全文获取类型
收费全文 | 787篇 |
免费 | 35篇 |
国内免费 | 20篇 |
专业分类
耳鼻咽喉 | 5篇 |
儿科学 | 1篇 |
妇产科学 | 12篇 |
基础医学 | 225篇 |
口腔科学 | 2篇 |
临床医学 | 28篇 |
内科学 | 140篇 |
皮肤病学 | 1篇 |
神经病学 | 151篇 |
特种医学 | 5篇 |
外科学 | 24篇 |
综合类 | 48篇 |
预防医学 | 24篇 |
眼科学 | 4篇 |
药学 | 103篇 |
中国医学 | 6篇 |
肿瘤学 | 63篇 |
出版年
2023年 | 3篇 |
2022年 | 10篇 |
2021年 | 14篇 |
2020年 | 8篇 |
2019年 | 4篇 |
2018年 | 8篇 |
2017年 | 15篇 |
2016年 | 12篇 |
2015年 | 13篇 |
2014年 | 38篇 |
2013年 | 38篇 |
2012年 | 41篇 |
2011年 | 64篇 |
2010年 | 46篇 |
2009年 | 60篇 |
2008年 | 67篇 |
2007年 | 48篇 |
2006年 | 37篇 |
2005年 | 33篇 |
2004年 | 27篇 |
2003年 | 29篇 |
2002年 | 15篇 |
2001年 | 14篇 |
2000年 | 14篇 |
1999年 | 13篇 |
1998年 | 20篇 |
1997年 | 11篇 |
1996年 | 18篇 |
1995年 | 21篇 |
1994年 | 9篇 |
1993年 | 9篇 |
1992年 | 10篇 |
1991年 | 11篇 |
1990年 | 7篇 |
1989年 | 8篇 |
1988年 | 4篇 |
1987年 | 3篇 |
1986年 | 7篇 |
1985年 | 5篇 |
1984年 | 2篇 |
1983年 | 2篇 |
1982年 | 2篇 |
1981年 | 6篇 |
1980年 | 2篇 |
1979年 | 3篇 |
1978年 | 2篇 |
1977年 | 6篇 |
1974年 | 1篇 |
1973年 | 1篇 |
1971年 | 1篇 |
排序方式: 共有842条查询结果,搜索用时 15 毫秒
51.
Keiji Shimada Mitsutoshi Nakamura Eiwa Ishida Munehiro Kishi Shin Yonehara Noboru Konishi 《Cancer science》2002,93(10):1164-1174
Fas-associated death domain (FADD) plays an important role as an adapter molecule in Fas (CD95/APO-l)-mediated apoptosis and contributes to anticancer drug-induced cytotoxicity. We treated three human prostate cancer cell lines with etoposide, a toposiomerase II inhibitor with activity against various tumors including prostate cancer. We found that the overexpression of FADD sensitizes etoposide-induced apoptosis through a rapid activation of c-Jun NH2 -terminal kinase (JNK) and, subsequently, of caspase 3. In addition, phosphorylation of FADD at serine 194 coincided with this sensitization. Treatment with the caspase 3 inhibitor, N-acetyl-Asp-Glu-Val-Asp-aldehyde (DEVD-CHO), or overexpression of either mitogen-activated protein kinase kinase (MKK) 7 or Bcl-xL canceled FADD-mediated sensitization to etoposide-induced apoptosis. Moreover, treatment with the caspase 8 inhibitor, benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone (z-IETD-fmk), or overexpression of viral FLICE/caspase-8-inhibitory protein (FLIP) from equine herpesvirus type 2 E8 also had an inhibitory effect, supporting a major involvement of a caspase 8-dependent mitochondrial pathway. Interestingly, FADD was phosphorylated, and etoposide-induced JNK/caspase activation and apoptosis were enhanced in the cells arrested at G2/M transition, but not in those overexpressing mutant FADD, in which 194 serine was replaced by alanine. Our results demonstrate that phosphorylated FADD-dependent activation of the JNK/caspase pathway plays a pivotal role in sensitization to etoposide-induced apoptosis in prostate cancer cells. 相似文献
52.
Regulation of the ATP-sensitive K+ (K-ATP) channel was examined in cell-attached and inside-out membrane patches of freshly isolated neurons from the ventromedial hypothalamic nucleus (VMN) of 7–14 day old male Sprague–Dawley rats. When inside-out patches were exposed to symmetrical K+, the reversal potential was −2.85±1.65 mV, the single channel conductance 46 pS, and the total conductance varied as a multiple of this value. Glucose (10 mM) reversibly inhibited channel activity in cell-attached preparations by 81%. In the presence of 0.1 mM ADP, 10, 5, and 1 mM ATP reversibly inhibited VMN K-ATP channels in inside-out patches by 88, 83, and 60%, respectively. This inhibition was not dependent on phosphorylation since 5 mM AMPPNP, the non-hydrolyzable analog of ATP, reversibly inhibited channel activity by 67%. Relatively high concentrations of glibenclamide (100 μM) also reversibly inhibited VMN K-ATP channel activity in cell attached and inside-out patches by 67 and 79%, respectively. Finally, the non-specific kinase inhibitor H7 (200 μM) decreased channel activity by 53% while the non-specific phosphatase inhibitor microcystin (250 nM) increased channel activity by 218%. These data suggest that while the inhibitory effect of ATP is not phosphorylation dependent, phosphorylation state is an important regulator of the VMN K-ATP channel. 相似文献
53.
Mitigation of Ectopic Calcification in Osteopontin-Deficient Mice by Exogenous Osteopontin 总被引:3,自引:0,他引:3
Ectopic calcification is a major cause of bioprosthetic heart valve failure. New therapeutic opportunities are offered by the growing understanding that ectopic calcification is an actively regulated process involving several key gene products. One of these products, osteopontin (OPN), is a glycosylated phosphoprotein previously shown to inhibit apatite crystal formation, induce carbonic anhydrase II, and promote mineral resorption. In this study, OPN-deficient mice (OPN−/−) were utilized as an in vivo model to stimulate the ectopic calcification of glutaraldehyde-fixed bovine pericardium (GFBP) tissue and to examine OPN delivery and structure-function relationships with respect to its anti-calcific activity. Significant calcification of GFBP tissue was obtained within 7 days of subcutaneous implantation in OPN−/− mice. Direct rescue of the calcification phenotype was achieved by the administration of exogenous recombinant rat, histidine-fused OPN (rat His-OPN) to the implant site via soluble injection (up to 72% mitigation achieved) or adsorption onto the implant materials (up to 91% mitigation achieved). Effects were specific, since neither fibronectin nor polyhistidine alone could mitigate calcification of GFBP. The maximum anti-calcific effect was achieved only when rat His-OPN was adequately phosphorylated and contained a functional arginine-glycine-aspartate (RGD) cell adhesive domain. Furthermore, CAII levels in host cells surrounding GFBP were greatest when phosphorylated, RGD-containing rat His-OPN was adsorbed. These data suggest that both physical inhibition, mediated by phosphorylation sites in OPN, as well as the induction of CAII and mineral regression, mediated by the RGD domain, contribute to the unique ability of OPN to mitigate ectopic calcification of bioprosthetic valve tissue. 相似文献
54.
The Pho85–Pho80 cyclin–CDK (cyclin-dependent protein kinase) complex of Saccharomyces cerevisiae functions as a key regulator of the phosphate-repressible acid phosphatase system. We have further characterized the Pho85–Pho80 kinase complex and identified the Pho80 cyclin subunit and the Pho81 CDK inhibitor as substrates of the Pho85 protein kinase. The phosphorylation sites within Pho80 have been identified at Ser234 and Ser267. Of the two sites, phosphorylation of Ser234 is required for Pho80 function, to form an active kinase complex and repress acid phosphatase expression. Evidence suggests that the activity of Pho81 is regulated by a post-translational modification and therefore that Pho85-mediated phosphorylation of Pho81 may alter its ability to function as a CDK inhibitor. Thus, the control of acid phosphatase expression involves the phosphorylation of several of the regulatory components of the system. 相似文献
55.
Bovine leukemia virus (BLV) causes persistent lymphocytosis, a preneoplastic, polyclonal expansion of B lymphocytes. The expansion increases viral transmission to new hosts, but the mechanisms of this expansion have not been determined. We hypothesized that BLV infection contributes to B-cell expansion by signaling initiated via viral transmembrane protein motifs undergoing tyrosine phosphorylation. Viral mimicry of host cell proteins is a well-demonstrated mechanism by which viruses may increase propagation or decrease recognition by the host immune system. The cytoplasmic tail of BLV transmembrane protein gp30 (TM) has multiple areas of homology to motifs of host cell signaling proteins, including two immunoreceptor tyrosine-based activation motifs (ITAMs) and two immunoreceptor tyrosine-based inhibition motifs (ITIMs), which are homologous to B-cell receptor and inhibitory co-receptor motifs. Signaling by these motifs in B cells typically relies on tyrosine phosphorylation, followed by interactions with Src-homology-2 (SH2) domains of nonreceptor protein tyrosine kinases or phosphatases. Phosphorylation of tyrosine residues in the cytoplasmic tail of TM was tested in four systems including ex vivo cultured peripheral blood mononuclear cells from BLV infected cows, BLV-expressing fetal lamb kidney cell and bat lung cell lines, and DT40 B cells transfected with a fusion of mouse extracellular CD8 and cytoplasmic TM. No phosphorylation of TM was detected in our experiments in any of the cell types utilized, or with various stimulation methods. Detection was attempted by immunoblotting for phosphotyrosines, or by metabolic labeling of cells. Thus BLV TM is not likely to modify host signal pathways through interactions between phosphorylated tyrosines of the ITAM or ITIM motifs and host-cell tyrosine kinases or phosphatases. 相似文献
56.
Metabotropic glutamate receptors (mGluR) serve important neuromodulatory roles at glutamatergic synapses to shape excitatory neurotransmission. Recent evidence indicates that the desensitization of mGluRs is an important determinant in regulating the functions of these receptors. The present results demonstrate that G protein-coupled receptor kinases (GRKs), which are known to regulate the desensitization of many G protein-coupled receptors, regulate both the expression and function of mGluR5 in a heterologous expression system. This regulatory event is limited to members of the GRK2 family since GRK4 family members do not elicit the same effects on mGluR5. Kinase activity is shown to be required for GRK-mediated regulation of mGluR5. Furthermore, the ability of GRK2 to regulate mGluR5 is dependent, at least in part, on the presence of threonine 840 in the carboxyl terminus of mGluR5. These studies identify novel roles for GRKs in regulating mGluR5 that may serve to further shape the function of these receptors in neurotransmission. 相似文献
57.
Induction of cell cycle arrest and apoptosis in human cervix carcinoma cells during therapy by cisplatin 总被引:1,自引:0,他引:1
The aim of the therapy of human malignancies is the inhibition of cell proliferation and/or induction of apoptosis. We studied the kinetics of the morphological and biochemical changes in HeLa cells during chemotherapy by cisplatin (CP). Apoptosis was evaluated by scoring of cells exhibiting changes characteristic for early and late stages of apoptosis as determined by Hoechst 33258 staining and by examination of positive reaction for activated caspase-3. Expression and intracellular localization of distinct proteins was analyzed by immunoblotting of subcellular fractions and segregation of nucleoli by immunocytochemistry. Chromatin fragmentation characteristic for apoptosis was observed in single cells after 3h cisplatin. A strong cytoplasmic accumulation of cytochrome C detected by immunoblotting 6h post-treatment was accompanied by an activation of caspase-9. Neither inhibition of cell division nor blocking of DNA replication preceded the onset of apoptosis. Our results show that after short treatment by CP, cell proliferation and apoptosis concomitantly occurred. 相似文献
58.
Purified bovine neurofilament (NF) subunit proteins were reassembled in vitro to form either homopolymeric or heteropolymeric intermediate-sized filaments using single or paired combinations of NF triplet proteins. Using conditions established for the reassembly of bovine NF triplet proteins, we demonstrated that the low Mr NF subunit (NF-L) alone and in combination with the middle Mr NF subunit (NF-M) reassembled very efficiently, i.e. greater than 95% of these proteins formed filaments within 90 min from the start of reassembly. In contra-distinction, the high Mr NF subunit (NF-H) alone and in combination with NF-M or NF-L underwent reassembly to a lesser extent, i.e. 62-88% of these proteins reassembled within 90 min. Immunolabeling of the reassembled NF polymers revealed striking differences in the organization of rod domain determinants. Specifically, antibodies specific for epitopes in the rod domains of NF-H, NF-M and NF-L failed to bind heteropolymeric filaments but recognized rod domains in the homopolymers. In contrast, antibodies specific to head and tail domains of all NF proteins labeled the reassembled hetero- and homopolymeric NFs. Double-labeling of heteropolymers demonstrated that pairs of different NF subunits coassembled into intermediate-sized filaments. Our results also showed that only copolymeric filaments of NF-L and NF-M, but not NF-L/NF-H and NF-M/NF-H were able to form long and stable 10-nm wide filaments. These observations provide new insights into the requirements for stable filament formation from NF subunits. In particular, they support the notion that only NF-L/NF-M, but not NF-L/NF-H or NF-M/NF-H might assemble into a stable filamentous network in vivo. 相似文献
59.
Ming-Yan Hei Hui-Kang Tao Qin Tang Bo Yu Ling-Ling Zhao 《Brazilian journal of medical and biological research》2012,45(10):962-967
Our objective was to investigate the protein level of phosphorylated N-methyl-D-aspartate (NMDA) receptor-1 at serine 897 (pNR1 S897) in both NMDA-induced brain damage and hypoxic-ischemic brain damage (HIBD), and to obtain further evidence that HIBD in the cortex is related to NMDA toxicity due to a change of the pNR1 S897 protein level. At postnatal day 7, male and female Sprague-Dawley rats (13.12 ± 0.34 g) were randomly divided into normal control, phosphate-buffered saline (PBS) cerebral microinjection, HIBD, and NMDA cerebral microinjection groups. Immunofluorescence and Western blot (N = 10 rats per group) were used to examine the protein level of pNR1 S897. Immunofluorescence showed that control and PBS groups exhibited significant neuronal cytoplasmic staining for pNR1 S897 in the cortex. Both HIBD and NMDA-induced brain damage markedly decreased pNR1 S897 staining in the ipsilateral cortex, but not in the contralateral cortex. Western blot analysis showed that at 2 and 24 h after HIBD, the protein level of pNR1 S897 was not affected in the contralateral cortex (P > 0.05), whereas it was reduced in the ipsilateral cortex (P < 0.05). At 2 h after NMDA injection, the protein level of pNR1 S897 in the contralateral cortex was also not affected (P > 0.05). The levels in the ipsilateral cortex were decreased, but the change was not significant (P > 0.05). The similar reduction in the protein level of pNR1 S897 following both HIBD and NMDA-induced brain damage suggests that HIBD is to some extent related to NMDA toxicity possibly through NR1 phosphorylation of serine 897. 相似文献
60.
目的探讨大鼠窦状卵泡中磷酸化蛋白激酶B(PKB,又称Akt)的表达与卵泡颗粒细胞凋亡之间的关系。方法应用免疫组织化学法和Western blot法检测磷酸化Akt(phosphorylation-Akt,p-Akt)在大鼠不同发育阶段卵泡颗粒细胞中的表达,应用TUNEL法检测成年大鼠卵泡颗粒细胞的凋亡情况。结果在大鼠窦前及窦状卵泡的颗粒细胞中,p-Akt表达高的颗粒细胞凋亡程度较轻,而p-Akt低表达的颗粒细胞凋亡现象较明显。结论 p-Akt的高表达在一定程度上与颗粒细胞凋亡受抑制有关,并有可能参与了窦状卵泡阶段卵泡的闭锁的选择。 相似文献