全文获取类型
收费全文 | 2344篇 |
免费 | 185篇 |
国内免费 | 170篇 |
专业分类
2699篇 |
出版年
2024年 | 10篇 |
2023年 | 35篇 |
2022年 | 89篇 |
2021年 | 124篇 |
2020年 | 119篇 |
2019年 | 124篇 |
2018年 | 107篇 |
2017年 | 100篇 |
2016年 | 108篇 |
2015年 | 134篇 |
2014年 | 169篇 |
2013年 | 172篇 |
2012年 | 205篇 |
2011年 | 197篇 |
2010年 | 105篇 |
2009年 | 110篇 |
2008年 | 92篇 |
2007年 | 98篇 |
2006年 | 98篇 |
2005年 | 67篇 |
2004年 | 56篇 |
2003年 | 58篇 |
2002年 | 42篇 |
2001年 | 35篇 |
2000年 | 33篇 |
1999年 | 28篇 |
1998年 | 20篇 |
1997年 | 21篇 |
1996年 | 18篇 |
1995年 | 17篇 |
1994年 | 24篇 |
1993年 | 12篇 |
1992年 | 16篇 |
1991年 | 13篇 |
1990年 | 7篇 |
1989年 | 5篇 |
1988年 | 8篇 |
1987年 | 7篇 |
1986年 | 4篇 |
1985年 | 5篇 |
1983年 | 2篇 |
1982年 | 2篇 |
1979年 | 3篇 |
排序方式: 共有2699条查询结果,搜索用时 15 毫秒
31.
Liu Q Zhan J Chen X Zheng S 《Biochemical and biophysical research communications》2006,343(3):857-863
Ricin is a potent ribosome inactivating protein and now has been widely used for synthesis of immunotoxins. To target ribosome in the mammalian cytosol, ricin must firstly retrograde transport from the endomembrane system to reach the endoplasmic reticulum (ER) where the ricin A chain (RTA) is recognized by ER components that facilitate its membrane translocation to the cytosol. In the study, the fusion gene of enhanced green fluorescent protein (EGFP)-RTA was expressed with the pET-28a (+) system in Escherichia coli under the control of a T7 promoter. The fusion protein showed a green fluorescence. The recombinant protein can be purified by metal chelated affinity chromatography on a column of NTA. The rabbit anti-GFP antibody can recognize the fusion protein of EGFP-RTA just like the EGFP protein. The cytotoxicity of EGFP-RTA and RTA was evaluated by the MTT assay in HeLa and HEP-G2 cells following fluid-phase endocytosis. The fusion protein had a similar cytotoxicity of RTA. After endocytosis, the subcellular location of the fusion protein can be observed with the laser scanning confocal microscopy and the immuno-gold labeling Electro Microscopy. This study provided important evidence by a visualized way to prove that RTA does reach the endoplasmic reticulum. 相似文献
32.
33.
本文用凯氏定氮法、双缩脲法及福林-酚法测定了胸腺肽含量,并加以比较,说明三种方法对其含量测定没有显著差异.在实践中可以根据不同条件选择适当的方法进行实验. 相似文献
34.
35.
转基因白桦中GUS基因表达的定量分析 总被引:1,自引:0,他引:1
以转基因白桦(Betula platyphylla)为材料,采用单酶切结合Southern杂交的方法揭示不同转基因植株中GUS基因的整合拷贝数为1—4个。采用组织化学染色法定性分析不同整合方式转基因白桦植株中GUS基因的表达。结果表明,11个转基因植株中有2株出现了GUS基因沉默,其余植株均有不同水平的GUS表达。在此基础上应用分光光度法定量分析不同拷贝数的GUS转基因白桦中β-葡萄糖醛酸酶活性。结果表明,在11个转基因尢性系中除2个株系的GUS基因沉默外,其它9个转基因植株中GUS酶活力差异明显,但这种差异与GUS基因的拷贝数没有必然联系。 相似文献
36.
Jingcheng Wu Wenyi Zhao Binbin Zhou Zhixi Su Xun Gu Zhan Zhou Shuqing Chen 《基因组蛋白质组与生物信息学报(英文版)》2018,16(4):276-282
Tumor-specific neoantigens have attracted much attention since they can be used as biomarkers to predict therapeutic effects of immune checkpoint blockade therapy and as potential targets for cancer immunotherapy. In this study, we developed a comprehensive tumor-specific neoantigen database (TSNAdb v1.0), based on pan-cancer immunogenomic analyses of somatic mutation data and human leukocyte antigen (HLA) allele information for 16 tumor types with 7748 tumor samples from The Cancer Genome Atlas (TCGA) and The Cancer Immunome Atlas (TCIA). We predicted binding affinities between mutant/wild-type peptides and HLA class I molecules by NetMHCpan v2.8/v4.0, and presented detailed information of 3,707,562/1,146,961 potential neoantigens generated by somatic mutations of all tumor samples. Moreover, we employed recurrent mutations in combination with highly frequent HLA alleles to predict potential shared neoantigens across tumor patients, which would facilitate the discovery of putative targets for neoantigen-based cancer immunotherapy. TSNAdb is freely available at http://biopharm.zju.edu.cn/tsnadb. 相似文献
37.
In this study, a gold nanoparticle (Au-NP)-based detection method for sensitive and specific DNA-based diagnostic applications is described. A sandwich format consisting of Au-NPs/DNA/PMP (Streptavidin-coated MagnetSphere Para-Magnetic Particles) was fabricated. PMPs captured and separated target DNA while Au-NPs modified with oligonucleotide detection sequences played a role in recognition and signal production. Due to the much lower stability of mismatched DNA strands caused by unstable duplex structures in solutions of relatively low salt concentration, hybridization efficiency in the presence of different buffers was well investigated, and thus, the optimized salt concentration allowed for discrimination of single-mismatched DNA (MMT) from perfectly matched DNA (PMT). Therefore, quantitative information concerning the target analyte was translated into a colorimetric signal, which could easily and quantitatively measured by low-cost UV-vis spectrophotometric analysis. The results indicated this to be a very simple and economic strategy for detection of single-mismatched DNA strands. 相似文献
38.
Jiheng Zhan Xing Li Dan Luo Yu Hou Yonghui Hou Shudong Chen Zhifeng Xiao Jiyao Luan Dingkun Lin 《Journal of cellular and molecular medicine》2020,24(9):5317-5329
Bone marrow mesenchymal stem cell (BMSC) transplantation represents a promising repair strategy following spinal cord injury (SCI), although the therapeutic effects are minimal due to their limited neural differentiation potential. Polydatin (PD), a key component of the Chinese herb Polygonum cuspidatum, exerts significant neuroprotective effects in various central nervous system disorders and protects BMSCs against oxidative injury. However, the effect of PD on the neuronal differentiation of BMSCs, and the underlying mechanisms remain inadequately understood. In this study, we induced neuronal differentiation of BMSCs in the presence of PD, and analysed the Nrf2 signalling and neuronal differentiation markers using routine molecular assays. We also established an in vivo model of SCI and assessed the locomotor function of the mice through hindlimb movements and electrophysiological measurements. Finally, tissue regeneration was evaluated by H&E staining, Nissl staining and transmission electron microscopy. PD (30 μmol/L) markedly facilitated BMSC differentiation into neuron‐like cells by activating the Nrf2 pathway and increased the expression of neuronal markers in the transplanted BMSCs at the injured spinal cord sites. Furthermore, compared with either monotherapy, the combination of PD and BMSC transplantation promoted axonal rehabilitation, attenuated glial scar formation and promoted axonal generation across the glial scar, thereby enhancing recovery of hindlimb locomotor function. Taken together, PD augments the neuronal differentiation of BMSCs via Nrf2 activation and improves functional recovery, indicating a promising new therapeutic approach against SCI. 相似文献
39.
40.
欧美杂种山杨体细胞无性系变异的分析 总被引:1,自引:2,他引:1
以成年欧美杂种山杨(Populustremula×P.tremuloides)优良无性系为材料,通过组织培养方法获得体细胞无性系,利用细胞学和分子生物学方法对其发生的变异进行研究。结果表明:用3.0mg.L-12,4-D诱导的再生植株的细胞染色体稳定性较差,所检测的144个细胞中,变异的细胞中多数发生了染色体加倍,二倍体细胞数仅占36.81%。有些染色体还发生了形态变异,染色体加长,形成带有随体或长臂较长的大型染色体。用1.0mg.L-16-BA诱导再生植株中染色体数量稳定性介于对照和3.0mg.L-12,4-D诱导的再生植株之间,在观察的142个细胞中二倍体细胞占54.93%。再生植株的AFLP分析表明,由激素诱导的再生植株中,AFLP谱带发生了变化,表明发生了体细胞无性系分子水平变异。 相似文献