS) lines can transform from sterility to fertility under lower temperatures and short-day (SD) conditions during anther development. So far, little is known about the DNA methylation variation of PTGMS throughout the genome in rice. In this study, we investigated DNA cytosine methylation alterations in the young panicles of PTGMS line PA64S under two different conditions using methylation sensitive amplified polymorphism (MSAP) method. Compared with the DNA methylation level of PA64S under lower temperatures and SD conditions (fertility), higher methylation was observed in PA64S (sterility). The sequences of 25 differentially amplified fragments were successfully obtained and annotated. Three methylated fragments, which are homologous to D2, NAD7 and psaA, were confirmed by bisulfite sequencing and their expression levels were also evaluated by qPCR. Real time quantitative PCR analysis revealed that five of the six selected methylated genes were downregulated in PA64S (sterility). These results suggested that DNA methylation may be involved in the sterility–fertility transition of PA64S under two different environmental conditions. 相似文献
996.
997.
Sonya Cressman Ying Sun E. Jane Maxwell Ning Fang David D. Y. Chen Pieter R. Cullis 《International journal of peptide research and therapeutics》2009,15(1):49-59
The cyclic peptide, cRGDf[N(me)]V, binds to the α
v
β
3 integrin and can disrupt binding of the integrin to its natural ligands in the extracellular matrix. In this work, the ability
of a water-soluble, fluorescently labeled variant of the RGD-containing peptide (cRGDfK-488) to bind to integrins on human
umbilical vascular endothelial cells (HUVEC) and subsequently undergo endocytosis was characterized. This information was
compared to the binding and uptake properties of an α
v
β
3 integrin-specific monoclonal antibody, LM609X. The specificity of the RGD-containing peptide is assessed by comparison with
control peptide that does not bind to the α
v
β
3 integrin, cRADfK-488. Using a high purity construct, it is shown that the RGD ligand exhibits dissociation constants in the
micromolar range whereas LM609X exhibits dissociation constants in the nanomolar range. However, the RGD ligand showed greater
uptake following incubation at temperatures which permit endocytosis. A 7.4-fold increase in uptake of the RGD peptide was
observed following a 1 h incubation with HUVEC at 37°C (an endocytosis permissive temperature), as compared to that at 4°C
(an endocytosis prohibitive temperature). In contrast, only a 1.9-fold increase in cell-associated fluorescence was observed
for similar incubations with LM609X. Results from fluorescence microscopy supports the notion that the RGD peptide is rapidly
endocytosed at 37°C as compared to LM609X. These results are discussed with regard to previous work indicating that RGD ligands
enter cells by integrin-independent pathways. These studies provide well-controlled measures of how RGD ligands stimulate
endocytosis. This may be of considerable interest for intracellular delivery of ligand-associated drugs in anti-angiogenic
applications. 相似文献
998.
Yanming Sun Ruofeng Yan Charles I. Muleke Guangwei Zhao lixin Xu Xiangrui Li 《International journal of peptide research and therapeutics》2007,13(3):387-392
The effect of Haemonchus contortus galectin peptides rHco-gal-m/f to induce apoptosis in the peripheral blood lymphocytes (PBLCs) of goats was investigated. Analysis of apoptosis was
carried out with agarose gel electrophoresis, flow cytometry and transmission electron microscopy. The results indicated that
there were visible apoptosis bodies and typical DNA ladders by genomic DNA fragmentation. The quantitative analysis of apoptosis
by flow cytometry indicated that rHco-gal-m/f peptides induced apoptosis was time and dose dependent. Ultrastructural studies of the PBLCs revealed that a large
number of apoptotic cells were present in galectin-treated cells, which had the typical morphologic changes of apoptosis such
as reduction of the cytoplasmic volume, loss of cell surface microvilli, chromatin condensation and fragmentation of the apoptotic
cells into small apoptotic bodies. 相似文献
999.
Background
Profile hidden Markov models (profile-HMMs) are sensitive tools for remote protein homology detection, but the main scoring algorithms, Viterbi or Forward, require considerable time to search large sequence databases. 相似文献1000.
In this study, an aqueous extract of leaves from Melothria maderaspatana was tested for in vitro antioxidant activity. Free radical scavenging assays, such as hydroxyl radical, hydrogen peroxide, superoxide anion radical
and 2,2-diphenyl-1-picryl hydrazyl (DPPH), 2,2’-azinobis-(3-ethyl-enzothiazoline-6-sulfonic acid) (ABTS) radical scavenging,
and reducing power assay, were studied. The extract effectively scavenged hydroxyl radical, hydrogen peroxide and superoxide
anion radicals. It also scavenged DPPH and ABTS radicals. Furthermore, it was found to have reducing power. All concentrations
of leaf extract exhibited free radical scavenging and antioxidant power, and the preventive effects were in a dose-dependent
manner. The antioxidant activities of the above were compared to standard antioxidants such as butylated hydroxytoluene (BHT),
ascorbic acid, and α-tocopherol. The results obtained in the present study indicate that the M. maderaspatana extract could be considered a potential source of natural antioxidant. 相似文献