全文获取类型
收费全文 | 761篇 |
免费 | 16篇 |
国内免费 | 14篇 |
出版年
2024年 | 2篇 |
2023年 | 1篇 |
2022年 | 12篇 |
2021年 | 10篇 |
2020年 | 17篇 |
2019年 | 20篇 |
2018年 | 20篇 |
2017年 | 14篇 |
2016年 | 10篇 |
2015年 | 15篇 |
2014年 | 35篇 |
2013年 | 47篇 |
2012年 | 29篇 |
2011年 | 35篇 |
2010年 | 27篇 |
2009年 | 37篇 |
2008年 | 44篇 |
2007年 | 44篇 |
2006年 | 51篇 |
2005年 | 27篇 |
2004年 | 36篇 |
2003年 | 20篇 |
2002年 | 21篇 |
2001年 | 6篇 |
2000年 | 3篇 |
1999年 | 3篇 |
1998年 | 8篇 |
1997年 | 8篇 |
1996年 | 9篇 |
1995年 | 7篇 |
1994年 | 5篇 |
1993年 | 6篇 |
1992年 | 6篇 |
1991年 | 6篇 |
1990年 | 8篇 |
1989年 | 10篇 |
1988年 | 1篇 |
1987年 | 5篇 |
1986年 | 4篇 |
1985年 | 11篇 |
1984年 | 37篇 |
1983年 | 28篇 |
1982年 | 20篇 |
1981年 | 7篇 |
1980年 | 8篇 |
1979年 | 9篇 |
1978年 | 2篇 |
排序方式: 共有791条查询结果,搜索用时 15 毫秒
531.
Margaret M. Krahn Donald W. Brown Tracky K. Collier Andrew J. Firedman Rand G. Jenkins Donald C. Malins 《Journal of biochemical and biophysical methods》1980,2(4):233-246
A rapid procedure for the determination of naphthalene and its metabolites in bile of rainbow trout and mice is described. The integrated analytical techniques combine high-performance liquid chromatography/ultraviolet fluorescence detection and plasma desotption/chemical ionization mass spectrometry for identification and quantitation. After separation by reverse-phase liquid chromatography, naphthalene and its metablolites are detected and quantitated by ultraviolet fluoresence spectometry. Identification of two metabolites is confirmed by mass spectometry. A direct insertion probe tip for a conventional chemical ionization mass spectometer was modified to obtain spectra of thermally labile compounds. A spectrum of less than 100 ng of naphthyl glucuronide, a labile glucuronic acid conjugate of 1-naphthol, was obtained with this system. 相似文献
532.
Since 2-fluoro-β-alanine (FBAL) conjugates of bile acids (BA), the primary biliary metabolites of fluoropyrimidine (FP) drugs, have been suggested to be related to the hepatotoxicity which develops in patients receiving FP chemotherapy by intrahepatic arterial infusion (Proc. Natl. Acad. Sci. USA 84, 5439–5443, 1987), it was important to determine whether they undergo enterohepatic circulation and hence accumulate in the liver and biliary system. In initial studies, sensitivity of FBAL-BA conjugates to hydrolysis by pancreatic enzymes was examined. In subsequent in vivo studies, a model FBAL-BA conjugate, FBAL-chenodeoxycholate (FBAL-CDC), was introduced into the lumen of the small intestine of anesthetized rats with biliary fistulas to quantitate the intestinal absorption, metabolism and tissue distribution of the conjugate. The results indicated that: (1) FBAL-BA conjugates were resistant to hydrolysis by pancreatic enzymes (carboxypeptidase A, carboxypeptidase B and trypsin) and by human pancreatic juice, but were completely hydrolyzed by cholylglycine hydrolase. (2) At least one-half of the administered FBAL-CDC was deconjugeted during the process of intestinal absorption, as shown by HPLC analysis of the radioactivity in portal venous blood. (3) Deconjugated FBAL or CDC was reconjugated in liver with other bile acids or amino acids (glycine and taurine), respectively, as shown by radiochromatography of bile. (4) FBAL, formed as a result of hydrolysis of FBAL-CDC, had a wide tissue distribution. In conclusion, FBAL-CDC has a rapid turnover during its enterohepatic circulation due to deconjugation in the intestine and reconjugation in the liver. 相似文献
533.
Transforming growth factor type e (TGFe) is a heat- and acid-stable polypeptide with an apparent molecular weight of 22,000, which stimulates the proliferation of certain epithelial and mesenchymal cells in monolayer and soft agar. TGFe has been purified to homogeneity. Initial acid-ethanol extraction of bovine kidney was followed by batch ion-exchange chromatography utilizing Bio Rex 70 resin. The activity eluted from the Bio Rex 70 resin was concentrated and diafiltered using an Amicon concentrator equipped with an S1Y10 spiral membrane, then was further purified by Bio-Gel P-60 molecular sieve chromatography. Active fractions from molecular sieve chromatography were pooled and purified by heparin-Sepharose affinity chromatography, followed by reverse-phase high-performance liquid chromatography using a microbore C-8 column. The final purification step involved electro-elution of TGFe separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purity of TGFe was assessed to be greater than 90%. 相似文献
534.
Metabolism of halazepam [7-chloro-1,3-dihydro-5-phenyl-1-(2,2,2-trifluoroethyl)-2H-1,4-benzod iazepin- 2-one, HZ] was studied by incubation with liver microsomes prepared from untreated, phenobarbital (PB)-treated, and 3-methylcholanthrene (3MC)-treated male Sprague-Dawley rats. Metabolites of HZ were separated by normal-phase HPLC. Relative rates of HZ metabolism by liver microsomes prepared from untreated and treated rats were PB-treated much greater than untreated greater than 3MC-treated at low concentration of microsomal enzymes (0.25 mg protein per ml of incubation mixture) and PB-treated much greater than 3MC-treated approximately untreated at high concentration of microsomal enzymes (2 mg protein per ml of incubation mixture). The relative amounts of major metabolites were found to be 3-hydroxy-HZ (3-OH-HZ) greater than N-desalkylhalazepam (NDZ, also known as N-desmethyldiazepam and nordiazepam) much greater than oxazepam (OX) for all three rat liver microsomal preparations and the distribution of metabolites was independent of microsomal enzyme concentrations. Enantiomers of 3-OH-HZ were resolved by HPLC on a Chiralcel OC column (cellulose trisphenylcarbamate coated on silica gel, particle size 10 microns). 3-OH-HZ enantiomeres have racemization half-lives of approximately 150 min in pH 4, 7.5, and 10 aqueous solutions. 3-OH-HZ formed in the metabolism of HZ by liver microsomes prepared from untreated and treated rats were found to have 3R/3S enantiomer ratios of 37/63 (untreated), 55/45 (PB-treated), and 36/64 (3MC-treated), respectively. N-dealkylation of 3-OH-HZ by liver microsomes from PB-treated rats was substrate enantioselective; the 3R-enantiomer was N-dealkylated faster than 3S-enantiomer.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
535.
536.
Nisar A. Shaikh 《Molecular and cellular biochemistry》1990,96(1):43-55
Choline phosphoglycerides (CPG) represent the major fraction of heart phospholipids. Since depletion of membrane phospholipids and accumulation of lyso-compounds, particularly lysophosphatidylcholines, have been implicated in arrhythmogenesis, it was of great interest to study the composition of this major phospholipid fraction of the heart at a molecular level in an established animal model. The data presented here describe the first report on the detailed chemical examination of CPG and resolution, characterization and quantitative analysis of the molecular species of this phospholipid fraction from porcine heart by high performance liquid chromatography (HPLC). This fraction constitutes 37.5 ± 0.7% (n = 21) of the total phospholipids and upon successive mild acid and alkaline hydrolyses revealed the presence of essentially three subclasses: diacyl-, alkenylacyl-, and alkylacyl glycerophosphorylcholines, in a relative abundance of 57.7 ± 2.2% (n = 8), 37.3 ± 1.3% (n = 8) and 4.6 ± 0.2% (n = 8), respectively. The fourth subclass, dialkyl CPG was found only in minute amounts (0.43 ± 0.05%, n = 8) and the presence of dialkenyl and alkenylalkyl analogues could not be detected. Alternatively, by converting the CPG fraction to benzoate derivatives after phospholipase C digestion, it was possible to isolate and quantitate subclass composition by TLC/spectroscopy or both subclass compositions and molecular species analysis by HPLC directly by a UV detector online with the column. By these techniques, subclass composition was found to be very similar to that obtained by the chemical hydrolysis technique. By HPLC, up to 25 species can be identified and quantitated in each subclass, their identity being confirmed by gas-liquid chromatography, after their isolation from the column. The analyses showed that up to 74% of the diacyl moiety consisted of 16:0–18:2 (34%), 16:0–18:1 (27%), and 18:0–18:2 (13%) species, while the major species of the alkenylacyl moiety were 16:0–18:2 (44%) 16:0–18:1 (13%), 16:0–20:4 (12%) and 18:1–18:2 (9%) making up more than 75% of the total mass of this subclass. The major molecular species of the alkylacyl moiety was 16:0–18:2, constituting up to 47% of this fraction, while others constituted about 10% (16:0–18:1), 9% (18:1–18:2), 8% (16:0–20:4) and 6% (18:0–18:2), making up 80% of the total mass.The ether chain composition of alkylacyl CPG whether determined after isolation of this fraction by the chemical hydrolysis technique or by HPLC was indistinguishable. Similarly, the aliphatic moieties of diradylglycerols, and their subclasses, whether analysed directly or reconstituted from the molecular species data, were very similar in composition, confirming the accuracy of the data and the reproducibility of the technique devised. This also suggests that this method is suitable to distinguish minor changes in the molecular species of CPG in the heart during the early phase of ischemia and in arrhythmias, and should facilitate further studies on the metabolism of the individual species in health and disease. 相似文献
537.
采用海马内注射方法,探讨了精氨酸加压素对大鼠穿梭箱条件回避行为的影响及其与海马中去甲肾上腺素能末梢递质的关系;用高效液相色谱-电化学检测法研究了精氨酸加压素对海马中单胺类递质及其代谢产物含量的影响。结果显示:(1)两侧海马内注射精氨酸加压素(各50pg),可延缓动物条件回避行为的消退;(2)用6-羟多巴胺(各10μg)损毁两侧海马中的去甲肾上腺素能末梢,可阻断精氨酸加压素的促记忆效应;(3)精氨酸加压素可加速海马中去甲肾上腺素和多巴胺的消失。上述结果提示,加压素对记忆的易化作用,至少部分通过海马起作用;海马内的去甲肾上腺素能末梢可能参与加压素对记忆过程的调节;加压素的作用很可能是通过加强脑内去甲肾上腺素的降解代谢来实现的。 相似文献
538.
在用RP-HPLC法C18色谱柱和紫外检测器检测发酵液中丙氨酸时,由于发酵液中存在丙酮酸,而丙酮酸对紫外吸收过强,且两者峰之间有重叠,导致无法准确检测出丙氨酸含量。为解决此问题,本文在RP-HPLC检测前首先对丙氨酸发酵液进行了乙醇沉淀预处理,实现了丙酮酸和丙氨酸在检测进样前的分离,消除了丙酮酸信号对丙氨酸的影响,能准确测定出丙氨酸含量。达到有效分离的发酵液与无水乙醇最佳比例为1∶14;经氨基酸自动分析仪检测发酵液中丙氨酸实际含量为92.60g/L;经RP-HPLC检测,沉淀中丙氨酸的含量为为92.04 g/L,丙氨酸的回收率高达99.40%。本研究利用乙醇对丙氨酸发酵液进行预处理,使RP-HPLC法可以运用于检测发酵液中丙氨酸含量,适用于工厂发酵生产过程中丙氨酸的快速检测。 相似文献
539.
Mahidhar Tatineni Sandeep Somani Gunaretnam Rajagopal Jinquan Luo Gary L. Gilliland Anastassis Perrakis Robbie P. Joosten 《Protein science : a publication of the Protein Society》2018,27(3):798-808
The Protein Data Bank (PDB) is the global archive for structural information on macromolecules, and a popular resource for researchers, teachers, and students, amassing more than one million unique users each year. Crystallographic structure models in the PDB (more than 100,000 entries) are optimized against the crystal diffraction data and geometrical restraints. This process of crystallographic refinement typically ignored hydrogen bond (H‐bond) distances as a source of information. However, H‐bond restraints can improve structures at low resolution where diffraction data are limited. To improve low‐resolution structure refinement, we present methods for deriving H‐bond information either globally from well‐refined high‐resolution structures from the PDB‐REDO databank, or specifically from on‐the‐fly constructed sets of homologous high‐resolution structures. Refinement incorporating HOmology DErived Restraints (HODER), improves geometrical quality and the fit to the diffraction data for many low‐resolution structures. To make these improvements readily available to the general public, we applied our new algorithms to all crystallographic structures in the PDB: using massively parallel computing, we constructed a new instance of the PDB‐REDO databank ( https://pdb-redo.eu ). This resource is useful for researchers to gain insight on individual structures, on specific protein families (as we demonstrate with examples), and on general features of protein structure using data mining approaches on a uniformly treated dataset. 相似文献
540.
Parasites have developed a variety of strategies for invading hosts and escaping their immune response. A common mechanism by which parasites escape nitric oxide (NO) toxicity is the activation of host arginase. This activation leads to a depletion of l-arginine, which is the substrate for NO synthase, resulting in lower levels of NO and increased production of polyamines that are necessary for parasite growth and differentiation. For this reason, small molecule inhibitors for arginase show promise as new anti-parasitic chemotherapeutics. However, few arginase inhibitors have been reported. Here, we describe the discovery of novel irreversible arginase inhibitors, and their characterization using biochemical, kinetic, and structural studies. Importantly, we determined the site on human arginase that is labeled by one of the small molecule inhibitors. The tandem mass spectra data show that the inhibitor occupies the enzyme active site and forms a covalent bond with Thr135 of arginase. These findings pave the way for the development of more potent and selective irreversible arginase inhibitors. 相似文献