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941.
Kentaro Uchida Kouji Naruse Masashi Satoh Kenji Onuma Masaki Ueno Shotaro Takano Ken Urabe Masashi Takaso 《Experimental Animals》2013,62(3):255-265
Although recent studies suggest that hyperlipidemia is a risk factor for osteoarthritis
(OA), the link between OA and hyperlipidemia is not fully understood. As the number of
activated, circulating myeloid cells is increased during hyperlipidemia, we speculate that
myeloid cells contribute to the pathology of OA. Here, we characterized myeloid cells in
STR/Ort mice, a murine osteoarthritis model, under hyperlipidemic conditions. Ratios of
myeloid cells in bone marrow, the spleen, and peripheral blood were determined by flow
cytometry. To examine the influence of the hematopoietic environment, including abnormal
stem cells, on the hematopoietic profile of STR/Ort mice, bone marrow transplantations
were performed. The relationship between hyperlipidemia and abnormal hematopoiesis was
examined by evaluating biochemical parameters and spleen weight of F2 animals
(STR/Ort x C57BL/6J). In STR/Ort mice, the ratio of CD11b+Gr1+ cells
in spleens and peripheral blood was increased, and CD11b+Gr1+ cells
were also present in synovial tissue. Splenomegaly was observed and correlated with the
ratio of CD11b+Gr1+ cells. When bone marrow from GFP-expressing mice
was transplanted into STR/Ort mice, no difference in the percentage of
CD11b+Gr1+ cells was observed between transplanted and age-matched
STR/Ort mice. Analysis of biochemical parameters in F2 mice showed that spleen
weight correlated with serum total cholesterol. These results suggest that the increase in
circulating and splenic CD11b+Gr1+ cells in STR/Ort mice originates
from hypercholesterolemia. Further investigation of the function of
CD11b+Gr1+ cells in synovial tissue may reveal the pathology of OA
in STR/Ort mice. 相似文献
942.
P16蛋白和生精细胞凋亡对热压和11酸睾酮诱导的恒河猴无精子症和少精子症中的作用 总被引:6,自引:0,他引:6
探讨了P16蛋白和生精细胞凋亡在热压和11酸睾酮诱导恒河猴无精子症和少精子症中作用间的关系。3′末端标记分析(TUNEL)结果显示热应激和超生理剂量睾酮能够诱导生精细胞出现凋亡信号,它分别于处理后第5天和第30天达到最强,免疫组化结果显示,热压或TU主要诱导精原细胞和其它生精细胞以及Sertoli细胞P16的表达。P16蛋白的表达在生精细胞凋亡晚期,即隐睾手术第10天或注射TU第60天后迅速升高并维持在热压或11酸睾酮诱导的早期精母细胞和精子细胞的凋亡和在晚期对精原细胞有丝分裂的抑制,二者共同作用导致热压或TU诱导的恒河猴无精子症和少精子症。 相似文献
943.
Juliana Artier Steven C. Holland Neil T. Miller Minquan Zhang Robert L. Burnap 《BBA》2018,1859(10):1108-1118
The CO2-concentrating mechanism (CCM) in cyanobacteria supports high rates of photosynthesis by greatly increasing the concentration of CO2 around the major carbon fixing enzyme, Rubisco. However, the CCM remains poorly understood, especially in regards to the enigmatic CO2-hydration enzymes which couple photosynthetically generated redox energy to the hydration of CO2 to bicarbonate. This CO2-hydration reaction is catalysed by specialized forms of NDH-1 thylakoid membrane complexes that contain phylogenetically unique extrinsic proteins that appear to couple CO2 hydration to NDH-1 proton pumping. The development of the first molecular genetic system to probe structure-function relationships of this important enzyme system is described. A CO2-hydration deficient strain was constructed as a recipient for DNA constructs containing different forms of the CO2-hydration system. This was tested by introducing a construct to an ectopic location that gives constitutive expression, rather than native inducible expression, of the ndhF3-ndhD3-cupA-cupS, (cupA operon) encoding high affinity CO2-hydration complex, NDH-13. Uptake assays show the restoration of high affinity for CO2 uptake, but demonstrate that the CupA complex can drive only modest uptake fluxes, underlining the importance of its tandem operation with the CupB-containing complex NDH-14, the complementary high flux, low affinity CO2 hydration system. Experiments with the carbonic anhydrase inhibitor, ethoxyzolamide, indicate that the NDH-13 complex is strongly inhibited, yet the remaining NDH-14 activity in the wild-type is less so, suggesting structural differences between the low affinity and high affinity CO2–hydration systems. This new construct will be an important tool to study and better understand cyanobacterial CO2 uptake systems. 相似文献
944.
This paper provides quantitative information concerning the response of ostracods to environmental variability in order to reconstruct past environments. Ostracod faunas from modern sediments of Bolivian lakes and swamps were studied. Ostracod distribution is controlled by several ecological characteristics such as lake-level and water chemistry. Statistical results indicate that three transfer functions (on water depth, Total dissolved Salts and water in Mg/Ca ratio) can be developed, from ostracod species frequencies in lacustrine sediments, with some restrictions for the two last ones. 相似文献
945.
Coagulation factor VIIa (FVIIa) belongs to a family of proteases being part of the stepwise, self-amplifying blood coagulation cascade. To investigate the impact of the mutation Met(298{156})Lys in FVIIa, we replaced the Gly(283{140})-Met(298{156}) loop with the corresponding loop of factor Xa. The resulting variant exhibited increased intrinsic activity, concurrent with maturation of the active site, a less accessible N-terminus, and, interestingly, an altered macromolecular substrate specificity reflected in an increased ability to cleave factor IX (FIX) and a decreased rate of FX activation compared to that of wild-type FVIIa. In complex with tissue factor, activation of FIX, but not of FX, returned to normal. Deconvolution of the loop graft in order to identify important side chain substitutions resulted in the mutant Val(158{21})Asp/Leu(287{144})Thr/Ala(294{152})Ser/Glu(296{154}) Ile/Met(298{156})Lys-FVIIa with almost the same activity and specificity profile. We conclude that a lysine residue in position 298{156} of FVIIa requires a hydrophilic environment to be fully accommodated. This position appears critical for substrate specificity among the proteases of the blood coagulation cascade due to its prominent position in the macromolecular exosite and possibly via its interaction with the corresponding position in the substrate (i.e. FIX or FX). 相似文献
946.
K. Hancock D. E. Broughel I. N. S. Moura A. Khan N. J. Pieniazek A. E. Gonzalez H. H. Garcia R. H. Gilman V. C. W. Tsang 《International journal for parasitology》2001,31(14):1601-1607
We examined the genetic variability in the pig–human tapeworm, Taenia solium, by sequencing the genes for cytochrome oxidase I, internal transcribed spacer 1, and a diagnostic antigen, Ts14, from individual cysts isolated from Peru, Colombia, Mexico, India, China, and the Philippines. For these genes, the rate of nucleotide variation was minimal. Isolates from these countries can be distinguished based on one to eight nucleotide differences in the 396 nucleotide cytochrome oxidase I (COI) sequence. However, all of the 15 isolates from within Peru had identical COI sequences. The Ts14 sequences from India and China were identical and differed from the Peru sequence by three nucleotides in 333. These data indicate that there is minimal genetic variability within the species T. solium. Minimal variability was also seen in the ITS1 sequence, but this variation was observed within the individual. Twenty-two cloned sequences from six isolates sorted into 13 unique sequences. The variability observed within the sequences from individual cysts was as great as the variability between the isolates. 相似文献
947.
Abstract: In a previous study, protein kinase FA /glycogen synthase kinase-3 ( FA /GSK-3 ) was identified as a myelin basic protein (MBP) kinase associated with intact brain myelin. In this report, the phosphorylation sites of MBP by kinase FA /GSk-3 were further determined by two-dimensional electrophoresis/TLC, phosphoamino acid analysis, tryptic peptide mapping, Edman degradation, and direct sequencing. Kinase FA /GSK-3 phosphorylates MBP on both threonine and serine residues. Three tryptic phosphopeptide peaks were resolved by C18 reverse-phase HPLC. Sequential manual Edman degradation together with direct sequence analysis revealed that T(p)PPPSQGK is the phosphorylation site sequence for the first major phosphopeptide peak. When mapping with the bovine brain MBP sequence, we finally demonstrate Thr97 -Pro, one of the in vivo phosphorylation sites in MBP, as the major site phosphorylated by kinase FA /GSK-3, implicating a physiologically relevant role of FA /GSK-3 in the regulation of brain myelin function. By using the same approach, we also identified NIVT94 (p)PR as the phosphorylation site sequence in the second major tryptic phosphopeptide derived from [32 P]MBP phosphorylated by kinase FA /GSK-3, further indicating that kinase FA /GSK-3 represents a Thr-Pro motif-directed MBP kinase involved in the phosphorylation of brain myelin. 相似文献
948.
Christelle David Laurent Bischoff Hervé Meudal Catherine Llorens-Cortes Bernard Pierre Roques Marie-Claude Fournié-Zaluski 《Letters in Peptide Science》1997,4(4-6):411-414
In order to study the physiological role of aminopeptidase A (APA),several -mercapto--amino acyl dipeptides were synthesized toobtain compounds having a high affinity for APA and a high selectivityversus aminopeptidase N (APN). Sulfonamide and carboxylate moieties whichhave been shown to be recognized by the S1 subsite of theenzyme were introduced on the side chain of the -mercapto--aminoacyl sub-unit, the latter being coupled to dipeptides optimized to interactwith the S1 andS2 subsites by means of combinatorialchemistry. Good affinities (16 nM) were obtained, the selectivity factorsbeing up to 160-fold versus APN. 相似文献
949.
目的构建大肠埃希菌(Escherichia coli)嘌呤核苷磷酸化酶(purine nucleoside phosphorylase,PNP)基因表达载体,研究其生物活性,为肿瘤的基因治疗奠定基础。方法PCR扩增大肠埃希菌K12的PNP基因,T4连接酶将PNP连接人pMSCV逆转录病毒载体,构建重组逆转录病毒载体pMSCV/PNP。pM—SCV/PNP转化感受态大肠埃希菌XLI-Blue,提取pMSCV/PNP,酶切、PCR和测序鉴定。病毒包装细胞293产生重组逆转录病毒pMSCV/PNP,流式细胞仪测病毒滴度。pMSCV/PNP转染胰腺癌细胞BXPC-3,倒置荧光显微镜观察,FACS分离转染阳性细胞(GFP阳性)。RT—PCR检测PNPmRNA在胰腺癌细胞BXPC-3细胞中的表达,MTT法检测PNP基因的生物活性。结果PCR扩增出大肠埃希菌PNP基因(738bp),酶切和PCR的电泳条带显示pMSCV/PNP,测序结果正常。293包装细胞产生高滴度(3.6×10^7U/m1)重组逆转录病毒pMSCV/PNP。RT—PCR实验结果表明,pMSCV/PNP转染的胰腺癌细胞BXPC-3表达PNPmRNA。前药6-甲基嘌呤-2’-脱氧核苷(MePdR)作用72h浓度达1.00mg/L,BXPC-3/PNP细胞存活率为10.09%,随着MePdR浓度加大,BXPC-3/PNP细胞存活率继续下降直至为0。结论构建了pMSCV/PNP载体,获得了表达大肠埃希菌PNP基因的BXPC-3细胞克隆,PNP/MePdR自杀基因系统对胰腺癌细胞BXPC-3有较强的抑杀作用。 相似文献
950.
引起人类呼吸道感染的冠状病毒已多达5种.冠状病毒与宿主相互作用决定了其致病性和免疫特性.冠状病毒感染后宿主会立即启动抗病毒天然免疫反应,而人类冠状病毒往往会编码特定蛋白逃逸或抑制宿主的天然免疫反应.NL63冠状病毒是一种新型人类冠状病毒,其非结构蛋白nsp3编码2个木瓜样蛋白酶(PLP)核心结构域PLP1和PLP2.前期研究发现,人类冠状病毒PLP2是一种病毒编码的去泛素化酶(DUB),但是对其DUB特性和功能还不清楚.研究发现,NL63冠状病毒PLP1和PLP2两个核心结构域中只有PLP2具有DUB活性,而且,PLP2的DUB活性对K48和K63连接的多聚泛素化修饰不表现明显特异性.同时,蛋白酶活性催化位点C1678和H1836突变后对其DUB活性有明显抑制作用,而蛋白酶活性催化位点D1849突变后对DUB活性无影响.其次,PLP2而非PLP1核心结构域能够明显抑制仙台病毒和重要信号蛋白(RIG-I、ERIS/STING/MITA)激活的干扰素表达,表明PLP2是一种冠状病毒编码的干扰素拮抗剂,而且PLP2的干扰素拮抗作用不完全依赖其蛋白酶活性.机制研究表明,PLP2能够与干扰素表达通路中的重要调节蛋白RIG-I和ERIS发生相互作用,通过对RIG-I和ERIS的去泛素化负调控宿主抗病毒天然免疫反应.此外,PLP2除利用DUB活性抑制干扰素表达外,很可能存在不依赖自身催化活性的其他组分共同抑制干扰素的产生.以上研究对阐明人类新发冠状病毒免疫和致病机理以及抗病毒药物研发具有重要参考价值. 相似文献