全文获取类型
收费全文 | 838篇 |
免费 | 65篇 |
专业分类
903篇 |
出版年
2023年 | 1篇 |
2022年 | 12篇 |
2021年 | 20篇 |
2020年 | 6篇 |
2019年 | 7篇 |
2018年 | 14篇 |
2017年 | 15篇 |
2016年 | 21篇 |
2015年 | 22篇 |
2014年 | 32篇 |
2013年 | 56篇 |
2012年 | 45篇 |
2011年 | 65篇 |
2010年 | 37篇 |
2009年 | 23篇 |
2008年 | 52篇 |
2007年 | 46篇 |
2006年 | 57篇 |
2005年 | 56篇 |
2004年 | 71篇 |
2003年 | 52篇 |
2002年 | 44篇 |
2001年 | 8篇 |
2000年 | 17篇 |
1999年 | 21篇 |
1998年 | 3篇 |
1997年 | 4篇 |
1996年 | 4篇 |
1995年 | 5篇 |
1994年 | 3篇 |
1993年 | 5篇 |
1992年 | 11篇 |
1991年 | 6篇 |
1990年 | 14篇 |
1989年 | 9篇 |
1988年 | 8篇 |
1987年 | 5篇 |
1986年 | 6篇 |
1985年 | 5篇 |
1984年 | 3篇 |
1982年 | 1篇 |
1981年 | 1篇 |
1978年 | 4篇 |
1977年 | 1篇 |
1976年 | 4篇 |
1975年 | 1篇 |
排序方式: 共有903条查询结果,搜索用时 15 毫秒
91.
92.
93.
Torihashi S Ozaki H Hori M Kita M Ohota S Karaki H 《Histochemistry and cell biology》2000,113(2):73-80
A great number of macrophages is found to be evenly distributed in the muscle layer of the gastrointestinal tract. We investigated
their effects on smooth muscle contraction and the initiation of immune reactions such as inflammatory responses. Macrophages
were demonstrated by the uptake of FITC-dextran and their ultrastructural features were elucidated by electron microscopy.
Muscle layers of rats’ ileums were incubated with lipopolysaccharide (LPS) for 4–8 h and the force of smooth muscle contraction
was measured. The induction effect of inducible nitric oxide synthase (iNOS) on macrophages was then checked by immunohistochemistry.
The expression of major histocompatibility complex (MHC) class II was also examined. Macrophages in the muscle layer were
confirmed as resident macrophages and were different from a population of dendritic cells. After incubation with LPS, macrophages
began to express iNOS and produced NO, and it reduced smooth muscle contraction. iNOS-immunopositive cells increased in a
time-dependent manner. Macrophages also began to express MHC class II. The total number of macrophages did not alter after
incubation. Results indicate that resident macrophages in the muscle layer induced iNOS as an inflammatory reaction, affected
smooth muscle contraction, and initiated immune response in the smooth muscle layer of the gastrointestinal tract, when activated
by LPS.
Accepted: 24 November 1999 相似文献
94.
95.
Tomohiro Nishimura Kei Higuchi Yoshimichi Sai Yuki Sugita Yuko Yoshida Masatoshi Tomi Masami Wada Tomohiko Wakayama Atsushi Tamura Sachiko Tsukita Tomoyoshi Soga Emi Nakashima 《PloS one》2014,9(8)
Ezrin is a membrane-associated cytoplasmic protein that serves to link cell-membrane proteins with the actin-based cytoskeleton, and also plays a role in regulation of the functional activities of some transmembrane proteins. It is expressed in placental trophoblasts. We hypothesized that placental ezrin is involved in the supply of nutrients from mother to fetus, thereby influencing fetal growth. The aim of this study was firstly to clarify the effect of ezrin on fetal growth and secondly to determine whether knockout of ezrin is associated with decreased concentrations of serum and placental nutrients. Ezrin knockout mice (Ez−/−) were confirmed to exhibit fetal growth retardation. Metabolome analysis of fetal serum and placental extract of ezrin knockout mice by means of capillary electrophoresis–time-of-flight mass spectrometry revealed a markedly decreased concentration of hypotaurine, a precursor of taurine. However, placental levels of cysteine and cysteine sulfinic acid (precursors of hypotaurine) and taurine were not affected. Lack of hypotaurine in Ez−/− mice was confirmed by liquid chromatography with tandem mass spectrometry. Administration of hypotaurine to heterogenous dams significantly decreased the placenta-to-maternal plasma ratio of hypotaurine in wild-type fetuses but only slightly decreased it in ezrin knockout fetuses, indicating that the uptake of hypotaurine from mother to placenta is saturable and that disruption of ezrin impairs the uptake of hypotaurine by placental trophoblasts. These results indicate that ezrin is required for uptake of hypotaurine from maternal serum by placental trophoblasts, and plays an important role in fetal growth. 相似文献
96.
Ana Carolina Barros de Genaro Rosana Emi Tamagawa Adriano Rodrigues Azzoni Snia Maria Alves Bueno Everson Alves Miranda 《Process Biochemistry》2002,37(12):1413-1420
Aprotinin, a bovine protease inhibitor currently also produced in recombinant bacteria, yeast, and corn, has valuable applications as a human therapeutic and in tissue culture. The objective of this work was to develop the basis of a large-scale aprotinin purification process centered on immobilized metal ion affinity chromatography (IMAC). This technique uses ligands—metal ions—of a lower cost and higher stability than those traditionally used in affinity chromatography. Since aprotinin does not interact with IMAC ligands, collection is from the nonretained fractions (negative chromatography). Stirred-tank batch IMAC adsorption experiments indicated that one-step aprotinin purification could not be successful. Immobilized chymotrypsin chromatography was then used as a prepurification step, yielding a suitable feed for IMAC (with purification factors as high as 476). IMAC column fed with these prepurified materials produced purified aprotinin in the nonretained fractions with purification factors as high as 952. 相似文献
97.
Kubota NK Ohta E Ohta S Koizumi F Suzuki M Ichimura M Ikegami S 《Bioorganic & medicinal chemistry》2003,11(21):4569-4575
Piericidins C5 (1) and C6 (2), two new members of the piericidin family, were isolated from a Streptomyces sp. and a Nocardioides sp., together with known piericidins C1 (3), C2 (4), C3 (5), C4 (6), D1 (7), and A3 (8). The structures were determined on the basis of their spectroscopic data. Both new compounds inhibited cell division of fertilized starfish (Asterina pectinifera) eggs at the minimum inhibitory concentration of 0.09 microg/mL. 相似文献
98.
Characterization of the unfolding process of lipocalin-type prostaglandin D synthase 总被引:6,自引:0,他引:6
Inui T Ohkubo T Emi M Irikura D Hayaishi O Urade Y 《The Journal of biological chemistry》2003,278(5):2845-2852
We found that low concentrations of guanidine hydrochloride (GdnHCl, <0.75 M) or urea (<1.5 M) enhanced the enzyme activity of lipocalin-type prostaglandin (PG) D synthase (L-PGDS) maximally 2.5- and 1.6-fold at 0.5 M GdnHCl and 1 M urea, respectively. The catalytic constants in the absence of denaturant and in the presence of 0.5 M GdnHCl or 1 m urea were 22, 57, and 30 min(-1), respectively, and the K(m) values for the substrate, PGH(2), were 2.8, 8.3, and 2.3 microm, respectively, suggesting that the increase in the catalytic constant was mainly responsible for the activation of L-PGDS. The intensity of the circular dichroism (CD) spectrum at 218 nm, reflecting the beta-sheet content, was also increased by either denaturant in a concentration-dependent manner, with the maximum at 0.5 M GdnHCl or 1 M urea. By plotting the enzyme activities against the ellipticities at 218 nm of the CD spectra of L-PGDS in the presence or absence of GdnHCl or urea, we found two states in the reversible folding process of L-PGDS: one is an activity-enhanced state and the other, an inactive state. The NMR analysis of L-PGDS revealed that the hydrogen-bond network was reorganized to be increased in the activity-enhanced state formed in the presence of 0.5 M GdnHCl or 1 m urea and to be decreased but still remain in the inactive intermediate observed in the presence of 2 M GdnHCl or 4 M urea. Furthermore, binding of the nonsubstrate ligands, bilirubin or 13-cis-retinal, to L-PGDS changed from a multistate mode in the native form of L-PGDS to a simple two-state mode in the activity-enhanced form, as monitored by CD spectra of the bound ligands. Therefore, L-PGDS is a unique protein whose enzyme activity and ligand-binding property are biphasically altered during the unfolding process by denaturants. 相似文献
99.
Kim YS Kim B Karaki H Hori M Ozaki H 《Biochemical and biophysical research communications》2003,311(4):972-978
We investigated the role of Rnd1, a member of the small GTP-binding Rho protein family, in the change in Ca(2+) sensitivity of contractile element in rat myometrium at estrus, gestation, and postpartum stages. In the permeabilized muscles, GTPgammaS or carbachol with GTP increased Ca(2+) sensitivity of contractile force in non-pregnant myometrium at the estrus stage, whereas these stimuli were ineffective in pregnant myometrium at day 21. After postpartum, the reduced Ca(2+) sensitization was recovered. Semi-quantitative RT-PCR analysis indicated that the expressions of RhoA, ROCKI, and ROCKII were not significantly different between non-pregnant and pregnant myometria. In contrast, the expression of Rnd1 was increased during the course of pregnancy, reaching a maximal at day 21, and rapidly declined after the delivery. On the other hand, Ca(2+) sensitization of contractile elements was decreased during the progress in gestation. These results suggest that Rnd1 may have an important role as a negative-feedback control of uterine contraction during gestation through the inhibition of RhoA-mediated increase in the Ca(2+) sensitivity of contractile elements. 相似文献
100.
4-Hydroxy-2-nonenal (HNE), a major lipid peroxidation-derived reactive aldehyde, is a potent inhibitor of sulfhydryl enzymes, such as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). It has been suggested that HNE exerts an inhibitory effect on the enzyme due to the modification of the cysteine residue (Cys-149) at the catalytic site generating the HNE-cysteine Michael addition-type adduct [Uchida, K., and Stadtman, E. R. (1993) J. Biol. Chem. 268, 6388-6393]. In the study presented here, to elucidate the mechanism for the inactivation of GAPDH by HNE, we attempted to identify the modification sites of the enzyme by monitoring the formation of the HNE Michael adducts by mass spectrometric methods. Incubation of GAPDH (1 mg/mL) with 1 mM HNE in 50 mM sodium phosphate buffer (pH 7.4) at 37 degrees C resulted in a time-dependent loss of enzyme activity, which was associated with the covalent binding of HNE to the enzyme. To identify the site of modification of GAPDH by HNE, both the HNE-pretreated and untreated GAPDH were digested with trypsin and V8 protease, and the resulting peptides were subjected to electrospray ionization liquid chromatography-mass spectrometry (ESI-LC-MS). This technique identified five peptides, which contained the HNE adducts at His-164, Cys-244, Cys-281, His-327, and Lys-331 and revealed that both His-164 and Cys-281 were very rapidly modified at 5 min, followed by Cys-244 at 15 min and His-327 and Lys-331 at 30 min. These observations and the observation that the HNE modification of the catalytic center, Cys-149, was not observed suggest that the HNE inactivation of GAPDH is not due to the modification of the catalytic center but to the selective modification of amino acids primarily located in the surface of the GAPDH molecule. 相似文献