全文获取类型
收费全文 | 1414篇 |
免费 | 137篇 |
国内免费 | 86篇 |
出版年
2024年 | 5篇 |
2023年 | 11篇 |
2022年 | 43篇 |
2021年 | 73篇 |
2020年 | 35篇 |
2019年 | 55篇 |
2018年 | 39篇 |
2017年 | 27篇 |
2016年 | 47篇 |
2015年 | 87篇 |
2014年 | 104篇 |
2013年 | 100篇 |
2012年 | 126篇 |
2011年 | 114篇 |
2010年 | 59篇 |
2009年 | 53篇 |
2008年 | 61篇 |
2007年 | 67篇 |
2006年 | 63篇 |
2005年 | 43篇 |
2004年 | 41篇 |
2003年 | 56篇 |
2002年 | 39篇 |
2001年 | 32篇 |
2000年 | 26篇 |
1999年 | 22篇 |
1998年 | 15篇 |
1997年 | 15篇 |
1996年 | 10篇 |
1995年 | 16篇 |
1994年 | 13篇 |
1993年 | 12篇 |
1992年 | 21篇 |
1991年 | 10篇 |
1990年 | 9篇 |
1989年 | 9篇 |
1988年 | 14篇 |
1987年 | 8篇 |
1985年 | 7篇 |
1983年 | 4篇 |
1982年 | 6篇 |
1980年 | 6篇 |
1979年 | 4篇 |
1978年 | 5篇 |
1976年 | 2篇 |
1975年 | 2篇 |
1974年 | 4篇 |
1973年 | 4篇 |
1972年 | 4篇 |
1970年 | 2篇 |
排序方式: 共有1637条查询结果,搜索用时 15 毫秒
101.
102.
103.
Wing Ki Wong Terrenz Kelly Jingjing Li Hoi Tang Ma Randy YC Poon 《Cell cycle (Georgetown, Tex.)》2015,14(24):3965-3977
Shugoshin (SGO1) plays a pivotal role in sister chromatid cohesion during mitosis by protecting the centromeric cohesin from mitotic kinases and WAPL. Mammalian cells contain at least 6 alternatively spliced isoforms of SGO1. The relationship between the canonical SGO1A with shorter isoforms including SGO1C remains obscure. Here we show that SGO1C was unable to replace the loss of SGO1A. Instead, expression of SGO1C alone induced aberrant mitosis similar to depletion of SGO1A, promoting premature sister chromatid separation, activation of the spindle-assembly checkpoint, and mitotic arrest. In disagreement with previously published data, we found that SGO1C localized to kinetochores. However, the ability to induce aberrant mitosis did not correlate with its kinetochore localization. SGO1C mutants that abolished binding to kinetochores still triggered premature sister chromatid separation. We provide evidence that SGO1C-mediated mitotic arrest involved the sequestering of PP2A–B56 pool. Accordingly, SGO1C mutants that abolished binding to PP2A localized to kinetochores but did not induce aberrant mitosis. These studies imply that the expression of SGO1C should be tightly regulated to prevent dominant-negative effects on SGO1A and genome instability. 相似文献
104.
A purification process for heparin and precursor polysaccharides using the pH responsive behavior of chitosan 下载免费PDF全文
Ujjwal Bhaskar Anne M. Hickey Guoyun Li Ruchir V. Mundra Fuming Zhang Li Fu Chao Cai Zhimin Ou Jonathan S. Dordick Robert J. Linhardt 《Biotechnology progress》2015,31(5):1348-1359
The contamination crisis of 2008 has brought to light several risks associated with use of animal tissue derived heparin. Because the total chemical synthesis of heparin is not feasible, a bioengineered approach has been proposed, relying on recombinant enzymes derived from the heparin/HS biosynthetic pathway and Escherichia coli K5 capsular polysaccharide. Intensive process engineering efforts are required to achieve a cost‐competitive process for bioengineered heparin compared to commercially available porcine heparins. Towards this goal, we have used 96‐well plate based screening for development of a chitosan‐based purification process for heparin and precursor polysaccharides. The unique pH responsive behavior of chitosan enables simplified capture of target heparin or related polysaccharides, under low pH and complex solution conditions, followed by elution under mildly basic conditions. The use of mild, basic recovery conditions are compatible with the chemical N‐deacetylation/N‐sulfonation step used in the bioengineered heparin process. Selective precipitation of glycosaminoglycans (GAGs) leads to significant removal of process related impurities such as proteins, DNA and endotoxins. Use of highly sensitive liquid chromatography‐mass spectrometry and nuclear magnetic resonance analytical techniques reveal a minimum impact of chitosan‐based purification on heparin product composition. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1348–1359, 2015 相似文献
105.
Jieyao Cheng Jinlin Hou Huiguo Ding Guofeng Chen Qing Xie Yuming Wang Minde Zeng Xiaojuan Ou Hong Ma Jidong Jia 《PloS one》2015,10(12)
Background and Aims
Noninvasive models have been developed for fibrosis assessment in patients with chronic hepatitis B. However, the sensitivity, specificity and diagnostic accuracy in evaluating liver fibrosis of these methods have not been validated and compared in the same group of patients. The aim of this study was to verify the diagnostic performance and reproducibility of ten reported noninvasive models in a large cohort of Asian CHB patients.Methods
The diagnostic performance of ten noninvasive models (HALF index, FibroScan, S index, Zeng model, Youyi model, Hui model, APAG, APRI, FIB-4 and FibroTest) was assessed against the liver histology by ROC curve analysis in CHB patients. The reproducibility of the ten models were evaluated by recalculating the diagnostic values at the given cut-off values defined by the original studies.Results
Six models (HALF index, FibroScan, Zeng model, Youyi model, S index and FibroTest) had AUROCs higher than 0.70 in predicting any fibrosis stage and 2 of them had best diagnostic performance with AUROCs to predict F≥2, F≥3 and F4 being 0.83, 0.89 and 0.89 for HALF index, 0.82, 0.87 and 0.87 for FibroScan, respectively. Four models (HALF index, FibroScan, Zeng model and Youyi model) showed good diagnostic values at given cut-offs.Conclusions
HALF index, FibroScan, Zeng model, Youyi model, S index and FibroTest show a good diagnostic performance and all of them, except S index and FibroTest, have good reproducibility for evaluating liver fibrosis in CHB patients.Registration Number
ChiCTR-DCS-07000039. 相似文献106.
Xiao-Xia Hu Li Fu Yan Li Ze-Bang Lin Xiang Liu Jing-Feng Wang Yang-Xin Chen Zhi-Ping Wang Xi Zhang Zhi-Jun Ou Jing-Song Ou 《PloS one》2015,10(9)
Vitamin E (VitE) only prevented cardiovascular diseases in some patients and the mechanisms remain unknown. VitE levels can be affected by aging and gender. We hypothesize that age and gender can influence VitE’s cardioprotective effect. Mice were divided into 4 groups according to age and gender, and each group of mice were divided into a control group and a VitE group. The mice were administered water or VitE for 21 days; Afterward, the cardiac function and myocardial infarct size and cardiomyocyte apoptosis were measured after myocardial ischemia reperfusion(MI/R). VitE may significantly improved cardiac function in young male mice and aged female mice by enhancing ERK1/2 activity and reducing JNK activity. Enhanced expression of HSP90 and Bcl-2 were also seen in young male mice. No changes in cardiac function and cardiac proteins were detected in aged male mice and VitE was even liked to exert a reverse effect in cardiac function in young mice by enhancing JNK activity and reducing Bcl-2 expression. Those effects were in accordance with the changes of myocardial infarction size and cardiomyocyte apoptosis in each group of mice. VitE may reduce MI/R injury by inhibiting cardiomyocyte apoptosis in young male mice and aged female mice but not in aged male mice. VitE was possibly harmful for young female mice, shown as increased cardiomyocyte apoptosis after MI/R. Thus, we speculated that the efficacy of VitE in cardiac protection was associated with age and gender. 相似文献
107.
Jean Huang Horng-Yih Ou James Lin Rudruidee Karnchanasorn Wei Feng Raynald Samoa Lee-Ming Chuang Ken C. Chiu 《PloS one》2015,10(10)
Background
The liver plays a key role in fuel metabolism. It is well established that liver disease is associated with an increased risk for diabetes mellitus. Hepatitis C virus infection has been known to increase the risk of diabetes. However, much less is known about the role of hepatitis B virus (HBV) infection in diabetes. We examined the association of diabetes based on the vaccination status for HBV.Methods
In this cross-sectional study, we included adult subjects (≥20 y/o) with HBV serology available from the National Health and Nutrition Examination Survey 2005–2010. Diabetes was defined as established diabetes or fasting plasma glucose concentration ≥7.0 mmol/L, 2-hour plasma glucose concentration ≥11.1 mmol/L, or HbA1c ≥ 47.5 mmol/mol (6.5%). Vaccination was based on the reported history and immunization was determined by HBV serology. The odds ratio (OR) with 95% confidence intervals (95% CI) were calculated with consideration of the following covariates: age, gender, BMI, ethnic/racial group, current smoker, current alcohol consumption, family history of diabetes, poverty index, and education.Results
This study included 15,316 subjects. Among them, 2,320 subjects was immunized based the HBV serology. Among 4,063 subjects who received HBV vaccination, successful vaccination was only noted in 39% of subjects. The HBV vaccination was not associated with diabetes (OR: 1.08, 95%CI: 0.96–1.23). Serology evidence of HBV immunization was associated with a reduced OR of diabetes (0.75, 95%CI: 0.62–0.90). Successful HBV vaccination was also associated with a reduced OR of diabetes (0.67, 95%CI: 0.52–0.84).Conclusions
Although our study shows the association of HBV vaccination with the reduced odds of diabetes by 33%, a prospective study is warranted to confirm and examine the impact of HBV vaccination in prevention of diabetes. 相似文献108.
Cardiac atlases play an important role in the computer-aided diagnosis of cardiovascular diseases, in particular they need to deal with large and highly variable image datasets. In this paper, we propose a new nonrigid registration algorithm incorporating shape information, to produce comprehensive atlases. For one thing, the multiscale gradient orientation features of images are combined to form the construction of multifeature mutual information. Additionally, the shape information of multiple-objects in images is incorporated into the cost function for registration. We demonstrate the merits of the new registration algorithm on the 3D data sets of 15 patients. The experimental results show that the new registration algorithm can outperform the conventional intensity-based registration method. The obtained atlas can represent the cardiac structures more accurately. 相似文献
109.
Kua-Chun Ou Chih-Yang Wang Kuan-Ting Liu Yi-Ling Chen Yi-Chen Chen Ming-Derg Lai Meng-Chi Yen 《Biochemical and biophysical research communications》2014
Transfer RNA (tRNA) abundance is one of the critical factors for the enhancement of protein productivity in prokaryotic and eukaryotic hosts. Gene copy number of tRNA and tRNA codon usage bias are generally used to match tRNA abundance of protein-expressing hosts and to optimize the codons of recombinant proteins. Because sufficient concentration of intracellular tRNA and optimized codons of recombinant proteins enhanced translation efficiency, we hypothesized that sufficient supplement of host’s tRNA improved protein productivity in mammalian cells. First, the small tRNA sequencing results of CHO-K1 cells showed moderate positive correlation with gene copy number and codon usage bias. Modification of human interleukin-2 (IL-2) through codons with high gene copy number and high codon usage bias (IL-2 HH, modified on Leu, Thr, Glu) significantly increased protein productivity in CHO-K1 cells. In contrast, modification through codons with relatively high gene copy number and low codon usage bias (IL-2 HL, modified on Ala, Thr, Val), or relatively low gene copy number and low codon usage bias (IL-2 LH, modified on Ala, Thr, Val) did not increase IL-2 productivity significantly. Furthermore, supplement of the alanine tRNA or threonine tRNA increased IL-2 productivity of IL-2 HL. In summary, we revealed a potential strategy to enhance productivity of recombinant proteins, which may be applied in production of protein drug or design of DNA vaccine. 相似文献
110.
胡萝卜(Daucus carota L.)中主要胡萝卜素和番茄红素含量的QTL分析 总被引:2,自引:0,他引:2
以高胡萝卜素自交系P50006和HCM A.C.为亲本构建的F2群体为作图群体,对胡萝卜中α-胡萝卜素、α-胡萝卜素、总胡萝卜素和番茄红素含量进行QTL定位及遗传分析。结果表明,α、β-胡萝卜素、总胡萝卜素和番茄红素含量的广义遗传力分别为0.75、0.50、0.31和0.93。遗传图谱包含91个SRAP(Sequence-related amplified polymorphism)标记,分布于9个连锁群,总长度502.9cM,标记间平均距离5.5cM。除α-胡萝卜素含量外,α-胡萝卜素、总胡萝卜素和番茄红素含量分别检测到1个主效QTL,均为加性遗传效应,分别解释表型变异为12.79%、12.87%和14.61%。此外,α-胡萝卜素和番茄红素含量还分别检测到1对上位性QTL,最大遗传效应分别为显性×加性互作和显性×显性互作,分别解释表型变异为15.1%和6.5%。文章中与QTL连锁的分子标记可用于高胡萝卜素、番茄红素的种质筛选和聚合育种。 相似文献