全文获取类型
收费全文 | 48678篇 |
免费 | 3908篇 |
国内免费 | 2890篇 |
专业分类
55476篇 |
出版年
2024年 | 93篇 |
2023年 | 553篇 |
2022年 | 1329篇 |
2021年 | 2354篇 |
2020年 | 1464篇 |
2019年 | 1786篇 |
2018年 | 1785篇 |
2017年 | 1325篇 |
2016年 | 1895篇 |
2015年 | 2818篇 |
2014年 | 3320篇 |
2013年 | 3631篇 |
2012年 | 4357篇 |
2011年 | 3895篇 |
2010年 | 2410篇 |
2009年 | 2065篇 |
2008年 | 2361篇 |
2007年 | 2091篇 |
2006年 | 1919篇 |
2005年 | 1590篇 |
2004年 | 1426篇 |
2003年 | 1180篇 |
2002年 | 1074篇 |
2001年 | 975篇 |
2000年 | 927篇 |
1999年 | 892篇 |
1998年 | 542篇 |
1997年 | 455篇 |
1996年 | 488篇 |
1995年 | 456篇 |
1994年 | 455篇 |
1993年 | 305篇 |
1992年 | 445篇 |
1991年 | 339篇 |
1990年 | 371篇 |
1989年 | 314篇 |
1988年 | 244篇 |
1987年 | 202篇 |
1986年 | 183篇 |
1985年 | 150篇 |
1984年 | 142篇 |
1983年 | 107篇 |
1982年 | 85篇 |
1981年 | 63篇 |
1979年 | 76篇 |
1978年 | 55篇 |
1977年 | 53篇 |
1975年 | 60篇 |
1974年 | 47篇 |
1973年 | 50篇 |
排序方式: 共有10000条查询结果,搜索用时 62 毫秒
981.
Li H Jin G Qin J Tian M Shi J Yang W Tan X Zhang X Zou L 《Histochemistry and cell biology》2011,136(5):515-526
During the central nervous system (CNS) development, radial glia cells (RGCs) play at least two essential roles, they contribute
to neuronal production and the subsequent guidance of neuronal migration, whereas its precise distribution and contribution
to cerebral cortex remains less understood. In this research, we used Vimentin as an astroglial marker and Sox2 as a neural
progenitor marker to identify and investigate RGCs in rat cerebral cortex at embryonic day (E) 16.5. We found that the Sox2+
progenitor cells localized in the germinal zone (GZ) of E16.5 cerebral cortex, ~95% Sox2+ cells co-localized with Vimentin+
or Nestin+ radial processes which extended to the pial surface across the cortical plate (CP). In vitro, we obtained RG-like
cells from E16.5 cerebral cortex on adherent conditions, these Sox2+ Radial glia (RG)-like cells shared some properties with
RGCs in vivo, and these Sox2+ RG-like cells could differentiate into astrocytes, oligodendrocytes and presented the radial
glia—neuron lineage differentiation ability. Taken together, we identified and investigated some characterizations and properties
of Sox2+ RGCs derived from E16.5 cerebral cortex, we suggested that the embryonic Sox2+ progenitor cells which located in
the cortical GZ were mainly composed of Sox2+ RGCs, and the cortex-derived Sox2+ RG-like cells displayed the radial glia—neuron
lineage differentiation ability as neuronal progenitors in vitro. 相似文献
982.
Alcyonacean octocorals in tropical reefs are usually not considered as reef builders. Some Sinularia species, however, are capable of consolidating sclerites at the colony base to form spiculite. Nanwan Bay, southern Taiwan,
features both fossilized and recently formed boulders composed of spiculite, thus demonstrating the role of Sinularia in contributing to the reef structure. Section radiography of an 18.5 kg spiculite boulder demonstrated a regular density
banding of 3–6-mm intervals. Core survey indicated spiculite coverage of 25–30% on the live reef and of 30–40% on the uplifted
boulders. Cores taken from living Sinularia revealed a distinct transition from discrete sclerites to compact spiculite and amorphous calcium carbonate cementing the
sclerites. In the widespread S. gibberosa, sclerite formation appeared to start intracellularly, followed by a prolonged extracellular calcification process. At the
calcification site, multiple sclerocytes formed expanded pseudopod-like membranes that interconnected, forming multicellular
vesicles (MCVs) around the sclerites. The MCVs and the pseudopods disappeared at sclerite maturation, followed by degradation
of the sclerocytes around the mature sclerites. At the colony base, granular vesicles were distributed among the sclerites,
indicating a cementing process in progress. These findings suggest that colonies of Sinularia are able to cement sclerites and consolidate them at their base into spiculite, thus making them reef builders. 相似文献
983.
Cytoglobin, generated using genetic engineering method, is a kind of recombinant human stellate cell activation-associated
protein. We speculate that it could influence the development of hepatic fibrosis like Sellate cell activation-associated
protein which was discovered by Kawada et al. Therefore, we investigated its anti-fibrosis effect on liver both in vivo and
in vitro. During our research, we found that cytoglobin showed obvious effect compared with the control group on Thioacetamide-induced
liver fibrosis in SD rats, including significantly decrease in aspartate aminotransferase, Hyaluronic acid, laminin and collagen
I(Col I) levels in serum and hydroxyproline in livers, which are the important indices reflecting the degree of hepatic fibrosis.
Meanwhile, the viability of rat hepatic stellate cell line T6 (HSC-T6) cells was inhibited by cytoglobin and the apoptosis
induced by cytoglobin in HSC-T6 cells was detected by Annexin V/PI double staining. Activation of the caspase cascade including
caspase-3 for the intrinsic pathways was demonstrated. The results also showed that the expression of Bcl-2 protein decreased
whereas that of Bax protein increased, leading to an increase of the Bax/Bcl-2 ratio. Our results demonstrated that cytoglobin
exhibited anti-fibrosis activity on livers in vivo and in vitro, involving apoptosis induction. 相似文献
984.
Yao YW Shi Y Jia ZF Jiang YH Gu Z Wang J Aljofan M Sun ZG 《Histochemistry and cell biology》2011,136(2):205-215
To investigate the biological significance of ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) involvement in oocyte maturation,
we screened for proteins that bound to UCH-L1 in mouse ovaries, and we found that the prostate tumor overexpressed-1 (PTOV1)
protein was able to bind to UCH-L1. PTOV1 is highly expressed in prostate cancers and considered as a potential marker for
carcinogenesis and the progress of prostate cancer. It was reported that PTOV1 plays an important role in cell cycle regulation,
but its role in mammalian oocyte development and meiosis is still unclear. In this paper, it was found that the expression
levels of PTOV1 in mouse ovaries progressively increased from prepubescence to adulthood. And we found by immunohistochemistry
that PTOV1 spreaded in both the cytoplasm and nuclei of oocytes during prepuberty, but in normal adult mouse oocytes, it concentrated
not only in nuclei but also on the plasma membrane, though in some oocytes with abnormal shapes, PTOV1 did not display the
typical distribution patterns. In granulosa cells, however, it was found to locate in the cytoplasm at all the selected ages.
In postnatal mouse ovaries (28 days), estradiol treatment induced the adult-specific distribution pattern of PTOV1 in oocytes.
In addition, UCH-L1 was shown to be associated with CDK1, which participated in the regulation of cell cycle and oocyte maturation.
Therefore, we propose that the distribution changes of PTOV1 are age-dependent, and significant for mouse oocyte development
and maturation. Moreover, the discovery that PTOV1 is associated with UCH-L1 in mouse oocytes supports the explanations for
that UCH-L1 is involved in oocyte development and maturation, especially under the regulation of estrogen. 相似文献
985.
Liang Zhang Zhongyang Ding Peng Xu Yuhong Wang Zhenghua Gu Zhu Qian Guiyang Shi Kechang Zhang 《Biotechnology and Bioprocess Engineering》2011,16(3):457-461
Tyrosinase is a key enzyme in the biosynthesis of melanin, and the use of inhibitors against tyrosinase can prevent hyperpigmentation
by inhibiting enzymatic oxidation. However, the current use of tyrosine inhibitors is limited by their low activities and
high toxicities. The aim of the present research was to develop novel whitening agents, or tyrosinase-targeted medicine, from
a submerged culture of the fungus Ganoderma lucidum. Methyl lucidenate F was isolated from the ethanol-soluble-acidic components (ESACs) of G. lucidum, with the structure of ESACs elucidated via UV, LC-MS, and 13C-NMR spectral analysis. The tyrosinase inhibitory activity was measured using catechol as a substrate. Methyl lucidenate
F displayed uncompetitive inhibition of the potato tyrosinase activity, for which Lineweaver-Burk plots revealed a maximum
reaction rate (V
max) of 0.4367/min, Michaelis constant (K
m) of 6.765 mM and uncompetitive inhibition constant (K
i) of 19.22 μM. Meanwhile, methyl lucidenate F (tetra cyclic triterpenoid) exhibited high tyrosinase inhibitory activity, with
an IC50 of 32.23 μM. These results suggest that methyl lucidenate F may serve as a potential candidate for skin-whitening agents. 相似文献
986.
987.
Tamminga C Sedegah M Regis D Chuang I Epstein JE Spring M Mendoza-Silveiras J McGrath S Maiolatesi S Reyes S Steinbeiss V Fedders C Smith K House B Ganeshan H Lejano J Abot E Banania GJ Sayo R Farooq F Belmonte M Murphy J Komisar J Williams J Shi M Brambilla D Manohar N Richie NO Wood C Limbach K Patterson NB Bruder JT Doolan DL King CR Diggs C Soisson L Carucci D Levine G Dutta S Hollingdale MR Ockenhouse CF Richie TL 《PloS one》2011,6(10):e25868
Background
A protective malaria vaccine will likely need to elicit both cell-mediated and antibody responses. As adenovirus vaccine vectors induce both these responses in humans, a Phase 1/2a clinical trial was conducted to evaluate the efficacy of an adenovirus serotype 5-vectored malaria vaccine against sporozoite challenge.Methodology/Principal Findings
NMRC-MV-Ad-PfC is an adenovirus vector encoding the Plasmodium falciparum 3D7 circumsporozoite protein (CSP). It is one component of a two-component vaccine NMRC-M3V-Ad-PfCA consisting of one adenovector encoding CSP and one encoding apical membrane antigen-1 (AMA1) that was evaluated for safety and immunogenicity in an earlier study (see companion paper, Sedegah et al). Fourteen Ad5 seropositive or negative adults received two doses of NMRC-MV-Ad-PfC sixteen weeks apart, at particle units per dose. The vaccine was safe and well tolerated. All volunteers developed positive ELISpot responses by 28 days after the first immunization (geometric mean 272 spot forming cells/million[sfc/m]) that declined during the following 16 weeks and increased after the second dose to levels that in most cases were less than the initial peak (geometric mean 119 sfc/m). CD8+ predominated over CD4+ responses, as in the first clinical trial. Antibody responses were poor and like ELISpot responses increased after the second immunization but did not exceed the initial peak. Pre-existing neutralizing antibodies (NAb) to Ad5 did not affect the immunogenicity of the first dose, but the fold increase in NAb induced by the first dose was significantly associated with poorer antibody responses after the second dose, while ELISpot responses remained unaffected. When challenged by the bite of P. falciparum-infected mosquitoes, two of 11 volunteers showed a delay in the time to patency compared to infectivity controls, but no volunteers were sterilely protected.Significance
The NMRC-MV-Ad-PfC vaccine expressing CSP was safe and well tolerated given as two doses, but did not provide sterile protection.Trial Registration
ClinicalTrials.gov NCT00392015相似文献988.
Lee CY Kam YW Fric J Malleret B Koh EG Prakash C Huang W Lee WW Lin C Lin RT Renia L Wang CI Ng LF Warter L 《PLoS pathogens》2011,7(12):e1002390
Chikungunya virus (CHIKV) is an alphavirus responsible for numerous epidemics throughout Africa and Asia, causing infectious arthritis and reportedly linked with fatal infections in newborns and elderly. Previous studies in animal models indicate that humoral immunity can protect against CHIKV infection, but despite the potential efficacy of B-cell-driven intervention strategies, there are no virus-specific vaccines or therapies currently available. In addition, CHIKV has been reported to elicit long-lasting virus-specific IgM in humans, and to establish long-term persistence in non-human primates, suggesting that the virus might evade immune defenses to establish chronic infections in man. However, the mechanisms of immune evasion potentially employed by CHIKV remain uncharacterized. We previously described two human monoclonal antibodies that potently neutralize CHIKV infection. In the current report, we have characterized CHIKV mutants that escape antibody-dependent neutralization to identify the CHIKV E2 domain B and fusion loop "groove" as the primary determinants of CHIKV interaction with these antibodies. Furthermore, for the first time, we have also demonstrated direct CHIKV cell-to-cell transmission, as a mechanism that involves the E2 domain A and that is associated with viral resistance to antibody-dependent neutralization. Identification of CHIKV sub-domains that are associated with human protective immunity, will pave the way for the development of CHIKV-specific sub-domain vaccination strategies. Moreover, the clear demonstration of CHIKV cell-to-cell transmission and its possible role in the establishment of CHIKV persistence, will also inform the development of future anti-viral interventions. These data shed new light on CHIKV-host interactions that will help to combat human CHIKV infection and inform future studies of CHIKV pathogenesis. 相似文献
989.
Background
Cardiac stem cells (CSCs) promote myocardial recovery following ischemia through their regenerative properties. However, little is known regarding the implication of paracrine action by CSCs in the setting of myocardial ischemia/reperfusion (I/R) injury although it is well documented that non-cardiac stem cells mediate cardioprotection via the production of paracrine protective factors. Here, we studied whether CSCs could initiate acute protection following global myocardial I/R via paracrine effect and what component from CSCs is critical to this protection.Methodology/Principal Findings
A murine model of global myocardial I/R was utilized to investigate paracrine effect of Sca-1+ CSCs on cardiac function. Intracoronary delivery of CSCs or CSC conditioned medium (CSC CM) prior to ischemia significantly improved myocardial function following I/R. siRNA targeting of VEGF in CSCs did not affect CSC-preserved myocardial function in response to I/R injury. However, differentiation of CSCs to cardiomyocytes (DCSCs) abolished this protection. Through direct comparison of the protein expression profiles of CSCs and DCSCs, SDF-1 was identified as one of the dominant paracrine factors secreted by CSCs. Blockade of the SDF-1 receptor by AMD3100 or downregulated SDF-1 expression in CSCs by specific SDF-1 siRNA dramatically impaired CSC-induced improvement in cardiac function and increased myocardial damage following I/R. Of note, CSC treatment increased myocardial STAT3 activation after I/R, whereas downregulation of SDF-1 action by blockade of the SDF-1 receptor or SDF-1 siRNA transfection abolished CSC-induced STAT3 activation. In addition, inhibition of STAT3 activation attenuated CSC-mediated cardioprotection following I/R. Finally, post-ischemic infusion of CSC CM was shown to significantly protect I/R-caused myocardial dysfunction.Conclusions/Significance
This study suggests that CSCs acutely improve post-ischemic myocardial function through paracrine factor SDF-1 and up-regulated myocardial STAT3 activation. 相似文献990.
Cathelicidin-BF15 (BF-15) is a 15-mer peptide derived from Cathelicidin-BF (BF-30), which is found in the venom of the snake Bungarus fasciatus and exhibits broad antimicrobial activity. Since BF-15 retains most part of the antimicrobial activity of BF-30 but has significantly reduced haemolytic activity and a much shorter sequence length (and less cost), it is a particularly attractive template around which to design novel antimicrobial peptides. However, the structure–activity relationship of it is still unknown. We designed and synthesized a series of C-terminal amidated analogs of BF-15 based on its amphipathic α-helix structure. And we characterized their antimicrobial potency and haemolytic activity. We identified the amidated BF-15 (analog B1) with potent antimicrobial activity against several antibiotic-resistant bacteria (MICs between 1 and 64 μg/mL, 2–16-folds higher than BF-30) and much lower haemolytic activity. The subsequent circular dichroism study results showed a typical α-helix pattern of analog B1 and the content of the α-helix structure of it increased significantly comparing with BF-30, which indicates the peptide sequence of BF-15 may provide a major contribution to the α-helix content of the whole BF-30 sequence. The peptide induced chaotic membrane morphology and cell debris as determined by electron microscopy. This suggests that the antimicrobial activity of B1 is based on cytoplasmic membrane permeability. Taken together, our results suggested that peptide B1 should be considered as an excellent candidate for developing therapeutic drugs. 相似文献