全文获取类型
收费全文 | 45714篇 |
免费 | 3854篇 |
国内免费 | 4978篇 |
出版年
2024年 | 107篇 |
2023年 | 606篇 |
2022年 | 1139篇 |
2021年 | 2391篇 |
2020年 | 1619篇 |
2019年 | 2138篇 |
2018年 | 2011篇 |
2017年 | 1475篇 |
2016年 | 1990篇 |
2015年 | 2867篇 |
2014年 | 3519篇 |
2013年 | 3712篇 |
2012年 | 4431篇 |
2011年 | 3923篇 |
2010年 | 2556篇 |
2009年 | 2426篇 |
2008年 | 2642篇 |
2007年 | 2433篇 |
2006年 | 2016篇 |
2005年 | 1780篇 |
2004年 | 1396篇 |
2003年 | 1317篇 |
2002年 | 1085篇 |
2001年 | 783篇 |
2000年 | 649篇 |
1999年 | 645篇 |
1998年 | 408篇 |
1997年 | 356篇 |
1996年 | 339篇 |
1995年 | 267篇 |
1994年 | 216篇 |
1993年 | 158篇 |
1992年 | 206篇 |
1991年 | 167篇 |
1990年 | 137篇 |
1989年 | 113篇 |
1988年 | 97篇 |
1987年 | 78篇 |
1986年 | 71篇 |
1985年 | 95篇 |
1984年 | 21篇 |
1983年 | 32篇 |
1982年 | 21篇 |
1981年 | 22篇 |
1980年 | 10篇 |
1979年 | 9篇 |
1978年 | 6篇 |
1977年 | 7篇 |
1974年 | 6篇 |
1969年 | 13篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
961.
962.
Joseph B Song Youssef S Tanagho Mohammed Haseebuddin Brian M Benway Alana C Desai Sam B Bhayani Robert S Figenshau 《Reviews in urology》2013,15(2):84-91
Retrieval of foreign bodies from the genitourinary system, most commonly inserted for sexual satisfaction or as a result of a psychiatric illness, can pose a significant surgical challenge. Due to their breadth of size, shape, and location within the genitourinary system, endoscopic management can be difficult. Here, we review the management of four cases of foreign object insertion into the genitourinary system and their outcomes and management.Key words: Foreign body insertion, Sounding, Genitourinary foreign object, Endoscopic extractionForeign objects within the genitourinary tract present a challenging urologic finding due to the diversity and breadth of presentation. Although many objects are easily removed, more complex approaches may be required depending on the size, shape, and location of the object.1,2 In this case series, we discuss the endoscopic management of four patients who presented with foreign bodies in the urethra. Two patients inserted beads into their genitourinary tract for the purpose of sexual stimulation and two patients had a history of psychiatric illness with multiple insertions of a diverse range of foreign objects. In all four cases, endoscopic management was successful in removing the objects, with no need for an open approach such as perineal urethrotomy or open cystostomy. In case 2, in which an open approach was attempted at an outside hospital, this open approach was associated with intraoperative complications. 相似文献
963.
Carston R. Wagner Shu-Ling Chang George W. Griesgraber Heng Song Edward J. McIntee Cheryl L. Zimmerman 《Nucleosides, nucleotides & nucleic acids》2013,32(4-5):913-919
Abstract Stable and water soluble amino acid phosphomonoester amidates of AZT were synthesized and shown to have potent anti-HIV-1 activity. Intracellular and cell extract metabolism studies revealed that these compounds are likely to be enzymatically converted to the corresponding monophosphates. In addition, we have shown that the half life and tissue distribution of a phosphoramidate of AZT is 5 and 10-fold greater, respectively, than AZT. 相似文献
964.
You-Bin Si Yan Xiang Chao Tian Xiong-Yuan Si Jing Zhou Dong-Mei Zhou 《Soil & Sediment Contamination》2013,22(1):72-84
Glyphosate [N-(phosphonomethyl)-glycine] is a herbicide widely used in large quantities in agricultural applications. It is also known to form complexes with metal ions, although its influence on metal behavior, such as lead (Pb) in soil, is not well understood. In this study, the adsorption and co-adsorption of Pb and glyphosate were determined on two soils [a red (RS) soil, Udic Ferrisol, and a yellow-brown (YB) soil, Udic Luvisol] of distinctly different chemical characteristics at varying pH conditions. Results indicate that the adsorption of lead and glyphosate strongly depends on soil types: the RS soil, characterized by a relatively high iron/aluminum content but a low pH and organic matter content, shows a much lower adsorption capacity for Pb but a higher sorption for glyphosate than the YB soil. The co-existence of Pb and glyphosate in soils resulted in complex interactions among Pb, glyphosate, Pb-glyphosate complexes, and soil minerals. The presence of glyphosate decreased Pb adsorption on the two soils, which was attributed primarily to the formation of soluble Pb-glyphosate complexes having relatively low affinities to soil surfaces. On the other hand, addition of Pb increased the adsorption of glyphosate on both soils, which was attributed to: (1) a decreased solution pH due to the ion exchange between Pb2+ and H+ on soil surfaces; and (2) increased sorption sites where Pb was adsorbed and acted as a bridge between glyphosate and the soil. The present study illustrates that the complex interactions among glyphosate, Pb, and soil may have important implications for the mobility and bioavailability of Pb in soil and should thus be considered in future environmental risk assessments. 相似文献
965.
Chen Ding Jing Jiang Junying Wei Wanlin Liu Wei Zhang Mingwei Liu Tianyi Fu Tianyuan Lu Lei Song Wantao Ying Cheng Chang Yangjun Zhang Jie Ma Lai Wei Anna Malovannaya Lijun Jia Bei Zhen Yi Wang Fuchu He Xiaohong Qian Jun Qin 《Molecular & cellular proteomics : MCP》2013,12(8):2370-2380
The current in-depth proteomics makes use of long chromatography gradient to get access to more peptides for protein identification, resulting in covering of as many as 8000 mammalian gene products in 3 days of mass spectrometer running time. Here we report a fast sequencing (Fast-seq) workflow of the use of dual reverse phase high performance liquid chromatography - mass spectrometry (HPLC-MS) with a short gradient to achieve the same proteome coverage in 0.5 day. We adapted this workflow to a quantitative version (Fast quantification, Fast-quan) that was compatible to large-scale protein quantification. We subjected two identical samples to the Fast-quan workflow, which allowed us to systematically evaluate different parameters that impact the sensitivity and accuracy of the workflow. Using the statistics of significant test, we unraveled the existence of substantial falsely quantified differential proteins and estimated correlation of false quantification rate and parameters that are applied in label-free quantification. We optimized the setting of parameters that may substantially minimize the rate of falsely quantified differential proteins, and further applied them on a real biological process. With improved efficiency and throughput, we expect that the Fast-seq/Fast-quan workflow, allowing pair wise comparison of two proteomes in 1 day may make MS available to the masses and impact biomedical research in a positive way.The performance of mass spectrometry has been improved tremendously over the last few years (1–3), making mass spectrometry-based proteomics a viable approach for large-scale protein analysis in biological research. Scientists around the world are striving to fulfill the promise of identifying and quantifying almost all gene products expressed in a cell line or tissue. This would make mass spectrometry-based protein analysis an approach that is compatible to the second-generation mRNA deep-seq technique (4, 5).Two liquid chromatography (LC)-MS strategies have been employed to achieve deep proteome coverage. One is a single run with a long chromatography column and gradient to take advantage of the resolving power of HPLC to reduce the complexity of peptide mixtures; the other is a sequential run with two-dimensional separation (typically ion-exchange and reverse phase) to reduce peptide complexity. It was reported by two laboratories that 2761 and 4500 proteins were identified with a 10 h chromatography gradient on a dual pressure linear ion-trap orbitrap mass spectrometer (LTQ Orbitrap Velos)(6–8). Similarly, 3734 proteins were identified using a 8 h gradient on a 2 m long column with a hybrid triple quadrupole - time of flight (Q-TOF, AB sciex 5600 Q-TOF)(9) mass spectrometer. The two-dimensional approach has yielded more identification with longer time. For example, 10,006 proteins (representing over 9000 gene products, GPs)1 were identified in U2OS cell (10), and 10,255 proteins (representing 9207 GPs) from HeLa cells (11). It took weeks (for example, 2–3 weeks) of machine running time to achieve such proteome coverage, pushing proteome analysis to the level that is comparable to mRNA-seq. With the introduction of faster machines, human proteome coverage now has reached the level of 7000–8500 proteins (representing 7000–8000 GPs) in 3 days (12). Notwithstanding the impressive improvement, the current approach using long column and long gradient suffers from inherent limitations: it takes long machine running time and it is challenging to keep reproducibility among repeated runs. Thus, current throughput and reproducibility have hindered the application of in-depth proteomics to traditional biological researches. A timesaving approach is in urgent need.In this study, we used the first-dimension (1D) short pH 10 RP prefractionation to reduce the complexity of the proteome (13), followed by sequential 30 min second-dimension (2D) short pH 3 reverse phase-(RP)-LC-MS/MS runs for protein identification (14). The results demonstrated that it is possible to identify 8000 gene products from mammalian cells within 12 h of total MS measurement time by applying this dual-short 2D-RPLC-MS/MS strategy (Fast sequencing, Fast-seq). The robustness of the strategy was revealed by parallel testing on different MS systems including quadrupole orbitrap mass spectrometer (Q-Exactive), hybrid Q-TOF (Triple-TOF 5600), and dual pressure linear ion-trap orbitrap mass spectrometer (LTQ-Orbitrap Velos), indicating the inherent strength of the approach as to merely taking advantage of the better MS instruments. This strategy increases the efficiency of MS sequencing in unit time for the identification of proteins. We achieved identification of 2200 proteins/30 mins on LTQ-Orbitrap Velos, 2800 proteins/30 mins on Q-Exactive and Triple-TOF 5600 respectively. We further optimized Fast-seq and worked out a quantitative-version of the Fast-seq workflow: Fast-quantification (Fast-quan) and applied it for protein abundance quantification in HUVEC cell that was treated with a drug candidate MLN4924 (a drug in phase III clinical trial). We were able to quantify > 6700 GPs in 1 day of MS running time and found 99 proteins were up-regulated with high confidence. We expect this efficient alternative approach for in-depth proteome analysis will make the application of MS-based proteomics more accessible to biological applications. 相似文献
966.
Jun-Young Ahn Joon-Yeop Lee Eun-Ju Yang Young-Jin Lee Kyung-Bon Koo Kyung-Sik Song Kyeong-Yeoll Lee 《Journal of Asia》2013,16(3):317-320
Two anthraquinones were isolated from the symbiotic bacteria Photorhabdus temperata of entomopathogenic nematodes Heterorhabditis spp. by repeated column chromatography. They were abundantly present in the culture medium and identified as 1,3-dimethoxy-8-hydroxy-9,10-anthraquinone and 3-methoxychrysazine by spectral analysis. The isolated anthraquinones were highly lethal to larvae of Culex pipiens pallens. Our results suggest that anthraquinones might be useful as biopesticides for the biological control of mosquitoes. 相似文献
967.
968.
Xiang-Jun?Jiang Yi?Hu Ling?Jiang Bin?Zou Ping?Song He?HuangEmail author 《Biotechnology and Bioprocess Engineering》2013,18(2):350-357
Statistical experimental designs combined with solvent engineering for optimization of enzymatic synthesis of L-ascorbyl palmitate were developed. First, the composition of the solvent for co-dissolving polar and apolar substrates was determined. The co-solvent mixture of tert-pentanol: DMSO at a ratio of 9:1 (v/v) and the optimal biocatalyst were obtained. Then, the Plackett-Burman design was implemented to screen the variables that significantly influence the conversion. The method of steepest ascent was used to approach the proximity of optimum. After determining the Plackett-Burman and steepest ascent designs, the optimum values were determined by central composite design under response surface methodology. The statistical analysis showed that the optimum reaction conditions (temperature 50°C, enzyme concentration 5.8 g/L, and substrate molar ratio 11:1, stirring rate 160 rpm, amount of molecular sieve 50 g/L, time 18 h) led to the maximum conversion (66.44%) and production concentration (20.63 g/L). A very satisfactory conversion (64.74%) and production concentration (20.13 g/L) could be achieved in short time (6 h). 相似文献
969.
Pellamaoershanensis Song & Li, sp. n., collected from a colony of Lasius (Dendrolasius) spathepus in Maoershan Natural Reserve, Guangxi, is diagnosed, described and illustrated. The discovery represents the first record of the genus in South China. 相似文献
970.
Kristine Y. Crous Audrey G. Quentin Yan‐Shih Lin Belinda E. Medlyn David G. Williams Craig V. M. Barton David S. Ellsworth 《Global Change Biology》2013,19(12):3790-3807
Eucalyptus species are grown widely outside of their native ranges in plantations on all vegetated continents of the world. We predicted that such a plantation species would show high potential for acclimation of photosynthetic traits across a wide range of growth conditions, including elevated [CO2] and climate warming. To test this prediction, we planted temperate Eucalyptus globulus Labill. seedlings in climate‐controlled chambers in the field located >700 km closer to the equator than the nearest natural occurrence of this species. Trees were grown in a complete factorial combination of elevated CO2 concentration (eC; ambient [CO2] +240 ppm) and air warming treatments (eT; ambient +3 °C) for 15 months until they reached ca. 10 m height. There was little acclimation of photosynthetic capacity to eC and hence the CO2‐induced photosynthetic enhancement was large (ca. 50%) in this treatment during summer. The warming treatment significantly increased rates of both carboxylation capacity (Vcmax) and electron transport (Jmax) (measured at a common temperature of 25 °C) during winter, but decreased them significantly by 20–30% in summer. The photosynthetic CO2 compensation point in the absence of dark respiration (Γ*) was relatively less sensitive to temperature in this temperate eucalypt species than for warm‐season tobacco. The temperature optima for photosynthesis and Jmax significantly changed by about 6 °C between winter and summer, but without further adjustment from early to late summer. These results suggest that there is an upper limit for the photosynthetic capacity of E. globulus ssp. globulus outside its native range to acclimate to growth temperatures above 25 °C. Limitations to temperature acclimation of photosynthesis in summer may be one factor that defines climate zones where E. globulus plantation productivity can be sustained under anticipated global environmental change. 相似文献