全文获取类型
收费全文 | 7402篇 |
免费 | 470篇 |
国内免费 | 277篇 |
专业分类
8149篇 |
出版年
2024年 | 4篇 |
2023年 | 99篇 |
2022年 | 164篇 |
2021年 | 198篇 |
2020年 | 164篇 |
2019年 | 191篇 |
2018年 | 289篇 |
2017年 | 146篇 |
2016年 | 153篇 |
2015年 | 184篇 |
2014年 | 511篇 |
2013年 | 532篇 |
2012年 | 303篇 |
2011年 | 438篇 |
2010年 | 486篇 |
2009年 | 549篇 |
2008年 | 552篇 |
2007年 | 554篇 |
2006年 | 490篇 |
2005年 | 414篇 |
2004年 | 338篇 |
2003年 | 308篇 |
2002年 | 268篇 |
2001年 | 132篇 |
2000年 | 114篇 |
1999年 | 97篇 |
1998年 | 117篇 |
1997年 | 72篇 |
1996年 | 44篇 |
1995年 | 52篇 |
1994年 | 48篇 |
1993年 | 36篇 |
1992年 | 25篇 |
1991年 | 9篇 |
1990年 | 11篇 |
1989年 | 8篇 |
1988年 | 6篇 |
1987年 | 4篇 |
1986年 | 5篇 |
1985年 | 6篇 |
1984年 | 3篇 |
1983年 | 6篇 |
1982年 | 7篇 |
1981年 | 6篇 |
1980年 | 1篇 |
1979年 | 2篇 |
1976年 | 1篇 |
1975年 | 1篇 |
1973年 | 1篇 |
排序方式: 共有8149条查询结果,搜索用时 0 毫秒
91.
正常小鼠高频心电图时域值和功率谱的研究 总被引:4,自引:1,他引:4
本文用南京新博公司生产的NHE-1000型心电高频信息检测分析仪研究了正常小鼠(昆明种)高频心电图(HF-ECG)的时域值和QRS波群的功率谱。主要结果如下(以正导为例,-X±SD):心率603±88次/min(n=74);P-R间期相对较长。为34.9±4.7ms(n=58),占心动周期的34.9±4.9%,这与人类有很大的不同;QRS波宽9.2±1.2ms,占心动周期的9.2±1.4%(n=74),这一结果与以前的文献报道相差较大。T波宽10.3±3.2ms,占心动周期的10.3±3.2%;Q-T间期19.4±3.2ms,占心动周期的19.5±3.6%;QRS波群峰-峰值(Vp-p)为1.456±0.480mV;T波高0.336±0.115mV;73只动物Ⅱ导联高频切迹总数只有3个,扭挫26个。Ⅱ导联QRS波群的功率谱特点:0—80Hz的相对能量为45.48±15.32%;80—200Hz为43.97±9.95%;200—300Hz为8.89±7.38%;300—1000Hz为1.66±2.74%。 相似文献
92.
Pall GS Wallis J Axton R Brownstein DG Gautier P Buerger K Mulford C Mullins JJ Forrester LM 《Genomics》2004,84(6):204-1059
We have identified and characterized a gene, Mospd3 on mouse chromosome 5 using gene trapping in ES cells. MOSPD3 is part of a family of proteins, including MOSPD1, which is defined by the presence of a major sperm protein (MSP) domain and two transmembrane domains. Interestingly Mospd3 is mammalian specific and highly conserved between mouse and man. Insertion of the gene trap vector at the Mospd3 locus is mutagenic and breeding to homozygosity results in a characteristic right ventricle defect and neonatal lethality in 50% of mice. The phenotypic defect is dependent on the genetic background, indicating the presence of genetic modifier loci. We speculate that the further characterization of Mospd3 will shed light on the complex genetic interactions involved in cardiac development and disease. 相似文献
93.
94.
Haejin Yoon Seung-Hyun ShinDong Hoon Shin Yang-Sook ChunJong-Wan Park 《Biochemical and biophysical research communications》2014
Hypoxia-inducible factors 1α and 2α (HIF-1α and HIF-2α) determine cancer cell fate under hypoxia. Despite the similarities of their structures, HIF-1α and HIF-2α have distinct roles in cancer growth under hypoxia, that is, HIF-1α induces growth arrest whereas HIF-2α promotes cell growth. Recently, sirtuin 1 (Sirt1) was reported to fine-tune cellular responses to hypoxia by deacetylating HIF-1α and HIF-2α. Yet, the roles of Sirt1 in HIF-1α and HIF-2α functions have been controversial. We here investigated the precise roles of Sirt1 in HIF-1α and HIF-2α regulations. Immunological analyses revealed that HIF-1α K674 and HIF-2α K741 are acetylated by PCAF and CBP, respectively, but are deacetylated commonly by Sirt1. In the Gal4 reporter systems, Sirt1 was found to repress HIF-1α activity constantly in ten cancer cell-lines but to regulate HIF-2α activity cell type-dependently. Moreover, Sirt1 determined cell growth under hypoxia depending on HIF-1α and HIF-2α. Under hypoxia, Sirt1 promoted cell proliferation of HepG2, in which Sirt1 differentially regulates HIF-1α and HIF-2α. In contrast, such an effect of Sirt1 was not shown in HCT116, in which Sirt1 inactivates both HIF-1α and HIF-2α because conflicting actions of HIF-1α and HIF-2α on cell growth may be offset. Our results provide a better understanding of the roles of Sirt1 in HIF-mediated hypoxic responses and also a basic concept for developing anticancer strategy targeting Sirt1. 相似文献
95.
目的:利用枯草杆菌芽孢呈递技术制备表达SARS冠状病毒S蛋白受体结合区(RBD)的重组芽孢。方法:将枯草杆菌 CotB 基因构建到基因组整合质粒pDG1664中,再将 RBD 基因连接到 CotB 基因的下游,构建成重组质粒pDG1664-CotB-RBD,通过同源重组整合到PY-79枯草杆菌基因组中;利用红霉素抗性筛选重组菌并进行PCR和DNA测序鉴定,Western印迹鉴定重组菌芽孢表面RBD蛋白的表达情况;用表达RBD的重组芽孢以口服方式免疫小鼠,通过ELISA和流式细胞术检测重组芽孢的免疫原性。结果:制备出枯草杆菌基因组整合了RBD抗原基因的重组菌株RS1931,形成的重组芽孢表达相对分子质量约62×103的CotB-RBD融合蛋白;重组芽孢免疫的小鼠血清RBD抗原特异性IgG抗体滴度在末次免疫后2周可达1∶10880,重组芽孢初免后18周的小鼠脾细胞中IFN-γ+CD4^+、IL-4+CD4^+和IFN-γ+CD8^+T细胞比例上调,表明重组芽孢经口服免疫产生良好的体液免疫和细胞免疫应答。结论:针对SARS冠状病毒S蛋白RBD建立了枯草杆菌芽孢呈递技术方法,制备出在枯草杆菌芽孢表面稳定表达外源RBD蛋白的重组株,获得的重组芽孢具有良好的免疫原性,为开发芽孢呈递型SARS疫苗奠定了基础。 相似文献
96.
Zhou Y Drummond DC Zou H Hayes ME Adams GP Kirpotin DB Marks JD 《Journal of molecular biology》2007,371(4):934-947
To determine the importance of single-chain Fv (scFv) affinity on binding, uptake, and cytotoxicity of tumor-targeting nanoparticles, the affinity of the epidermal growth factor receptor (EGFR) scFv antibody C10 was increased using molecular evolution and yeast display. A library containing scFv mutants was created by error-prone PCR, displayed on the surface of yeast, and higher affinity clones selected by fluorescence activated cell sorting. Ten mutant scFv were identified that had a 3-18-fold improvement in affinity (KD=15-88 nM) for EGFR-expressing A431 tumor cells compared to C10 scFv (KD=264 nM). By combining mutations, higher affinity scFv were generated with KD ranging from 0.9 nM to 10 nM. The highest affinity scFv had a 280-fold higher affinity compared to that of the parental C10 scFv. Immunoliposome nanoparticles (ILs) were prepared using EGFR scFv with a 280-fold range of affinities, and their binding and uptake into EGFR-expressing tumor cells was quantified. At scFv densities greater than 148 scFv/IL, there was no effect of scFv affinity on IL binding and uptake into tumor cells, or on cytotoxicity. At lower scFv densities, there was less uptake and binding for ILs constructed from the very low affinity C10 scFv. The results show the importance of antibody fragment density on nanoparticle uptake, and suggest that engineering ultrahigh affinity scFv may be unnecessary for optimal nanoparticle targeting. 相似文献
97.
98.
Sara Kanje Raminta Venskutonytė Julia Scheffel Johan Nilvebrant Karin Lindkvist-Petersson Sophia Hober 《Journal of molecular biology》2018,430(18):3427-3438
Presented here is an engineered protein domain, based on Protein A, that displays a calcium-dependent binding to antibodies. This protein, ZCa, is shown to efficiently function as an affinity ligand for mild purification of antibodies through elution with ethylenediaminetetraacetic acid. Antibodies are commonly used tools in the area of biological sciences and as therapeutics, and the most commonly used approach for antibody purification is based on Protein A using acidic elution. Although this affinity-based method is robust and efficient, the requirement for low pH elution can be detrimental to the protein being purified. By introducing a calcium-binding loop in the Protein A-derived Z domain, it has been re-engineered to provide efficient antibody purification under mild conditions. Through comprehensive analyses of the domain as well as the ZCa–Fc complex, the features of this domain are well understood. This novel protein domain provides a very valuable tool for effective and gentle antibody and Fc-fusion protein purification. 相似文献
99.
Bradley HJ Liu X Collins V Owide J Goli GR Smith M Surprenant A White SJ Jiang LH 《FEBS letters》2010,584(23):4740-4744
We investigated membrane targeting of the P2X7 receptor (P2X7R) in polarized epithelial cells using immunofluorescent confocal imaging. The wild-type receptor was targeted to the basolateral membrane, independently of adaptor protein μ1B. Deletion of the majority of the intracellular C-terminus, or the last 26 residues (P570-Y595), conferred targeting of the protein to the apical membrane. Alanine substitution in the microdomain P582-Q587 caused similar apical membrane targeting without major effect on the receptor function and surface expression. Our results show basolateral membrane targeting of the P2X7R in epithelial cells and that the intracellular C-terminal microdomain P582-Q587 is crucial in this process.
Structured summary
MINT-8055849:Beta-catenin (uniprotkb:B6V8E6) and P2X7R (uniprotkb:Q64663) colocalize (MI:0403) by fluorescence microscopy (MI:0416) 相似文献100.
Intracellular antibody fragments that interfere with molecular interactions inside cells are valuable in investigation of interactomes and in therapeutics, but their application demands that they function in the reducing cellular milieu. We show here a 2.7-Å crystal structure of intracellular antibody folds based on scaffolds developed from intracellular antibody capture technology, and we reveal that there is no structural or functional difference with or without the intra-domain disulfide bond of the variable domain of heavy chain or the variable domain of light chain. The data indicate that, in the reducing in vivo environment, the absence of the intra-domain disulfide bond is not an impediment to correction of antibody folding or to interaction with antigen. Thus, the structural constraints for in-cell function are intrinsic to variable single-domain framework sequences, providing a generic scaffold for isolation of functional intracellular antibody single domains. 相似文献