全文获取类型
收费全文 | 10222篇 |
免费 | 918篇 |
国内免费 | 911篇 |
出版年
2024年 | 12篇 |
2023年 | 152篇 |
2022年 | 186篇 |
2021年 | 546篇 |
2020年 | 428篇 |
2019年 | 486篇 |
2018年 | 471篇 |
2017年 | 309篇 |
2016年 | 458篇 |
2015年 | 710篇 |
2014年 | 790篇 |
2013年 | 829篇 |
2012年 | 937篇 |
2011年 | 807篇 |
2010年 | 541篇 |
2009年 | 450篇 |
2008年 | 553篇 |
2007年 | 475篇 |
2006年 | 424篇 |
2005年 | 346篇 |
2004年 | 284篇 |
2003年 | 264篇 |
2002年 | 205篇 |
2001年 | 177篇 |
2000年 | 153篇 |
1999年 | 168篇 |
1998年 | 92篇 |
1997年 | 82篇 |
1996年 | 75篇 |
1995年 | 75篇 |
1994年 | 101篇 |
1993年 | 48篇 |
1992年 | 53篇 |
1991年 | 67篇 |
1990年 | 42篇 |
1989年 | 48篇 |
1988年 | 32篇 |
1987年 | 27篇 |
1986年 | 34篇 |
1985年 | 27篇 |
1984年 | 17篇 |
1983年 | 12篇 |
1982年 | 10篇 |
1981年 | 5篇 |
1980年 | 5篇 |
1978年 | 4篇 |
1973年 | 4篇 |
1970年 | 3篇 |
1969年 | 3篇 |
1968年 | 4篇 |
排序方式: 共有10000条查询结果,搜索用时 71 毫秒
81.
Wan-Jin Chen Xiang-Ping Yao Qi-Jie Zhang Wang Ni Jin He Hong-Fu Li Xin-Yi Liu Gui-Xian Zhao Shen-Xing Murong Ning Wang Zhi-Ying Wu 《Gene》2013
Idiopathic basal ganglia calcification (IBGC) is a rare neuropsychiatric disorder characterized by bilateral and symmetric cerebral calcifications. Recently, SLC20A2 was identified as a causative gene for familial IBGC, and three mutations were reported in a northern Chinese population. Here, we aimed to explore the mutation spectrum of SLC20A2 in a southern Chinese population. Sanger sequencing was employed to screen mutations within SLC20A2 in two IBGC families and 14 sporadic IBGC cases from a southern Han Chinese population. Four novel mutations (c.82G > A p.D28N, c.185T > C p.L62P, c.1470_1478delGCAGGTCCT p.Q491_L493del and c.935-1G > A) were identified in two families and two sporadic cases, respectively; none were detected in 200 unrelated controls. No mutation was found in the remaining 12 patients. Different mutations may result in varied phenotypes, including brain calcification and clinical manifestations. Our study supports the hypothesis that SLC20A2 is a causative gene of IBGC and expands the mutation spectrum of SLC20A2, which facilitates the understanding of the genotype–phenotype correlation of IBGC. 相似文献
82.
83.
Shengping Huang Jingjing Chen Quanjiao Chen Huadong Wang Yanfeng Yao Jianjun Chen Ze Chen 《Journal of virology》2013,87(2):767-778
Influenza A virus NS2 protein, also called nuclear export protein (NEP), is crucial for the nuclear export of viral ribonucleoproteins. However, the molecular mechanisms of NEP mediation in this process remain incompletely understood. A leucine-rich nuclear export signal (NES2) in NEP, located at the predicted N2 helix of the N-terminal domain, was identified in the present study. NES2 was demonstrated to be a transferable NES, with its nuclear export activity depending on the nuclear export receptor chromosome region maintenance 1 (CRM1)-mediated pathway. The interaction between NEP and CRM1 is coordinately regulated by both the previously reported NES (NES1) and now the new NES2. Deletion of the NES1 enhances the interaction between NEP and CRM1, and deletion of the NES1 and NES2 motifs completely abolishes this interaction. Moreover, NES2 interacts with CRM1 in the mammalian two-hybrid system. Mutant viruses containing NES2 alterations generated by reversed genetics exhibit reduced viral growth and delay in the nuclear export of viral ribonucleoproteins (vRNPs). The NES2 motif is highly conserved in the influenza A and B viruses. The results demonstrate that leucine-rich NES2 is involved in the nuclear export of vRNPs and contributes to the understanding of nucleocytoplasmic transport of influenza virus vRNPs. 相似文献
84.
Microsatellite genetic variation in the Chinese endemic Eucommia ulmoides (Eucommiaceae): implications for conservation 总被引:1,自引:0,他引:1
Jinju Zhang Chong Xing Hua Tian Xiaohong Yao 《Botanical journal of the Linnean Society. Linnean Society of London》2013,173(4):775-785
Eucommia ulmoides, a traditional Chinese medicinal plant, is endangered as a consequence of long‐term and widespread harvest in the late 20th century. It has been widely cultivated as a source of herbal medicine and for use in the organic chemical industry in China. In this study, eight microsatellite markers were applied to investigate genetic diversity in E. ulmoides. Three hundred individuals from one semi‐wild population and nine cultivated populations across its main production area were collected. A high level of genetic diversity at population levels (HE = 0.716) was observed. The highly outcrossed mating system, high longevity of E. ulmoides and seed admixture may be responsible for high genetic variation within populations. A genetic bottleneck was observed in one population. Populations were only slightly differentiated from one another (FST = 0.063); this was also supported by AMOVA, which revealed that 94.05% of the total variation resided within populations. This is probably attributable to long‐distance gene flow mediated by the exchange of seeds by local farmers. Implications of these results for the conservation of genetic resources of E. ulmoides are discussed. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 775–785. 相似文献
85.
Bin Wu Alys Peisley Claire Richards Hui Yao Xiaohui Zeng Cecilie Lin Feixia Chu Thomas Walz Sun Hur 《Cell》2013,152(1-2):276-289
Highlights? MDA5 forms an open, C-shaped ring around the viral dsRNA stem ? The CTD of MDA5 has a different orientation and flexibility compared to RIG-I ? MDA5 forms filaments by stacking monomers head-to-tail with a 70° turn per monomer ? The 2CARD domain assembles into oligomers that activate interferon signaling via MAVS 相似文献
86.
摘要:【目的】建立多粘类芽胞杆菌SC2 的基因敲除体系。【方法】利用电转化把温敏型自杀质粒pRN5101导入到多粘类芽胞杆菌SC2中。采用基因重组技术敲除SC2 中的多粘菌素基因E(pmxE),得到突变株SC2-E。利用抗细菌性能检测和高效液相色谱分析合成多粘菌素的能力,来证实pmxE基因是否被敲除。【结果】成功构建了多粘类芽胞杆菌SC2 的基因敲除体系。pRN5101转入SC2后能够在28℃复制,39℃自杀。突变株失去了合成多粘菌素的能力,成功敲除pmxE基因,验证了此体系的可用性。【结论】首次构建了多粘类芽胞杆菌的基因敲除体系,拓展了pRN5101的使用范围,为研究多粘类芽胞杆菌的基因功能提供了高效的遗传操作工具。 相似文献
87.
[目的]探讨ompH基因在禽多杀性巴氏杆菌致病过程中的作用.[方法]利用同源重组原理构建中间为四环素抗性基因,两侧为ompH基因上下游同源序列同源的敲除载体pWSK29△ompH,将敲除载体电击转入C48-3株感受态细胞中,通过四环素抗性和菌落PCR筛选ompH基因的敲除突变株,并通过组合PCR、逆转录PCR和DNA测序对突变株进行验证.用生物学功能实验比较野生株、互补株和突变株在生长速率、荚膜结构、粘着能力和致病性等方面的差异.[结果]组合PCR、逆转录PCR和DNA测序结果证实ompH基因的敲除突变株C48-3△ompH构建成功,电镜观察结果证实ompH基因的缺陷影响细菌的荚膜合成能力,粘附实验结果显示与野生株C48-3和互补株C48-3C相比突变株C48-3△ompH对CEF细胞的粘附能力显著降低(P<0.01),而小鼠毒力实验结果表明突变株C48-3△ompH的致病性相对减弱.[结论]本实验构建的突变株C48-3△ompH,为进一步研究多杀性巴氏杆菌的致病机理奠定基础. 相似文献
88.
Jiongyuan Xu Lizhi Niu Feng Mu Shupeng Liu Yin Leng Mengtian Liao Jianying Zeng Fei Yao Jibing Chen Jialiang Li Kecheng Xu 《Cryobiology》2013
Esophageal cancer is common in China. There is a lack of treatment strategies for metastatic esophageal cancer (MEC) after radical surgery on the primary tumor. Cryoablation is an attractive option because tumor necrosis can be safely induced in a minimally invasive manner. This study assessed its therapeutic effect in MEC after failure of radical surgery. One hundred and forty patients met the inclusion criteria from May, 2003 to March, 2011. Comprehensive cryotherapy of multiple metastases was performed on 105 patients; 35 received chemotherapy. No severe complications occurred during or after cryoablation. Overall survival (OS) was assessed according to therapeutic protocol, pathologic type, treatment timing and number of procedures. The OS of patients who received comprehensive cryoablation (44 ± 20 months) was significantly longer than that of those who underwent chemotherapy (23 ± 24 months; P = 0.0006). In the cryotherapy group, the OS for squamous cell carcinoma (45 ± 19 months) was longer than that for adenocarcinoma (33 ± 18 months; P = 0.0435); the OS for timely cryoablation (46 ± 19 months) was longer than that for delayed cryoablation (33 ± 20 months; P = 0.0193); the OS for multiple cryoablation (50 ± 17 months) was longer than that for single cryoablation (37 ± 20 months; P = 0.0172); and the OS for cryo-immunotherapy (56 ± 17 months) was longer than that for cryoablation alone (39 ± 19 months; P = 0.0011). Thus, comprehensive cryotherapy may have advantages over chemotherapy in the treatment of MEC and, in patients with squamous cell carcinoma, supplementary immunotherapy and timely and multiple cryoablation may be associated with a better prognosis. 相似文献
89.
Dongbing Cui Lujiang Zhang Zhiqiang Yao Xu Liu Jinping Lin Y. Adam Yuan Dongzhi Wei 《Journal of biotechnology》2013
Short-chain dehydrogenase Gox2181 from Gluconobacter oxydans catalyzes the reduction of 2,3-pentanedione by using NADH as the physiological electron donor. To realize its synthetic biological application for coenzyme recycling use, computational design and site-directed mutagenesis have been used to engineer Gox2181 to utilize not only NADH but also NADPH as the electron donor. Single and double mutations at residues Q20 and D43 were made in a recombinant expression system that corresponded to Gox2181-D43Q and Gox2181-Q20R&D43Q, respectively. The design of mutant Q20R not only resolved the hydrogen bond interaction and electrostatic interaction between R and 2′-phosphate of NADPH, but also could enhance the binding with 2′-phophated of NADPH by combining with D43Q. Molecular dynamics simulation has been carried out to testify the hydrogen bond interactions between mutation sites and 2′-phosphate of NADPH. Steady-state turnover measurement results indicated that Gox2181-D43Q could use both NADH and NADPH as its coenzyme, and so could Gox2181-Q20R&D43Q. Meanwhile, compared to the wild-type enzyme, Gox2181-D43Q exhibited dramatically reduced enzymatic activity while Gox2181-Q20R&D43Q successfully retained the majority of enzymatic activity. 相似文献
90.
Boyang Cao Fangfang Yao Xiangqian Liu Lu Feng Lei Wang 《Applied and environmental microbiology》2013,79(21):6647-6654
Legionella is ubiquitous in many environments. At least 50 species and 70 serogroups of the Gram-negative bacterium have been identified. Of the 50 species, 20 are pathogenic, and Legionella pneumophila is responsible for the great majority (approximately 90%) of the Legionnaires'' disease cases that occur. Furthermore, of the 15 L. pneumophila serogroups identified, O1 alone causes more than 84% of the Legionnaires'' disease cases that occur worldwide. Rapid and reliable assays for the detection and identification of L. pneumophila in water, environmental, and clinical samples are in great demand. L. pneumophila bacteria are traditionally identified by their O antigens by immunological methods. We have recently developed an O serogroup-specific DNA microarray for the detection of all 15 distinct O-antigen forms of L. pneumophila, including serogroups O1 to O15. A total of 35 strains were used to verify the specificity of the microarray, including 15 L. pneumophila O-antigen standard reference strains and seven L. pneumophila clinical isolates as target strains, seven reference strains of other non-pneumophila Legionella species as closely related strains, and six non-Legionella bacterial species as nonrelated strains. The detection sensitivity was 1 ng of genomic DNA or 0.4 CFU/ml in water samples with filter enrichment and plate culturing. This study demonstrated that the microarray allows specific, sensitive, and reproducible detection of L. pneumophila serogroups. To the best of our knowledge, this is the first report of a microarray serotyping method for all 15 distinct O-antigen forms of L. pneumophila. 相似文献