全文获取类型
收费全文 | 18506篇 |
免费 | 917篇 |
国内免费 | 27篇 |
专业分类
19450篇 |
出版年
2024年 | 31篇 |
2023年 | 164篇 |
2022年 | 375篇 |
2021年 | 703篇 |
2020年 | 414篇 |
2019年 | 441篇 |
2018年 | 643篇 |
2017年 | 603篇 |
2016年 | 786篇 |
2015年 | 890篇 |
2014年 | 1131篇 |
2013年 | 1550篇 |
2012年 | 1630篇 |
2011年 | 1435篇 |
2010年 | 832篇 |
2009年 | 730篇 |
2008年 | 842篇 |
2007年 | 816篇 |
2006年 | 681篇 |
2005年 | 618篇 |
2004年 | 501篇 |
2003年 | 415篇 |
2002年 | 372篇 |
2001年 | 308篇 |
2000年 | 279篇 |
1999年 | 255篇 |
1998年 | 102篇 |
1997年 | 76篇 |
1996年 | 82篇 |
1995年 | 78篇 |
1994年 | 57篇 |
1993年 | 56篇 |
1992年 | 152篇 |
1991年 | 143篇 |
1990年 | 113篇 |
1989年 | 85篇 |
1988年 | 127篇 |
1987年 | 98篇 |
1986年 | 80篇 |
1985年 | 81篇 |
1984年 | 81篇 |
1983年 | 46篇 |
1982年 | 40篇 |
1981年 | 47篇 |
1980年 | 43篇 |
1979年 | 57篇 |
1978年 | 35篇 |
1977年 | 43篇 |
1976年 | 32篇 |
1974年 | 29篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
991.
Jyoti Kushwah Puneet Kumar Veena Garg Vishal Singh Somvanshi 《Indian journal of microbiology》2017,57(1):125-128
Photorhabdus is an insect-pathogenic Gram negative enterobacterium found in the gut of Heterorhabditis nematodes. Photorhabdus is highly virulent to insects, and can kill insects rapidly upon injection at very low concentrations of one to few cells. We characterized the virulence of Photorhabdus symbionts isolated from the Heterorhabditis nematodes collected from various parts of India by injecting different concentrations of bacterial cells into fourth instar larval stage of insect Galleria mellonella. Photorhabdus luminescens ssp. akhurstii strain IARI-SGMG3 from Meghalaya was identified as the most virulent of all the tested strains on the basis of LT50 and LC50 values. This study forms a basis for further investigations on the genetic basis of virulence in Photorhabdus bacteria. 相似文献
992.
993.
Neeraj J Agrawal Bernhard Helk Sandeep Kumar Neil Mody Hasige A. Sathish Hardeep S. Samra 《MABS-AUSTIN》2016,8(1):43-48
Highly concentrated antibody solutions often exhibit high viscosities, which present a number of challenges for antibody-drug development, manufacturing and administration. The antibody sequence is a key determinant for high viscosity of highly concentrated solutions; therefore, a sequence- or structure-based tool that can identify highly viscous antibodies from their sequence would be effective in ensuring that only antibodies with low viscosity progress to the development phase. Here, we present a spatial charge map (SCM) tool that can accurately identify highly viscous antibodies from their sequence alone (using homology modeling to determine the 3-dimensional structures). The SCM tool has been extensively validated at 3 different organizations, and has proved successful in correctly identifying highly viscous antibodies. As a quantitative tool, SCM is amenable to high-throughput automated analysis, and can be effectively implemented during the antibody screening or engineering phase for the selection of low-viscosity antibodies. 相似文献
994.
Udaykumar Kage Arun Kumar Dhananjay Dhokane Shailesh Karre 《Critical reviews in biotechnology》2016,36(5):917-930
A tremendous decline in cultivable land and resources and a huge increase in food demand calls for immediate attention to crop improvement. Though molecular plant breeding serves as a viable solution and is considered as “foundation for twenty-first century crop improvement”, a major stumbling block for crop improvement is the availability of a limited functional gene pool for cereal crops. Advancement in the next generation sequencing (NGS) technologies integrated with tools like metabolomics, proteomics and association mapping studies have facilitated the identification of candidate genes, their allelic variants and opened new avenues to accelerate crop improvement through development and use of functional molecular markers (FMMs). The FMMs are developed from the sequence polymorphisms present within functional gene(s) which are associated with phenotypic trait variations. Since FMMs obviate the problems associated with random DNA markers, these are considered as “the holy grail” of plant breeders who employ targeted marker assisted selections (MAS) for crop improvement. This review article attempts to consider the current resources and novel methods such as metabolomics, proteomics and association studies for the identification of candidate genes and their validation through virus-induced gene silencing (VIGS) for the development of FMMs. A number of examples where the FMMs have been developed and used for the improvement of cereal crops for agronomic, food quality, disease resistance and abiotic stress tolerance traits have been considered. 相似文献
995.
The ability to feed an expanding world population poses one of the greatest challenges to mankind in the future. Accompanying
the increased demand for food by the expected nine billion inhabitants of Earth in 2050 will be a continual decrease in arable
land area, together with a decline in crop yield due to a variety of stresses. For these formidable challenges to be met,
future crops should not only by high-yielding, but also stress-tolerant and disease-resistant. In this review, we highlight
the importance of genetic engineering as an indispensable tool to generate just such future crops. We briefly discuss strategies
and available tools for biotechnological crop improvement and identify selected examples of candidate genes that may be manipulated
so that current biological maxima in yield may be surpassed by comfortable margins. Future prospects and the necessity for
basic research aimed at identifying novel target genes are also discussed. 相似文献
996.
Anantha R. Nookala Junhao Li Anusha Ande Lei Wang Naveen K. Vaidya Weihua Li Santosh Kumar Anil Kumar 《PloS one》2016,11(1)
Cytochrome P450 3A4 (CYP3A4) is the major drug metabolic enzyme, and is involved in the metabolism of antiretroviral drugs, especially protease inhibitors (PIs). This study was undertaken to examine the effect of methamphetamine on the binding and metabolism of PIs with CYP3A4. We showed that methamphetamine exhibits a type I spectral change upon binding to CYP3A4 with δAmax and KD of 0.016±0.001 and 204±18 μM, respectively. Methamphetamine-CYP3A4 docking showed that methamphetamine binds to the heme of CYP3A4 in two modes, both leading to N-demethylation. We then studied the effect of methamphetamine binding on PIs with CYP3A4. Our results showed that methamphetamine alters spectral binding of nelfinavir but not the other type I PIs (lopinavir, atazanavir, tipranavir). The change in spectral binding for nelfinavir was observed at both δAmax (0.004±0.0003 vs. 0.0068±0.0001) and KD (1.42±0.36 vs.2.93±0.08 μM) levels. We further tested effect of methamphetamine on binding of 2 type II PIs; ritonavir and indinavir. Our results showed that methamphetamine alters the ritonavir binding to CYP3A4 by decreasing both the δAmax (0.0038±0.0003 vs. 0.0055±0.0003) and KD (0.043±0.0001 vs. 0.065±0.001 nM), while indinavir showed only reduced KD in presence of methamphetamine (0.086±0.01 vs. 0.174±0.03 nM). Furthermore, LC-MS/MS studies in high CYP3A4 human liver microsomes showed a decrease in the formation of hydroxy ritonavir in the presence of methamphetamine. Finally, CYP3A4 docking with lopinavir and ritonavir in the absence and presence of methamphetamine showed that methamphetamine alters the docking of ritonavir, which is consistent with the results obtained from spectral binding and metabolism studies. Overall, our results demonstrated differential effects of methamphetamine on the binding and metabolism of PIs with CYP3A4. These findings have clinical implication in terms of drug dose adjustment of antiretroviral medication, especially with ritonavir-boosted antiretroviral therapy, in HIV-1-infected individuals who abuse methamphetamine. 相似文献
997.
Glucose deprivation induced upregulation of phosphoenolpyruvate carboxykinase modulates virulence in Leishmania donovani 下载免费PDF全文
Savita Saini Ayan Kumar Ghosh Ruby Singh Sushmita Das Kumar Abhishek Ajay Kumar Sudha Verma Abhishek Mandal Abul Hasan Sardar Bidyut Purkait Ashish Kumar Kislay Kumar Sinha Pradeep Das 《Molecular microbiology》2016,102(6):1020-1042
Various physiological stimuli trigger the conversion of noninfective Leishmania donovani promastigotes to the infective form. Here, we present the first evidence of the effect of glucose starvation, on virulence and survival of these parasites. Glucose starvation resulted in a decrease in metabolically active parasites and their proliferation. However, this was reversed by supplementation of gluconeogenic amino acids. Glucose starvation induced metacyclogenesis and enhanced virulence through protein kinase A regulatory subunit (LdPKAR1) mediated autophagy. Glucose starvation driven oxidative stress upregulated the antioxidant machinery, culminating in increased infectivity and greater parasitic load in primary macrophages. Interestingly, phosphoenolpyruvate carboxykinase (LdPEPCK), a gluconeogenic enzyme, exhibited the highest activity under glucose starvation to regulate growth of L. donovani by alternatively utilising amino acids. Deletion of LdPEPCK (Δpepck) decreased virulent traits and parasitic load in primary macrophages but increased autophagosome formation in the mutant parasites. Furthermore, Δpepck parasites failed to activate the Pentose Phosphate Pathway shunt, abrogating NADPH/NADP+ homoeostasis, conferring increased susceptibility towards oxidants following glucose starvation. In conclusion, this study showed that L. donovani undertakes metabolic rearrangements via gluconeogenesis under glucose starvation for acquiring virulence and its survival in the hostile environment. 相似文献
998.
Allergic diseases affect more than 25% of the world population and result from a complex interplay between genetic and environmental factors. Recent evidence has shown that BDNF (Brain Derived Neurotrophic Factor) could serve as an important marker of allergic disease. Increased levels of BDNF in blood, bronchoalveolar lavage fluid and nasal lavage fluid positively correlate with disease activity and severity in patients with allergic rhinitis (AR), asthma and atopic eczema. However, reports on the association between genetic variation in BDNF and allergic disease have been controversial. This study therefore aims to clarify the relationship between single nucleotide polymorphisms (SNPs) in BDNF and a genetic predisposition to AR and asthma in an ethnic Chinese population of Singapore. Volunteers with a self-reported history of asthma (718 subjects) or a history of AR as determined by a researcher-administered questionnaire (795 subjects) were used in this study, alongside controls with no personal or family history of allergy (717 subjects). The association results identified a significant association for the tagSNP rs10767664 with a significant PDominant = 0.0007 and OR = 1.3 for AR and PDominant = 0.0005 and OR = 1.3 for asthma (using a dominant model of association). The haplotype based analysis also identified a significant association further confirming the single SNP association. The SNP rs10767664 is strongly linked (r2 = 0.95) to the functional polymorphism rs6265 (Val66Met), which has previously been reported to be associated to allergic phenotypes and also shown to affect BDNF expression. BDNF is a therefore a key molecular player in allergy. Further studies on polymorphisms within BDNF may shed light on its role in the pathogenesis of allergic diseases and potentially serve as biomarkers for allergic disease. 相似文献
999.
1000.
Chitobiase (CHB) is an important enzyme for the production of N-acetyl-d-glucosamine from the chitin biopolymer in the series of chitinolytic enzymes. Majority of over-expressed CHB (58 %) in E. coli expression system led to formation of inclusion bodies. The production and soluble yield of active CHB was enhanced by co-expression with GroEL/ES chaperonin, optimizing culture conditions and solubilization followed by refolding of remaining inactive chitobiase present in the form of inclusion bodies. The growth of recombinant E. coli produced 42 % CHB in soluble form and the rest (~58 %) as inclusion bodies. The percentage of active CHB was enhanced to 71 % by co-expression with GroEL/ES chaperonin system and optimizing culture conditions (37 °C, 200 rpm, IPTG—0.5 mM, l-arabinose—13.2 mM). Of the remaining inactive CHB present in inclusion bodies, 37 % could be recovered in active form using pulsatile dilution method involving denaturants (2 M urea, pH 12.5) and protein refolding studies (1.0 M l-arginine, 5 % glycerol). Using combinatorial approach, 80 % of the total CHB expressed, could be recovered from cells grown in one litre of LB medium is a step forward in replacing hazardous chemical technology by biotechnological process for the production of NAG from chitinous waste. 相似文献