全文获取类型
收费全文 | 7698篇 |
免费 | 527篇 |
国内免费 | 3篇 |
专业分类
8228篇 |
出版年
2023年 | 48篇 |
2022年 | 94篇 |
2021年 | 181篇 |
2020年 | 109篇 |
2019年 | 128篇 |
2018年 | 173篇 |
2017年 | 190篇 |
2016年 | 268篇 |
2015年 | 420篇 |
2014年 | 435篇 |
2013年 | 522篇 |
2012年 | 625篇 |
2011年 | 531篇 |
2010年 | 354篇 |
2009年 | 317篇 |
2008年 | 401篇 |
2007年 | 442篇 |
2006年 | 384篇 |
2005年 | 358篇 |
2004年 | 340篇 |
2003年 | 318篇 |
2002年 | 297篇 |
2001年 | 88篇 |
2000年 | 52篇 |
1999年 | 88篇 |
1998年 | 83篇 |
1997年 | 65篇 |
1996年 | 51篇 |
1995年 | 61篇 |
1994年 | 55篇 |
1993年 | 57篇 |
1992年 | 50篇 |
1991年 | 50篇 |
1990年 | 53篇 |
1989年 | 36篇 |
1988年 | 38篇 |
1987年 | 18篇 |
1986年 | 30篇 |
1985年 | 31篇 |
1984年 | 30篇 |
1983年 | 19篇 |
1982年 | 28篇 |
1981年 | 24篇 |
1980年 | 14篇 |
1979年 | 18篇 |
1977年 | 12篇 |
1976年 | 19篇 |
1975年 | 13篇 |
1974年 | 21篇 |
1970年 | 16篇 |
排序方式: 共有8228条查询结果,搜索用时 0 毫秒
971.
Viggiano E Passavanti MB Pace MC Sansone P Spaziano G Viggiano A Aurilio C Monda M Viggiano A Pota V De Luca B De Luca E 《Journal of cellular physiology》2012,227(5):1988-1991
Glutamine (gln) is the most abundant free amino acid in the blood. It is involved in important metabolic and biochemical processes, like cell proliferation and oxidative stress. Previous studies have demonstrated that gln concentration in human plasma decreases in several conditions such as sepsis, ischemia-reperfusion, trauma, major surgery and burn. The aim of the present work was to compare the acute effects of different types of surgical interventions and of anesthetization on blood gln concentration. Plasma samples from 88 subjects (30 males and 58 females) were collected before and after major or minor surgery and the gln concentration was analyzed with high-performance liquid chromatography. The results showed that plasma gln concentration after surgery was lower than pre-surgery values and that in major surgery the decrease of gln was higher than in minor surgery. No significant effect was shown for sex or type of anesthesia. These results demonstrate the importance of a gln supplementation before a surgical intervention and show that the amount of gln supplementation should also be adjusted based on the type of surgery. 相似文献
972.
We show via single-molecule mechanical unfolding experiments that the osmolyte glycerol stabilizes the native state of the human cardiac I27 titin module against unfolding without shifting its unfolding transition state on the mechanical reaction coordinate. Taken together with similar findings on the immunoglobulin-binding domain of streptococcal protein G (GB1), these experimental results suggest that osmolytes act on proteins through a common mechanism that does not entail a shift of their unfolding transition state. We investigate the above common mechanism via an Ising-like model for protein mechanical unfolding that adds worm-like-chain behavior to a recent generalization of the Wako-Saitô-Muñoz-Eaton model with support for group-transfer free energies. The thermodynamics of the model are exactly solvable, while protein kinetics under mechanical tension can be simulated via Monte Carlo algorithms. Notably, our force-clamp and velocity-clamp simulations exhibit no shift in the position of the unfolding transition state of GB1 and I27 under the effect of various osmolytes. The excellent agreement between experiment and simulation strongly suggests that osmolytes do not assume a structural role at the mechanical unfolding transition state of proteins, acting instead by adjusting the solvent quality for the protein chain analyte. 相似文献
973.
Grimaldi S Arias-Cartin R Lanciano P Lyubenova S Szenes R Endeward B Prisner TF Guigliarelli B Magalon A 《The Journal of biological chemistry》2012,287(7):4662-4670
Escherichia coli nitrate reductase A (NarGHI) is a membrane-bound enzyme that couples quinol oxidation at a periplasmically oriented Q-site (QD) to proton release into the periplasm during anaerobic respiration. To elucidate the molecular mechanism underlying such a coupling, endogenous menasemiquinone-8 intermediates stabilized at the QD site (MSQD) of NarGHI have been studied by high-resolution pulsed EPR methods in combination with 1H2O/2H2O exchange experiments. One of the two non-exchangeable proton hyperfine couplings resolved in hyperfine sublevel correlation (HYSCORE) spectra of the radical displays characteristics typical from quinone methyl protons. However, its unusually small isotropic value reflects a singularly low spin density on the quinone carbon α carrying the methyl group, which is ascribed to a strong asymmetry of the MSQD binding mode and consistent with single-sided hydrogen bonding to the quinone oxygen O1. Furthermore, a single exchangeable proton hyperfine coupling is resolved, both by comparing the HYSCORE spectra of the radical in 1H2O and 2H2O samples and by selective detection of the exchanged deuterons using Q-band 2H Mims electron nuclear double resonance (ENDOR) spectroscopy. Spectral analysis reveals its peculiar characteristics, i.e. a large anisotropic hyperfine coupling together with an almost zero isotropic contribution. It is assigned to a proton involved in a short ∼1.6 Å in-plane hydrogen bond between the quinone O1 oxygen and the Nδ of the His-66 residue, an axial ligand of the distal heme bD. Structural and mechanistic implications of these results for the electron-coupled proton translocation mechanism at the QD site are discussed, in light of the unusually high thermodynamic stability of MSQD. 相似文献
974.
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
975.
Pierdonato Bruno Giovanna Gentile Rita ManciniClaudia De Vitis Maria Cristina Esposito Davide Scozzi Mario MastrangeloAlberto Ricci Ibrahim Mohsen Gennaro CilibertoMaurizio Simmaco Salvatore Mariotta 《Biochemical and biophysical research communications》2012,426(3):306-309
Background
CpG island hypermethylation of gene promoters and regulatory regions is a well-known mechanism of epigenetic silencing of tumor suppressors and is directly linked to carcinogenesis. Wilm’s tumor gene (WT1) is a tumor suppressor protein involved in the regulation of human cell growth and differentiation and a modulator of oncogenic K Ras signaling in lung cancer. Changes in the pattern of methylation of the WT1 gene have not yet been studied in detail in human lung cancer. In this study we compared the methylation profile of WT1 gene in samples of neoplastic and non-neoplastic lung tissue taken from the same patients.Methods
DNA was extracted from neoplastic and normal lung tissue obtained from 16 patients with non small cell lung cancer (NSCLC). The methylation status of 29 CpG islands in the 5′ region of WT1 was determined by pyrosequencing. Statistical analysis was carried out by T test and Mann Whitney test.Results
The mean percentage of methylation, considering all CpG islands of WT1 in the neoplastic tissues of the 16 NSCLC patients, was 16.2 ± 3.4, whereas in the normal lung tissue from the same patients it was 5.6 ± 1.7 (p < 0.001). Adenocarcinomas presented higher methylation levels than squamous cell carcinomas (p < 0,001).Conclusions
Methylation of WT1 gene is significantly increased in NSCLC. Both histotype and exposure to cigarette smoke heavily influence the pattern of CpG islands which undergo hypermethylation. 相似文献976.
977.
Emmanuel Richaud Ludmila Audouin Bruno Fayolle Jacques Verdu Lyda Matisová-Rychlá Jozef Rychlý 《Chemistry and physics of lipids》2012,165(7):753-759
This study deals with oxidation kinetics of three unsaturated fatty esters: methyl oleate, methyl linoleate and methyl linolenate at temperatures ranging from 90 to 150 °C. The reaction was monitored by chemiluminescence. A kinetic model was derived from a simple mechanistic scheme, in which initiation is due to hydroperoxides decomposition, whereas propagation results from the abstraction of the most labile hydrogen and termination results only from the bimolecular combination of peroxyl radicals. Analysis of induction period duration indicated that hydroperoxides mainly decompose by a bimolecular process. The model well predicts the main features of the experimental chemiluminescence curves. Kinetic parameters of the three unsaturated fatty esters were assessed from inverse method and discussed. 相似文献
978.
The phase diagram of mixed monolayers composed of dimyristoyl phosphatidylcholine (DMPC) and stearic acid (SA) on different subphases was previously reported. It was observed that on acid subphases, liquid-condensed domains with shapes that depend on the SA proportion are formed. For mixtures with 40-45mole% of SA, the domain shape changes from flower-like to circular domains. In this work, we carried out a detailed study of the driving force for the shape change. We find that it is related to the domain density which, in turn, is driven by the domain nucleation process and thus by oversaturation of the system leading to phase segregation. This could be a way of self-regulating the local electrostatics and mechanical properties in membrane surfaces with segregated phase domains. 相似文献
979.
The organization of lipids and proteins into domains in cell membranes is currently an established subject within biomembrane research. Fluorescent probes have been used to detect and characterize these membrane lateral heterogeneities. However, a comprehensive understanding of the link between the probes' fluorescence features and membrane lateral organization can only be achieved if their photophysical properties are thoroughly defined. In this work, a systematic characterization of N-(lyssamine Rhodamine B sulfonyl)-1,2-dioleoyl-sn-3-phosphatidylehanolamine (Rhod-DOPE) absorption and fluorescence behavior in gel, liquid-ordered (l(o)) and liquid-disordered (l(d)) model membranes was performed. In agreement with a previous study, it was found that Rhod-DOPE fluorescence lifetimes present a strong sensitivity to lipid phases, becoming significantly shorter in l(o) membranes as the probe membrane concentration increases. The sensitivity of Rhod-DOPE absorption and fluorescence properties to the membrane phase was further explored. In particular, the fluorescence lifetime sensitivity was shown to be a consequence of the enhanced Rhod-DOPE fluorescence dynamic self-quenching, due to the formation of probe-rich membrane domains in these condensed phases that cannot be considered as typical probe aggregates, as excitonic interaction is not observed. The highly efficient dynamic self-quenching was shown to be specific to l(o) phases, pointing to an important effect of membrane dipole potential in this process. Altogether, this work establishes how to use Rhod-DOPE fluorescence properties in the study of membrane lipid lateral heterogeneities, in particular cholesterol-enriched lipid rafts. 相似文献
980.
Non-alcoholic staetohepatitis (NASH) is associated with fat deposition in the liver favoring inflammatory processes and development of fibrosis, cirrhosis and finally hepatocellular cancer. In Western lifestyle countries, NASH has reached a 20% prevalence in the obese population with escalating tendency in the future. Very often, liver transplantation is the only therapeutic option. Recently, transplantation of hepatocyte-like cells differentiated from mesenchymal stem cells was suggested a feasible alternative to whole organ transplantation to ameliorate donor organ shortage. Hence, in the present work an animal model of NASH was established in immunodeficient mice to investigate the feasibility of human stem cell-derived hepatocyte-like cell transplantation. NASH was induced by feeding a methionine/choline-deficient diet (MCD-diet) for up to 5 weeks. Animals developed a fatty liver featuring fibrosis and elevation of the proinflammatory markers serum amyloid A (SAA) and tumor necrosis factor alpha (TNFα). Hepatic triglycerides were significantly increased as well as alanine aminotransferase demonstrating inflammation-linked hepatocyte damage. Elevation of αSMA mRNA and collagen I as well as liver architecture deterioation indicated massive fibrosis. Both short- and long-term post-transplantation human hepatocyte-like cells resided in the mouse host liver indicating parenchymal penetration and most likely functional engraftment. Hence, the NASH model in the immunodeficient mouse is the first to allow for the assessment of the therapeutic impact of human stem cell-derived hepatocyte transplantation. 相似文献