全文获取类型
收费全文 | 22482篇 |
免费 | 3060篇 |
国内免费 | 445篇 |
出版年
2023年 | 78篇 |
2022年 | 181篇 |
2021年 | 396篇 |
2020年 | 308篇 |
2019年 | 362篇 |
2018年 | 474篇 |
2017年 | 391篇 |
2016年 | 555篇 |
2015年 | 700篇 |
2014年 | 877篇 |
2013年 | 966篇 |
2012年 | 1116篇 |
2011年 | 1194篇 |
2010年 | 689篇 |
2009年 | 667篇 |
2008年 | 835篇 |
2007年 | 772篇 |
2006年 | 748篇 |
2005年 | 646篇 |
2004年 | 580篇 |
2003年 | 586篇 |
2002年 | 543篇 |
2001年 | 2318篇 |
2000年 | 2161篇 |
1999年 | 1534篇 |
1998年 | 459篇 |
1997年 | 478篇 |
1996年 | 402篇 |
1995年 | 379篇 |
1994年 | 287篇 |
1993年 | 242篇 |
1992年 | 749篇 |
1991年 | 608篇 |
1990年 | 521篇 |
1989年 | 408篇 |
1988年 | 320篇 |
1987年 | 252篇 |
1986年 | 188篇 |
1985年 | 145篇 |
1984年 | 92篇 |
1983年 | 67篇 |
1982年 | 48篇 |
1981年 | 45篇 |
1980年 | 31篇 |
1979年 | 33篇 |
1978年 | 28篇 |
1976年 | 33篇 |
1974年 | 24篇 |
1973年 | 32篇 |
1970年 | 25篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
981.
David A. X. Nayagam Ceara McGowan Joel Villalobos Richard A. Williams Cesar Salinas-LaRosa Penny McKelvie Irene Lo Meri Basa Justin Tan Chris E. Williams 《Journal of visualized experiments : JoVE》2013,(78)
With the recent development of retinal prostheses, it is important to develop reliable techniques for assessing the safety of these devices in preclinical studies. However, the standard fixation, preparation, and automated histology procedures are not ideal. Here we describe new procedures for evaluating the health of the retina directly adjacent to an implant. Retinal prostheses feature electrode arrays in contact with eye tissue. Previous methods have not been able to spatially localize the ocular tissue adjacent to individual electrodes within the array. In addition, standard histological processing often results in gross artifactual detachment of the retinal layers when assessing implanted eyes. Consequently, it has been difficult to assess localized damage, if present, caused by implantation and stimulation of an implanted electrode array. Therefore, we developed a method for identifying and localizing the ocular tissue adjacent to implanted electrodes using a (color-coded) dye marking scheme, and we modified an eye fixation technique to minimize artifactual retinal detachment. This method also rendered the sclera translucent, enabling localization of individual electrodes and specific parts of an implant. Finally, we used a matched control to increase the power of the histopathological assessments. In summary, this method enables reliable and efficient discrimination and assessment of the retinal cytoarchitecture in an implanted eye. 相似文献
982.
Wei-Meng Woo Scott X. Atwood Hanson H. Zhen Anthony E. Oro 《Journal of visualized experiments : JoVE》2013,(72)
Hair follicle morphogenesis, a complex process requiring interaction between epithelia-derived keratinocytes and the underlying mesenchyme, is an attractive model system to study organ development and tissue-specific signaling. Although hair follicle development is genetically tractable, fast and reproducible analysis of factors essential for this process remains a challenge. Here we describe a procedure to generate targeted overexpression or shRNA-mediated knockdown of factors using lentivirus in a tissue-specific manner. Using a modified version of a hair regeneration model 5, 6, 11, we can achieve robust gain- or loss-of-function analysis in primary mouse keratinocytes or dermal cells to facilitate study of epithelial-mesenchymal signaling pathways that lead to hair follicle morphogenesis. We describe how to isolate fresh primary mouse keratinocytes and dermal cells, which contain dermal papilla cells and their precursors, deliver lentivirus containing either shRNA or cDNA to one of the cell populations, and combine the cells to generate fully formed hair follicles on the backs of nude mice. This approach allows analysis of tissue-specific factors required to generate hair follicles within three weeks and provides a fast and convenient companion to existing genetic models. 相似文献
983.
X.‐R. Liu M. Wu N. He H. Meng L. Wen J.‐L. Wang M.‐P. Zhang W.‐B. Li X. Mao J.‐M. Qin B.‐M. Li B. Tang Y.‐H. Deng Y.‐W. Shi T. Su Y.‐H. Yi B.‐S. Tang W.‐P. Liao 《Genes, Brain & Behavior》2013,12(2):234-240
Paroxysmal dyskinesias (PDs) are a group of episodic movement disorders with marked variability in clinical manifestation and potential association with epilepsy. PRRT2 has been identified as a causative gene for PDs, but the phenotypes and inheritance patterns of PRRT2 mutations need further clarification. In this study, 10 familial and 21 sporadic cases with PDs and PDs‐related phenotypes were collected. Genomic DNA was screened for PRRT2 mutations by direct sequencing. Seven PRRT2 mutations were identified in nine (90.0%) familial cases and in six (28.6%) sporadic cases. Five mutations are novel: two missense mutations (c.647C>G/p.Pro216Arg and c.872C>T/p.Ala291Val) and three truncating mutations (c.117delA/p.Val41TyrfsX49, c.510dupT/p.Leu171SerfsX3 and c.579dupA/p.Glu194ArgfsX6). Autosomal dominant inheritance with incomplete penetrance was observed in most of the familial cases. In the sporadic cases, inheritance was heterogeneous including recessive inheritance with compound heterozygous mutations, inherited mutations with incomplete parental penetrance and de novo mutation. Variant phenotypes associated with PRRT2 mutations, found in 36.0% of the affected cases, included febrile convulsions, epilepsy, infantile non‐convulsive seizures (INCS) and nocturnal convulsions (NC). All patients with INCS or NC, not reported previously, displayed abnormalities on electroencephalogram (EEG). No EEG abnormalities were recorded in patients with classical infantile convulsions and paroxysmal choreoathetosis (ICCA)/paroxysmal kinesigenic dyskinesia (PKD). Our study further confirms that PRRT2 mutations are the most common cause of familial PDs, displaying both dominant and recessive inheritance. Epilepsy may occasionally occur in ICCA/PKD patients with PRRT2 mutations. Variant phenotypes INCS or NC differ from classical ICCA/PKD clinically and electroencephalographically. They have some similarities with, but not identical to epilepsy, possibly represent an overlap between ICCA/PKD and epilepsy . 相似文献
984.
Dormancy is a condition that delays or inhibits growth in seed, vegetative buds, and floral buds. In peach, seed germination occurs when seed accumulate sufficient stratification and growing degree hours to break dormancy and begin growing. Correlations have been reported between mean seed stratification requirements and mean bud chilling requirements among Prunus families, but an individual seed’s germination date and subsequent vegetative and floral bud break date are not correlated. Prior to this study, the genetic factors involved in regulating seed dormancy and their location on the peach genomic map were unknown. Segregating F2 seed were collected from a high?×?low chill F1 peach hybrid in 2005, 2006, and 2008. Germination date and growth habit was measured after the stratification requirement of the 2005 seed was fully met. The seed collected in 2006 and 2008 received varying amounts of stratification, which enabled data on stratification requirement, heat requirement, and growth habit to be collected. Genomic DNA was extracted from seedling leaf tissue and screened with SSR markers selected from the Prunus reference map at an average resolution of 20 cM. Seed dormancy quantitative trait loci (QTLs) were detected on G1, G4, G6/8, and G7. The QTLs detected on G6/8 and G7 were discovered in the same region as QTLs associated with floral bud chilling requirement and bloom time in peach. 相似文献
985.
986.
Zuhu Yu Liangchao Ni Duqun Chen Qiang Zhang Zhengming Su Yadong Wang Wenshui Yu Xionghui Wu Jiongxian Ye Shangqi Yang Yongqing Lai Xianxin Li 《Journal of molecular histology》2013,44(6):669-677
MicroRNA-7 (miR-7) has been described as a tumor suppressor in several human cancers, but the results of a study to identify miRNAs associated with metastatic capability in breast cancer suggested that miR-7 may be characterized as an oncogene. The present study was to determine the expression and function of miR-7 in renal cell carcinoma. Quantitative real-time polymerase chain reaction was used to validate the expressions of miR-7 in 48 paired renal cell carcinomas (RCC) and normal tissues, based on the preliminary sequencing results of miRNAs. Furthermore, the impacts of miR-7 on cell migration, proliferation and apoptosis were analyzed using wound scratch assay, MTT and flow cytometry, respectively. The results demonstrated that miR-7 was up-regulated in RCC compared with normal tissues (p = 0.001). Down-regulation of miR-7 with synthesized inhibitor inhibited cell migration in vitro, suppressed cell proliferation and induced renal cancer cell apoptosis, prompting that miR-7 could be characterized as an oncogene in RCC. The present study was the first to reveal that miR-7 was up-regulated in RCC and it played an important role in RCC by affecting cellular migration, proliferation and apoptosis. Further researches should be conducted to explore the roles and target genes of miR-7 in RCC and other cancers. 相似文献
987.
Jian Chen Feihui Zou Hongran Fu Hui Mao Mingjie Gong Lanchun Ni Xide Xu Jinlong Shi Kaifu Ke Maohong Cao Fei Zhou Wei Shi 《Journal of molecular histology》2013,44(3):271-283
Traumatic brain injury (TBI) triggers a complex series of neurochemical and signaling changes that lead to neuronal dysfunction and overreactive astrocytes. In the current study, we showed that interactions between SCYL1-bp1 and Pirh2 are involved in central nervous system (CNS) injury and repair. Western blot and immunohistochemical analysis of an acute traumatic brain injury model in adult rats revealed significantly increased levels of SCYL1-bp1 and Pirh2 in the ipsilateral brain cortex, compared to contralateral cerebral cortex. Immunofluorescence double-labeling analyses further revealed that SCYL1-bp1 is mainly co-expressed with NeuN. Terminal deoxynucleotidyl transferase-mediated biotinylated-dUTP nick-end labeling staining data supported the involvement of SCYL1-bp1 and Pirh2 in neuronal apoptosis after brain injury. We additionally examined the expression profiles of active caspase-3, which were altered in correlation with the levels of SCYL1-bp1 and Pirh2. Notably, both SCYL1-bp1 and Pirh2 were colocalized with active caspase-3, and all three proteins participated in neuronal apoptosis. Immunoprecipitation experiments further revealed interactions of these proteins with each other in the pathophysiology process. To our knowledge, this is the first study to report interactions between SCYL1-bp1 and Pirh2 in traumatic brain. Our data collectively indicate that SCYL1-bp1 and Pirh2 play important roles in CNS pathophysiology after TBI. 相似文献
988.
989.
Qinghong Wang Jijun Zhou Bei Zhang Zhiqiang Tian Jun Tang Yanhua Zheng Zemin Huang Yi Tian Zhengcai Jia Yan Tang Jennifer C. van Velkinburgh Qing Mao Xiuwu Bian Yifang Ping Bing Ni Yuzhang Wu 《PLoS pathogens》2013,9(6)
IL-23 regulates myriad processes in the innate and adaptive immune systems, and is a critical mediator of the proinflammatory effects exerted by Th17 cells in many diseases. In this study, we investigated whether and how hepatitis B virus (HBV) causes liver damage directly through the IL-23 signaling pathway. In biopsied liver tissues from HBV-infected patients, expression of both IL-23 and IL-23R was remarkably elevated. In vivo observations also indicated that the main sources of IL-23 were myeloid dendritic cells (mDCs) and macrophages. Analysis of in vitro differentiated immature DCs and macrophages isolated from healthy donors revealed that the HBV surface antigen (HBsAg) efficiently induces IL-23 secretion in a mannose receptor (MR)-dependent manner. Culture with an endosomal acidification inhibitor and the dynamin inhibitor showed that, upon binding to the MR, the HBsAg is taken up by mDCs and macrophages through an endocytosis mechanism. In contrast, although the HBV core antigen (HBcAg) can also stimulate IL-23 secretion from mDCs, the process was MR- and endocytosis-independent. In addition, IL-23 was shown to be indispensible for HBsAg-stimulated differentiation of naïve CD4+ T cells into Th17 cells, which were determined to be the primary source of IL-17 in HBV-infected livers. The cognate receptor, IL-17R, was found to exist on the hepatic stellate cells and mDCs, both of which might represent the potential target cells of IL-17 in hepatitis B disease. These data provide novel insights into a yet unrecognized mechanism of HBV-induced hepatitis, by which increases in IL-23 expression, through an MR/endocytosis-dependent or -independent manner, produce liver damage through the IL-23/IL-17 axis. 相似文献
990.
Alexandra M. Lopes Kenneth I. Aston Emma Thompson Filipa Carvalho Jo?o Gon?alves Ni Huang Rune Matthiesen Michiel J. Noordam Inés Quintela Avinash Ramu Catarina Seabra Amy B. Wilfert Juncheng Dai Jonathan M. Downie Susana Fernandes Xuejiang Guo Jiahao Sha António Amorim Alberto Barros Angel Carracedo Zhibin Hu Matthew E. Hurles Sergey Moskovtsev Carole Ober Darius A. Paduch Joshua D. Schiffman Peter N. Schlegel Mário Sousa Douglas T. Carrell Donald F. Conrad 《PLoS genetics》2013,9(3)
Gonadal failure, along with early pregnancy loss and perinatal death, may be an important filter that limits the propagation of harmful mutations in the human population. We hypothesized that men with spermatogenic impairment, a disease with unknown genetic architecture and a common cause of male infertility, are enriched for rare deleterious mutations compared to men with normal spermatogenesis. After assaying genomewide SNPs and CNVs in 323 Caucasian men with idiopathic spermatogenic impairment and more than 1,100 controls, we estimate that each rare autosomal deletion detected in our study multiplicatively changes a man''s risk of disease by 10% (OR 1.10 [1.04–1.16], p<2×10−3), rare X-linked CNVs by 29%, (OR 1.29 [1.11–1.50], p<1×10−3), and rare Y-linked duplications by 88% (OR 1.88 [1.13–3.13], p<0.03). By contrasting the properties of our case-specific CNVs with those of CNV callsets from cases of autism, schizophrenia, bipolar disorder, and intellectual disability, we propose that the CNV burden in spermatogenic impairment is distinct from the burden of large, dominant mutations described for neurodevelopmental disorders. We identified two patients with deletions of DMRT1, a gene on chromosome 9p24.3 orthologous to the putative sex determination locus of the avian ZW chromosome system. In an independent sample of Han Chinese men, we identified 3 more DMRT1 deletions in 979 cases of idiopathic azoospermia and none in 1,734 controls, and found none in an additional 4,519 controls from public databases. The combined results indicate that DMRT1 loss-of-function mutations are a risk factor and potential genetic cause of human spermatogenic failure (frequency of 0.38% in 1306 cases and 0% in 7,754 controls, p = 6.2×10−5). Our study identifies other recurrent CNVs as potential causes of idiopathic azoospermia and generates hypotheses for directing future studies on the genetic basis of male infertility and IVF outcomes. 相似文献