全文获取类型
收费全文 | 1452篇 |
免费 | 113篇 |
国内免费 | 74篇 |
出版年
2024年 | 7篇 |
2023年 | 32篇 |
2022年 | 38篇 |
2021年 | 50篇 |
2020年 | 59篇 |
2019年 | 67篇 |
2018年 | 51篇 |
2017年 | 50篇 |
2016年 | 39篇 |
2015年 | 46篇 |
2014年 | 82篇 |
2013年 | 98篇 |
2012年 | 61篇 |
2011年 | 82篇 |
2010年 | 68篇 |
2009年 | 91篇 |
2008年 | 78篇 |
2007年 | 88篇 |
2006年 | 74篇 |
2005年 | 70篇 |
2004年 | 65篇 |
2003年 | 48篇 |
2002年 | 30篇 |
2001年 | 31篇 |
2000年 | 15篇 |
1999年 | 22篇 |
1998年 | 14篇 |
1997年 | 21篇 |
1996年 | 13篇 |
1995年 | 14篇 |
1994年 | 15篇 |
1993年 | 14篇 |
1992年 | 15篇 |
1991年 | 5篇 |
1990年 | 8篇 |
1989年 | 8篇 |
1988年 | 3篇 |
1987年 | 9篇 |
1986年 | 2篇 |
1985年 | 11篇 |
1984年 | 9篇 |
1983年 | 10篇 |
1982年 | 4篇 |
1981年 | 6篇 |
1980年 | 6篇 |
1977年 | 2篇 |
1974年 | 3篇 |
1973年 | 2篇 |
1971年 | 1篇 |
1970年 | 1篇 |
排序方式: 共有1639条查询结果,搜索用时 15 毫秒
101.
Shay Bracha Michael McNamara Ian Hilgart Milan Milovancev Jan Medlock Cheri Goodall Samanthi Wickramasekara Claudia S. Maier 《Analytical biochemistry》2014
Transitional cell carcinoma (TCC), the most common cancer of the urinary bladder in dogs, is usually diagnosed at an advanced disease stage with limited response to chemotherapy. Commercial screening tests lack specificity and current diagnostic procedures are invasive. A proof of concept pilot project for analyzing the canine urinary proteome as a noninvasive diagnostic tool for TCC identification was conducted. Urine was collected from 12 dogs in three cohorts (healthy, urinary tract infection, TCC) and analyzed using liquid chromatography tandem mass spectrometry. The presence of four proteins (macrophage capping protein, peroxiredoxin 5, heterogeneous nuclear ribonucleoproteins A2/B, and apolipoprotein A1) was confirmed via immunoblot. Of the total 379 proteins identified, 96 were unique to the TCC group. A statistical model, designed to evaluate the accuracy of this multiplex biomarker approach for diagnosis of TCC, predicted the presence of disease with 90% accuracy. 相似文献
102.
Vijaya Saradhi UV Ling Y Wang J Chiu M Schwartz EB Fuchs JR Chan KK Liu Z 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(30):3045-3051
Curcumin and tetrahydrocurcumin (THC) have been found as potent DNMT1 inhibitors, but they suffer from low oral bioavailability and rapid metabolism in vivo. To circumvent these problems, two curcumin analogs: 1,7-bis(3,4-dimethoxyphenyl)-4,4-dimethyl-1,6-heptadiene-3,5-dione (TMC) and 1,7-bis(3,4-dimethoxyphenyl)-4-cyclohexyl-1,6-heptadiene-3,5-dione (DMCHC) have been synthesized to enhance their stability by blocking the two metabolic sites, the phenolic and C4 methylene moieties. Both compounds have shown inhibitory activity on M. SssI similar to that of curcumin and THC (Poster, M1114, AAPS, 2009). Preclinical pharmacokinetics has yet to be performed. In this paper, a simple liquid chromatography-tandem mass spectrometric method was developed for the determination of these four curcuminoids in cell medium and mouse plasma. The method showed linearity from 1 to 1000 ng/mL with the lower limit of quantification of 1 ng/mL in cell medium, and 5 ng/mL in mouse plasma for all test curcuminoids. The within-day coefficients of variation were found to be below 15% and the accuracy was in the range of 85-115%. This method was subsequently used to evaluate their stability in these matrices and a pilot pharmacokinetics of curcumin, DMCHC and TMC in mice after an intraperitoneal (i.p.) cassette dosing of 10mg/kg each. Curcuminoids degraded in two phases with terminal half lives of 186, 813, 724, and 2000 min for curcumin, THC, TMC, and DMCHC, respectively, in cell culture medium. In plasma, their respective half lives were 111, 232, 1202 and 3000 min. These data demonstrated that their stability is in the order curcumin相似文献
103.
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
104.
Samantha A. Nelis Catherine Sievers Mark Jarrett Lisa M. Nissen Carl M.J. Kirkpatrick P. Nicholas Shaw 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(22):2018-2022
In this paper, a method for the sensitive and reproducible analysis of lignocaine and its four principal metabolites, monoethylxylidide (MEGX), glycylxylidide (GX), 3-hydroxylignocaine (3-HO-LIG), 4-hydroxylignocaine (4-HO-LIG) in equine urine and plasma samples is presented. The method uses liquid chromatography coupled to tandem mass spectrometry operating in electrospray ionisation positive ion mode (+ESI) via multiple reaction monitoring (MRM). Sample preparation involved solid-phase extraction using a mixed-mode phase. The internal standard adopted was lignocaine-d10. Lignocaine and its metabolites were successfully resolved using an octadecylsilica reversed-phase column using a gradient mobile phase of acetonitrile and 0.1% (v/v) aqueous formic acid at a flow rate of 300 μL/min. Target analytes and the internal standard were determined by using the following transitions; lignocaine, 235.2 > 86.1; 3-HO-LIG and 4-HO-LIG, 251.2 > 86.1; MEGX, 207.1 > 58.1; GX, 179.1 > 122.1; and lignocaine-d10, 245.2 > 96.1. Calibration curves were generated over the range 1–100 ng/mL for plasma samples and 1–1000 ng/mL for urine samples. The method was validated for instrument linearity, repeatability and detection limit (IDL), method linearity, repeatability, detection limit (MDL), quantitation limit (LOQ) and recovery. The method was successfully used to analyse both plasma and urine samples following a subcutaneous administration of lignocaine to a thoroughbred horse. 相似文献
105.
Dezhi Kong Sanni Li Xiaowei Zhang Jianmin Gu Man Liu Yan Meng Yan Fu Xiaojin La Gangqiang Xue Lantong Zhang Qiao Wang 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(29):2989-2996
A simple, sensitive and selective liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for the simultaneous determination of m-nisoldipine and its three metabolites in rat plasma has been developed using nitrendipine as an internal standard (IS). Following liquid–liquid extraction, the analytes were separated using an isocratic mobile phase on a reverse phase C18 column and analyzed by MS in the multiple reaction monitoring (MRM) mode. To avoid contamination by residual sample in the injection syringe, a special injection protocol was developed. We found that m-nisoldipine, metabolite M1 and IS could be ionized under positive or negative electrospray ionization conditions, whereas metabolite M and M2 could only be ionized in the positive mode. The mass spectrometry fragmentation pathways for these analytes are analyzed and discussed herein. The total analysis time required less than 5 min per sample. We employed this method successfully to study the metabolism of m-nisoldipine when it was orally administered to rats at a dose of 9 mg/kg. Three metabolites of m-nisoldipine and an unknown compound of molecular weight 386 were found for the first time in rat plasma. The concentration of the potentially active metabolite was approximately equal to its parent compound concentration. 相似文献
106.
尖孢镰刀菌生产蒽醌色素的液体发酵条件研究 总被引:2,自引:0,他引:2
优化了尖孢镰刀菌液体发酵生产蒽醌类红色素的发酵条件。通过单因素实验和正交优化实验,确定最佳产色素发酵培养基为:可溶性淀粉30%,(NH4)2SO4 3%,MgSO4 0.3%,KH2PO4 4%,pH 6.0。产色素最适培养条件为:初始pH6.0,装液量20%,接种量10%,吐温-80添加量1%,温度28℃,摇床转速200r/min,发酵周期120h。此条件下,色素效价即可达到8.184U/ml,比优化前提高了1.8倍。国内首次对尖孢镰刀菌所产蒽醌色素进行研究,为其进一步应用奠定基础。 相似文献
107.
Tocopherols (vitamin E) are lipid soluble antioxidants synthesized by plants and some cyanobacteria. We have earlier reported that overexpression of the γ-tocopherol methyl transferase (γ-TMT) gene from Arabidopsis thaliana in transgenic Brassica juncea plants resulted in an over six-fold increase in the level of α-tocopherol, the most active form of all the tocopherols. Tocopherol levels have been shown to increase in response to a variety of abiotic stresses. In the present study on Brassica juncea, we found that salt, heavy metal and osmotic stress induced an increase in the total tocopherol levels. Measurements of seed germination, shoot growth and leaf disc senescence showed that transgenic Brassica juncea plants overexpressing the γ-TMT gene had enhanced tolerance to the induced stresses. Analysis of the chlorophyll a fluorescence rise kinetics, from the initial “O” level to the “P” (the peak) level, showed that there were differential effects of the applied stresses on different sites of the photosynthetic machinery; further, these effects were alleviated in the transgenic (line 16.1) Brassica juncea plants. We show that α-tocopherol plays an important role in the alleviation of stress induced by salt, heavy metal and osmoticum in Brassica juncea. 相似文献
108.
Seema Joshi Gopal Singh Bisht Diwan S. Rawat Rita Kumar Santosh Pasha 《生物化学与生物物理学报:生物膜》2010,1798(10):1864-1875
Cationic antimicrobial peptides (CAMPs) are novel candidates for drug development. Here we describe design of six short and potent CAMPs (SA-1 to SA-6) based on a minimalist template of 12 residues H+HHG+HH+HH+NH2 (where H: hydrophobic amino acid and +: charged hydrophilic amino acid). Designed peptides exhibit good antibacterial activity in micro molar concentration range (1-32 μg/ml) and rapid clearance of Gram-positive and Gram-negative bacterial strains at concentrations higher than MIC. For elucidating mode of action of designed peptides various biophysical studies including CD and Trp fluorescence were performed using model membranes. Further based on activity, selectivity and membrane bound structure; modes of action of Trp rich peptide SA-3 and template based peptide SA-4 were compared. Calcein dye leakage and transmission electron microscopic studies with model membranes exhibited selective membrane active mode of action for peptide SA-3 and SA-4. Extending our work from model membranes to intact E. coli ATCC 11775 in scanning electron micrographs we could visualize different patterns of surface perturbation caused by peptide SA-3 and SA-4. Further at low concentration rapid translocation of FITC-tagged peptide SA-3 into the cytoplasm of E. coli cells without concomitant membrane perturbation indicates involvement of intracellular targeting mechanism as an alternate mode of action as was also evidenced in DNA retardation assay. For peptide SA-4 concentration dependent translocation into the bacterial cytoplasm along with membrane perturbation was observed. Establishment of a non specific membrane lytic mode of action of these peptides makes them suitable candidates for drug development. 相似文献
109.
110.
Villalba MM Litchfield VJ Smith RB Franklin AM Livingstone C Davis J 《Journal of biochemical and biophysical methods》2007,70(5):797-802
Quinones are well established as key players in the production of reactive oxygen species within cellular environments. Many factors govern their cytotoxicity but most studies have been restricted to a few, core, derivatives. A new strategy for the in situ production of quinone derivatives has been developed such that libraries of diverse functionality can be rapidly created without recourse to extensive synthetic procedures. The approach relies upon nucleophilic addition by reduced thiol derivatives to the quinone core within a pre-culture assay mixture and provides a generic strategy that exploits the large reservoir of commercial thiols currently available. A readily accessible chromatographic method has been developed that allows the derivatisation process to be easily monitored and the purity of the resulting one pot preparation to be assessed. The viability of the combinatorial approach has been fully validated through comparison with a range of quinone-S-conjugates prepared using conventional bench synthesis. The latter have been fully characterised. 相似文献