全文获取类型
收费全文 | 14730篇 |
免费 | 1523篇 |
国内免费 | 1486篇 |
专业分类
17739篇 |
出版年
2024年 | 45篇 |
2023年 | 215篇 |
2022年 | 525篇 |
2021年 | 833篇 |
2020年 | 608篇 |
2019年 | 767篇 |
2018年 | 757篇 |
2017年 | 551篇 |
2016年 | 733篇 |
2015年 | 928篇 |
2014年 | 1132篇 |
2013年 | 1125篇 |
2012年 | 1315篇 |
2011年 | 1202篇 |
2010年 | 715篇 |
2009年 | 687篇 |
2008年 | 745篇 |
2007年 | 642篇 |
2006年 | 514篇 |
2005年 | 465篇 |
2004年 | 479篇 |
2003年 | 492篇 |
2002年 | 438篇 |
2001年 | 375篇 |
2000年 | 284篇 |
1999年 | 235篇 |
1998年 | 155篇 |
1997年 | 114篇 |
1996年 | 120篇 |
1995年 | 75篇 |
1994年 | 77篇 |
1993年 | 61篇 |
1992年 | 59篇 |
1991年 | 64篇 |
1990年 | 52篇 |
1989年 | 34篇 |
1988年 | 28篇 |
1987年 | 27篇 |
1986年 | 18篇 |
1985年 | 24篇 |
1984年 | 5篇 |
1983年 | 6篇 |
1982年 | 5篇 |
1981年 | 4篇 |
1980年 | 2篇 |
1979年 | 1篇 |
1950年 | 1篇 |
排序方式: 共有10000条查询结果,搜索用时 9 毫秒
71.
ABSTRACT: BACKGROUND: Lipase from Rhizopus chinensis is a versatile biocatalyst for various bioconversions and has been expressed at high-level in Pichia pastoris. However, the use of R. chinensis lipase in industrial applications is restricted by its low thermostability. Directed evolution has been proven to be a powerful and efficient protein engineering tool for improvement of biocatalysts. The present work describes improvement of the thermostability of R. chinensis lipase by directed evolution using P. pastoris as the host. RESULTS: An efficient, fast and highly simplified method was developed to create a mutant gene library in P. pastoris based on in vivo recombination, whose recombination efficiency could reach 2.3 x 105 /mug DNA. The thermostability of r27RCL was improved significantly by two rounds of error-prone PCR and two rounds of DNA shuffling in P. pastoris. The S4-3 variant was found to be the most thermostable lipase, under the conditions tested. Compared with the parent, the optimum temperature of S4-3 was two degrees higher, Tm was 22 degrees higher and half-lives at 60degreesC and 65degreesC were 46- and 23- times longer. Moreover, the catalytic efficiency kcat/Km of S4-3 was comparable to the parent. Stabilizing mutations probably increased thermostability by increasing the hydrophilicity and polarity of the protein surface and creating hydrophobic contacts inside the protein. CONCLUSIONS: P. pastoris was shown to be a valuable cell factory to improve thermostability of enzymes by directed evolution and it also could be used for improving other properties of enzymes. In this study, by using P. pastoris as a host to build mutant pool, we succeeded in obtaining a thermostable variant S4-3 without compromising enzyme activity and making it a highly promising candidate for future applications at high temperatures. 相似文献
72.
A purified polysaccharide ACDP-2 was isolated from water extract of the stems of Cistanche deserticola. Chemical and spectroscopic analyses indicated that ACDP-2 is a highly branched arabinogalactan polymer that composes of linked d-galactopyranose and d-glucopyranose, which contains predominantly a branching point at the 6-carbon. The branched side-chains compose of terminal-, 1,5-, and 1,3,5-linked arabinofuranosyl residues. ACDP-2 showed an effect in stimulating the immune response, which when applied onto the cultured mouse lymphocytes induced the cell proliferation in a dose-dependent manner. 相似文献
73.
The splicing of many alternative exons in the precursor messenger RNA (pre-mRNA) is regulated by extracellular factors but the underlying molecular bases remain unclear. Here we report the differential regulation of Bcl-x pre-mRNA splicing by extracellular factors and their distinct requirements for pre-mRNA elements. In K562 leukemia cells, treatment with interleukin-6 (IL-6) or granulocyte-macrophage colony stimulating factor (GM-CSF) reduced the proportion of the Bcl-xL variant mRNA while treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) had no effect. In U251 glioma cells, however, TPA efficiently increased the Bcl-xL level. These regulations were also seen for a transfected splicing reporter mini-gene. Further analyses of deletion mutants indicate that nucleotides 1-176 of the downstream intron are required for the IL-6 effect, whereas additional nucleotides 177-284 are essential for the GM-CSF effect. As for the TPA effect, only nucleotides 1-76 are required in the downstream intron. Thus, IL-6, GM-CSF and TPA differentially regulate Bcl-x splicing and require specific intronic pre-mRNA sequences for their respective effects. 相似文献
74.
Yi Tan Lingchao Miao Jianbo Xiao Wai San Cheang 《Current issues in molecular biology》2022,44(5):2175
The potential anti-diabetic effect of resveratrol derivative, 3,3′,4,5′-tetramethoxy-trans-stilbene (3,3′,4,5′-TMS) and its underlying mechanism in high glucose (HG) and dexamethasone (DXMS)-stimulated insulin-resistant HepG2 cells (IR-HepG2) were investigated. 3,3′,4,5′-TMS did not reduce the cell viability of IR-HepG2 cells at the concentrations of 0.5–10 µM. 3,3′,4,5′-TMS increased the potential of glucose consumption and glycogen synthesis in a concentration-dependent manner in IR-HepG2 cells. 3,3′,4,5′-TMS ameliorated insulin resistance by enhancing the phosphorylation of glycogen synthase kinase 3 beta (GSK3β), inhibiting phosphorylation of insulin receptor substrate-1 (IRS-1), and activating phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in IR-HepG2 cells. Furthermore, 3,3′,4,5′-TMS significantly suppressed levels of reactive oxygen species (ROS) with up-regulation of nuclear factor erythroid 2-related factor 2 (Nrf2) expression. To conclude, the beneficial effect of 3,3′,4,5′-TMS against insulin resistance to increase glucose consumption and glycogen synthesis was mediated through activation of IRS/PI3K/Akt signaling pathways in the IR-HepG2 cells, accomplished with anti-oxidative activity through up-regulation of Nrf2. 相似文献
75.
76.
77.
Jiajia Ma Zijia Ren Yang Ma Lu Xu Ying Zhao Chaogu Zheng Yinghui Fang Ting Xue Baolin Sun Weihua Xiao 《The Journal of biological chemistry》2009,284(50):34600-34606
78.
贵州产五步蛇蛇毒磷脂酶A2的抗凝血及抗血栓作用研究 总被引:1,自引:0,他引:1
目的研究贵州产五步蛇蛇毒磷脂酶A2的抗凝血及抗血栓作用。方法对大鼠静脉注射不同剂量的PLA2,测定大鼠的凝血时间、凝血酶原时间、抗凝效价和血栓抑制率。结果PLA2浓度达到1.0mg/kg以上,能显著延长大鼠凝血时间及凝血酶原时间,其抗凝效价相当于肝素钠的24.8%~31.5%。PLA2能显著抑制大鼠实验性动脉及静脉血栓的形成。结论贵州产五步蛇PLA2具有较强的抗凝血和抗血栓作用。 相似文献
79.
80.
Quercetin and other flavonoids have been reported to exhibit both antioxidant and pro‐oxidant properties. Most studies about the pro‐oxidative ability were conducted in the presence of metal ions, and the essential functional moiety of quercetin responsible for the pro‐oxidative effect is still unclear. In this study, we evaluated the pro‐oxidative abilities in the absence of metal ions of two quercetin derivatives, i.e., quercetin‐3′‐O‐β‐D ‐glucoside ( 1 ) and quercetin‐3‐O‐β‐D ‐glucoside ( 2 ), by assessing DNA cleavage and HO.‐radical production. The binding mode between these compounds and DNA was studied by fluorescence and viscometric titrations. The results showed that 1 can efficiently induce oxidative damage to plasmid DNA, while 2 shows poor activity. Both 1 and 2 bind to DNA via groove‐binding. These results proved that the α‐hydroxy‐β‐oxo‐α,β‐enone moiety contributes to the pro‐oxidative activity of quercetin. 相似文献