全文获取类型
收费全文 | 511篇 |
免费 | 46篇 |
专业分类
557篇 |
出版年
2024年 | 2篇 |
2022年 | 3篇 |
2021年 | 4篇 |
2020年 | 5篇 |
2019年 | 4篇 |
2018年 | 6篇 |
2017年 | 6篇 |
2016年 | 11篇 |
2015年 | 16篇 |
2014年 | 23篇 |
2013年 | 17篇 |
2012年 | 43篇 |
2011年 | 36篇 |
2010年 | 35篇 |
2009年 | 24篇 |
2008年 | 41篇 |
2007年 | 43篇 |
2006年 | 37篇 |
2005年 | 20篇 |
2004年 | 33篇 |
2003年 | 35篇 |
2002年 | 30篇 |
2001年 | 2篇 |
2000年 | 5篇 |
1999年 | 6篇 |
1998年 | 8篇 |
1997年 | 10篇 |
1996年 | 10篇 |
1995年 | 4篇 |
1994年 | 5篇 |
1993年 | 6篇 |
1992年 | 4篇 |
1991年 | 5篇 |
1990年 | 2篇 |
1989年 | 1篇 |
1988年 | 2篇 |
1987年 | 2篇 |
1986年 | 1篇 |
1985年 | 1篇 |
1984年 | 3篇 |
1983年 | 1篇 |
1982年 | 1篇 |
1981年 | 3篇 |
1975年 | 1篇 |
排序方式: 共有557条查询结果,搜索用时 15 毫秒
81.
Benjamin S. Walsh Steven R. Parratt Natasha L. M. Mannion Rhonda R. Snook Amanda Bretman Tom A. R. Price 《Ecology and evolution》2021,11(24):18238
The impact of rising global temperatures on survival and reproduction is putting many species at risk of extinction. In particular, it has recently been shown that thermal effects on reproduction, especially limits to male fertility, can underpin species distributions in insects. However, the physiological factors influencing fertility at high temperatures are poorly understood. Key factors that affect somatic thermal tolerance such as hardening, the ability to phenotypically increase thermal tolerance after a mild heat shock, and the differential impact of temperature on different life stages are largely unexplored for thermal fertility tolerance. Here, we examine the impact of high temperatures on male fertility in the cosmopolitan fruit fly Drosophila virilis. We first determined whether temperature stress at either the pupal or adult life history stage impacts fertility. We then tested the capacity for heat‐hardening to mitigate heat‐induced sterility. We found that thermal stress reduces fertility in different ways in pupae and adults. Pupal heat stress delays sexual maturity, whereas males heated as adults can reproduce initially following heat stress, but become sterile within seven days. We also found evidence that while heat‐hardening in D. virilis can improve high temperature survival, there is no significant protective impact of this same hardening treatment on fertility. These results suggest that males may be unable to prevent the costs of high temperature stress on fertility through heat‐hardening, which limits a species’ ability to quickly and effectively reduce fertility loss in the face of short‐term high temperature events. 相似文献
82.
Karen Plaut Rhonda Maple Erika Ginsburg Barbara Vonderhaar 《Journal of cellular physiology》1999,180(2):298-304
The objective of this study was to determine whether sustained progesterone (P) use in the absence of estrogen could influence mammary development in mice. Three-week-old intact or ovariectomized mice were primed with subcutaneous (s.c.) cholesterol (C), estrogen (E), P, or estrogen and progesterone (E/P) together. Nine days after priming, mammary glands were removed and incubated as a whole organ in media supplemented with various combinations of lactogenic hormones. After 5 days in whole organ culture, glands were removed and end buds, alveolar buds and lobulo-alveoli were quantified. Glands from mice primed with C or E developed significantly less lobulo-alveoli than glands from mice primed with P or E/P. While the development was greater in animals treated with E/P compared to those treated with P, it was clear that P in the absence of E could still induce lobulo-alveolar development. We have shown in this paper that P, in the absence of E, can stimulate cell proliferation during priming. Subsequently, the P primed glands can differentiate in response to lactogenic hormones. J. Cell. Physiol. 180:298–304, 1999. © 1999 Wiley-Liss, Inc. 相似文献
83.
84.
Lisec J Meyer RC Steinfath M Redestig H Becher M Witucka-Wall H Fiehn O Törjék O Selbig J Altmann T Willmitzer L 《The Plant journal : for cell and molecular biology》2008,53(6):960-972
Plant growth and development are tightly linked to primary metabolism and are subject to natural variation. In order to obtain an insight into the genetic factors controlling biomass and primary metabolism and to determine their relationships, two Arabidopsis thaliana populations [429 recombinant inbred lines (RIL) and 97 introgression lines (IL), derived from accessions Col-0 and C24] were analyzed with respect to biomass and metabolic composition using a mass spectrometry-based metabolic profiling approach. Six and 157 quantitative trait loci (QTL) were identified for biomass and metabolic content, respectively. Two biomass QTL coincide with significantly more metabolic QTL (mQTL) than statistically expected, supporting the notion that the metabolic profile and biomass accumulation of a plant are linked. On the same basis, three out the six biomass QTL can be simulated purely on the basis of metabolic composition. QTL based on analysis of the introgression lines were in substantial agreement with the RIL-based results: five of six biomass QTL and 55% of the mQTL found in the RIL population were also found in the IL population at a significance level of P ≤ 0.05, with >80% agreement on the allele effects. Some of the differences could be attributed to epistatic interactions. Depending on the search conditions, metabolic pathway-derived candidate genes were found for 24–67% of all tested mQTL in the database AraCyc 3.5. This dataset thus provides a comprehensive basis for the detection of functionally relevant variation in known genes with metabolic function and for identification of genes with hitherto unknown roles in the control of metabolism. 相似文献
85.
Genital transmission of HPV in a mouse model is potentiated by nonoxynol-9 and inhibited by carrageenan 总被引:2,自引:0,他引:2
Roberts JN Buck CB Thompson CD Kines R Bernardo M Choyke PL Lowy DR Schiller JT 《Nature medicine》2007,13(7):857-861
Genital human papillomavirus (HPV) infection is the most common sexually transmitted infection, and virtually all cases of cervical cancer are attributable to infection by a subset of HPVs (reviewed in ref. 1). Despite the high incidence of HPV infection and the recent development of a prophylactic vaccine that confers protection against some HPV types, many features of HPV infection are poorly understood. It remains worthwhile to consider other interventions against genital HPVs, particularly those that target infections not prevented by the current vaccine. However, productive papillomavirus infection is species- and tissue-restricted, and traditional models use animal papillomaviruses that infect the skin or oral mucosa. Here we report the development of a mouse model of cervicovaginal infection with HPV16 that recapitulates the establishment phase of papillomavirus infection. Transduction of a reporter gene by an HPV16 pseudovirus was characterized by histology and quantified by whole-organ, multispectral imaging. Disruption of the integrity of the stratified or columnar genital epithelium was required for infection, which occurred after deposition of the virus on the basement membrane underlying basal keratinocytes. A widely used vaginal spermicide, nonoxynol-9 (N-9), greatly increased susceptibility to infection. In contrast, carrageenan, a polysaccharide present in some vaginal lubricants, prevented infection even in the presence of N-9, suggesting that carrageenan might serve as an effective topical HPV microbicide. 相似文献
86.
The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis 总被引:11,自引:0,他引:11
Wang S Aurora AB Johnson BA Qi X McAnally J Hill JA Richardson JA Bassel-Duby R Olson EN 《Developmental cell》2008,15(2):261-271
Endothelial cells play essential roles in maintenance of vascular integrity, angiogenesis, and wound repair. We show that an endothelial cell-restricted microRNA (miR-126) mediates developmental angiogenesis in vivo. Targeted deletion of miR-126 in mice causes leaky vessels, hemorrhaging, and partial embryonic lethality, due to a loss of vascular integrity and defects in endothelial cell proliferation, migration, and angiogenesis. The subset of mutant animals that survives displays defective cardiac neovascularization following myocardial infarction. The vascular abnormalities of miR-126 mutant mice resemble the consequences of diminished signaling by angiogenic growth factors, such as VEGF and FGF. Accordingly, miR-126 enhances the proangiogenic actions of VEGF and FGF and promotes blood vessel formation by repressing the expression of Spred-1, an intracellular inhibitor of angiogenic signaling. These findings have important therapeutic implications for a variety of disorders involving abnormal angiogenesis and vascular leakage. 相似文献
87.
88.
Membrane-associated guanylate kinases (MAGUKs) regulate the formation and
function of molecular assemblies at specialized regions of the membrane.
Allosteric regulation of an intramolecular interaction between the Src
homology 3 (SH3) and guanylate kinase (GK) domains of MAGUKs is thought to
play a central role in regulating MAGUK function. Here we show that a mutant
of the Drosophila MAGUK Discs large (Dlg), dlgsw,
encodes a form of Dlg that disrupts the intramolecular association while
leaving the SH3 and GK domains intact, providing an excellent model system to
assess the role of the SH3-GK intramolecular interaction in MAGUK function.
Analysis of asymmetric cell division of maternal-zygotic
dlgsw embryonic neuroblasts demonstrates that the
intramolecular interaction is not required for Dlg localization but is
necessary for cell fate determinant segregation to the basal cortex and
mitotic spindle alignment with the cortical polarity axis. These defects
ultimately result in improper patterning of the embryonic central nervous
system. Furthermore, we demonstrate that the sw mutation of Dlg results in
unregulated complex assembly as assessed by GukHolder association with the
SH3-GK versus PDZ-SH3-GK modules of Dlgsw. From these
studies, we conclude that allosteric regulation of the SH3-GK intramolecular
interaction is required for regulation of MAGUK function in asymmetric cell
division, possibly through regulation of complex assembly.The membrane-associated guanylate kinase
(MAGUK)2 superfamily
consists of ubiquitous scaffolding proteins that are composed of a common core
of contiguously linked modular domains (protein-protein interaction domains,
PDZ and SH3 domains, and a domain with homology to the yeast guanylate kinase,
GK domain). MAGUKs are concentrated at sites of cell-cell contact
(1) and organize a variety of
cell adhesion molecules, cytoskeletal proteins, receptors, ion channels, and
associated signaling molecules at specialized regions of the membrane
(2). Protein complex
organization by MAGUKs has been thought to occur, at least in part, through
allosteric regulation that arises from an intramolecular interaction between
the SH3 and GK domains. This interaction has been shown to regulate binding of
numerous MAGUK ligands in vitro
(3–5).
However, whereas the SH3-GK interaction has been extensively characterized
biochemically
(6–8),
very little data exists regarding its physiological relevance. Here we
investigate the role of the SH3-GK intramolecular interaction of Discs large
(Dlg), a prototype of the MAGUK superfamily.Loss of dlg activity results in overgrowth of imaginal discs and
tumor formation (9). Dlg
localizes to septate and neuromuscular junctions and is essential for
establishing and maintaining apicobasal polarity of Drosophila
epithelia (10). Dlg also plays
an important role in regulating the process of asymmetric cell division (ACD)
(11–13).
ACD is a mechanism for generating cellular diversity via unequal mitotic
divisions of progenitor cells. For instance, in wild-type Drosophila
neuroblasts, cell signaling networks interact to allow for asymmetric
segregation of basal cell fate determinants, followed by alignment of the
mitotic spindle along the apical-basal cortical polarity axis (see Refs.
14 and
15) for review). ACD results
in the formation of a self-renewing, stem-cell like neuroblast and a smaller
ganglion mother cell, which has neuronal or glial fate. In dlg
germline clone (dlgGLC) embryonic metaphase neuroblasts,
ganglion mother cell fate determinants are not restricted to the basal cortex
(11,
12) and the mitotic spindle
does not reliably align with the apical-basal cortical polarity axis
(13). Defects in neurogenesis
have also been observed for dlgGLC embryos
(16), as well as embryos
treated with RNA interference against an alternatively spliced isoform of Dlg
(17), providing evidence for
the function of Dlg in neuronal differentiation and axon guidance. Such
defects in neurogenesis are thought to be attributed to defective localization
of basal cell fate determinants during ACD
(17). Although Dlg function is
important in a broad range of dynamic cellular processes, the role of the Dlg
SH3-GK intramolecular interaction in Dlg activity is poorly understood.One potential role for the SH3-GK intramolecular association is to regulate
MAGUK complex assembly. In vitro binding assays demonstrated that
mutations disrupting this intramolecular interaction allowed mutant SH3-GK
modules to associate with SH3 or GK domains of various MAGUK proteins in
trans, providing a mechanism of regulating oligomerization of MAGUKs
in vivo (6,
8). A role in clustering of ion
channels was also observed as mutations that disrupted the intramolecular
association, whereas having no effect on association with the potassium
channel KV1.4 or homo-oligomerization of PSD-95, resulted in loss
of channel clustering in vivo
(18). Furthermore, multiple
in vitro studies support its regulation of binding of protein ligands
with the GK domain of MAGUKs: examples include interaction of GK domains of
Dlg with GukHolder (GukH) (3),
SAP97 with guanylate kinase-associated protein
(4), and PSD-93 with the
microtubule-associated protein 1A
(5). These studies suggest that
allosteric modulation of the SH3-GK intramolecular interaction is important
for regulation of complex assembly, yet little evidence exists that such
regulation of MAGUKs is required for their function in vivo.In crystal structures of the SH3-GK module of PSD-95, two β-strands
that emerge from the SH3 and GK domains appear to mediate the interaction
between the domains (Fig.
1A) (19,
20). Functional studies of the
interaction of these β-strands demonstrate that a COOH-terminal
truncation of the strand following the GK domain abrogates the intramolecular
association of the SH3-GK module. This 13-amino acid truncation of PSD-95 is
comparable with a dlg allele that had previously been identified in a
genetic screen, dlgsw
(21). Drosophila
containing only the sw form of Dlg (refer to COOH-terminal sequences in
Fig. 1B) die in the
latter stages of embryonic development due to failure of dorsal closure and
terminal defects (21). We
hypothesized that this mutation disrupts the intramolecular interaction while
leaving the SH3 and GK domains largely intact, making the
dlgsw allele an excellent model system for assessing the
role of this interaction in MAGUK function.Open in a separate windowFIGURE 1.The dlgsw allele disrupts the SH3-GK intramolecular
interaction formed by interacting E and Fβ-strands. A, a
ribbon diagram of the SH3-GK module of PSD-95 (Protein Data Bank 1KJW
(19)) created using MOLSCRIPT
(58) and RASTER3D
(59), highlighting the SH3
(red), HOOK (gray), and GK (blue) domains. The
intramolecular interaction between the SH3 and GK domains is mediated by the
interaction of two β-strands (shown in black), one strand is
contributed by the linker region following the SH3 domain (strand E), whereas
the second strand is contributed by a COOH-terminal strand that follows the GK
domain (strand F). B, sequence comparison of the COOH-terminal
regions of WT Dlg versus Dlgsw. C, GST pull-downs
of the Dlg SH3 domain using GST fusions of WT and mutant Dlg GK domains.
Association of protein components was detected by an anti-His antibody that
recognized an NH2-terminal on the SH3 domain of Dlg. D,
circular dichroism wavelength scans, monitoring the change in ellipticity as a
function of wavelength. Changes in α-helical content are indicated by
the change in ellipticity at 222 nm (see black dashed line).
Comparison of CD spectra of WT PDZ-SH3-GK (green circles) and
PDZ-SH3-GK(sw) (red squares) indicate no significant change in
α helical character. E, Sedfit analysis of sedimentation
velocity scans of WT PDZ-SH3-GK (green solid line) and
PDZ-SH3-GKsw (red dashed line). Note that the estimated
molecular weights provided are based upon the assumption that the protein
exists in a globular conformation.In the studies presented here, we utilized the dlgsw
mutant allele to explore the role of the intramolecular interaction in
regulating complex assembly and furthermore, to assess the in vivo,
physiological significance of this interaction for Dlg function in neuroblast
asymmetric cell division. We find that disruption of the SH3-GK intramolecular
interaction results in unregulated complex assembly. Such a loss in regulated
complex assembly results in loss of Dlg function in the process of asymmetric
cell division as assessed by localization of cell fate determinants in a
dividing neuroblast and alignment of the mitotic spindle along the
apical-basal cortical polarity axis, ultimately resulting in defects in
neurogenesis. Thus, these data contribute to our understanding of how the
SH3-GK intramolecular interaction regulates MAGUK function in formation and
regulation of membrane specializations. 相似文献
89.
Optimization of primer-specific filter metrics for the assessment of mitochondrial DNA sequence data
Filter metrics are used as a quick assessment of sequence trace files in order to sort data into different categories (i.e. high quality, review, and low quality) without human intervention. The filter metrics consist of two numerical parameters for sequence quality assessment: trace score (TS) and contiguous read length (CRL). Primer-specific settings for the TS and CRL were established using a calibration dataset of 2817 traces and validated using a concordance dataset of 5617 traces. Prior to optimization, 57% of the traces required manual review before import into a sequence analysis program, whereas after optimization only 28% of the traces required manual review. After optimization of primer-specific filter metrics for mitochondrial DNA sequence data, an overall reduction of review of trace files translates into increased throughput of data analysis and decreased time required for manual review. 相似文献
90.