全文获取类型
收费全文 | 2975篇 |
免费 | 238篇 |
国内免费 | 1篇 |
出版年
2023年 | 20篇 |
2022年 | 37篇 |
2021年 | 74篇 |
2020年 | 31篇 |
2019年 | 42篇 |
2018年 | 75篇 |
2017年 | 52篇 |
2016年 | 91篇 |
2015年 | 157篇 |
2014年 | 154篇 |
2013年 | 228篇 |
2012年 | 229篇 |
2011年 | 250篇 |
2010年 | 165篇 |
2009年 | 147篇 |
2008年 | 182篇 |
2007年 | 171篇 |
2006年 | 148篇 |
2005年 | 149篇 |
2004年 | 149篇 |
2003年 | 135篇 |
2002年 | 110篇 |
2001年 | 17篇 |
2000年 | 19篇 |
1999年 | 24篇 |
1998年 | 27篇 |
1997年 | 19篇 |
1996年 | 27篇 |
1995年 | 21篇 |
1994年 | 15篇 |
1993年 | 21篇 |
1992年 | 15篇 |
1991年 | 11篇 |
1990年 | 11篇 |
1989年 | 8篇 |
1988年 | 7篇 |
1987年 | 12篇 |
1986年 | 8篇 |
1985年 | 10篇 |
1984年 | 12篇 |
1983年 | 14篇 |
1982年 | 15篇 |
1981年 | 11篇 |
1980年 | 14篇 |
1979年 | 14篇 |
1978年 | 9篇 |
1977年 | 6篇 |
1976年 | 11篇 |
1975年 | 7篇 |
1973年 | 5篇 |
排序方式: 共有3214条查询结果,搜索用时 31 毫秒
21.
22.
Gloria del Solar Gabriela Kramer Sara Ballester Manuel Espinosa 《Molecular & general genetics : MGG》1993,241(1-2):97-105
Deletion of a region of the promiscuous plasmid pLS1 encompassing the initiation signals for the synthesis of the plasmid lagging strand led to plasmid instability in Streptococcus pneumoniae and Bacillus subtilis. This defect could not be alleviated by increasing the number of copies (measured as double-stranded plasmid DNA) to levels similar to those of the wild-type plasmid pLS1. Our results indicate that in the vicinity of, or associated with the single-stranded origin region of pLS1 there is a plasmid component involved in its stable inheritance. Homology was found between the DNA gyrase binding site within the par region of plasmid pSC101 and the pLS1 specific recombination site RSR. 相似文献
23.
The phytogeography of the genus Allium in Siberia and Mongolia is described, based on the numerical classification of a matrix of 56 species and 769 Operational Geographic Uniis (OGUs). Two main diversity centers can be detected, the Altai-Tuva region and southeastern Siberia, which can be further subdivided into 4 subcenters: Altai Mts., Tuva Mts., southern Baikal and Dahuria. The first three subcenters. located in southern Siberia, are rich in endemic species, which are mostly bound to semi-arid environments such as montane steppes and alpine vegetation. These old, isolated mountain ranges constitute the main refugial centers for the Allium flora of Siberia and Mongolia. The Tuva subcenter, rich in endemics and poor in polyploid species, seems to be the most conservative area; the south Baikal region, much richer in polyploid species, appears as an important center af speciation. 相似文献
24.
The requirement of inorganic phosphate (Pi) for oxidative phosphorylation in eukaryotic cells is fulfilled through specific Pi transport systems. The mitochondrial proton/phosphate symporter (Pic) is a membrane-embedded protein which translocates Pi from the cytosol into the mitochondrial matrix. Pic is responsible for the very rapid transport of most of the Pi used in ATP synthesis. During the past five years there have been advances on several fronts. Genomic and cDNA clones for yeast, bovine, rat, and human Pic have been isolated and sequenced. Functional expression of yeast Pic in yeast strains deficient in Pi transport and expression inEscherichia coli of a chimera protein involving Pic and ATP synthase subunit have been accomplished. Pic, in contrast to other members of the family of transporters involved in energy metabolism, was demonstrated to have a presequence, which optimizes the import of the precursor protein into mitochondria. Six transmembrane segments appear to be a structural feature shared between Pic and other mitochondrial anion carriers, and recent-site directed mutagenesis studies implicate structure-functional relationships to bacteriorhodopsin. These recent advances on Pic will be assessed in light of a more global interpretation of transport mechanism across the inner mitochondrial membrane. 相似文献
25.
Presymptomatic direct detection of adenomatous polyposis coli (APC) gene mutations in familial adenomatous polyposis 总被引:5,自引:0,他引:5
The recent identification of the familial adenomatous polyposis (FAP) gene (designated as APC) enables conclusive genetic testing of at-risk family members for the specific mutation in families in which the germline gene mutation has been characterized. Presymptomatic molecular diagnosis of FAP was performed by direct direction of mutations in lymphocyte DNA in four families. Each of the families has a different mutation of the APC gene. Twenty-seven offspring of affected individuals (a priori risk of 50%) were tested. Ten of the 27 had already developed clinical features of FAP. Of the remaining seventeen, two had had a negative colon exam at an early age, and nine had never had colon exams (mean age, 12.1±3.1 SD years). Six children from this group (54%) were found to carry their affected parent's mutation. No change in the conventional FAP colon screening regimen is recommended for these children. In contrast, when direct tests indicate that an individual does not have the FAP mutation, we recommended that screening be decreased. Reduction of uncertainty for at-risk FAP family members is an important benefit of genetic testing. 相似文献
26.
Fluorescent detection-polymerase chain reaction (FD-PCR) assay on microwell plates as a screening test for salmonellas in foods 总被引:7,自引:1,他引:6
R.J. Cano S.R. Rasmussen Gloria Sánchez Fraga J.C. Palomares 《Journal of applied microbiology》1993,75(3):247-253
This study evaluates a polymerase chain reaction assay coupled with a fluorescent detection in microwell plates for salmonellas in food samples. Chelex 100-extracted cultures and bulk and processed food samples were used as templates for a PCR assay in microwell plates, with a primer pair that amplifies a 206 bp segment of IS 200 . The PCR products were then denatured by heat and transferred to CovaLink NH plates (Nunc) to which capture oligonucleotides were covalently bound. Hybridization was performed for 1 h at 55°C, the microwells were washed and an alkaline phosphatase-labelled probe, complementary of an internal sequence of the PCR product, was added. After stringent washes, 100 μl of 1 mmol 1-1 AttoPhosTM (JBL Scientific) was then added to the wells and the fluorescence measurement system (Millipore). The level of detection of the assay was as low as 1–10 cfu. A total of 172 food samples were tested, both by culture and FD-PCR. Of these 53 were culture positive and 119 culture negative. The sensitivity of the FD-PCR assay was 100% and the specificity was 90.1%. Positive and negative predictive values were 82.8 and 100%, respectively. Based on the results obtained in this study it appears that the FD-PCR. assay described here can be useful to screen a large number of food samples for contamination by salmonellas. 相似文献
27.
Matteo Zurlo Francesco Nicoli Davide Proietto Beatrice Dallan Cristina Zuccato Lucia Carmela Cosenza Jessica Gasparello Chiara Papi Elisabetta d'Aversa Monica Borgatti Chiara Scapoli Alessia Finotti Roberto Gambari 《Journal of cellular and molecular medicine》2023,27(3):353-364
Inhibitors of the mammalian target of rapamycin (mTOR) have been proposed to improve vaccine responses, especially in the elderly. Accordingly, testing mTOR inhibitors (such as Sirolimus) and other geroprotective drugs might be considered a key strategy to improve overall health resilience of aged populations. In this respect, Sirolimus (also known as rapamycin) is of great interest, in consideration of the fact that it is extensively used in routine therapy and in clinical studies for the treatment of several diseases. Recently, Sirolimus has been considered in laboratory and clinical studies aimed to find novel protocols for the therapy of hemoglobinopathies (e.g. β-Thalassemia). The objective of the present study was to analyse the activity of CD4+ and CD8+ T cells in β-Thalassemia patients treated with Sirolimus, taking advantages from the availability of cellular samples of the NCT03877809 clinical trial. The approach was to verify IFN-γ releases following stimulation of peripheral blood mononuclear cells (PBMCs) to stimulatory CEF and CEFTA peptide pools, stimulatory for CD4+ and CD8+ T cells, respectively. The main results of the present study are that treatment of β-Thalassemia patients with Sirolimus has a positive impact on the biological activity and number of memory CD4+ and CD8+ T cells releasing IFN-γ following stimulation with antigenic stimuli present in immunological memory. These data are to our knowledge novel and in our opinion of interest, in consideration of the fact that β-Thalassemia patients are considered prone to immune deficiency. 相似文献
28.
29.
Summary A buffer system consisting of 50 mM Tris-HCl-TRIZMA base plus 10 mM EDTA was used to rapidly dissolve gellan gels used for maintaining transformed carrot root cultures. The optimum conditions of pH 7.5 in the presence of 10 mM EDTA for dissolving gellan were first worked out on a model test system containing 0.4% gellan, 0.025% MgSO4·7H2O, and blue dye. The conditions were then tested on gellan gels (0.2% gellan plus nutrients) containing carrot roots. This gel dissolution system was rapid (18 to 20 min), did not require heating, and could also be efficiently performed at 4 °C. Furthermore, the buffer system used for gel dissolution is a standard one used for plant cell fractionation studies. 相似文献
30.
Diana Villegas-Coronado Jesús Adriana Soto-Guzman Juan Manuel Martínez-Soto Nayelli Guadalupe Teran-Saavedra Ana Maria Guzman-Partida Luz Vazquez-Moreno Ana Gloria Villalba-Villalba Amir Maldonado Irlanda Lagarda-Diaz 《化学与生物多样性》2023,20(7):e202300051
Acute monocytic leukemia is a type of myeloid leukemia that develops in monocytes. The current clinical therapies for leukemia are unsatisfactory due to their side effects and nonspecificity toward target cells. Some lectins display antitumor activity and may specifically recognize cancer cells by binding to carbohydrate structures on their surface. Therefore, this study evaluated the response of the human monocytic leukemia cell lines THP-1 to the Olneya tesota PF2 lectin. The induction of apoptosis and reactive oxygen species production in PF2-treated cells was evaluated by flow cytometry, and the lectin-THP-1 cell interaction and mitochondrial membrane potential were evaluated by confocal fluorescence microscopy. PF2 genotoxicity was evaluated by DNA fragmentation analysis via gel electrophoresis. The results showed that PF2 binds to THP-1 cells, triggers apoptosis and DNA degradation, changes the mitochondrial membrane potential, and increases reactive oxygen species levels in PF2-treated THP-1 cells. These results suggest the potential use of PF2 for developing alternative anticancer treatments with enhanced specificity. 相似文献