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Ahn K Yeyeodu S Collette J Madden V Arthur J Li L Erickson AH 《Traffic (Copenhagen, Denmark)》2002,3(2):147-159
In transformed mouse fibroblasts, a significant proportion of the lysosomal cysteine protease cathepsin L remains in cells as an inactive precursor which associates with membranes by a mannose phosphate-independent interaction. When microsomes prepared from these cells were resolved on sucrose gradients, this procathepsin L was localized in dense vesicles distinct from those enriched for growth hormone, which is secreted constitutively when expressed in fibroblasts. Ultrastructural studies using antibodies directed against the propeptide to avoid detection of the mature enzyme in lysosomes revealed that the proenzyme was concentrated in dense cores within small vesicles and multivesicular endosomes which labeled with antibodies specific for CD63. Consistent with the resemblance of these cores to those of regulated secretory granules, secretion of procathepsin L from fibroblasts was modestly stimulated by phorbol, 12-myristate, 13-acetate. When protein synthesis was blocked with cycloheximide and lysosomal proteolysis inhibited with leupeptin, procathepsin L was found to gradually convert to the active single-chain protease. The data suggest that when synthesis levels are high, a portion of the procathepsin L is packaged in dense cores within multivesicular endosomes localized near the plasma membrane. Gradual activation of this proenzyme achieves targeting of the proenzyme to lysosomes by a mannose phosphate receptor-independent pathway. 相似文献
54.
The saliva of ticks contains a complex mixture of bioactive molecules including proteins that modulate host responses ensuring
successful feeding. The limited amount of saliva that can be obtained from ticks has hampered characterization of salivary
proteins using traditional protein chemistry. Recent improvements in two-dimensional gel electrophoresis, mass spectrometry,
and bioinformatics provide new tools to characterize small amounts of protein. These methods were employed to characterize
salivary proteins from Amblyomma americanum and Amblyomma maculatum. Salivation was induced by injection of dopamine and theophylline. It was necessary to desalt and concentrate saliva before
analysis by 2-D electrophoresis. Comparison of 1-D and 2-D gel patterns revealed that the major protein component of saliva
did not appear on 2-D gels. Characterization of this protein showed that it was identical to the major protein present in
the hemolymph of both tick species. Protein profiles obtained by 1-D and 2-D gel electrophoresis were similar for both tick
species, however, higher concentrations of lower molecular weight proteins were present in A. maculatum. Protein analysis by MALDI-TOF mass spectrometry and western blot analysis showed that except for the most abundant protein
with a molecular weight of 95 kDa, all of the proteins detected were of host origin. It is not known if this is an artifact
of the collection method or has physiological significance. In either case, in these species of ticks, host proteins will
have to be removed from saliva samples prior to 2-D analysis in order to characterize lower abundance proteins of tick origin.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
55.
Madden SF O'Donovan B Furney SJ Brady HR Silvestre G Doran PP 《Bioinformatics (Oxford, England)》2003,19(12):1594-1595
Digital Extractor is a program for the high-throughput processing of data sets derived from digital differential display-based comparisons of EST libraries. These comparisons can be utilized to identify discrete subsets of genes whose expression is restricted to distinct tissue types. The program facilitates these investigations by permitting parallel annotation of genes identified as being differentially expressed. 相似文献
56.
Chintamani Sharma R Badran R Singhal V Saxena S Bansal A 《World journal of surgical oncology》2003,1(1):13
Background
Sweat gland adenocarcinoma is a rare malignancy with high metastatic potential seen more commonly in later years of life. Scalp is the most common site of occurrence and it usually spreads to lymph nodes. Liver, lung and bones are the distant sites of metastasis with fatal results. The differentiation between apocrine and eccrine metastatic sweat gland carcinoma is often difficult. The criteria's are inadequate to be of any practical utility. 相似文献57.
1. The rostral ventrolateral medulla (RVLM) is essential for the normal control of arterial pressure. This region of the brainstem is heterogeneous, and contains spinally projecting neurons that contain phenylethanolamine-N-methyltransferase (C1 neurons) and non-C1 neurons.2. The availability of a toxin, a dopamine--hydroxylase antibody conjugated to saporin, that can selectively destroy C1 neurons within the RVLM allows for the study of cardiovascular regulation in rats following depletion of the C1 neuronal population.3. C1 neurons are not essential for the maintenance of resting blood pressure in anesthetized rats, but do contribute to many cardiovascular responses mediated through the RVLM.4. The depressor response elicited by clonidine injected into the RVLM is dependent upon C1 neurons.5. Studies in rats with selective toxin-induced destruction of RVLM C1 neurons demonstrate that C1 neurons contribute importantly to cardiovascular regulation. 相似文献
58.
Wheeler DL Church DM Lash AE Leipe DD Madden TL Pontius JU Schuler GD Schriml LM Tatusova TA Wagner L Rapp BA 《Nucleic acids research》2001,29(1):11-16
In addition to maintaining the GenBank nucleic acid sequence database, the National Center for Biotechnology Information (NCBI) provides data analysis and retrieval resources that operate on the data in GenBank and a variety of other biological data made available through NCBI's Web site. NCBI data retrieval resources include Entrez, PubMed, LocusLink and the Taxonomy Browser. Data analysis resources include BLAST, Electronic PCR, OrfFinder, RefSeq, UniGene, HomoloGene, Database of Single Nucleotide Polymorphisms (dbSNP), Human Genome Sequencing, Human MapViewer, GeneMap'99, Human-Mouse Homology Map, Cancer Chromosome Aberration Project (CCAP), Entrez Genomes, Clusters of Orthologous Groups (COGs) database, Retroviral Genotyping Tools, Cancer Genome Anatomy Project (CGAP), SAGEmap, Gene Expression Omnibus (GEO), Online Mendelian Inheri-tance in Man (OMIM), the Molecular Modeling Database (MMDB) and the Conserved Domain Database (CDD). Augmenting many of the Web applications are custom implementations of the BLAST program optimized to search specialized data sets. All of the resources can be accessed through the NCBI home page at: http://www.ncbi.nlm.nih. gov. 相似文献
59.
60.
Halla TR Madden JA Gordon JB 《American journal of physiology. Lung cellular and molecular physiology》2000,278(5):L968-L973
Pulmonary venous constriction leads to significant pulmonary hypertension and increased edema formation in several models using newborns. Although alkalosis is widely used in treating neonatal and pediatric pulmonary hypertension, its effects on pulmonary venous tone have not previously been directly measured. This study sought to determine whether alkalosis caused pulmonary venous relaxation and, if so, to identify the mediator(s) involved. Pulmonary venous rings (500-microm external diameter) were isolated from 1-wk-old piglets and precontracted with the thromboxane mimetic U-46619. Responses to hypocapnic alkalosis were then measured under control conditions after inhibition of endothelium-derived modulator activity or K(+) channels. In control rings, alkalosis caused a 34.4 +/- 4.8% decrease in the U-46619-induced contraction. This relaxation was significantly blunted in rings without functional endothelium and in rings treated with nitric oxide synthase or guanylate cyclase inhibitors. However, neither cyclooxygenase inhibition nor voltage-dependent, calcium-dependent, or ATP-dependent K(+)-channel inhibitors altered alkalosis-induced relaxation. These data suggest that alkalosis caused significant dilation of piglet pulmonary veins that was mediated by the nitric oxide-cGMP pathway. 相似文献