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61.
Porro D Gasser B Fossati T Maurer M Branduardi P Sauer M Mattanovich D 《Applied microbiology and biotechnology》2011,89(4):939-948
Recombinant DNA (rDNA) technologies allow the production of a wide range of peptides, proteins and metabolites from naturally
non-producing cells. Since human insulin was the first heterologous compound produced in a laboratory in 1977, rDNA technology
has become one of the most important technologies developed in the 20th century. Recombinant protein and metabolites production
is a multi-billion dollar market. The development of a new product begins with the choice of the cell factory. The final application
of the compound dictates the main criteria that should be taken into consideration: (1) quality, (2) quantity, (3) yield and
(4) space time yield of the desired product. Quantity and quality are the most predominant requirements that must be considered
for the commercial production of a protein. Quantity and yield are the requirements for the production of a metabolite. Finally,
space time yield is crucial for any production process. It therefore becomes clear why the perfect host does not exist yet,
and why—despite important advances in rDNA applications in higher eukaryotic cells—microbial biodiversity continues to represent
a potential source of attractive cell factories. In this review, we compare the advantages and limitations of the principal
yeast and bacterial workhorse systems. 相似文献
62.
Xiangming Li Yujian Zhang Li Jing Zongming Fu Ou Ma Jishna Ganguly Nilesh Vaidya Richard Sisson Jennifer Naginskaya Avinash Chinthala Minggang Cui Ryan Yamagata Mark Wilson Matthew Sanders Zihao Wang Paola Lo Surdo Marcin Bugno 《Biotechnology progress》2020,36(2):e2914
Mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. Individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. Automated cell culture and analytics systems have been developed to enable high complexity clone selection workflows; while ensuring traceability, safety, and quality of cell lines intended for biopharmaceutical applications. Here we show that comprehensive and quantitative assessment of cell growth, productivity, and product quality attributes are feasible at the 200–1,200 cell colony stage, within 14 days of the single cell cloning in static 96-well plate culture. The early cell line characterization performed prior to the clone expansion in suspension culture can be used for a single-step, direct selection of high quality clones. Such clones were comparable, both in terms of productivity and critical quality attributes (CQAs), to the top-ranked clones identified using an established iterative clone screening approach. Using a complex, multi-subunit antigen as a model protein, we observed stable CQA profiles independently of the cell culture format during the clonal expansion as well as in the batch and fed-batch processes. In conclusion, we propose an accelerated clone selection approach that can be readily incorporated into various cell line development workstreams, leading to significant reduction of the project timelines and resource requirements. 相似文献
63.
64.
Marcella Barbarino Daniele Cesari Maria Bottaro Luca Luzzi Asadoor Namagerdi Franca Maria Bertolino Cristiana Bellan Fabrizio Proietti Pasquale Somma Mariacarolina Micheli Maria Margherita de Santi Raffaella Guazzo Luciano Mutti Luigi Pirtoli Piero Paladini Paola Indovina Antonio Giordano 《Journal of cellular and molecular medicine》2020,24(10):5565-5577
Malignant mesothelioma (MM) is an aggressive asbestos-related cancer of the serous membranes. Despite intensive treatment regimens, MM is still a fatal disease, mainly due to the intrinsic resistance to current therapies and the lack of predictive markers and new valuable molecular targets. Protein arginine methyltransferase 5 (PRMT5) inhibition has recently emerged as a potential therapy against methylthioadenosine phosphorylase (MTAP)-deficient cancers, in which the accumulation of the substrate 5'-methylthioadenosine (MTA) inhibits PRMT5 activity, thus sensitizing the cells to further PRMT5 inhibition. Considering that the MTAP gene is frequently codeleted with the adjacent cyclin-dependent kinase inhibitor 2A (CDKN2A) locus in MM, we assessed whether PRMT5 could represent a therapeutic target also for this cancer type. We evaluated PRMT5 expression, the MTAP status and MTA content in normal mesothelial and MM cell lines. We found that both administration of exogenous MTA and stable PRMT5 knock-down, by short hairpin RNAs (shRNAs), selectively reduced the growth of MTAP-deleted MM cells. We also observed that PRMT5 knock-down in MTAP-deficient MM cells reduced the expression of E2F1 target genes involved in cell cycle progression and of factors implicated in epithelial-to-mesenchymal transition. Therefore, PRMT5 targeting could represent a promising new therapeutic strategy against MTAP-deleted MMs. 相似文献
65.
66.
MC Pansani PS Azevedo BP Rafacho MF Minicucci F Chiuso-Minicucci SG Zorzella-Pezavento JS Marchini GJ Padovan AA Fernandes BB Matsubara LS Matsubara LA Zornoff SA Paiva 《PloS one》2012,7(7):e41439
Introduction
Micronutrient deficiency is observed in heart failure patients. Taurine, for example, represents 50% of total free amino acids in the heart, and in vivo studies have linked taurine deficiency with cardiomyopathy.Methods
Thirty-four male Wistar rats (body weight = 100 g) were weighed and randomly assigned to one of two groups: Control (C) or taurine-deficient (T (-)). Beta-alanine at a concentration of 3% was added to the animals’ water to induce taurine deficiency in the T (-) group. On day 30, the rats were individually submitted to echocardiography; morphometrical and histopathological evaluation and metalloproteinase activity, oxidative stress and inflammation evaluation were performed. Tissue samples were collected to determine the taurine concentration in the heart.Results
Taurine deficiency led to decreases in: ventricular wall thickness, left ventricle dry weight, myocyte sectional area, left ventricle posterior wall thickness and ventricular geometry. With regard to heart function, the velocity of the A wave, the ratio between the E and A wave, the ejection fraction, fractional shortening and cardiac output values were decreased in T (-) rats, suggesting abnormal diastolic and systolic function. Increased fibrosis, inflammation and increased activation of metalloproteinases were not observed. Oxidative stress was increased in deficient animals.Conclusions
These data suggest that taurine deficiency promotes structural and functional cardiac alterations with unique characteristics. 相似文献67.
Mayi TH Daoudi M Derudas B Gross B Bories G Wouters K Brozek J Caiazzo R Raverdi V Pigeyre M Allavena P Mantovani A Pattou F Staels B Chinetti-Gbaguidi G 《The Journal of biological chemistry》2012,287(26):21904-21913
Obesity is associated with a significantly increased risk for cancer suggesting that adipose tissue dysfunctions might play a crucial role therein. Macrophages play important roles in adipose tissue as well as in cancers. Here, we studied whether human adipose tissue macrophages (ATM) modulate cancer cell function. Therefore, ATM were isolated and compared with monocyte-derived macrophages (MDM) from the same obese patients. ATM, but not MDM, were found to secrete factors inducing inflammation and lipid accumulation in human T47D and HT-29 cancer cells. Gene expression profile comparison of ATM and MDM revealed overexpression of functional clusters, such as cytokine-cytokine receptor interaction (especially CXC-chemokine) signaling as well as cancer-related pathways, in ATM. Comparison with gene expression profiles of human tumor-associated macrophages showed that ATM, but not MDM resemble tumor-associated macrophages. Indirect co-culture experiments demonstrated that factors secreted by preadipocytes, but not mature adipocytes, confer an ATM-like phenotype to MDM. Finally, the concentrations of ATM-secreted factors related to cancer are elevated in serum of obese subjects. In conclusion, ATM may thus modulate the cancer cell phenotype. 相似文献
68.
Bandi E Jevsek M Mars T Jurdana M Formaggio E Sciancalepore M Fumagalli G Grubic Z Ruzzier F Lorenzon P 《American journal of physiology. Cell physiology》2008,294(1):C66-C73
The aim of this study was to elucidate the mechanisms responsible for the effects of innervation on the maturation of excitation-contraction coupling apparatus in human skeletal muscle. For this purpose, we compared the establishment of the excitation-contraction coupling mechanism in myotubes differentiated in four different experimental paradigms: 1) aneurally cultured, 2) cocultured with fetal rat spinal cord explants, 3) aneurally cultured in medium conditioned by cocultures, and 4) aneurally cultured in medium supplemented with purified recombinant chick neural agrin. Ca(2+) imaging indicated that coculturing human muscle cells with rat spinal cord explants increased the fraction of cells showing a functional excitation-contraction coupling mechanism. The effect of spinal cord explants was mimicked by treatment with medium conditioned by cocultures or by addition of 1 nM of recombinant neural agrin to the medium. The treatment with neural agrin increased the number of human muscle cells in which functional ryanodine receptors (RyRs) and dihydropyridine-sensitive L-type Ca(2+) channels were detectable. Our data are consistent with the hypothesis that agrin, released from neurons, controls the maturation of the excitation-contraction coupling mechanism and that this effect is due to modulation of both RyRs and L-type Ca(2+) channels. Thus, a novel role for neural agrin in skeletal muscle maturation is proposed. 相似文献
69.
Foods and liver health 总被引:1,自引:0,他引:1
Morisco F Vitaglione P Amoruso D Russo B Fogliano V Caporaso N 《Molecular aspects of medicine》2008,29(1-2):144-150
Chronic liver damage is a worldwide common pathology, characterised by an inflammatory and fibrotic process that leads to a progressive evolution from chronic hepatitis to cirrhosis and hepatocellular carcinoma (HCC). A major role for fats and oxidative stress has been recently demonstrated in the pathogenesis of liver diseases. In the clinical practice, dietary recommendations in the management of chronic diseases often rely on denying patients certain foods, which results in a severe reduction of quality of life. In this paper a new perspective based on the development of Food intended for Specific Medical Purposes (FSMP) containing highly bioavailable antioxidant compounds or polyunsaturated-fatty acids, has been highlighted as a tool for preventive and curative medicine, to be associated to pharmacological treatments. 相似文献
70.
The Dof protein DAG1 mediates PIL5 activity on seed germination by negatively regulating GA biosynthetic gene AtGA3ox1 总被引:2,自引:0,他引:2