全文获取类型
收费全文 | 1377篇 |
免费 | 119篇 |
专业分类
1496篇 |
出版年
2023年 | 9篇 |
2022年 | 18篇 |
2021年 | 30篇 |
2020年 | 12篇 |
2019年 | 28篇 |
2018年 | 26篇 |
2017年 | 38篇 |
2016年 | 35篇 |
2015年 | 57篇 |
2014年 | 67篇 |
2013年 | 82篇 |
2012年 | 104篇 |
2011年 | 79篇 |
2010年 | 51篇 |
2009年 | 41篇 |
2008年 | 59篇 |
2007年 | 64篇 |
2006年 | 60篇 |
2005年 | 55篇 |
2004年 | 69篇 |
2003年 | 41篇 |
2002年 | 49篇 |
2001年 | 47篇 |
2000年 | 31篇 |
1999年 | 34篇 |
1998年 | 10篇 |
1997年 | 13篇 |
1996年 | 11篇 |
1995年 | 14篇 |
1994年 | 14篇 |
1993年 | 8篇 |
1992年 | 26篇 |
1991年 | 14篇 |
1990年 | 16篇 |
1989年 | 13篇 |
1988年 | 11篇 |
1987年 | 12篇 |
1986年 | 17篇 |
1985年 | 15篇 |
1984年 | 15篇 |
1983年 | 11篇 |
1982年 | 11篇 |
1975年 | 7篇 |
1974年 | 5篇 |
1973年 | 11篇 |
1972年 | 6篇 |
1971年 | 7篇 |
1970年 | 5篇 |
1969年 | 7篇 |
1968年 | 6篇 |
排序方式: 共有1496条查询结果,搜索用时 15 毫秒
91.
Héctor Flores-Villase?or Adrian Canizalez-Román Magda Reyes-Lopez Kamram Nazmi Mireya de la Garza Jorge Zazueta-Beltrán Nidia León-Sicairos Jan G. M. Bolscher 《Biometals》2010,23(3):569-578
Increased prevalence of antibiotic-resistant bacteria has become a major threat to the health sector worldwide due to their
virulence, limited therapeutic options and distribution in both hospital and community settings. Discovery and development
of new agents to combat antibiotic-resistant bacteria is thus needed. This study therefore aimed to evaluate the ability of
bovine lactoferrin (LF), peptides from two antimicrobial domains lactoferricin B (LFcin17-30) and lactoferrampin (LFampin265-284)
and a chimeric construct (LFchimera) containing both peptides, as potential bactericidal agents against clinical isolates
of antibiotic-resistant Staphylococcus aureus and Escherichia coli. Results in kinetics of growth show that LF chimera and peptides inhibited the growth of both bacterial species. By confocal
microscopy and flow cytometry it was observed that LF and FITC-labeled peptides are able to interact with these bacteria and
cause membrane permeabilization, as monitored by propidium iodide staining, these effects were decreased by preincubation
with lipopolysaccharide in E. coli. By electron microscopy, a clear cellular damage was observed in bacteria after treatments with LFchimera and peptides, suggesting
that interaction and membrane disruption are probably involved as a mechanism of action. In conclusion, results show that
LFchimera, LF and peptides have potential as bactericidal agents in the antibiotic-resistant strains of S.
aureus and E. coli and also the work strongly suggest that LFcin17-30 and LFampin265-284 acts synergistically with antibiotics against multidrug
resistant EPEC and MRSA in vitro. 相似文献
92.
Rojas NL Ortiz GE Baruque DJ Cavalitto SF Ghiringhelli PD 《Journal of industrial microbiology & biotechnology》2011,38(9):1437-1447
The pg1 gene from the filamentous fungus Aspergillus kawachii, which codifies for an acid polygalacturonase, was cloned into the pYES2 expression vector, giving rise to the pYES2:pg1ΔI construct. Engineered Saccharomyces cerevisiae, transformed with pYES2:pg1ΔI construct, both expressed and exported an active polygalacturonase with a MW of ~60 kDa and an isoelectric point of 3.7, similar to those reported for the wild-type enzyme. The recombinant enzyme has the ability to hydrolyze polygalacturonic acid at pH 2.5. Heterologous PG1 production was studied under controlled conditions in batch and fed-batch systems. A simultaneous addition of glucose and galactose was found to be the most suitable feeding strategy assayed, resulting in a final PG1 production of 50 U/ml. The production process proposed in this study could be applied for the industrial production of a novel and useful polygalacturonase. 相似文献
93.
Ajit G. Thomas Cliona M. O’Driscoll Joseph Bressler Walter E. Kaufmann Camilo J. Rojas Barbara S. Slusher 《Biochemical and biophysical research communications》2014
Glutaminase plays a critical role in the generation of glutamate, a key excitatory neurotransmitter in the CNS. Excess glutamate release from activated macrophages and microglia correlates with upregulated glutaminase suggesting a pathogenic role for glutaminase. Both glutaminase siRNA and small molecule inhibitors have been shown to decrease excess glutamate and provide neuroprotection in multiple models of disease, including HIV-associated dementia (HAD), multiple sclerosis and ischemia. Consequently, inhibition of glutaminase could be of interest for treatment of these diseases. Bis-2-(5-phenylacetimido-1,2,4-thiadiazol-2-yl)ethyl sulfide (BPTES) and 6-diazo-5-oxo-l-norleucine (DON), two most commonly used glutaminase inhibitors, are either poorly soluble or non-specific. Recently, several new BPTES analogs with improved physicochemical properties were reported. To evaluate these new inhibitors, we established a cell-based microglial activation assay measuring glutamate release. Microglia-mediated glutamate levels were significantly augmented by tumor necrosis factor (TNF)-α, phorbol 12-myristate 13-acetate (PMA) and Toll-like receptor (TLR) ligands coincident with increased glutaminase activity. While several potent glutaminase inhibitors abrogated the increase in glutamate, a structurally related analog devoid of glutaminase activity was unable to block the increase. In the absence of glutamine, glutamate levels were significantly attenuated. These data suggest that the in vitro microglia assay may be a useful tool in developing glutaminase inhibitors of therapeutic interest. 相似文献
94.
Hamel B Monaghan-Benson E Rojas RJ Temple BR Marston DJ Burridge K Sondek J 《The Journal of biological chemistry》2011,286(14):12141-12148
SmgGDS is an atypical guanine nucleotide exchange factor (GEF) that promotes both cell proliferation and migration and is up-regulated in several types of cancer. SmgGDS has been previously shown to activate a wide variety of small GTPases, including the Ras family members Rap1a, Rap1b, and K-Ras, as well as the Rho family members Cdc42, Rac1, Rac2, RhoA, and RhoB. In contrast, here we show that SmgGDS exclusively activates RhoA and RhoC among a large panel of purified GTPases. Consistent with the well known properties of GEFs, this activation is catalytic, and SmgGDS preferentially binds to nucleotide-depleted RhoA relative to either GDP- or GTPγS-bound forms. However, mutational analyses indicate that SmgGDS utilizes a distinct exchange mechanism compared with canonical GEFs and in contrast to known GEFs requires RhoA to retain a polybasic region for activation. A homology model of SmgGDS highlights an electronegative surface patch and a highly conserved binding groove. Mutation of either area ablates the ability of SmgGDS to activate RhoA. Finally, the in vitro specificity of SmgGDS for RhoA and RhoC is retained in cells. Together, these results indicate that SmgGDS is a bona fide GEF that specifically activates RhoA and RhoC through a unique mechanism not used by other Rho family exchange factors. 相似文献
95.
A complex formation between hemin and a congruous oligonucleotide not only greatly enhances the former’s peroxidative activity
but also results in a biocatalyst (DNAzyme) with a novel specificity. Herein substrate, regio-, enantiomeric, and diastereomeric
selectivities of heme, the DNAzyme, and the enzyme horseradish peroxidase are comparatively examined. 相似文献
96.
Leung SM Rojas R Maples C Flynn C Ruiz WG Jou TS Apodaca G 《Molecular biology of the cell》1999,10(12):4369-4384
Efficient postendocytic membrane traffic in polarized epithelial cells is thought to be regulated in part by the actin cytoskeleton. RhoA modulates assemblies of actin in the cell, and it has been shown to regulate pinocytosis and phagocytosis; however, its effects on postendocytic traffic are largely unexplored. To this end, we expressed wild-type RhoA (RhoAWT), dominant active RhoA (RhoAV14), and dominant inactive RhoA (RhoAN19) in Madin-Darby canine kidney (MDCK) cells expressing the polymeric immunoglobulin receptor. RhoAV14 expression stimulated the rate of apical and basolateral endocytosis, whereas RhoAN19 expression decreased the rate from both membrane domains. Polarized basolateral recycling of transferrin was disrupted in RhoAV14-expressing cells as a result of increased ligand release at the apical pole of the cell. Degradation of basolaterally internalized epidermal growth factor was slowed in RhoAV14-expressing cells. Although apical recycling of immunoglobulin A (IgA) was largely unaffected in cells expressing RhoAV14, transcytosis of basolaterally internalized IgA was severely impaired. Morphological and biochemical analyses demonstrated that a large proportion of IgA internalized from the basolateral pole of RhoAV14-expressing cells remained within basolateral early endosomes and was slow to exit these compartments. RhoAN19 and RhoAWT expression had little effect on these postendocytic pathways. These results indicate that in polarized MDCK cells activated RhoA may modulate endocytosis from both membrane domains and postendocytic traffic at the basolateral pole of the cell. 相似文献
97.
Tatiana L. Hernández-Palma Luis Alberto Rueda-Solano Janne K. Valkonen Bibiana Rojas 《Journal of evolutionary biology》2023,36(7):1040-1049
Deimatic displays, where sudden changes in prey appearance elicit aversive predator reactions, have been suggested to occur in many taxa. These (often only putative) displays frequently involve different components that may also serve antipredator functions via other mechanisms (e.g., mimicry, warning signalling, body inflation). The Colombian four-eyed frog, Pleurodema brachyops, has been suggested to gain protection against predation through putative deimatic displays where they inflate and elevate the posterior part of their body revealing eye-like colour markings. We exposed stationary artificial frogs to wild predators to test whether the two components (eyespot/colour markings, defensive posture) of their putative deimatic display, and their combination, provide protection from predation without the sudden change in appearance. We did not detect any obvious additive effect of defensive posture and eyespots/colour markings on predation risk, but found a marginally significant trend for model frogs in the resting posture to be less attacked when displaying eyespots/colour markings than when they were not, suggesting that the presence of colour markings/eyespots may provide some protection on its own. Additionally, we found that models in a resting posture were overall more frequently attacked on the head than models in a defensive posture, indicating that a defensive posture alone could help redirect predator attacks to non-vital parts of the body. The trends found in our study suggest that the different components of P. brachyops' coloration may serve different functions during a deimatic display, but further research is needed to elucidate the role of each component when accompanied by sudden prey movement. 相似文献
98.
Reproductive biology of Philodryas patagoniensis (Snakes: Dipsadidae) in south Brazil: Female reproductive cycle 下载免费PDF全文
Luiza Loebens Claudio Augusto Rojas Selma Maria Almeida‐Santos Sonia Zanini Cechin 《Acta zoologica》2018,99(2):105-114
In this study, we describe the female reproductive cycle of Philodryas patagoniensis in south Brazil, which was described through morpho‐anatomical and histological analyses. The peak of secondary vitellogenesis occurred during winter–spring (July–December), ovulation in spring (October–December), mating and fertilization in spring–summer (October–February), oviposition in spring–autumn (October–May) and births from late spring to autumn (December–July). The diameter of vitellogenic follicles/eggs was larger in winter–spring than in other seasons. The diameter of the shell glands was also larger in winter–spring. In spite of the clear reproductive peak, gonads only showed reduced activity in the autumn. Therefore, at the individual level, females have a discontinuous cyclical reproduction; in the populational level, the reproductive cycle is seasonal semisynchronous. We support the hypothesis that P. patagoniensis have the ability to produce multiple clutches with long‐term stored sperm. Sexual dimorphism in body size was evident, and females are significantly larger and heavier than males. Larger females were able to produce follicles and eggs in larger amount and size. The maternal body size was positively related to the reproductive effort and fecundity. To conclude, we deliberated about the proximal and distal causes that influence the reproductive traits and patterns of P. patagoniensis. 相似文献
99.
Simian virus 40 large T-antigen G-quadruplex DNA helicase inhibition by G-quadruplex DNA-interactive agents 总被引:1,自引:0,他引:1
On the basis of growing evidence for G-quadruplex DNA structures in genomic DNA and the presumed need to resolve these structures for DNA replication, the G-quadruplex DNA unwinding ability of a prototypical replicative helicase, SV40 large T-antigen (T-ag), was investigated. Here, we demonstrate that this G-quadruplex helicase activity is robust and comparable to the duplex helicase activity of T-ag. Analysis of the SV40 genome demonstrates the presence of sequences that may form intramolecular G-quadruplexes, which are the presumed natural substrates for the G-quadruplex helicase activity of T-ag. A number of G-quadruplex-interactive agents as well as new perylene diimide (PDI) derivatives have been investigated as inhibitors of both the G-quadruplex and the duplex DNA helicase activities of T-ag. A unique subset of these G-quadruplex-interactive agents inhibits the G-quadruplex DNA unwinding activity of T-ag, relative to those reported to inhibit G-quadruplex DNA unwinding by RecQ-family helicases. We also find that certain PDIs are both potent and selective inhibitors of the G-quadruplex DNA helicase activity of T-ag. Surface plasmon resonance and fluorescence spectroscopic G-quadruplex DNA binding studies of these T-ag G-quadruplex helicase inhibitors have been carried out, demonstrating the importance of attributes in addition to binding affinity for G-quadruplex DNA that may be important for inhibition. The identification of potent and selective inhibitors of the G-quadruplex helicase activity of T-ag provides tools for probing the specific role of this activity in SV40 replication. 相似文献
100.