全文获取类型
收费全文 | 48719篇 |
免费 | 6093篇 |
国内免费 | 21055篇 |
出版年
2024年 | 400篇 |
2023年 | 1339篇 |
2022年 | 2369篇 |
2021年 | 2864篇 |
2020年 | 2495篇 |
2019年 | 2945篇 |
2018年 | 2308篇 |
2017年 | 1949篇 |
2016年 | 2170篇 |
2015年 | 3044篇 |
2014年 | 4240篇 |
2013年 | 3802篇 |
2012年 | 5139篇 |
2011年 | 4956篇 |
2010年 | 3786篇 |
2009年 | 3878篇 |
2008年 | 4110篇 |
2007年 | 3919篇 |
2006年 | 3546篇 |
2005年 | 3002篇 |
2004年 | 2395篇 |
2003年 | 2060篇 |
2002年 | 1836篇 |
2001年 | 1666篇 |
2000年 | 1504篇 |
1999年 | 1017篇 |
1998年 | 519篇 |
1997年 | 389篇 |
1996年 | 307篇 |
1995年 | 283篇 |
1994年 | 254篇 |
1993年 | 172篇 |
1992年 | 191篇 |
1991年 | 155篇 |
1990年 | 127篇 |
1989年 | 97篇 |
1988年 | 91篇 |
1987年 | 72篇 |
1986年 | 70篇 |
1985年 | 99篇 |
1984年 | 68篇 |
1983年 | 48篇 |
1982年 | 85篇 |
1981年 | 23篇 |
1980年 | 8篇 |
1978年 | 5篇 |
1973年 | 5篇 |
1968年 | 3篇 |
1965年 | 10篇 |
1950年 | 13篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
921.
922.
923.
924.
甲基营养菌MP688萄糖脱氢酶基因分离鉴定及性质研究 总被引:1,自引:0,他引:1
目的:鉴定甲基营养菌MP688中的葡萄糖脱氢酶基因。方法:对甲基营养菌MP688基因组序列进行比对和分析,找到与已知细菌葡萄糖脱氢酶同源性最高的基因序列mpq_2164,且该基因所编码蛋白经分析具有跨膜结构域。设计51物扩增mpq_2164和缺失跨膜区域序列的s-mpq_2164,将PCR产物克隆到表达载雄pET-15b上,在大肠杆菌BL21中完成异源重组表达,然后通过组氨酸标签镍柱亲和层析纯化,采用DCIP法测定葡萄糖脱氢酶的活力。结果:分离了甲基营养菌MP688中的葡糖糖脱氢酶基因,并实现了s-mpq_2164的高效异源重组表达;MPQ2164的氯基酸序列与已知的葡萄糖脱氢酶相似性很低,但酶活测定结果表明S-MPQ-2164具有很高的葡糖糖脱氢酶活性。结论:MPQ_2164是-个依赖于吡咯喹啉醌的葡萄糖脱氢酶,去掉跨膜结构域有利于该蛋白的异源嘉{大, 相似文献
925.
926.
927.
目的:通过DNA重组技术表达肠出血性大肠杆菌(EHEC)0157:H7的EspA和EspB蛋白,并分析它们的免疫保护性。方法:采用PCR技术从EHEC0157:H7基因组中扩增espA和espB基因,连接至pET-22b(4-)载体上,转化至宿主细胞大肠杆菌BL21(DE3),经IPTG诱导表达,用亲和层析纯化目的蛋白,SDS-PAGE测定其相对分子质量,免疫小鼠分析其免疫保护性。结果:重组espA和espB基因片段的测序结果与GenBank中的相应基因序列完全一致,一致性均为100%;得到了纯度为95%以上的重组EspA和EspB蛋白,免疫小鼠所得到的抗体效价均为10^6。结论:重组EspA和EspB蛋白获得了可溶性表达,表达的蛋白具有良好的免疫保护性,为进一步制备疫苗奠定了基础。 相似文献
928.
Feng Wang Li Zuo Bo Hong Dongyi Han Ethan M Range Lingyun Zhao Yanan Sui Weiwei Guo Liangfa Liu 《Journal of biomedical science》2013,20(1):91
Background
Noise induced injury of the cochlea causes shifts in activation thresholds and changes of frequency response in the inferior colliculus (IC). Noise overexposure also induces pathological changes in the cochlea, and is highly correlated to hearing loss. However, the underlying mechanism has not been fully elucidated. In this study, we hypothesized that overexposure to noise induces substantial electrophysiological changes in the IC of guinea pigs.Results
During the noise exposure experiment, the animals were undergoing a bilateral exposure to noise. Additionally, various techniques were employed including confocal microscopy for the detection of cochlea hair cells and single neuron recording for spontaneous firing activity measurement. There were alterations among three types of frequency response area (FRA) from sound pressure levels, including V-, M-, and N-types. Our results indicate that overexposure to noise generates different patterns in the FRAs. Following a short recovery (one day after the noise treatment), the percentage of V-type FRAs considerably decreased, whereas the percentage of M-types increased. This was often caused by a notch in the frequency response that occurred at 4 kHz (noise frequency). Following a long recovery from noise exposure (11–21 days), the percentage of V-types resumed to a normal level, but the portion of M-types remained high. Interestingly, the spontaneous firing in the IC was enhanced in both short and long recovery groups.Conclusion
Our data suggest that noise overexposure changes the pattern of the FRAs and stimulates spontaneous firing in the IC in a unique way, which may likely relate to the mechanism of tinnitus. 相似文献929.
Guohua Feng Ji-Yuan Zhang Qing-Lei Zeng Lei Jin Junliang Fu Bin Yang Ying Sun Tianjun Jiang Xiangsheng Xu Zheng Zhang Jinhong Yuan Liyuan Wu Fu-Sheng Wang 《Molecules and cells》2013,36(4):362-367
Interleukin-21 (IL-21)+CD4+ T cells are involved in the immune response against hepatitis B virus (HBV) by secreting IL-21. However, the role of IL-21+CD4+ T cells in the immune response against chronic hepatitis C (CHC) virus infection is poorly understood. This study aimed to investigate the role of IL-21+CD4+ T cells in CHC patients and the potential mechanisms. The study subjects included nineteen CHC patients who were grouped by viral load (low, < 106 RNA copies/ml, n = 8; high, > 106 RNA copies/ml, n = 11). The peripheral frequency of HCV-specific IL-21+CD4+ T cells was higher in the low viral load group and was negatively correlated with the serum HCV RNA viral load in all CHC patients. Meanwhile, IL-21+ cells accumulated in the liver in the low viral load group. In vitro, IL-21 treatment increased the expression of proliferation markers and cytolytic molecules on HCV-specific CD8+ T cells. In summary, these findings suggest that HCV-specific IL-21+CD4+ T cells might contribute to HCV control by rescuing HCV-specific CD8+ T cells in CHC patients. 相似文献
930.
Feng Xue Alex B. Lennon Katey K. McKayed Veronica A. Campbell 《Computer methods in biomechanics and biomedical engineering》2013,16(5):468-476
A finite element model of a single cell was created and used to compute the biophysical stimuli generated within a cell under mechanical loading. Major cellular components were incorporated in the model: the membrane, cytoplasm, nucleus, microtubules, actin filaments, intermediate filaments, nuclear lamina and chromatin. The model used multiple sets of tensegrity structures. Viscoelastic properties were assigned to the continuum components. To corroborate the model, a simulation of atomic force microscopy indentation was performed and results showed a force/indentation simulation with the range of experimental results. A parametric analysis of both increasing membrane stiffness (thereby modelling membrane peroxidation with age) and decreasing density of cytoskeletal elements (thereby modelling reduced actin density with age) was performed. Comparing normal and aged cells under indentation predicts that aged cells have a lower membrane area subjected to high strain as compared with young cells, but the difference, surprisingly, is very small and may not be measurable experimentally. Ageing is predicted to have a more significant effect on strain deep in the nucleus. These results show that computation of biophysical stimuli within cells are achievable with single-cell computational models; correspondence between computed and measured force/displacement behaviours provides a high-level validation of the model. Regarding the effect of ageing, the models suggest only small, although possibly physiologically significant, differences in internal biophysical stimuli between normal and aged cells. 相似文献