全文获取类型
收费全文 | 27458篇 |
免费 | 1538篇 |
国内免费 | 4172篇 |
出版年
2024年 | 50篇 |
2023年 | 380篇 |
2022年 | 599篇 |
2021年 | 769篇 |
2020年 | 719篇 |
2019年 | 913篇 |
2018年 | 745篇 |
2017年 | 801篇 |
2016年 | 909篇 |
2015年 | 1049篇 |
2014年 | 1313篇 |
2013年 | 1946篇 |
2012年 | 1127篇 |
2011年 | 1300篇 |
2010年 | 1062篇 |
2009年 | 1453篇 |
2008年 | 1525篇 |
2007年 | 1632篇 |
2006年 | 1536篇 |
2005年 | 1484篇 |
2004年 | 1359篇 |
2003年 | 1305篇 |
2002年 | 1082篇 |
2001年 | 858篇 |
2000年 | 712篇 |
1999年 | 672篇 |
1998年 | 690篇 |
1997年 | 564篇 |
1996年 | 527篇 |
1995年 | 538篇 |
1994年 | 509篇 |
1993年 | 406篇 |
1992年 | 384篇 |
1991年 | 291篇 |
1990年 | 243篇 |
1989年 | 230篇 |
1988年 | 212篇 |
1987年 | 165篇 |
1986年 | 122篇 |
1985年 | 163篇 |
1984年 | 191篇 |
1983年 | 122篇 |
1982年 | 125篇 |
1981年 | 79篇 |
1980年 | 60篇 |
1979年 | 62篇 |
1978年 | 47篇 |
1977年 | 26篇 |
1976年 | 40篇 |
1974年 | 26篇 |
排序方式: 共有10000条查询结果,搜索用时 937 毫秒
1.
Plant somatic cells have the capability to switch their cell fates from differentiated to undifferentiated status under proper
culture conditions, which is designated as totipotency. As a result, plant cells can easily regenerate new tissues or organs
from a wide variety of explants. However, the mechanism by which plant cells have such remarkable regeneration ability is
still largely unknown. In this study, we used a set of meristem-specific marker genes to analyze the patterns of stem cell
differentiation in the processes of somatic embryogenesis as well as shoot or root organogenesis in vitro. Our studies furnish preliminary and important information on the patterns of the de novo stem cell differentiation during various types of in vitro organogenesis. 相似文献
2.
Wen-Yeh Hsieh Tang-Ching Kuan Kun-Shan Cheng Yan-Chiou Liao Mu-Yuan Chen Pei-Heng Lin Yuan-Chang Hsu Chen-Yi Huang Wei-Hua Hsu Sheng-Yao Yu Chih-Sheng Lin 《International journal of biological sciences》2012,8(8):1197-1205
Objective: Pleural effusion is common problem, but the rapid and reliable diagnosis for specific pathogenic effusions are lacking. This study aimed to identify the diagnosis based on clinical variables to differentiate pleural tuberculous exudates from other pleural effusions. We also investigated the role of renin-angiotensin system (RAS) and matrix metalloproteinase (MMPs) in the pathogenesis of pleural exudates.Experimental design: The major components in RAS and extracellular matrix metabolism, including angiotensin converting enzyme (ACE), ACE2, MMP-2 and MMP-9 activities, were measured and compared in the patients with transudative (n = 45) and exudative (n = 80) effusions. The exudative effusions were come from the patients with tuberculosis (n = 20), pneumonia (n = 32), and adenocarcinoma (n = 28).Results: Increased ACE and equivalent ACE2 activities, resulting in a significantly increased ACE/ACE2 ratio in exudates, were detected compared to these values in transudates. MMP-9 activity in exudates was significantly higher than that in transudates. The significant correlation between ACE and ACE2 activity that was found in transudates was not found in exudates. Advanced analyses showed significantly increased ACE and MMP-9 activities, and decreased ACE2 activity in tuberculous pleural effusions compared with those in pneumonia and adenocarcinoma effusions. The results indicate that increased ACE and MMP-9 activities found in the exudates were mainly contributed from a higher level of both enzyme activities in the tuberculous pleural effusions.Conclusion: Interplay between ACE and ACE2, essential functions in the RAS, and abnormal regulation of MMP-9 probably play a pivotal role in the development of exudative effusions. Moreover, the ACE/ACE2 ratio combined with MMP-9 activity in pleural fluid may be potential biomarkers for diagnosing tuberculous pleurisy. 相似文献
3.
4.
5.
Sandrine Poncet Armelle Delecluse Guido Anello ré Klier Georges Rapoport 《FEMS microbiology letters》1994,117(1):91-95
Abstract The genes encoding the CryIVB and CryIVD crystal polypeptides of B. thuringiensis subsp. israelensis were cloned indepently on a stable shuttle vector, and transfered into B. sphaericus 2297. Recombinant cells expressed the B. thuringiensis toxins during sporulation and were shown to be toxic to Aedes aegypti fourth instar larvae, whereas the parental strain was not. 相似文献
6.
Steven J. Tucker David McClelland Kristina Sep?i? Roderick H. Scott 《生物化学与生物物理学报:生物膜》2003,1614(2):171-181
The ability of two alkyl pyridinium sponge toxin preparations (poly-APS and halitoxin) to form transient pores/lesions in cell membranes and allow transfection of plasmid cDNA have been investigated using HEK 293 cells. Poly-APS and halitoxin preparations caused a collapse in membrane potential, reductions in input resistance and increased Ca2+ permeability. At least partial recovery was observed after poly-APS application but recovery was more rarely seen with halitoxin. The transfection with plasmid cDNAs for an enhanced green fluorescent protein (EGFP) and human tumour necrosis factor receptor 2 (TNFR2) was assessed for both toxin preparations and compared with lipofectamine. Stable transfection was achieved with poly-APS although it was less efficient than lipofectamine. These results show that viable cells transfected with alien cDNA can be obtained using novel transient pore-forming alkyl pyridinium sponge toxins and a simple pre-incubation protocol. This provides the first proof of principle that pore-forming alkyl pyridinium compounds can be used to deliver cDNA to the intracellular environment without permanently compromising the plasma membrane. 相似文献
7.
8.
We re-engineered a classic tool for mutagenesis and gene expression studies in Gram-negative bacteria. Our modified Tn5-based transposon contains multiple features that allow rapid selection for mutants, direct quantification of gene expression and straightforward cloning of the inactivated gene. The promoter-less gfp-km cassette provides selection and reporter assay depending on the activity of the promoter upstream of the transposon insertion site. The cat gene facilitates positive antibiotic selection for mutants, while the narrow R6Kγ replication origin forces transposition in recipient strains lacking the pir gene and enables cloning of the transposon flanked with the disrupted gene from the chromosome. The suicide vector pCKD100, a plasmid that could be delivered into recipient cells through biparental mating or electroporation, harbours the modified transposon. We used the transposon to mutagenize Pectobacterium versatile KD100, Pseudumonas coronafaciens PC27R and Escherichia coli 35150N. The fluorescence intensities of mutants expressing high GFP could be quantified and detected qualitatively. Transformation efficiency from conjugation ranged from 1600 to 1900 CFU per ml. We sequenced the upstream flanking regions, identified the putative truncated genes and demonstrated the restoration of the GFP phenotype through marker exchange. The mini-Tn5 transposon was also utilized to construct mutant a library of P. versatile for forward genetic screens. 相似文献
9.
D E Bowman 《Biochemical and biophysical research communications》1979,87(1):78-84
The oxidized B chain of insulin was used as a simple model for further consideration of limited proteolysis with low substrate:enzyme ratios. With low B chain:trypsin ratios, the ordinarily slower cleavage rate of the -Lys29-Ala30 bond essentially equaled the cleavage saturation rate of the -Arg22-Gly23 bond. This led to the disappearance of octapeptide which ordinarily forms most rapidly. Heptapeptide and alanine, formed mainly by cleavage of the octapeptide, decreased somewhat at high enzyme relative levels. Trypsin added to B chain formed a single chromatographic peak. 相似文献
10.
The psbQ gene encoding a 16-kDa polypeptide of the oxygen-evolving complex of photosystem II has been isolated from Arabidopsis thaliana and characterized. The gene consists of a 28 nucleotide long leader sequence, two introns and three exons encoding a 223-amino-acid precursor polypeptide. The first 75 amino acids act as a transit peptide for the translocation of the polypeptide into the thylakoid lumen. Expression studies show that the gene is light-inducible and expresses only in green tissues with high steady-state mRNA levels in leaves. Using this gene as a probe, restriction fragment length polymorphism between two ecotypes, Columbia and Estland, has also been detected. 相似文献