全文获取类型
收费全文 | 920篇 |
免费 | 65篇 |
专业分类
985篇 |
出版年
2022年 | 8篇 |
2021年 | 15篇 |
2020年 | 9篇 |
2019年 | 6篇 |
2018年 | 17篇 |
2017年 | 7篇 |
2016年 | 24篇 |
2015年 | 39篇 |
2014年 | 49篇 |
2013年 | 51篇 |
2012年 | 68篇 |
2011年 | 70篇 |
2010年 | 39篇 |
2009年 | 27篇 |
2008年 | 57篇 |
2007年 | 50篇 |
2006年 | 43篇 |
2005年 | 57篇 |
2004年 | 36篇 |
2003年 | 44篇 |
2002年 | 55篇 |
2001年 | 15篇 |
2000年 | 9篇 |
1999年 | 7篇 |
1998年 | 11篇 |
1997年 | 6篇 |
1996年 | 9篇 |
1995年 | 5篇 |
1994年 | 13篇 |
1993年 | 8篇 |
1992年 | 14篇 |
1991年 | 4篇 |
1990年 | 9篇 |
1989年 | 4篇 |
1988年 | 14篇 |
1987年 | 4篇 |
1986年 | 8篇 |
1985年 | 3篇 |
1984年 | 5篇 |
1983年 | 6篇 |
1981年 | 6篇 |
1980年 | 7篇 |
1979年 | 6篇 |
1978年 | 5篇 |
1977年 | 5篇 |
1976年 | 7篇 |
1974年 | 3篇 |
1971年 | 5篇 |
1970年 | 3篇 |
1954年 | 2篇 |
排序方式: 共有985条查询结果,搜索用时 0 毫秒
101.
Marklund M Landberg R Åman P Kamal-Eldin A 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2011,879(9-10):647-651
Alkylresorcinols (AR) are amphiphilic compounds present at high concentrations in the outer parts of wheat and rye kernels. Due to their specificity to whole grain and bran products of these cereals, AR and their metabolites have been proposed as biomarkers for intake of such foods. Two alkylresorcinol metabolites, 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA), have previously been quantified in human urine using two different methodologies: high-performance liquid chromatography coupled to a coulometric electrode array detector (HPLC-CEAD) and gas chromatography in combination with mass spectrometry (GC-MS). In this study, these two methodologies were compared by analysing 114 urine samples from free-living Swedish subjects consuming their habitual diet. Data were evaluated by graphical investigation of difference-plots and statistical inference of agreement was assessed by weighted Deming regression analysis. The median DHBA concentrations were 11 μM (GC-MS) and 13 μM (HPLC-CEAD), respectively. Both difference-plot and regression analysis showed a small but statistically significant additive bias, with HPLC-CEAD resulting in a slightly higher DHBA concentration than GC-MS. The median concentration of DHPPA was 18 μM for both methods. Examination of the difference-plot of DHPPA did not indicate any systematic difference between the methods, but regression analysis showed small but statistically significant constant and proportional biases. The conclusion was that the two methodologies are equally suitable for analysis of alkylresorcinol metabolites in human urine and that any small systematic differences observed are most likely of limited practical importance. 相似文献
102.
Guglielmetti S Karp M Mora D Tamagnini I Parini C 《Applied microbiology and biotechnology》2007,74(5):1053-1061
In this study, we performed molecular characterization and sequence analysis of three plasmids from the human intestinal isolate
Bifidobacterium longum biovar longum NAL8 and developed a novel vector screening system. Plasmids pNAL8H (10 kb) and pNAL8M (4.9 kb) show close sequence similarity
to and the same gene organization as the already characterized B. longum plasmids. The B. longum plasmid pNAC1 was identified as being most closely related to pNAL8L (3.5 kb). However, DNA sequence analysis suggested that
direct repeat-rich sites could have promoted several recombination events to diversify the two plasmid molecules. We verified
the likely rolling circle replication of plasmid pNAL8L and studied the phylogenetic relationship in all the Bifidobacterium plasmids fully sequenced to date based on in silico comparative sequence analysis of their replication proteins and iteron regions. Our transformation experiments confirmed
that the ColE1 replication origin from high-copy-number pUC vectors could interfere with the replication apparatus of Bifidobacterium plasmids and give rise to false positive clones. As a result, we developed a system suitable for avoiding possible interference
by other functional replication modules on the vector and for screening functional replicons from wild-type plasmids.
Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users only. 相似文献
103.
104.
We describe a supervised prediction method for diagnosis of acute myeloid leukemia (AML) from patient samples based on flow cytometry measurements. We use a data driven approach with machine learning methods to train a computational model that takes in flow cytometry measurements from a single patient and gives a confidence score of the patient being AML-positive. Our solution is based on an regularized logistic regression model that aggregates AML test statistics calculated from individual test tubes with different cell populations and fluorescent markers. The model construction is entirely data driven and no prior biological knowledge is used. The described solution scored a 100% classification accuracy in the DREAM6/FlowCAP2 Molecular Classification of Acute Myeloid Leukaemia Challenge against a golden standard consisting of 20 AML-positive and 160 healthy patients. Here we perform a more extensive validation of the prediction model performance and further improve and simplify our original method showing that statistically equal results can be obtained by using simple average marker intensities as features in the logistic regression model. In addition to the logistic regression based model, we also present other classification models and compare their performance quantitatively. The key benefit in our prediction method compared to other solutions with similar performance is that our model only uses a small fraction of the flow cytometry measurements making our solution highly economical. 相似文献
105.
Early succession of bacterial biofilms in paper machines 总被引:1,自引:0,他引:1
Marja Tiirola Tomi Lahtinen Matti Vuento Christian Oker-Blom 《Journal of industrial microbiology & biotechnology》2009,36(7):929-937
Formation of biofilms causes severe problems in paper machines, and hence financial costs. It would be preferable to prevent
attachment of the primary-colonizing bacteria than to control the growth of secondary communities, which are sheltered by
exopolysaccharide slime layers. We have therefore investigated the early succession of paper-machine biofilms by incubating
stainless-steel test coupons in the process water-flow lines in two paper machines operating in slightly alkaline conditions
in temperatures (45 and 49°C) supporting thermophilic microbes. Microbial succession was profiled using length heterogeneity
analysis of PCR-amplified 16S rRNA genes (LH-PCR) and linking the sequence data of the created 16S rRNA gene libraries to
the dominant LH-PCR peaks. Although the bacterial fingerprints obtained from the attached surface communities varied slightly
in different samples, the biomarker signals of the dominating primary-colonizing bacterial groups remained high over time
in each paper machine. Most of the 16S rRNA gene copies in the early biofilms were assigned to the genera Rhodobacter, Tepidimonas, and Cloacibacterium. The dominance of these sequence types decreased in the developing biofilms. Finally, as phylogenetically identical primary-colonizers
were detected in the two different paper mills, the machines evidently had similar environmental conditions for bacterial
growth and potentially a common source of contamination. 相似文献
106.
Denis V. Abramochkin Eugenia I. Alekseeva Matti Vornanen 《Comparative biochemistry and physiology. Toxicology & pharmacology : CBP》2013,158(3):181-186
KB-R7943 (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea) was developed as a specific inhibitor of the sarcolemmal sodium–calcium exchanger (NCX) with potential experimental and therapeutic use. However, KB-R7943 is shown to be a potent blocker of several ion currents including inward and delayed rectifier K+ currents of cardiomyocytes. To further characterize KB-R7943 as a blocker of the cardiac inward rectifiers we compared KB-R7943 sensitivity of the background inward rectifier (IK1) and the carbacholine-induced inward rectifier (IKACh) currents in mammalian (Rattus norvegicus; rat) and fish (Carassius carassius; crucian carp) cardiac myocytes. The basal IK1 of ventricular myocytes was blocked with apparent IC50-values of 4.6 × 10− 6 M and 3.5 × 10− 6 M for rat and fish, respectively. IKACh was almost an order of magnitude more sensitive to KB-R7943 than IK1 with IC50-values of 6.2 × 10− 7 M for rat and 2.5 × 10− 7 M for fish. The fish cardiac NCX current was half-maximally blocked at the concentration of 1.9–3 × 10− 6 M in both forward and reversed mode of operation. Thus, the sensitivity of three cardiac currents to KB-R7943 block increases in the order IK1 ~ INCX < IKACh. Therefore, the ability of KB-R7943 to block inward rectifier potassium currents, in particular IKACh, should be taken into account when interpreting the data with this inhibitor from in vivo and in vitro experiments in both mammalian and fish models. 相似文献
107.
Role of recycling endosomes and lysosomes in dynein-dependent entry of canine parvovirus 总被引:5,自引:5,他引:5 下载免费PDF全文
Suikkanen S Sääjärvi K Hirsimäki J Välilehto O Reunanen H Vihinen-Ranta M Vuento M 《Journal of virology》2002,76(9):4401-4411
Canine parvovirus (CPV) is a nonenveloped virus with a 5-kb single-stranded DNA genome. Lysosomotropic agents and low temperature are known to prevent CPV infection, indicating that the virus enters its host cells by endocytosis and requires an acidic intracellular compartment for penetration into the cytoplasm. After escape from the endocytotic vesicles, CPV is transported to the nucleus for replication. In the present study the intracellular entry pathway of the canine parvovirus in NLFK (Nordisk Laboratory feline kidney) cells was studied. After clustering in clathrin-coated pits and being taken up in coated vesicles, CPV colocalized with coendocytosed transferrin in endosomes resembling recycling endosomes. Later, CPV was found to enter, via late endosomes, a perinuclear vesicular compartment, where it colocalized with lysosomal markers. There was no indication of CPV entry into the trans-Golgi or the endoplasmic reticulum. Similar results were obtained both with full and with empty capsids. The data thus suggest that CPV or its DNA was released from the lysosomal compartment to the cytoplasm to be then transported to the nucleus. Electron microscopy analysis revealed endosomal vesicles containing CPV to be associated with microtubules. In the presence of nocodazole, a microtubule-disrupting drug, CPV entry was blocked and the virus was found in peripheral vesicles. Thus, some step(s) of the entry process were dependent on microtubules. Microinjection of antibodies to dynein caused CPV to remain in pericellular vesicles. This suggests an important role for the motor protein dynein in transporting vesicles containing CPV along the microtubule network. 相似文献
108.
Matti Uusitupa Markku Peltonen Jaana Lindstr?m Sirkka Aunola Pirjo Ilanne-Parikka Sirkka Kein?nen-Kiukaanniemi Timo T. Valle Johan G. Eriksson Jaakko Tuomilehto for the Finnish Diabetes Prevention Study Group 《PloS one》2009,4(5)
Background
The Finnish Diabetes Prevention Study (DPS) was a randomized controlled trial, which showed that it is possible to prevent type 2 diabetes by lifestyle changes. The aim of the present study was to examine whether the lifestyle intervention had an effect on the ten-year mortality and cardiovascular morbidity in the DPS participants originally randomized either into an intervention or control group. Furthermore, we compared these results with a population-based cohort comprising individuals of varying glucose tolerance states.Methods and Findings
Middle-aged, overweight people with IGT (n = 522) were randomized into intensive intervention (including physical activity, weight reduction and dietary counseling), or control “mini-intervention” group. Median length of the intervention period was 4 years and the mean follow-up was 10.6 years. The population-based reference study cohort included 1881 individuals (1570 with normal glucose tolerance, 183 with IGT, 59 with screen-detected type 2 diabetes, 69 with previously known type 2 diabetes) with the mean follow-up of 13.8 years. Mortality and cardiovascular morbidity data were collected from the national Hospital Discharge Register and Causes of Death Register. Among the DPS participants who consented for register linkage (n = 505), total mortality (2.2 vs. 3.8 per 1000 person years, hazard ratio HR = 0.57, 95% CI 0.21–1.58) and cardiovascular morbidity (22.9 vs. 22.0 per 1000 person years, HR = 1.04, 95% CI 0.72–1.51) did not differ significantly between the intervention and control groups. Compared with the population-based cohort with impaired glucose tolerance, adjusted HRs were 0.21 (95% CI 0.09–0.52) and 0.39 (95% CI 0.20–0.79) for total mortality, and 0.89 (95% CI 0.62–1.27) and 0.87 (0.60–1.27) for cardiovascular morbidity in the intervention and control groups of the DPS, respectively. The risk of death in DPS combined cohort was markedly lower than in FINRISK IGT cohort (adjusted HR 0.30, 95% CI 0.17–0.54), but there was no significant difference in the risk of CVD (adjusted HR 0.88, 95% CI 0.64–1.21).Conclusions
Lifestyle intervention among persons with IGT did not decrease cardiovascular morbidity during the first 10 years of follow-up. However, the statistical power may not be sufficient to detect small differences between the intervention and control groups. Low total mortality among participants of the DPS compared with individuals with IGT in the general population could be ascribed to a lower cardiovascular risk profile at baseline and regular follow-up.Trial Registration
ClinicalTrials.gov NCT00518167相似文献109.
Billaud JN Peterson D Barr M Chen A Sallberg M Garduno F Goldstein P McDowell W Hughes J Jones J Milich D 《Journal of virology》2005,79(21):13656-13666
The particulate hepatitis core protein (HBcAg) represents an efficient carrier platform with many of the characteristics uniquely required for the delivery of weak immunogens to the immune system. Although the HBcAg is highly immunogenic, the existing HBcAg-based platform technology has a number of theoretical and practical limitations, most notably the "preexisting immunity" and "assembly" problems. To address the assembly problem, we have developed the core protein from the woodchuck hepadnavirus (WHcAg) as a new particulate carrier platform system. WHcAg appears to tolerate insertions of foreign epitopes at a greater number of positions than HBcAg. For example, both within the external loop region and outside the loop region a total of 17 insertion sites were identified on WHcAg. Importantly, the identification of an expanded number of insertion sites was dependent on additional modifications to the C terminus that appear to stabilize the various internal insertions. Indeed, 21 separate C-terminal modifications have been generated that can be used in combination with the 17 insertion sites to ensure efficient hybrid WHcAg particle assembly. This combinatorial technology is also dependent on the sequence of the heterologous insert. Therefore, the three variables of insert position, C terminus, and epitope sequence are relevant in the design of hybrid WHcAg particles for vaccine purposes. 相似文献
110.
Borisovska M Zhao Y Tsytsyura Y Glyvuk N Takamori S Matti U Rettig J Südhof T Bruns D 《The EMBO journal》2005,24(12):2114-2126
SNARE proteins (soluble NSF-attachment protein receptors) are thought to be central components of the exocytotic mechanism in neurosecretory cells, but their precise function remained unclear. Here, we show that each of the vesicle-associated SNARE proteins (v-SNARE) of a chromaffin granule, synaptobrevin II or cellubrevin, is sufficient to support Ca(2+)-dependent exocytosis and to establish a pool of primed, readily releasable vesicles. In the absence of both proteins, secretion is abolished, without affecting biogenesis or docking of granules indicating that v-SNAREs are absolutely required for granule exocytosis. We find that synaptobrevin II and cellubrevin differentially control the pool of readily releasable vesicles and show that the v-SNARE's amino terminus regulates the vesicle's primed state. We demonstrate that dynamics of fusion pore dilation are regulated by v-SNAREs, indicating their action throughout exocytosis from priming to fusion of vesicles. 相似文献