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981.
Murphy G Knäuper V Lee MH Amour A Worley JR Hutton M Atkinson S Rapti M Williamson R 《Biochemical Society symposium》2003,(70):65-80
Pericellular proteolysis represents one of the key modes by which the cell can modulate its environment, involving not only turnover of the extracellular matrix but also the regulation of cell membrane proteins, such as growth factors and their receptors. The metzincins are active players in such proteolytic events, and their mode of regulation is therefore of particular interest and importance. The TIMPs (tissue inhibitors of metalloproteinases) are established endogenous inhibitors of the matrix metalloproteinases (MMPs), and some have intriguing abilities to associate with the pericellular environment. It has been shown that TIMP-2 can bind to cell surface MT1-MMP (membrane-type 1 MMP) to act as a 'receptor' for proMMP-2 (progelatinase A), such that the latter can be activated efficiently in a localized fashion. We have examined the key structural features of TIMP-2 that determine this unique function, showing that Tyr36 and Glu192-Asp193 are vital for specific interactions with MT1-MMP and proMMP-2 respectively, and hence activation of proMMP-2. TIMP-3 is sequestered at the cell surface by association with the glycosaminoglycan chains of proteoglycans, especially heparan sulphate, and we have shown that it may play a role in the regulation of some ADAMs (a disintegrin and metalloproteinases), including tumour necrosis factor alpha-converting enzyme (TACE; ADAM17). We have established that key residues in TIMP-3 determine its interaction with TACE. Further studies of the features of TIMP-3 that determine specific binding to both ADAM and glycosaminoglycan are required in order to understand these unique properties. 相似文献
982.
The nucleic acid stain SYBR Green I was evaluated for use with solid-phase laser cytometry to obtain total bacterial cell counts from several water sources with small bacterial numbers. Results were obtained within 30 min and exceeded or equaled counts on R2A agar plates incubated for 14 days at room temperature. 相似文献
983.
Hong Y Berrang ME Liu T Hofacre CL Sanchez S Wang L Maurer JJ 《Applied and environmental microbiology》2003,69(6):3492-3499
Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 x 10(2) and 4 x 10(1) CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100- to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa = 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for SALMONELLA: With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry. 相似文献
984.
Paddick JS Brailsford SR Kidd EA Gilbert SC Clark DT Alam S Killick ZJ Beighton D 《Applied and environmental microbiology》2003,69(11):6475-6480
The genotypic diversity of Actinomyces naeslundii genospecies 2 (424 isolates) and Streptococcus oralis (446 isolates) strains isolated from two sound approximal sites in all subjects who were either caries active (seven subjects) or caries free (seven subjects) was investigated by using the repetitive extragenic palindromic PCR. The plaque from the caries-active subjects harbored significantly greater proportions of mutans streptococci and lactobacilli and a smaller proportion of A. naeslundii organisms than the plaque sampled from the caries-free subjects. These data confirmed that the sites of the two groups of subjects were subjected to different environmental stresses, probably determined by the prevailing or fluctuating acidic pH values. We tested the hypothesis that the microfloras of the sites subjected to greater stresses (the plaque samples from the caries-active subjects) would exhibit reduced genotypic diversity since the sites would be less favorable. We found that the diversity of A. naeslundii strains did not change (chi2 = 0.68; P = 0.41) although the proportional representation of A. naeslundii was significantly reduced (P < 0.05). Conversely, the diversity of the S. oralis strains increased (chi2 = 11.71; P = 0.0006) and the proportional representation of S. oralis did not change. We propose that under these environmental conditions the diversity and number of niches within the oral biofilm that could be exploited by S. oralis increased, resulting in the increased genotypic diversity of this species. Apparently, A. naeslundii was not able to exploit the new niches since the prevailing conditions within the niches may have been deleterious and not supportive of its proliferation. These results suggest that environmental stress may modify a biofilm such that the diversity of the niches is increased and that these niches may be successfully exploited by some, but not necessarily all, members of the microbial community. 相似文献
985.
De La Fuente J Golsteyn Thomas EJ van den Bussche RA Hamilton RG Tanaka EE Druhan SE Kocan KM 《Applied and environmental microbiology》2003,69(8):5001-5005
Anaplasma marginale (Rickettsiales: Anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world. Although serologic tests have identified American bison, Bison bison, as being infected with A. marginale, the present study was undertaken to confirm A. marginale infection and to characterize isolates obtained from naturally infected bison in the United States and Canada. Major surface protein (MSP1a and MSP4) sequences of bison isolates were characterized in comparison with New World cattle isolates. Blood from one U.S. bison was inoculated into a susceptible, splenectomized calf, which developed acute anaplasmosis, demonstrating infectivity of this A. marginale bison isolate for cattle. The results of this study showed that these A. marginale isolates obtained from bison were similar to ones from naturally infected cattle. 相似文献
986.
987.
The effects of endogenous and exogenous C2H4 and C2H4 inhibitors on the postharvest leaf and flower quality of Oriental lily Stargazer were investigated. Endogenous C2H4 was not produced by freshly harvested excised leaves or flowers. Treatment of freshly harvested excised flowers, buds, leaves, and intact cut stems with C2H4 concentrations as high as 10 µl·l–1 did not affect bud opening or longevity or the development of leaf yellowing. Therefore, treatment with anti-C2H4 compounds, such as silver thiosulfate (STS) and 1-methylcyclopropene (1-MCP), did not improve the quality of the flowers. Data thus indicate that freshly harvested Stargazer were not sensitive to C2H4. Sensitivity of Stargazer to C2H4, however, increased dramatically following cold storage, as exposure of cold-stored stems to C2H4 concentrations as low as 0.3 µl·l–1 significantly affected bud opening. The development of leaf yellowing on cold-stored stems was not affected by the exogenous C2H4. Pretreating cold-stored stems with 1-MCP significantly reduced blasting of small buds that failed to develop due to carbohydrate depletion and reduced the percentage of buds that did not fully open. Concurrently, 1-MCP did not affect the quality of the leaves. These data indicate that sensitivity of cut lilies to C2H4 differs following cold storage and that 1-MCP is a more suitable anti-C2H4 compound than STS. Furthermore, studies on endogenous C2H4 production revealed that, while C2H4 was not detected in freshly harvested buds and leaves, it was produced by both following cold storage. The latter produced C2H4 at a higher rate than the former. Results of this study clearly indicate that there are two situations in which lilies will benefit from pretreatment with an anti-C2H4 compound (1) when cut stems contain buds that are marginally small for opening and (2) when cut stems will be cold stored before marketing. 相似文献
988.
The distributions of the times to turbidity for wells inoculated with single cells of Listeria innocua were determined in different environmental conditions (pH 4.5 to 7 and with 0.5% to 8% of NaCl at 30 degrees C). It was established by statistical analysis that the main source of the variability of the detection times, T, is the variability of individual lag times. A linear relation dev(T) approximately T was observed between the detection times and their standard deviation. At slow growth, other sources of variability became increasingly significant. 相似文献
989.
Bakirtzis G Jamieson S Aasen T Bryson S Forrow S Tetley L Finbow M Greenhalgh D Hodgins M 《Cell communication & adhesion》2003,10(4-6):359-364
To elucidate the mode of action of dominant mutant connexins in causing inherited skin diseases, transgenic mice were produced that express the true Vohwinkel syndrome-associated mutant Cx26 (D66H), from a keratin 10 promoter, specifically in the suprabasal epidermal keratinocytes. Following birth, the transgenic mice developed keratoderma similar to that of human carriers of Cx26 (D66H). Expression of the transgene resulted in a loss of Cx26 and Cx30 at intercellular junctions of epidermal keratinocytes and accumulation of these connexins in the cytoplasm. Injection of primary mouse keratinocytes with Lucifer Yellow showed no difference in terms of dye spreading between transgenic and non transgenic keratinocytes in vitro. Expression of the mutant Cx26 (D66H) did not interfere with the formation of the epidermal water barrier during late embryonic development. Attempts to produce transgenic mice expressing the wild type form of Cx26 from the K10 promoter failed to produce viable animals although transgenic embryos were recovered at days 9 and 12 of gestation, suggesting that the transgene might be embryonic lethal. 相似文献
990.