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991.
992.
Subnuclear Trafficking of Glucocorticoid Receptors In Vitro: Chromatin Recycling and Nuclear Export 总被引:1,自引:1,他引:0 下载免费PDF全文
We have used digitonin-permeabilized cells to examine in vitro nuclear export of glucocorticoid receptors (GRs). In situ biochemical extractions in this system revealed a distinct subnuclear compartment, which collects GRs that have been released from chromatin and serves as a nuclear export staging area. Unliganded nuclear GRs within this compartment are not restricted in their subnuclear trafficking as they have the capacity to recycle to chromatin upon rebinding hormone. Thus, GRs that release from chromatin do not require transit through the cytoplasm to regain functionality. In addition, chromatin-released receptors export from nuclei of permeabilized cells in an ATP- and cytosol-independent process that is stimulated by sodium molybdate, other group VI-A transition metal oxyanions, and some tyrosine phosphatase inhibitors. The stimulation of in vitro nuclear export by these compounds is not unique to GR, but is restricted to other proteins such as the 70- and 90-kD heat shock proteins, hsp70 and hsp90, respectively, and heterogeneous nuclear RNP (hnRNP) A1. Under analogous conditions, the 56-kD heat shock protein, hsp56, and hnRNP C do not export from nuclei of permeabilized cells. If tyrosine kinase inhibitors genistein and tyrphostin AG126 are included to prevent increased tyrosine phosphorylation, in vitro nuclear export of GR is inhibited. Thus, our results are consistent with the involvement of a phosphotyrosine system in the general regulation of nuclear protein export, even for proteins such as GR and hnRNP A1 that use distinct nuclear export pathways. 相似文献
993.
Rotavirus RNA polymerase requires the core shell protein to synthesize the double-stranded RNA genome. 总被引:11,自引:6,他引:5 下载免费PDF全文
Rotavirus cores contain the double-stranded RNA (dsRNA) genome, RNA polymerase VP1, and guanylyltransferase VP3 and are enclosed within a lattice formed by the RNA-binding protein VP2. Analysis of baculovirus-expressed core-like particles (CLPs) has shown that VP1 and VP2 assemble into the simplest core-like structures with replicase activity and that VP1, but not VP3, is essential for replicase activity. To further define the role of VP1 and VP2 in the synthesis of dsRNA from viral mRNA, recombinant baculoviruses containing gene 1 (rBVg1) and gene 2 (rBVg2) of SA11 rotavirus were generated and used to express recombinant VP1 (rVP1) and rVP2, respectively. After purification, the proteins were assayed individually and together for the ability to catalyze the synthesis of dsRNA in a cell-free replication system. The results showed that dsRNA was synthesized only in assays containing rVP1 and rVP2, thus establishing that both proteins are essential for replicase activity. Even in assays containing a primer-linked mRNA template, neither rVP1 nor rVP2 alone directed RNA synthesis. Characterization of the cis-acting replication signals in mRNA recognized by the replicase of rVP1 and rVP2 showed that they were the same as those recognized by the replicase of virion-derived cores, thus excluding a role for VP3 in recognition of the mRNA template by the replicase. Analysis of RNA-protein interactions indicated that the mRNA template binds strongly to VP2 in replicase assays but that the majority of the dsRNA product neither is packaged nor stably associates with VP2. The results of replicase assays performed with mutant VP2 containing a deletion in its RNA-binding domain suggests that the essential role for VP2 in replication is linked to the protein's ability to bind the mRNA template for minus-strand synthesis. 相似文献
994.
Possible role of macrophage-derived soluble mediators in the pathogenesis of encephalomyocarditis virus-induced diabetes in mice. 总被引:4,自引:1,他引:3 下载免费PDF全文
K Hirasawa H S Jun K Maeda Y Kawaguchi S Itagaki T Mikami H S Baek K Doi J W Yoon 《Journal of virology》1997,71(5):4024-4031
Pancreatic islets from DBA/2 mice infected with the D variant of encephalomyocarditis (EMC-D) virus revealed lymphocytic infiltration with moderate to severe destruction of pancreatic beta cells. Our previous studies showed that the major population of infiltrating cells at the early stages of infection is macrophages. The inactivation of macrophages prior to viral infection resulted in the prevention of diabetes, whereas activation of macrophages prior to viral infection resulted in the enhancement of beta-cell destruction. This investigation was initiated to determine whether macrophage-produced soluble mediators play a role in the destruction of pancreatic beta cells in mice infected with a low dose of EMC-D virus. When we examined the expression of the soluble mediators interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and inducible nitric oxide synthase (iNOS) in the pancreatic islets, we found that these mediators were clearly expressed at an early stage of insulitis and that this expression was evident until the development of diabetes. We confirmed the expression of these mediators by in situ hybridization with digoxigenin-labelled RNA probes or immunohistochemistry in the pancreatic islets. Mice treated with antibody against IL-1beta or TNF-alpha or with the iNOS inhibitor aminoguanidine exhibited a significant decrease in the incidence of diabetes. Mice treated with a combination of anti-IL-1beta antibody, anti-TNF-alpha antibody, and aminoguanidine exhibited a greater decrease in the incidence of disease than did mice treated with one of the antibodies or aminoguanidine. On the basis of these observations, we conclude that macrophage-produced soluble mediators play an important role in the destruction of pancreatic beta cells, resulting in the development of diabetes in mice infected with a low dose of EMC-D virus. 相似文献
995.
将ADH2基因的UAS与带有不同长度缺失上游区的SUC3基因融合,构建成4种具有不同融合启动子的SUC2基因的表达质粒YRD1101.YFD110△9.YFD110△17、YFD110△11。将这些质粒及对照表达质粒YFD26△1.YFD25转化酵母菌Y33,在阻遏与去阻遏培养条件下,对各种转化子所产生的蔗糖酶进行了活性测定和组分分析。结果表明:在葡萄糖去阻遏生长条件下,YFD110△1的启动子组合中UASsuc2和UASADH2对SUC2基因的表达有协同激活作用。在阻遏条件下Y33/YFD110△1与Y33/YFD110△9、Y33/YFD26△1、Y33/YFD25一样,均表达很低的糖基化蔗糖酶,3种去阻遏培养条件比较说明,在低糖培养基中对糖基化蔗糖酶表达的去阻遏效果最佳 相似文献
996.
A novel cytokine fusion protein was constructed by fusing granulocyte macrophage colony stimulat-ing factor (GM-CSF) with monocyte chemotactic activating factor (MCAF), which acts as a factor directing effector cells (monocytes) to a target site. The recombinant human GM-CSF/MCAF fusion protein could sustain the growth of GM-CSF-dependent cell line TF1 and was chemotactic for monocytes. The in vitro antitumor effect showed that rhGM-CSF/MCAF could activate monocytes to inhibit the growth of several human tumor cell lines, including a promyelocyte leukemia cell line HL-60, a lung adenocarcinoma cell line A549, a hepatoma cell line SMMC-7721 and a melanoma cell line Bowes. Furthermore, the cytotoxicity of monocytes activated by rhGM-CSF/MCAF against HL-60 and A549 was greater than that activated by GM-CSF or MCAF alone, even greater than that activated by a combina-tion of GM-CSF and MCAF, suggesting that the fusion protein has synergistic or enhanced effects. The in vivo anti-tumor effect indicated that 相似文献
997.
Construction of a trivalent candidate Shigella vaccine strain with host-vector balanced-lethal system 总被引:1,自引:0,他引:1
A trivalent liveShigella vaccine candidate FSD01 against S.flexneri 2a, S.sonnei and S.dysenteriae I was constructed. This candidate strain was based on the S.flexneri 2a vaccine T32. By homologous recombination exchange, the chromosomalasd gene of T32 was site-specifically inactivated, resulting in the strain unable to grow normally in LB broth, while anotherasd gene of S.mutans was employed to construct an Asd+ complementary vector. This combination ofasd
- host/Asd+ vector formed a balanced-lethal expression system in T32 strain. By use of this system, two important protective antigen
genes coding for S.sonnei Form I antigen and Shiga toxin B subunit were cloned and expressed in T32, which led to the construction of trivalent candidate
vaccine FSD01. Experimental results showed that this strain was genetically stable, but its recombinant plasmid was non-resistant.
Moreover, it was able to effectively express trivalent antigens in one host and induce protective responses in mice against
the challenges of the above threeShigella strains. 相似文献
998.
YANG Jianbo WU Lijun LI Li WU JiadaoYU Zengliang XU Zhihong 《中国科学:生命科学英文版》1997,40(1):107-112
While M13mp18 double-stranded DNA was irradiated with ion beam, and transfected intoE. coli JM103, a decrease of transfecting activity was discovered. The lacZ- mutation frequency at 20% survival could reach (3.6–16.8) × 104, about 2, 3–10 times that of unirradiated M13DNA. Altogether, 27 IacZ-mutants were selected, 10 of which were used for sequencing. 7 of the sequenced mutants show base changes in 250-bp region
examined (the remaining 3 mutants probably have base changes outside the regions sequenced). 5 of the base-changed mutants
contain more than one mutational base sites (some of them even have 5–6 mutational base sites in 250-bp region examined);
this dense distribution of base changes in polysites has seldom been seen in X-rays, Y-rays or UV induced DNA mutations. Our
experiments also showed that the types of base changes include transitions(50%), transversions (45%) and deletion (5%); no
addition or duplication was observed. The transitions were mainly C→T and A→G; the transversions were mainly C→A and C→G.
The mutations involving cytosine residue (in the template strand) constitute about 60% of all the base changes observed. In
comparison with the surrounding sequences of mutational base sites, the base located between TG and CT is found to be easily
substituted. 相似文献
999.
Sequence variation and genetic diversity in the giant panda 总被引:3,自引:0,他引:3
ZHANG YapingOliver A. RyderFAN Zhiyong ZHANG HemingHE TingmeiHE Guangxin ZHANG Anju FEI LisongZHONG Shunlong CHEN HongZHANG Chenglin YANG Minghai ZHU Feibing PENG Zhenxin PU Tianchun CHEN Yucun YAO OMinda GUO Wei 《中国科学:生命科学英文版》1997,40(2):210-216
About 336–444 bp mitochondrial D-loop region and tRNA gene were sequenced for 40 individuals of the giant panda which were
collected from Mabian, Meigu, Yuexi, Baoxing, Pingwu, Qingchuan, Nanping and Baishuijiang, respectively. 9 haplotypes were
found in 21 founders. The results showed that the giant panda has low genetic variations, and that there is no notable genetic
isolation among geographical populations. The ancestor of the living giant panda population perhaps appeared in the late Pleistocene,
and unfortunately, might have suffered bottleneck attacks. Afterwards, its genetic diversity seemed to recover to some extent.
Project supported by the “8.5” Key Project of Chinese Academy of Sciences, the Chairman Foundation of Chinese Academy of Sciences,
K. C. Wang Education Foundation, the Applied Basic Research Foundation of Yunnan, the National Natural Science Foundation
of China, the Special Foundation for Returned Chinese Scientists, and Zoological Society of San Diego. 相似文献
1000.
母兔配种后10小时血清中若干生理指标与子代性比的相关 总被引:1,自引:0,他引:1
测定日本大耳白兔母兔配种后10小时血清中的10项生理指标,并与母兔所产每窝仔兔的性比(雄性个体所占比率)进行对应分析(窝仔数<6的数据未参与此项分析)。结果表明:母兔血清中FSH、T(睾酮)、 Na+和M2+的浓度在高、低两个性比组间有显著差异, T3(三碘甲状腺原氨酸)的差异接近显著水平(P<0.1);并且,FSH和T3与子代性比里显著负相关(r分别为-0.50和-0.46),Mg2+与子代性比里显著正相关(r=0.39);同时,Mg2+/Ca2+、Mg2+×Na+、Mg2+×K+和T3×FSH与子代性比的相关分别为0.40、0.43、0.39和-0.53(P<0.05)。 相似文献