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61.
从江浙蝮蛇中分离纯化的碱性磷脂酶A2在pH9.5,0.05mmol/LCHES缓冲液中,用汽相悬滴扩散的方法,获得了适用于高分辨率X射线结构分析的单晶.经X200B面探测器分析,表明该晶体属于正交晶系,P2I2I2I空间群,晶胞参数为a=97.13,b=103.69,c=23.27.并收集了一套衍射数据,独立衍射点数12001个,数据完整度为86.2%,Rmerge为0.0459.最高分辨率达2.0,根据分子量与晶胞体积估算,一个不对称单位含两个分子. 相似文献
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Meng Sun Jing Ren Hui Du Yanmin Zhang Jie Zhang Sicen Wang Langchong He 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(28):2712-2718
We have developed an online analytical method that combines A431 cell membrane chromatography (A431/CMC) with high performance liquid chromatography and mass spectrometry (LC/MS) for identifying active components from Radix Caulophylli acting on human EGFR. Retention fractions on A431/CMC model were captured onto an enrichment column and the components were directly analyzed by combining a 10-port column switcher with an LC/MS system for separation and preliminary identification. Using Sorafenib tosylate as a positive control, taspine and caulophine from Radix Caulophylli were identified as the active molecules which could act on the EGFR. This A431/CMC-online-LC/MS method can be applied for screening active components acting on EGFR from traditional Chinese medicines exemplified by Radix Caulophylli and will be of great utility in drug discovery using natural medicinal herbs as a source of novel compounds. 相似文献
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Hang Yang Huijun Yuan Xiaohui Zhao Meng Xun Shangrui Guo Nan Wang Bing Liu Hongliang Wang 《中国病毒学》2022,37(3):380-389
The recent COVID-19 pandemic poses a global health emergency. Cellular entry of the causative agent SARS-CoV-2 is mediated by its spike protein interacting with cellular receptor-human angiotensin converting enzyme 2 (ACE2). Here, by using lentivirus based pseudotypes bearing spike protein, we demonstrated that entry of SARS-CoV-2 into host cells was dependent on clathrin-mediated endocytosis, and phosphoinositides played essential roles during this process. In addition, we showed that the intracellular domain and the catalytic activity of ACE2 were not required for efficient virus entry. Finally, we showed that the current predominant Delta variant, although with high infectivity and high syncytium formation, also entered cells through clathrin-mediated endocytosis. These results provide new insights into SARS-CoV-2 cellular entry and present proof of principle that targeting viral entry could be an effective way to treat different variant infections. 相似文献
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The p38 mitogen-activated protein kinase (MAPK) is involved in various processes, including stress responses, development, and differentiation. However, little information on p38 MAPK in insects is available. In this study, a p38 MAPK gene, Accp38b, was isolated from Apis cerana cerana and characterized. The quantitative real-time PCR (Q-PCR) analysis revealed that Accp38b was induced by multiple stressors. Notably, the expression of Accp38b was relatively higher in the pupae phase than in other developmental phases. During the pupae phase, Accp38b expression was higher in the thorax than in the head and abdomen and higher in the fat body than in the muscle and midgut. Immunohistochemisty showed significant positive staining of Accp38b in sections from the brain, eyes, fat body, and midgut of A. cerana cerana. These results suggest that Accp38b may play a crucial role in stress responses and have multiple aspects function during development. 相似文献
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Renal cell carcinoma (RCC) is one of the most common malignancies in the urinary system. Due to the lack of early symptoms, diagnosis of RCC usually occurs at late stages or after cancer metastasis leading to poor prognosis. Therefore, it is crucial to study early molecular mechanisms and biomarkers. Previous studies have suggested that microRNAs are involved in RCC initiation and development, making them a good candidate for early diagnosis and therapy. MiR146b-5P plays important roles in the progression of multiple cancers including thyroid cancer, pancreatic cancer, cervical cancer. However, it is not clear whether and how miR146b-5P is involved in RCC. In this study, we aimed to investigate the function of miR146b-5P in RCC. We examined the expression levels of miR146b-5p in renal cancer tissue and cell lines. We also explored the effects of blocking miR146b-5p in renal tumor growth and inflammatory signaling. Finally, we determined if miR146b-5p regulates tumorigenesis through TRAF6. We found that miR146b-5p levels were significantly increased in renal cancer tissue and renal cancer cells. Blocking miR146b-5p suppressed renal tumor growth and enhanced inflammatory response through increased TRAF6 expression. These effects were eliminated in TRAF6 knockout mice. Our results suggest that enhanced miR146b-5p expression may be a biomarker for RCC and modulating miR146b-5p and TRAF6 levels represent a potential therapeutic strategy for RCC. 相似文献
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用改良的MTT法测定rhG-CSF活性 总被引:1,自引:1,他引:1
MTT测定法是根据线粒体脱氢酶催化MTT形成蓝色甲■的多少来检测活细胞数和功能状态的,但原始方法中存在着一些问题,如敏感性偏低、有机溶剂产生蛋白质沉淀以及产物的溶解度偏低等。为了摸索测定 rhG-CSF活性的最适条件,我们以 NFS-60细胞为对象,比较了多种溶解缓冲液,并且对细胞数、MTT浓度及保留时间、溶解液用量等条件进行了选择。结果表明,DMF-20%SDS和 20%SDS的效果最好,测定时细胞数为每孔 1000个细胞,所加 MTT浓度为 1mg/ml,保留时间为 4 h,溶解液的用量为每孔 100μl。 相似文献