首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   21005篇
  免费   1688篇
  国内免费   1720篇
  24413篇
  2024年   55篇
  2023年   315篇
  2022年   712篇
  2021年   1154篇
  2020年   765篇
  2019年   975篇
  2018年   912篇
  2017年   614篇
  2016年   911篇
  2015年   1339篇
  2014年   1517篇
  2013年   1579篇
  2012年   1934篇
  2011年   1699篇
  2010年   995篇
  2009年   920篇
  2008年   1088篇
  2007年   904篇
  2006年   827篇
  2005年   649篇
  2004年   508篇
  2003年   448篇
  2002年   371篇
  2001年   323篇
  2000年   321篇
  1999年   324篇
  1998年   208篇
  1997年   242篇
  1996年   191篇
  1995年   189篇
  1994年   162篇
  1993年   129篇
  1992年   181篇
  1991年   143篇
  1990年   146篇
  1989年   98篇
  1988年   90篇
  1987年   87篇
  1986年   63篇
  1985年   65篇
  1984年   43篇
  1983年   47篇
  1982年   21篇
  1981年   16篇
  1980年   14篇
  1979年   12篇
  1978年   10篇
  1969年   9篇
  1968年   8篇
  1965年   16篇
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
131.
近自然化改造作为森林新增碳汇的最有希望的选择之一,将如何通过改变林分结构影响林分生物量和生产力进而影响林分固碳能力和潜力目前尚不清楚,因此,了解近自然化改造对人工林生物量及其分配的影响,对人工林生态系统碳管理具有重要意义。以马尾松近自然化改造林(P(CN))、马尾松未改造纯林(P(CK))、杉木近自然改造林(C(CN))和杉木未改造纯林(C(CK))4种人工林为研究对象,采用样方调查和生物量实测的方法,分析4种林分生物量差异,旨在揭示近自然化改造对马尾松和杉木人工林生物量及其分配的影响。结果表明:马尾松杉木人工林近自然化改造通过调整林分结构显著提升马尾松和杉木人工林生物量和生产力,8a后马尾松和杉木林分生物量分别增加46.71%和37.24%。乔木层生物量在林分生物量总量中占主导地位(95.48%-98.82%),并对林分生态系统总生物量变化起决定性作用。林分生物量和生产力的增加主要因为近自然化改造改变了林分群落结构,进而提高了乔木层生产力。研究结果表明,合理的经营措施不仅可以改善林分结构,提升林分生产力,并可为增强植被固碳能力创造有利条件。  相似文献   
132.
Huang FS  Hu Q  Shi YL 《生理学报》1998,50(2):145-152
抗疟新药青蒿素的某些含氮衍生物在动物实验中还显示中局部麻醉效应。本工作利用分化的NC108-15细胞,在全细胞电压箝位下,比较了五种青蒿素 生物和普鲁卡因对电压门控钠流和延迟整汉钾流的作用。所观察的五种衍生物均部分可逆地和剂量依赖地抑制INa,其中含芳香环的SM541作用最强,效应与普鲁卡因的相近。300μmol/L普卡因对IK只有微弱的抑制作用;而五种衍生物在相同浓度下均明显抑制IK,并加快它的  相似文献   
133.
You J  Hu FQ  Du YZ  Yuan H 《Biomacromolecules》2007,8(8):2450-2456
Herein, polymeric micelles with glycolipid-like structure and about 40 nm diameter are prepared by self-aggregation from stearate-grafted chitosan oligosaccharides in aqueous medium. The micelles, with high degree of substitution (DS), present specific spatial structure with multiple hydrophobic "minor cores", and thus obtain excellent internalization into cancer cells and accumulation in cytoplasm. Furthermore, the micelles showed pH-sensitive properties, thus favoring intracellular delivery of encapsulated drug via endocytosis. The cell cytotoxicity of paclitaxel encapsulated in micelles was improved sharply and contributed to the increased intracellular delivery of the drug. The present micelles are a promising carrier candidate for targeting therapy of antitumor drugs with a cytoplasmic molecule target.  相似文献   
134.
用不同剂量的60Co-γ射线诱变钝顶螺旋藻Spirulinaplatensis出发株(Sp)IS-90010,筛选获得两株抗高光抑制突变株(Sp)AIp-90010和(Sp)AIp-90011,然后比较出发株和突变株的一般形态和生理生化特性。出发株和突变株的一般形态有较大的差异,与出发株相比,两个突变株藻丝体显著变短,螺旋数目大大减小。出发株是对高光敏感的品系,而突变株表现明显的抗高光抑制,13000lx光强下出发株和两个突变株的代时分别为29.4、20.8和22.2h。(Sp)AIp-90011的光合作用和呼吸作用与出发株相似,而(Sp)AIp-90010明显地表现出高光合、低呼吸作用。突变株和出发株都属于中温品系,最适温度为28℃,具有较广的温度适应范围(23~35℃)以及相同的耐盐性,但突变株的生长速率比出发株快、代时短。此外三者的蛋白质含量、氨基酸组成差别不大:(Sp)AIp-90011的可溶性多糖比出发株减少40%。而在(Sp)AIp-90010中其含量提高60%。  相似文献   
135.
Genetic manipulation of antibiotic producers, such as Streptomyces species, is a rational approach to improve the properties of biologically active molecules. However, this can be a slow and sometimes problematic process. Red/ET recombination in an Escherichia coli host has permitted rapid and more versatile engineering of geldanamycin biosynthetic genes in a complementation plasmid, which can then be readily transferred into the Streptomyces host from which the corresponding wild type gene(s) has been removed. With this rapid Red/ET recombination and gene complementation approach, efficient gene disruptions and gene replacements in the geldanamycin biosynthetic gene cluster have been successfully achieved. As an example, we describe here the creation of a ketoreductase 6 null mutation in an E. coli high-copy-number plasmid carrying gdmA2A3 from Streptomyces hygroscopicus NRRL3602 and the subsequent complementation of a gdmA2A3 deletion host with this plasmid to generate a novel geldanamycin analog.  相似文献   
136.
Virus-like particles (VLPs) are empty particles consisting of virus capsid proteins that closely resemble native virus but are devoid of the native viral nucleic acids and therefore have attracted significant attention as noninfectious vaccines. A recombinant baculovirus, vIBD-7, which encodes the structural proteins (VP2, VP3, and VP4) of infectious bursal disease virus (IBDV), produces native IBD VLPs in infected Spodoptera frugiperda insect cells. Another baculovirus, vEDLH-22, encodes VP2 that is fused with a histidine affinity-tag (VP2H) at the C-terminus. By co-infection with these two baculoviruses, hybrid VLPs with histidine tags were formed and purified by immobilized metal affinity chromatography (Hu et al., 1999). Also, we demonstrated that varying the MOI ratio of these infecting viruses altered the extent of VP2H incorporated into the particles. A dynamic mathematical model that described baculovirus infection and VLP synthesis (Hu and Bentley, 2000) was adapted here for co-infection and validated by immunofluorescence labeling. It was shown to predict the VLP composition as a dynamic function of MOI. A constraint in the VP2H content incorporated into the particles was predicted and shown by experiments. Also, the MOI ratio of both infecting viruses was shown to be the major factor influencing the composition of the hybrid particles and an important factor in determining the overall yield. ELISA results confirmed that VP2H was exhibited to a varied extent on the outer surface of the particles. This model provides insight on the use of virus co-infection in virus-mediated recombinant protein expression systems and aids in the optimization of chimeric VLP synthesis.  相似文献   
137.
Liu YQ  Hu XY  Lu T  Cheng YN  Young CY  Yuan HQ  Lou HX 《PloS one》2012,7(5):e38000
Previously, we reported that retigeric acid B (RB), a natural pentacyclic triterpenic acid isolated from lichen, inhibited cell growth and induced apoptosis in androgen-independent prostate cancer (PCa) cells. However, the mechanism of action of RB remains unclear. In this study, we found that using PC3 and DU145 cells as models, RB inhibited phosphorylation levels of IκBα and p65 subunit of NF-κB in a time- and dosage-dependent manner. Detailed study revealed that RB blocked the nuclear translocation of p65 and its DNA binding activity, which correlated with suppression of NF-κB-regulated proteins including Bcl-2, Bcl-x(L), cyclin D1 and survivin. NF-κB reporter assay suggested that RB was able to inhibit both constitutive activated-NF-κB and LPS (lipopolysaccharide)-induced activation of NF-κB. Overexpression of RelA/p65 rescued RB-induced cell death, while knockdown of RelA/p65 significantly promoted RB-mediated inhibitory effect on cell proliferation, suggesting the crucial involvement of NF-κB pathway in this event. We further analyzed antitumor activity of RB in in vivo study. In C57BL/6 mice carrying RM-1 homografts, RB inhibited tumor growth and triggered apoptosis mainly through suppressing NF-κB activity in tumor tissues. Additionally, DNA microarray data revealed global changes in the gene expression associated with cell proliferation, apoptosis, invasion and metastasis in response to RB treatment. Therefore, our findings suggested that RB exerted its anti-tumor effect by targeting the NF-κB pathway in PCa cells, and this could be a general mechanism for the anti-tumor effect of RB in other types of cancers as well.  相似文献   
138.
Gao Y  Chen S  Hu M  Hu Q  Luo J  Li Y 《PloS one》2012,7(6):e38137
Chlorpyrifos is of great environmental concern due to its widespread use in the past several decades and its potential toxic effects on human health. Thus, the degradation study of chlorpyrifos has become increasing important in recent years. A fungus capable of using chlorpyrifos as the sole carbon source was isolated from organophosphate-contaminated soil and characterized as Cladosporium cladosporioides Hu-01 (collection number: CCTCC M 20711). A novel chlorpyrifos hydrolase from cell extract was purified 35.6-fold to apparent homogeneity with 38.5% overall recovery by ammoniumsulfate precipitation, gel filtration chromatography and anion-exchange chromatography. It is a monomeric structure with a molecular mass of 38.3 kDa. The pI value was estimated to be 5.2. The optimal pH and temperature of the purified enzyme were 6.5 and 40°C, respectively. No cofactors were required for the chlorpyrifos-hydrolysis activity. The enzyme was strongly inhibited by Hg2?, Fe3?, DTT, β-mercaptoethanol and SDS, whereas slight inhibitory effects (5-10% inhibition) were observed in the presence of Mn2?, Zn2?, Cu2?, Mg2?, and EDTA. The purified enzyme hydrolyzed various organophosphorus insecticides with P-O and P-S bond. Chlorpyrifos was the preferred substrate. The Km and Vmax values of the enzyme for chlorpyrifos were 6.7974 μM and 2.6473 μmol·min?1, respectively. Both NH2-terminal sequencing and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometer (MALDI-TOF-MS) identified an amino acid sequence MEPDGELSALTQGANS, which shared no similarity with any reported organophosphate-hydrolyzing enzymes. These results suggested that the purified enzyme was a novel hydrolase and might conceivably be developed to fulfill the practical requirements to enable its use in situ for detoxification of chlorpyrifos. Finally, this is the first described chlorpyrifos hydrolase from fungus.  相似文献   
139.
Cycloocygenases 2 (COX2)-prostanoid pathway plays important and complex roles in the pathogenesis of various liver diseases. Most studies indicated that COX2-prostanoid pathway might suppress hepatic fibrogenesis by decreasing proliferation, migration, and contractility of hepatic stellate cells (HSCs). In animal model, COX2-prostanoid pathway increases portal hypertension, which can be reduced by treatment with COX2 inhibitor. In cirrhosis, COX2-prostanoid pathway may reduce formation of ascites by enhancing free water excretion, and protect gastric mucosa from ulcerative insults. Aberrant expression of COX2 has been well associated with hepatocarcinogenesis. COX2 inhibitors can effectively suppress proliferation of hepatocellular carcinoma (HCC) cells. This provided rationale for further testing COX2 inhibitors as clinical agents for HCC chemoprovention. Further studies will be needed to examine how COX2 inhibitors affect pathogenesis of various liver diseases.  相似文献   
140.
Despite the publication of several software tools for analysis of glycopeptide tandem mass spectra, there remains a lack of consensus regarding the most effective and appropriate methods. In part, this reflects problems with applying standard methods for proteomics database searching and false discovery rate calculation. While the analysis of small post-translational modifications (PTMs) may be regarded as an extension of proteomics database searching, glycosylation requires specialized approaches. This is because glycans are large and heterogeneous by nature, causing glycopeptides to exist as multiple glycosylated variants. Thus, the mass of the peptide cannot be calculated directly from that of the intact glycopeptide. In addition, the chemical nature of the glycan strongly influences product ion patterns observed for glycopeptides. As a result, glycopeptidomics requires specialized bioinformatics methods. We summarize the recent progress towards a consensus for effective glycopeptide tandem mass spectrometric analysis.  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号