全文获取类型
收费全文 | 1374篇 |
免费 | 165篇 |
国内免费 | 52篇 |
出版年
2024年 | 9篇 |
2023年 | 57篇 |
2022年 | 51篇 |
2021年 | 68篇 |
2020年 | 57篇 |
2019年 | 62篇 |
2018年 | 42篇 |
2017年 | 37篇 |
2016年 | 35篇 |
2015年 | 60篇 |
2014年 | 72篇 |
2013年 | 85篇 |
2012年 | 59篇 |
2011年 | 83篇 |
2010年 | 48篇 |
2009年 | 58篇 |
2008年 | 70篇 |
2007年 | 57篇 |
2006年 | 63篇 |
2005年 | 66篇 |
2004年 | 40篇 |
2003年 | 38篇 |
2002年 | 33篇 |
2001年 | 18篇 |
2000年 | 18篇 |
1999年 | 26篇 |
1998年 | 15篇 |
1997年 | 15篇 |
1996年 | 18篇 |
1995年 | 18篇 |
1994年 | 19篇 |
1993年 | 19篇 |
1992年 | 19篇 |
1991年 | 11篇 |
1990年 | 11篇 |
1989年 | 9篇 |
1988年 | 10篇 |
1987年 | 5篇 |
1986年 | 5篇 |
1985年 | 10篇 |
1984年 | 15篇 |
1983年 | 9篇 |
1982年 | 9篇 |
1981年 | 17篇 |
1980年 | 9篇 |
1979年 | 11篇 |
1978年 | 12篇 |
1974年 | 4篇 |
1973年 | 3篇 |
1972年 | 2篇 |
排序方式: 共有1591条查询结果,搜索用时 46 毫秒
111.
An alternative method for serum protein depletion/enrichment by precipitation at mildly acidic pH values and low ionic strength 下载免费PDF全文
Ann‐Kristin Henning Dirk Albrecht Katharina Riedel Thomas C. Mettenleiter Axel Karger 《Proteomics》2015,15(11):1935-1940
Serum proteome analysis is severely hampered by the extreme dynamic range of protein concentrations, but tools for the specific depletion of highly abundant serum proteins lack for most farm and companion animals. A well‐established alternative strategy to reduce the dynamic range of plasma protein concentrations, treatment with combinatorial peptide ligand libraries (CPLL), is generally applicable but requires large amounts of sample. Therefore, additional depletion/enrichment protocols for plasma and serum samples from animals are desirable. In this respect, we have tested a protein precipitate that formed after withdrawal of salt from human, bovine, or porcine serum at pH 4.2. The bovine sample was composed of over 300 proteins making it a potential source for biomarker discovery. Precipitation was highly reproducible and the concentrations of albumin and other highly abundant serum proteins were strongly reduced. In comparison to the CPLL treatment, precipitation did not introduce any selection bias based on hydrophathy or pI. However, the composition of both preparations was partially complementary. Salt withdrawal at pH 4.2 is suggested as additional depletion/enrichment strategy for serum samples. Also, we point out that the removal of precipitates from serum samples under the described conditions bears the risk of losing a valuable protein fraction. 相似文献
112.
Torben Breindahl Ole Simonsen Kirsten Andreasen 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(1):76-82
A high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS–MS) method, using back-flush column-switching was developed for total drug concentrations of ropivacaine in serum and drainage blood in the measuring range 0.1–10 μg/mL. Samples were diluted with internal standard (2H7-ropivacaine) and extraction buffer, centrifuged and injected directly onto a BioTrap 500 MS extraction column. Using a time programmed six-port valve switch, ropivacaine was back-flushed onto a Zorbax SB-Aq analytical column, gradient eluted and finally detected after electro spray ionisation and multiple reaction monitoring (MRM) of the transitions m/z 275 → m/z 126 and m/z 282 → m/z 133 for ropivacaine and 2H7-ropivacaine, respectively. Accuracy (bias-%) was −1.5 to 5.8% and intermediate precision (C.V.) was 1.4–3.1%. The low sample amount required (10 μL), high specificity and short run time (6 min) makes it very suitable for determination of ropivacaine. Using the same methodology as described above and 200 μL ultrafiltrate, the free drug concentrations of ropivacaine in serum could be precisely determined with a C.V. below 3%. The method was used to investigate the safety of reinfusion of drainage blood after knee and hip arthroplasty when ropivacaine (Naropin®) was used for local analgesia. Data for 30 patients are summarised. 相似文献
113.
Linsheng Liu Jiye Aa Guangji Wang Bei Yan Xinwen Wang Bei Cao Mengjie Li Yuanting Zheng Fang Zhou Zimei Wu 《Analytical biochemistry》2010,406(2):105-6697
In metabolomic research, blood plasma and serum have been considered to possess similar compositions and properties. Their perceived equivalence has resulted in researchers choosing arbitrarily between serum and plasma for analysis. Here, routine serum and plasma were prepared and their low-molecular-weight compounds were determined using gas chromatography/time-of-flight mass spectrometry. Principal components analysis was applied to process the acquired data, and marked differences in metabolite profiles were observed between serum and plasma. Of the 72 identified compounds, 36 (50%) discriminate serum from plasma, with 29 and 7 metabolites showing a significantly higher abundance (t test, P < 0.05) in serum and plasma, respectively. Incubation of blood had distinct effects on the analyte peak areas, with the effects being more pronounced for plasma than for serum and more pronounced for a shorter incubation than for a longer incubation. These results highlight the importance in choosing serum or plasma as the analytical sample and in stipulating the incubation time. Because incubation affected the analyte peak areas less in serum than in plasma, we recommend serum as the sample of choice in metabolomic studies. 相似文献
114.
Serum or plasma can be utilized in a variety of studies targeted toward the discovery of disease biomarkers. In this study, the proteome profiles of plasma samples prepared using various anticoagulants (EDTA, heparin or citrate), were compared with those of serum using two-dimensional electrophoresis (2-DE). Proteins which evidenced different levels in the plasma and serum were screened and identified using ESI-Q-TOF MS/MS. The proteins which became detectable after the removal of fibrinogen from serum were identified as pigment epithelial differentiating factor (four spots), fetuin-like protein, and the hemopexin precursor. In particular, three proteins, pre-serum amyloid P component, plasma glutathione peroxidase precursor, and tetranectin, evidenced increased volume intensity only in the plasma samples prepared with EDTA. 相似文献
115.
Chromium(III) is often claimed to have a positive effect on body composition, while the responses in researches with supplementation
of different chemical form of chromium are various and inconsistent. We have studied the effects of 6 weeks of treatment with
three different forms of chromium (300 μg/kg) as chromium chloride, chromium tripicolinate, and chromium nanocomposite (CrNano)
on growth, body composition, serum parameters, and tissue chromium in rats. The supplementataion of CrNano significantly increased
average daily gain, food efficiency, and lean body mass and decreased fat mass and body fat proportion and serum levels of
glucose, urea nitrogen, triglyceride, and insulin. Chromium contents in liver, kidney, and hind leg muscle were increased
significantly with the addition of CrNano in diet. The results indicate that chromium nanocomposite has higher efficacy on
growth and body composition compared to the traditional chromium agents. 相似文献
116.
Zoffmann S Bertrand S Do QT Bertrand D Rognan D Hibert M Galzi JL 《Journal of neurochemistry》2007,101(2):506-516
Neurokinin A stimulates physiological responses in the peripheral and central nervous systems upon interacting primarily with the tachykinin NK2 receptor (NK2R). In this study, the structure of NKA bound to the NK2R is characterised by use of fluorescence resonance energy transfer. Four fluorescent NKA analogues with Texas red introduced at amino acid positions 1, 4, 7 and 10 were prepared. When bound to a NK2R carrying enhanced green fluorescent protein at the N-terminus, all peptides reduce green fluorescent protein fluorescence from 10% to 50% due to energy transfer. The derived donor-acceptor distances are 46, 55, 59 and 69 A for the fluorophore linked to positions 1-10, respectively. The monotonic increase in distance clearly indicates that the peptide adopts an extended structure when bound to its receptor. The present data are used, in combination with rhodopsin structure, fluorescence studies, photoaffinity labelling and site-directed mutagenesis data to design a computer model of the NKA-NK2R complex. We propose that the N-terminus of NKA is exposed and accessible to the extracellular medium. Subsequent amino acids of the NKA peptide become progressively more buried residues up to approximately one-third of the transmembrane-spanning domain. 相似文献
117.
Tokumura A Kume T Fukuzawa K Tahara M Tasaka K Aoki J Arai H Yasuda K Kanzaki H 《Life sciences》2007,80(18):1641-1649
Levels of lysophosphatidic acid (LPA), an important phospholipid mediator, in serum and ascitic fluid from ovarian cancer patients were shown to be higher than those from healthy women and from patients with other type of cancer, respectively. Although LPA in human serum seems mainly to be generated by lysophospholipase D (lysoPLD), the source and pathway for LPA in the ascitic fluid remain still obscure. In this study, we examined whether lysoPLD activity producing bioactive LPA in human peritoneal fluid was significantly elevated under pathological statuses. Lysophospholipase D activity in human peritoneal fluids was measured by quantifying choline released from exogenous lysophosphatidylcholine on their incubation at 37 degrees C. We also compared the activity of lysoPLD in sera from patients with different gynecologic diseases. We found relatively high lysoPLD activity in peritoneal fluids from patients with ovarian cancer, dermoid cyst or mucinous cystadenoma, whereas there were no significant differences in the serum lysoPLD activity among clinical groups and healthy subjects. The lysoPLD in the peritoneal fluid was found to have similar substrate specificity and metal ion requirement to those of serum lysoPLD, that has been identified as autotaxin, a tumor cell-motility stimulating protein. Our results suggest that increased lysoPLD activity in peritoneal fluid from patients with certain gynecologic tumors might be relevant to its potential of tumor progression. 相似文献
118.
This article describes a multicommutated flow injection-solid phase spectroscopy system implemented with photochemically induced fluorescence for the determination of flufenamic acid (FFA). A strongly fluorescent photoproduct is generated when FFA is irradiated online under UV light in a strong sulfuric medium. The photoproduct generated is retained on C(18) silica gel (which fills the detection area of the flow cell) and directly monitored on the active solid support at 258/442 nm (lambda(ex)/lambda(em)). After maximum signal recording, the sensing zone is regenerated by eluting the retained photoproduct with an appropriate H(2)SO(4)/MeOH solution. The sensor, completely automated, is based on the use of three-way solenoid valves conveniently operated by a homemade multicommutation software written in Java language. The system is calibrated at 10 and 60s for sampling time, showing detection limits of 1.28 x 10(-9) and 5.33 x 10(-10) molL(-1) and sampling rates of 38 and 28 h(-1), respectively, with relative standard deviations of 0.9 and 1.2%. The applicability of the method is demonstrated for the determination of FFA in human serum, human urine, and a pharmaceutical preparation without any pre-treatment. Good recovery levels were achieved between 90.5 and 103.7%. 相似文献
119.
120.
苦瓜的营养价值、化学成分以及药理作用研究进展 总被引:1,自引:0,他引:1
本文总结了半个多世纪来国内外学者对苦瓜的研究,为苦瓜进一步开发提供了借鉴。 相似文献