全文获取类型
收费全文 | 900篇 |
免费 | 43篇 |
出版年
2023年 | 4篇 |
2022年 | 12篇 |
2021年 | 30篇 |
2020年 | 10篇 |
2019年 | 16篇 |
2018年 | 18篇 |
2017年 | 15篇 |
2016年 | 36篇 |
2015年 | 48篇 |
2014年 | 48篇 |
2013年 | 91篇 |
2012年 | 80篇 |
2011年 | 73篇 |
2010年 | 54篇 |
2009年 | 47篇 |
2008年 | 51篇 |
2007年 | 58篇 |
2006年 | 55篇 |
2005年 | 34篇 |
2004年 | 42篇 |
2003年 | 40篇 |
2002年 | 25篇 |
2001年 | 9篇 |
2000年 | 5篇 |
1999年 | 2篇 |
1998年 | 2篇 |
1997年 | 1篇 |
1996年 | 1篇 |
1995年 | 1篇 |
1992年 | 2篇 |
1991年 | 3篇 |
1990年 | 1篇 |
1989年 | 4篇 |
1988年 | 2篇 |
1986年 | 3篇 |
1985年 | 2篇 |
1984年 | 4篇 |
1983年 | 1篇 |
1982年 | 3篇 |
1981年 | 2篇 |
1980年 | 1篇 |
1977年 | 1篇 |
1976年 | 1篇 |
1975年 | 2篇 |
1973年 | 1篇 |
1972年 | 2篇 |
排序方式: 共有943条查询结果,搜索用时 31 毫秒
61.
Claudin-16 is involved in the paracellular reabsorption of Mg2+ in the thick ascending limb of Henle. Little is known about the mechanism regulating the tight junctional localization of claudin-16. Here, we examined the effect of Mg2+ deprivation on the distribution and function of claudin-16 using Madin-Darby canine kidney (MDCK) cells expressing FLAG-tagged claudin-16. Mg2+ deprivation inhibited the localization of claudin-16 at tight junctions, but did not affect the localization of other claudins. Re-addition of Mg2+ induced the tight junctional localization of claudin-16, which was inhibited by U0126, a MEK inhibitor. Transepithelial permeability to Mg2+ was also inhibited by U0126. The phosphorylation of ERK was reduced by Mg2+ deprivation, and recovered by re-addition of Mg2+. These results suggest that the MEK/ERK-dependent phosphorylation of claudin-16 affects the tight junctional localization and function of claudin-16. Mg2+ deprivation decreased the phosphothreonine levels of claudin-16. The phosphothreonine levels of T225A and T233A claudin-16 were decreased in the presence of Mg2+ and these mutants were widely distributed in the plasma membrane. Furthermore, TER and transepithelial Mg2+ permeability were decreased in the mutants. We suggest that the tight junctional localization of claudin-16 requires a physiological Mg2+ concentration and the phosphorylation of threonine residues via a MEK/ERK-dependent pathway. 相似文献
62.
Akira Ikari Kosuke Atomi Keishi Kinjo Yohei Sasaki Junko Sugatani 《Life sciences》2010,86(19-20):766-773
AimsLoss of magnesium (Mg2+) inhibits cell proliferation and augments nephrotoxicant-induced renal injury, but the role of Mg2+ has not been clarified in detail. We examined the effect of extracellular Mg2+ deprivation on a MEK–ERK cascade and cell proliferation using a renal epithelial cell line, Madin-Darby canine kidney (MDCK) cells.Main methodsMDCK cells were cultured in Mg2+-containing or Mg2+-free media. A HA-tagged constitutively active (CA)-MEK1 and a dominant negative (DN)-MEK1 were transfected into MDCK cells. The level of protein was examined by Western blotting. The intracellular free Mg2+ concentration ([Mg2+]i) was measured using a fluorescent dye, mag-fura 2. Cell proliferation was determined by WST-1 assay. Dead cells were identified by staining with annexin V-FITC and propidium iodide.Key findingsIn the presence of fetal calf serum (FCS), Mg2+ deprivation decreased phosphorylated-ERK1/2 (p-ERK1/2) levels and [Mg2+]i. Re-addition of Mg2+ increased p-ERK1/2 levels, which were inhibited by U0126, a specific inhibitor of a MEK–ERK cascade. Glutathione-S-transferase pull-down and coimmunoprecipitation assays showed that CA-MEK1 and DN-MEK1 binds with ERK1/2 in the presence of Mg2+. In contrast, neither CA-MEK1 nor DN-MEK1 bound to ERK1/2 in the absence of Mg2+. These results indicate that the MEK–ERK cascade is regulated by [Mg2+]i. Cell proliferation was increased by the treatment with FCS or the expression of CA-MEK1 in the presence of Mg2+, but was inhibited by Mg2+ deprivation. Mg2+ deprivation did not increase the number of dead cells.SignificanceMg2+ is involved in the regulation of the MEK–ERK cascade and cell proliferation in MDCK cells. 相似文献
63.
Kumai T Takeba Y Matsumoto N Nakaya S Tsuzuki Y Yanagida Y Hayashi M Kobayashi S 《Life sciences》2007,81(15):1193-1198
We investigated the effects of fasudil, a Rho kinase inhibitor, on hypertension in spontaneously hypertensive rats and on the catecholamine synthetic pathway. Ten-week-old male SHR and Wistar-Kyoto rats were administered fasudil (10 mg/kg/day s.c.) for 4 days. Systolic blood pressure was measured using the tail-cuff method. Catecholamine levels were measured with high-performance liquid chromatography-ECD methods. Tyrosine hydroxylase protein levels were measured in Western blot analysis. The tyrosine hydroxylase mRNA level was measured using real-time PCR methods. Fasudil significantly decreased systolic blood pressure in spontaneously hypertensive rats, but not in Wistar-Kyoto rats. Fasudil also significantly decreased catecholamine, tyrosine hydroxylase protein, and tyrosine hydroxylase mRNA levels in the adrenal medulla of spontaneously hypertensive rats. These results suggest that the depressor effects of fasudil on hypertension in spontaneously hypertensive rats may be related to inhibition of the catecholamine synthetic pathway. 相似文献
64.
Watanabe Y Sakihara T Mukuda T Ando M 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》2007,177(8):867-873
The effects of isotocin (IT) and vasotocin (VT), which are fish analogues of mammalian oxytocin and vasopressin respectively,
were examined in the isolated upper esophageal sphincter (UES) muscle. IT relaxed and VT constricted the UES muscle in a concentration-dependent
manner. The relaxation by IT and the contraction by VT were completely blocked by H-9405 (an oxytocin receptor antagonist)
and by H-5350 (a V1-receptor antagonist), respectively, suggesting that the eel UES possesses both IT and VT receptors. Truncated fragments of
VT did not show any significant effects, indicating that all nine residues are essential for the VT and IT actions. IT may
relax the UES muscle through enhancing cAMP production, since similar relaxation was also observed after treatment with 3-isobutyl-1-methylxantine,
forskolin and 8-bromoadenosine, 3′, 5′-cyclic mono-phosphate (8BrcAMP). Although 8-bromoguanosine, 3′, 5′-cyclic monophosphate
also relaxed the UES, its effect was less than 1/3 of that 8BrcAMP, suggesting minor contribution of nitric oxide (NO) in
the relaxation of the UES muscle. Both peptides seem to act directly on the UES muscle, not through release of other substances
from the epithelial cells, since similar relaxation and contraction were observed even in the scraped UES preparations. When
IT and VT were intravenously administrated (in vivo experiments), the drinking rate of the seawater eel was enhanced by IT
and was inhibited by VT. These effects correspond to the in vitro results described above, relaxation by IT and contraction
by VT in the UES muscle. The significance of the relaxing effect by IT is discussed with respect to controlling the drinking
behavior of the eel. 相似文献
65.
Overwintering freeze-tolerant larvae of Chilo suppressalis can survive at -25 degrees C, but non-diapausing larvae cannot. We reported earlier that to prevent intracellular freezing, which causes death in overwintering larvae of the Saigoku ecotype distributed in southwestern Japan, water leaves and glycerol enters fat body cells through water channels during freezing. However, it is still unclear how diapause and low-temperature exposure are related to the acquisition of freeze tolerance. We compared the extent of tissue damage, accumulation of glycerol, and transport of glycerol and water in fat body tissues between cold-acclimated and non-acclimated non-diapausing and diapausing larvae. The tissue from cold-acclimated diapausing larvae could survive only when frozen in Grace's insect medium with 0.25 M glycerol at -20 degrees C. The protection provided by glycerol was offset by mercuric chloride, which is a water-channel inhibitor. Fat body tissue isolated from non-acclimated diapausing larvae was injured by freezing even though glycerol was added to the medium, but the level of freezing injury was significantly lower than in non-diapausing larvae. Radiotracer assays in cold-acclimated diapausing larvae showed that during freezing, water left the cells into the medium and glycerol entered the cells from the medium at the same time. Therefore, in Saigoku ecotype larvae of the rice stem borer, both diapause and cold-acclimation are essential to accumulate glycerol and activate aquaporin for the avoidance of freezing injury. 相似文献
66.
Nakamichi Y Udagawa N Kobayashi Y Nakamura M Yamamoto Y Yamashita T Mizoguchi T Sato M Mogi M Penninger JM Takahashi N 《Journal of immunology (Baltimore, Md. : 1950)》2007,178(1):192-200
Osteoprotegerin (OPG) is a decoy receptor for receptor activator of NF-kappaB ligand (RANKL). We previously reported that OPG deficiency elevated the circulating level of RANKL in mice. Using OPG(-/-) mice, we investigated whether OPG is involved in the shedding of RANKL by cells expressing RANKL. Osteoblasts and activated T cells in culture released a large amount of RANKL in the absence of OPG. OPG or a soluble form of receptor activator of NF-kappaB (the receptor of RANKL) suppressed the release of RANKL from those cells. OPG- and T cell-double-deficient mice showed an elevated serum RANKL level equivalent to that of OPG(-/-) mice, indicating that circulating RANKL is mainly derived from bone. The serum level of RANKL in OPG(-/-) mice was increased by ovariectomy or administration of 1alpha,25-dihydroxyvitamin D(3). Expression of RANKL mRNA in bone, but not thymus or spleen, was increased in wild-type and OPG(-/-) mice by 1alpha,25-dihydroxyvitamin D(3). These results suggest that OPG suppresses the shedding of RANKL from osteoblasts and that the serum RANKL in OPG(-/-) mice exactly reflects the state of bone resorption. 相似文献
67.
Borna disease virus matrix protein is an integral component of the viral ribonucleoprotein complex that does not interfere with polymerase activity 下载免费PDF全文
Chase G Mayer D Hildebrand A Frank R Hayashi Y Tomonaga K Schwemmle M 《Journal of virology》2007,81(2):743-749
We have recently shown that the matrix protein M of Borna disease virus (BDV) copurifies with the affinity-purified nucleoprotein (N) from BDV-infected cells, suggesting that M is an integral component of the viral ribonucleoprotein complex (RNP). However, further studies were hampered by the lack of appropriate tools. Here we generated an M-specific rabbit polyclonal antiserum to investigate the intracellular distribution of M as well as its colocalization with other viral proteins in BDV-infected cells. Immunofluorescence analysis revealed that M is located both in the cytoplasm and in nuclear punctate structures typical for BDV infection. Colocalization studies indicated an association of M with nucleocapsid proteins in these nuclear punctate structures. In situ hybridization analysis revealed that M also colocalizes with the viral genome, implying that M associates directly with viral RNPs. Biochemical studies demonstrated that M binds specifically to the phosphoprotein P but not to N. Binding of M to P involves the N terminus of P and is independent of the ability of P to oligomerize. Surprisingly, despite P-M complex formation, BDV polymerase activity was not inhibited but rather slightly elevated by M, as revealed in a minireplicon assay. Thus, unlike M proteins of other negative-strand RNA viruses, BDV-M seems to be an integral component of the RNPs without interfering with the viral polymerase activity. We propose that this unique feature of BDV-M is a prerequisite for the establishment of BDV persistence. 相似文献
68.
Matsuki Y Ohmura-Hoshino M Goto E Aoki M Mito-Yoshida M Uematsu M Hasegawa T Koseki H Ohara O Nakayama M Toyooka K Matsuoka K Hotta H Yamamoto A Ishido S 《The EMBO journal》2007,26(3):846-854
The presence of post-translational regulation of MHC class II (MHC II) under physiological conditions has been demonstrated recently in dendritic cells (DCs) that potently function as antigen-presenting cells (APCs). Here, we report that MARCH-I, an E3 ubiquitin ligase, plays a pivotal role in the post-translational regulation of MHC II in B cells. MARCH-I expression was particularly high in B cells, and the forced expression of MARCH-I induced the ubiquitination of MHC II. In B cells from MARCH-I-deficient mice (MARCH-I KO), the half-life of surface MHC II was prolonged and the ubiquitinated form of MHC II completely disappeared. In addition, MARCH-I-deficient B cells highly expressed exogenous antigen-loaded MHC II on their surface and showed high ability to present exogenous antigens. These results suggest that the function of MHC II in B cells is regulated through ubiquitination by MARCH-I. 相似文献
69.
Ishikawa T Terai S Urata Y Marumoto Y Aoyama K Murata T Mizunaga Y Yamamoto N Nishina H Shinoda K Sakaida I 《Cell and tissue research》2007,327(3):463-470
We previously reported that fibroblast growth factor 2 (FGF2) facilitated the differentiation of transplanted bone marrow
cells (BMCs) into hepatocytes. Our earlier study also demonstrated that administration of FGF2 in combination with bone marrow
transplantation (BMT) synergistically activated tumor necrosis factor-alpha signaling and significantly improved liver function
and prognosis more than BMT alone. However, the way that it affected the extracellular matrix remained unclear. Here, we investigated
the effect of FGF2 treatment together with BMT on liver fibrosis in mice treated with carbon tetrachloride (CCl4). Transplantation of BMCs and concurrent treatment with FGF2 caused a statistically significant reduction in CCl4-induced liver fibrosis that was accompanied by strong expression of matrix metalloproteinase 9 as compared with FGF2-only
treatment or BMT alone. Moreover, in this process, the proliferation of bone-marrow-derived cells was accelerated without
causing apoptosis. Thus, the administration of FGF2 in combination with BMT synergistically improves CCl4-induced liver fibrosis in mice. This treatment has the potential of being an effective therapy for patients with liver cirrhosis.
This study was supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (nos. 16390211
and 16590597) and for translational research from the Ministry of Health, Labor and Welfare (H-trans-5 and H17-Special-015). 相似文献
70.
Design and synthesis of new potent dipeptidyl peptidase IV inhibitors with enhanced ex vivo duration
Kondo T Nekado T Sugimoto I Ochi K Takai S Kinoshita A Tajima Y Yamamoto S Kawabata K Nakai H Toda M 《Bioorganic & medicinal chemistry》2007,15(7):2631-2650
A series of 5beta-methylprolyl-2-cyanopyrrolidine analogs were synthesized and evaluated as DPP-IV inhibitors, and the duration of their ex vivo activity was assessed. Comparison of their potency and duration of action was done among three different species. The mode of binding was investigated, and the effect on the plasma glucose level was evaluated. Structure-activity relationships are also presented. 相似文献