全文获取类型
收费全文 | 2266篇 |
免费 | 171篇 |
出版年
2024年 | 3篇 |
2023年 | 10篇 |
2022年 | 12篇 |
2021年 | 59篇 |
2020年 | 31篇 |
2019年 | 53篇 |
2018年 | 78篇 |
2017年 | 52篇 |
2016年 | 92篇 |
2015年 | 117篇 |
2014年 | 156篇 |
2013年 | 169篇 |
2012年 | 222篇 |
2011年 | 213篇 |
2010年 | 141篇 |
2009年 | 99篇 |
2008年 | 152篇 |
2007年 | 150篇 |
2006年 | 114篇 |
2005年 | 111篇 |
2004年 | 107篇 |
2003年 | 75篇 |
2002年 | 67篇 |
2001年 | 7篇 |
2000年 | 9篇 |
1999年 | 18篇 |
1998年 | 18篇 |
1997年 | 17篇 |
1996年 | 11篇 |
1995年 | 9篇 |
1994年 | 6篇 |
1993年 | 10篇 |
1992年 | 7篇 |
1991年 | 5篇 |
1990年 | 5篇 |
1989年 | 4篇 |
1988年 | 6篇 |
1987年 | 2篇 |
1985年 | 4篇 |
1983年 | 2篇 |
1981年 | 6篇 |
1980年 | 2篇 |
1977年 | 2篇 |
1972年 | 2篇 |
1969年 | 1篇 |
1909年 | 1篇 |
排序方式: 共有2437条查询结果,搜索用时 15 毫秒
131.
Antonio Bevilacqua Monica Mischitelli Valeria Pietropaolo Emanuela Ciuffreda Milena Sinigaglia Maria Rosaria Corbo 《PloS one》2015,10(10)
Alicyclobacillus acidoterrestris is the main cause of most spoilage problems in fruit juices and acidic products. Since soil borne species often contaminate fruit juices and do not need strict extreme requirements for survival, it is a great concern to investigate whether and how soil species could evolve from their ecological niches in microbial community to new environments as fruit juices. In this study, 23 isolates of thermo-acidophilic, spore-forming bacteria from soil were characterized by cultural and molecular methods. In addition, 2 strains isolated from a spoilage incident in pear juice were typed. Strains phenotyping showed that they could be grouped into 3 different clusters, and some isolates showed identical or quite similar patterns. Analyzing pH and temperature ranges for growth, the majority of strains were able to grow at values described for many species of Alicyclobacillus. Qualitative utilization of lysine, arginine and indole production from tryptophan revealed, for the first time, deamination of lysine and decarboxylation of arginine. Resistance to 5% NaCl as well as the ability to hydrolyze starch and gelatin, nitrate reduction, catalase and oxidase activities confirmed literature evidences. Examining of 16S rRNA, showed that isolates were divided into three blocks represented by effectively soil species and strains that are moving from soil to other possible growing source characterized by parameters that could strongly influence bacterial survival. RAPD PCR technique evidenced a great variability in banding patterns and, although it was not possible to obtain genotypically well-distinguished groups, it was feasible to appreciate genetic similarity between some strains. In conclusion, the investigation of a microbial community entails a combination of metagenomic and classic culture-dependent approaches to expand our knowledge about Alicyclobacillus and to look for new subspecies. 相似文献
132.
Eira Valeria Barrón Edgar Roman-Bassaure Ana Laura Sánchez-Sandoval Ana María Espinosa Mariano Guardado-Estrada Ingrid Medina Eligia Juárez Ana Alfaro Miriam Bermúdez Rubén Zamora Carlos García-Ruiz Juan Carlos Gomora Susana Kofman E. Martha Pérez-Armendariz Jaime Berumen 《PloS one》2015,10(9)
The cyclin-dependent kinase inhibitor 3 (CDKN3) gene, involved in mitosis, is upregulated in cervical cancer (CC). We investigated CDKN3 mRNA as a survival biomarker and potential therapeutic target for CC. CDKN3 mRNA was measured in 134 CC and 25 controls by quantitative PCR. A 5-year survival study was conducted in 121 of these CC patients. Furthermore, CDKN3-specific siRNAs were used to investigate whether CDKN3 is involved in proliferation, migration, and invasion in CC-derived cell lines (SiHa, CaSki, HeLa). CDKN3 mRNA was on average 6.4-fold higher in tumors than in controls (p = 8 x 10−6, Mann-Whitney). A total of 68.2% of CC patients over expressing CDKN3 gene (fold change ≥ 17) died within two years of diagnosis, independent of the clinical stage and HPV type (Hazard Ratio = 5.0, 95% CI: 2.5–10, p = 3.3 x 10−6, Cox proportional-hazards regression). In contrast, only 19.2% of the patients with lower CDKN3 expression died in the same period. In vitro inactivation of CDKN3 decreased cell proliferation on average 67%, although it had no effect on cell migration and invasion. CDKN3 mRNA may be a good survival biomarker and potential therapeutic target in CC. 相似文献
133.
134.
Samuele Raccosta Mauro Manno Donatella Bulone Daniela Giacomazza Valeria Militello Vincenzo Martorana Pier Luigi San Biagio 《European biophysics journal : EBJ》2010,39(6):1007-1017
The formation of protein aggregates is important in many fields of life science and technology. The morphological and mechanical
properties of protein solutions depend upon the molecular conformation and thermodynamic and environmental conditions. Non-native
or unfolded proteins may be kinetically trapped into irreversible aggregates and undergo precipitation or gelation. Here,
we study the thermal aggregation of lysozyme in neutral solutions. We characterise the irreversible unfolding of lysozyme
by differential scanning calorimetry. The structural properties of aggregates and their mechanisms of formation with the eventual
gelation are studied at high temperature by spectroscopic, rheological and scattering techniques. The experiments show that
irreversible micron-sized aggregates are organised into larger clusters according to a classical mechanism of diffusion and
coagulation, which leads to a percolative transition at high concentrations. At a smaller length scale, optical and atomic
force microscopy images reveal the existence of compact aggregates, which are the origin of the aggregation irreversibility. 相似文献
135.
Franciscanas Pontoporia blainvillei are small dolphins endemic to Brazil, Uruguay and Argentina. During routine pathologic examinations, chronic pneumonia associated with cholesterol deposits was found in 16.7% of stranded and incidentally bycaught franciscanas (n = 60), and was more frequent in dolphins from the Brazilian state of S?o Paulo (Franciscana Management Area II) and frequently accompanied by splenic lymphoid hyperplasia. It is unclear why these otherwise uncommon lesions were relatively frequent in the studied dolphins, and further research is advised. 相似文献
136.
Valeria Sabaj Mario Galindo Daniela Silva Lea Sandoval Juan C. Rodríguez 《Molecular biology reports》2010,37(6):2927-2933
Toxoplasma gondii is one of the most successful protozoan parasites given its ability to manipulate the immune system and establish a chronic infection. It is a parasite with a significant impact on human health, mainly in immunocompromised patients. In Europe and North America, only a few clonal genotypes (I, II and III) seem to be responsible for the vast majority of Toxoplasma infections. Surface antigen 2 gene (SAG2) has been extensively used for genotyping T. gondii isolates. The analysis of this locus reveals that in Northern hemisphere, human disease causing isolates are mainly type II, whereas T. gondii isolated from different animals are both type II and III. Since the immune response depends on parasite genotype, it seems relevant to characterize parasites producing human toxoplasmosis in different geographical areas. The growing information about the prevalent T. gondii genotypes in South America mostly refers to domestic animals. This is the first report of genetic characterization of T. gondii isolates from clinical samples in Chile, South America. All the samples analyzed corresponded to SAG2 type I isolates, and they differ from classic SAG2 type I by genetic polymorphisms. This study contributes to the scarce available information on T. gondii at South America, and reinforces an emerging concept suggesting that SAG2 type I, rather than II, parasites are a frequent cause of clinical toxoplasmosis in this continent. 相似文献
137.
Valeria Castelletto Ian W. Hamley Teck Lim Matias B. De Tullio Eduardo M. Castaño 《Journal of peptide science》2010,16(9):443-450
We study the complex formation of a peptide βAβAKLVFF, previously developed by our group, with Aβ(1–42) in aqueous solution. Circular dichroism spectroscopy is used to probe the interactions between βAβAKLVFF and Aβ(1–42), and to study the secondary structure of the species in solution. Thioflavin T fluorescence spectroscopy shows that the population of fibers is higher in βAβAKLVFF/Aβ(1–42) mixtures compared to pure Aβ(1–42) solutions. TEM and cryo‐TEM demonstrate that co‐incubation of βAβAKLVFF with Aβ(1–42) causes the formation of extended dense networks of branched fibrils, very different from the straight fibrils observed for Aβ(1–42) alone. Neurotoxicity assays show that although βAβAKLVFF alters the fibrillization of Aβ(1–42), it does not decrease the neurotoxicity, which suggests that toxic oligomeric Aβ(1–42) species are still present in the βAβAKLVFF/Aβ(1–42) mixtures. Our results show that our designed peptide binds to Aβ(1–42) and changes the amyloid fibril morphology. This is shown to not necessarily translate into reduced toxicity. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
138.
Maria Luisa Balestrieri Alfonso Giovane Lara Milone Francesca Felice Carmela Fiorito Valeria Crudele Annaclaudia Esposito Raffaele Rossiello Pellegrino Biagio Minucci Bartolomeo Farzati Luigi Servillo Claudio Napoli 《Journal of biochemical and molecular toxicology》2010,24(6):351-360
Exposure of human endothelial progenitor cells (EPCs) to tumor necrosis factor‐α (TNF‐α) reduced their number and biological activity. Yet, signal transduction events linked to TNF‐α action are still poorly understood. To address this issue, we examined the possible effect of fasudil and Y27632, two inhibitors of Rho kinase pathway, which is involved in endothelial dysfunction, atherosclerosis, and in‐ flammation. Results demonstrated that incubation with fasudil starting from 50 μM but not Y27632 determined a dose‐dependent improvement of EPC number during exposure to TNF‐α (P < 0.05 vs. TNF‐α alone). Analysis of the signal transduction pathway activated by TNF‐α revealed that the increased expression of p‐p38 was not significantly altered by fasudil. Instead, fasudil blocked the TNF‐α induced phosphorylation of Erk1/2 (P < 0.05 vs. TNF‐α) as well as the inhibitor of Erk1/2‐specific phosphorylated form, i.e., PD98059 (P < 0.05 vs. TNF‐α). These results were confirmed by analysis of these kinases by confocal microscopy. Finally, 2D‐DIGE and MALDI‐TOF/TOF analysis of EPCs treated with fasudil revealed increased expression levels of an actin‐related protein and an adenylyl cyclase associated protein and decreased expression levels of proteins related to radical scavenger and nucleotide metabolism. These findings suggest that fasudil positively affects EPC number and that other major signals might take part to this complex pathway. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:351–360, 2010; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.20345 相似文献
139.
Brevis P Morales E Bravo JC Monasterio V Mánques B Zaror L Abaca P Toloza L 《Revista iberoamericana de micología》2010,27(4):183-185
BackgroundRhinosporidiosis is a chronic, granulomatous, and non-contagious infection, in which highly vascularized polyps (mainly present in the nasal cavity) appear. These polyps usually bleed easily.AimsTo present the case of a 14 year-old male suffering from an obstruction and injury of the right nostril due to a polypoid shaped-lesion with a raspberry-like appearance.MethodsA wide surgery resection of the base of the lesion was performed, as well as a standard histopathology procedure, including microscopic analysis with haematoxylin-eosin and Grocott staining.Results and conclusionsThe histopathology report indicated that the chronic inflammatory polyp was compatible with rhinosporidiosis. 相似文献
140.
Angela Chambery Valeria Severino Antimo Di Maro Antimo D’Aniello Menotti Ruvo Augusto Parente 《Amino acids》2010,38(4):1031-1041
Thyrotropin-releasing hormone (TRH) is involved in a wide range of biological responses. It has a central role in the endocrine
system and regulates several neurobiological activities. In the present study, a rapid, sensitive and selective liquid chromatography–mass
spectrometry method for the identification and quantification of TRH has been developed. The methodology takes advantage of
the specificity of the selected-ion monitoring acquisition mode with a limit of detection of 1 fmol. Furthermore, the MS/MS
fragmentation pattern of TRH has been investigated to develop a selected reaction monitoring (SRM) method that allows the
detection of a specific b2 product ion at m/z 249.1, corresponding to the N-terminus dipeptide pyroglutamic acid–histidine. The method has been tested on rat hypothalami to evaluate its suitability
for the detection within very complex biological samples. 相似文献