全文获取类型
收费全文 | 52620篇 |
免费 | 3838篇 |
国内免费 | 13篇 |
专业分类
56471篇 |
出版年
2023年 | 356篇 |
2022年 | 497篇 |
2021年 | 961篇 |
2020年 | 812篇 |
2019年 | 920篇 |
2018年 | 1477篇 |
2017年 | 1340篇 |
2016年 | 1977篇 |
2015年 | 2781篇 |
2014年 | 2801篇 |
2013年 | 3549篇 |
2012年 | 4526篇 |
2011年 | 4219篇 |
2010年 | 2360篇 |
2009年 | 1954篇 |
2008年 | 3174篇 |
2007年 | 3045篇 |
2006年 | 2903篇 |
2005年 | 2513篇 |
2004年 | 2453篇 |
2003年 | 2258篇 |
2002年 | 2053篇 |
2001年 | 1054篇 |
2000年 | 1097篇 |
1999年 | 685篇 |
1998年 | 348篇 |
1997年 | 224篇 |
1996年 | 243篇 |
1995年 | 243篇 |
1994年 | 216篇 |
1993年 | 207篇 |
1992年 | 284篇 |
1991年 | 217篇 |
1990年 | 188篇 |
1989年 | 191篇 |
1988年 | 165篇 |
1987年 | 158篇 |
1986年 | 143篇 |
1985年 | 154篇 |
1984年 | 172篇 |
1983年 | 107篇 |
1982年 | 116篇 |
1981年 | 91篇 |
1980年 | 91篇 |
1979年 | 110篇 |
1978年 | 82篇 |
1976年 | 97篇 |
1975年 | 87篇 |
1974年 | 79篇 |
1973年 | 82篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
181.
Cairns L Cirò M Minuzzo M Morlé F Starck J Ottolenghi S Ronchi A 《Journal of cellular physiology》2003,195(1):38-49
Erythropoiesis requires the stepwise action on immature progenitors of several growth factors, including stem cell factor (SCF), interleukin 3 (IL-3), and erythropoietin (Epo). Epo is required to sustain proliferation and survival of committed progenitors and might further modulate the level of expression of several erythroid genes, including globin genes. Here we report a new SCF-dependent immortalized mouse progenitor cell line (GATA-1 ts SCF) that can also grow in either Epo or IL-3 as the sole growth factor. When grown in SCF, these cells show an "open" chromatin structure of the beta-globin LCR, but do not significantly express globin. However, Epo or IL-3 induce globin expression and are required for its maintainance. This effect of IL-3 is unexpected as IL-3 was previously reported either to be unable to induce hemoglobinization, or even to antagonize it. This suggests that GATA-1 ts SCF cells may have progressed to a stage in which globin genes are already poised for expression and only require signal(s) that can be elicited by either Epo or IL-3. Through the use of inhibitors, we suggest that p38 may be one of the molecules modulating induction and maintenance of globin expression. 相似文献
182.
183.
A simple and efficient method for the oligodeoxyribonucleotide-directed mutagenesis of double-stranded plasmid DNA. 总被引:4,自引:0,他引:4
A method for the oligodeoxyribonucleotide-directed mutagenesis of double-stranded DNA without the necessity for phenotypic selection is described. Plasmids denatured with alkali and purified by adsorption to and elution from nitrocellulose have single-stranded regions where primers can hybridize and serve as templates for a T7 DNA polymerase-catalyzed synthesis of complementary mutant DNA strands. When this procedure was carried out such that the original nonmutant strand contained uracil [method of Kunkel, Proc. Natl. Acad. Sci. USA 82(1985)488-492], mutation frequencies of between 30% and 40% were obtained. The technique has been used to generate mutant genes in plasmids of a wide variety of sizes. The largest plasmid manipulated and successfully mutagenized was 22 kb. The method is rapid and efficient and is not dependent upon either f1 phage vectors or the presence of restriction sites in the vicinity of the sequence targeted for mutation. 相似文献
184.
Functional interaction between PARP-1 and PARP-2 in chromosome stability and embryonic development in mouse 总被引:8,自引:0,他引:8
Ménissier de Murcia J Ricoul M Tartier L Niedergang C Huber A Dantzer F Schreiber V Amé JC Dierich A LeMeur M Sabatier L Chambon P de Murcia G 《The EMBO journal》2003,22(9):2255-2263
The DNA damage-dependent poly(ADP-ribose) polymerases, PARP-1 and PARP-2, homo- and heterodimerize and are both involved in the base excision repair (BER) pathway. Here, we report that mice carrying a targeted disruption of the PARP-2 gene are sensitive to ionizing radiation. Following alkylating agent treatment, parp-2(-/-)-derived mouse embryonic fibroblasts exhibit increased post-replicative genomic instability, G(2)/M accumulation and chromosome mis-segregation accompanying kinetochore defects. Moreover, parp-1(-/-)parp-2(-/-) double mutant mice are not viable and die at the onset of gastrulation, demonstrating that the expression of both PARP-1 and PARP-2 and/or DNA-dependent poly(ADP-ribosyl) ation is essential during early embryogenesis. Interestingly, specific female embryonic lethality is observed in parp-1(+/-)parp-2(-/-) mutants at E9.5. Meta phase analyses of E8.5 embryonic fibroblasts highlight a specific instability of the X chromosome in those females, but not in males. Together, these results support the notion that PARP-1 and PARP-2 possess both overlapping and non-redundant functions in the maintenance of genomic stability. 相似文献
185.
Milton Maciel Jr. Fábia da Silva Pereira Cruz Marli Tenório Cordeiro Márcia Archer da Motta Klécia Marília Soares de Melo Cassemiro Rita de Cássia Carvalho Maia Regina Célia Bressan Queiroz de Figueiredo Ricardo Galler Marcos da Silva Freire Joseph Thomas August Ernesto T. A. Marques Rafael Dhalia 《PLoS neglected tropical diseases》2015,9(4)
Attenuated yellow fever (YF) virus 17D/17DD vaccines are the only available protection from YF infection, which remains a significant source of morbidity and mortality in the tropical areas of the world. The attenuated YF virus vaccine, which is used worldwide, generates both long-lasting neutralizing antibodies and strong T-cell responses. However, on rare occasions, this vaccine has toxic side effects that can be fatal. This study presents the design of two non-viral DNA-based antigen formulations and the characterization of their expression and immunological properties. The two antigen formulations consist of DNA encoding the full-length envelope protein (p/YFE) or the full-length envelope protein fused to the lysosomal-associated membrane protein signal, LAMP-1 (pL/YFE), aimed at diverting antigen processing/presentation through the major histocompatibility complex II precursor compartments. The immune responses triggered by these formulations were evaluated in H2b and H2d backgrounds, corresponding to the C57Bl/6 and BALB/c mice strains, respectively. Both DNA constructs were able to induce very strong T-cell responses of similar magnitude against almost all epitopes that are also generated by the YF 17DD vaccine. The pL/YFE formulation performed best overall. In addition to the T-cell response, it was also able to stimulate high titers of anti-YF neutralizing antibodies comparable to the levels elicited by the 17DD vaccine. More importantly, the pL/YFE vaccine conferred 100% protection against the YF virus in intracerebrally challenged mice. These results indicate that pL/YFE DNA is an excellent vaccine candidate and should be considered for further developmental studies. 相似文献
186.
Julio Cesar Madureira de-Freitas-Junior Sandra Carvalho Ana M. Dias Patrícia Oliveira Joana Cabral Raquel Seruca Carla Oliveira José Andrés Morgado-Díaz Celso A. Reis Salomé S. Pinho 《PloS one》2013,8(11)
Changes in glycosylation are considered a hallmark of cancer, and one of the key targets of glycosylation modifications is E-cadherin. We and others have previously demonstrated that E-cadherin has a role in the regulation of bisecting GlcNAc N-glycans expression, remaining to be determined the E-cadherin-dependent signaling pathway involved in this N-glycans expression regulation. In this study, we analysed the impact of E-cadherin expression in the activation profile of receptor tyrosine kinases such as insulin receptor (IR) and IGF-I receptor (IGF-IR). We demonstrated that exogenous E-cadherin expression inhibits IR, IGF-IR and ERK 1/2 phosphorylation. Stimulation with insulin and IGF-I in MDA-MD-435 cancer cells overexpressing E-cadherin induces a decrease of bisecting GlcNAc N-glycans that was accompanied with alterations on E-cadherin cellular localization. Concomitantly, IR/IGF-IR signaling activation induced a mesenchymal-like phenotype of cancer cells together with an increased tumor cell invasion capability. Altogether, these results demonstrate an interplay between E-cadherin and IR/IGF-IR signaling as major networking players in the regulation of bisecting N-glycans expression, with important effects in the modulation of epithelial characteristics and tumor cell invasion. Here we provide new insights into the role that Insulin/IGF-I signaling play during cancer progression through glycosylation modifications. 相似文献
187.
Felipe Lira Pedro S. Perez José A. Baranauskas Sérgio R. Nozawa 《Applied and environmental microbiology》2013,79(10):3156-3159
Antimicrobial resistance is a persistent problem in the public health sphere. However, recent attempts to find effective substitutes to combat infections have been directed at identifying natural antimicrobial peptides in order to circumvent resistance to commercial antibiotics. This study describes the development of synthetic peptides with antimicrobial activity, created in silico by site-directed mutation modeling using wild-type peptides as scaffolds for these mutations. Fragments of antimicrobial peptides were used for modeling with molecular modeling computational tools. To analyze these peptides, a decision tree model, which indicated the action range of peptides on the types of microorganisms on which they can exercise biological activity, was created. The decision tree model was processed using physicochemistry properties from known antimicrobial peptides available at the Antimicrobial Peptide Database (APD). The two most promising peptides were synthesized, and antimicrobial assays showed inhibitory activity against Gram-positive and Gram-negative bacteria. Colossomin C and colossomin D were the most inhibitory peptides at 5 μg/ml against Staphylococcus aureus and Escherichia coli. The methods described in this work and the results obtained are useful for the identification and development of new compounds with antimicrobial activity through the use of computational tools. 相似文献
188.
Erpan Ahat Sarah Bui Jianchao Zhang Felipe da Veiga Leprevost Lisa Sharkey Whitney Reid Alexey I. Nesvizhskii Henry L. Paulson Yanzhuang Wang 《The Journal of biological chemistry》2022,298(8)
Recent studies demonstrated that the Golgi reassembly stacking proteins (GRASPs), especially GRASP55, regulate Golgi-independent unconventional secretion of certain cytosolic and transmembrane cargoes; however, the underlying mechanism remains unknown. Here, we surveyed several neurodegenerative disease–related proteins, including mutant huntingtin (Htt-Q74), superoxide dismutase 1 (SOD1), tau, and TAR DNA–binding protein 43 (TDP-43), for unconventional secretion; our results show that Htt-Q74 is most robustly secreted in a GRASP55-dependent manner. Using Htt-Q74 as a model system, we demonstrate that unconventional secretion of Htt is GRASP55 and autophagy dependent and is enhanced under stress conditions such as starvation and endoplasmic reticulum stress. Mechanistically, we show that GRASP55 facilitates Htt secretion by tethering autophagosomes to lysosomes to promote autophagosome maturation and subsequent lysosome secretion and by stabilizing p23/TMED10, a channel for translocation of cytoplasmic proteins into the lumen of the endoplasmic reticulum–Golgi intermediate compartment. Moreover, we found that GRASP55 levels are upregulated by various stresses to facilitate unconventional secretion, whereas inhibition of Htt-Q74 secretion by GRASP55 KO enhances Htt aggregation and toxicity. Finally, comprehensive secretomic analysis identified novel cytosolic cargoes secreted by the same unconventional pathway, including transgelin (TAGLN), multifunctional protein ADE2 (PAICS), and peroxiredoxin-1 (PRDX1). In conclusion, this study defines the pathway of GRASP55-mediated unconventional protein secretion and provides important insights into the progression of Huntington’s disease. 相似文献
189.
Julie A. Hawkins Laura White Olascoaga Colin E. Hughes José-Luis R. Contreras Jiménez Pedro Mercado Ruaro 《Plant Systematics and Evolution》1999,216(1-2):49-68
Morphometric, cytogenetic, geographical and ecological evidence for hybridization betweenParkinsonia aculeata andCercidium praecox is presented. Morphometric investigation using the character count procedure and cytogenetic observations confirm hybrid status. All diagnostic morphometric characters were intermediate in the hybrid. Both parents (2n = 28) show regular tetrad formation and pollen fertility greater than 94%. Hybrids have a chromosome number of 2n = 28 or 2n = 30, and display meiotic abnormalities including lagging chromosomes and micronucleus formation; less than 21% of hybrid pollen was fertile. Ecological and geographical information suggests that hybridization is occurring at increasing frequency due to the expanding range ofP. aculeata associated with cultivation as an ornamental, coupled with ecological disturbance and weediness, and the cultivation ofC. praecox and hybrids as fodder, ornamental and shade trees. Hybrid fertility and phenological observations, in conjunction with F-weighted principal component analysis, suggest that the progeny of F1 hybrids are established. The hybrid is formally described asP. ×carterae. 相似文献
190.
Chern-En Chiang Lisa Naditch-Br?lé Sandrine Brette José Silva-Cardoso Habib Gamra Jan Murin Oleg J. Zharinov Philippe Gabriel Steg 《PloS one》2016,11(1)