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31.
Nitric oxide detoxification systems enhance survival of Neisseria meningitidis in human macrophages and in nasopharyngeal mucosa 下载免费PDF全文
Nitric oxide (NO) contributes to mammalian host defense by direct microbicidal activity and as a signaling molecule of innate immune responses. Macrophages produce NO via the inducible NO synthase (iNOS). The genome of Neisseria meningitidis includes two genes, norB (encoding nitric oxide reductase) and cycP (encoding cytochrome c'), both of which detoxify NO in pure cultures of N. meningitidis. We show here that norB, and to a lesser extent cycP, enhance survival of N. meningitidis within primary human macrophages. Furthermore, accumulation of lysosome-associated membrane protein 1 (LAMP-1) is modified in phagosomes containing an isogenic norB mutant of N. meningitidis compared to the wild type. The survival enhancement conferred by norB and cycP is ablated by pretreatment of macrophages with the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA). Despite this evidence that NO detoxification confers advantage, we find, using a highly sensitive chemiluminescence technique, that human macrophage-associated [NO] is low even after activation by lipopolysaccharide and interferon alpha. Furthermore, wild-type N. meningitidis further depletes cell-associated NO during phagocytosis by an active mechanism and survives relatively poorly in the presence of L-NMMA, suggesting that the wild-type organism may utilize NO for optimal survival during intracellular life. The natural habitat of N. meningitidis is the human nasopharynx. Using a nasopharyngeal mucosa organ culture system, we show that mutants lacking norB and cycP also survive poorly in nasopharyngeal tissue compared to wild-type N. meningitidis. These findings indicate that the meningococcus requires active NO detoxification systems for optimal survival during experimental nasopharyngeal colonization and processing by human phagocytic cells. 相似文献
32.
Labeling of cells with ferumoxides-protamine sulfate complexes does not inhibit function or differentiation capacity of hematopoietic or mesenchymal stem cells 总被引:12,自引:0,他引:12
Arbab AS Yocum GT Rad AM Khakoo AY Fellowes V Read EJ Frank JA 《NMR in biomedicine》2005,18(8):553-559
Two FDA-approved agents, ferumoxides (Feridex), a suspension of superparamagnetic iron oxide (SPIO) nanoparticles and protamine sulfate, a drug used to reverse heparin anticoagulation, can be complexed and used to label cells magnetically ex vivo. Labeling stem cells with ferumoxides-protamine sulfate (FePro) complexes allows for non-invasive monitoring by MRI. However, in order for stem cell trials or therapies to be effective, this labeling technique must not inhibit the ability of cells to differentiate. In this study, we examined the effect of FePro labeling on stem cell differentiation. Viability, phenotypic expression and differential capacity of FePro labeled CD34 + hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC) were compared with unlabeled control cells. Colony-forming unit (CFU) assays showed that the capacity to differentiate was equivalent for labeled and unlabeled HSC. Furthermore, labeling did not alter expression of surface phenotypic markers (CD34, CD31, CXCR4, CD20, CD3 and CD14) on HSC, as measured by flow cytometry. SDF-1-induced HSC migration and HSC differentiation to dendritic cells were also unaffected by FePro labeling. Both FePro-labeled and unlabeled MSC were cultured in chondrogenesis-inducing conditions. Alcian blue staining for proteoglycans revealed similar chondrogenic differentiation for both FePro-labeled and unlabeled cells. Furthermore, collagen X proteins, indicators of cartilage formation, were detected at similar levels in both labeled and unlabeled cell pellets. Prussian blue staining confirmed that cells in labeled pellets contained iron oxide, whereas cells in unlabeled pellets did not. It is concluded that FePro labeling does not alter the function or differentiation capacity of HSC and MSC. These data increase confidence that MRI studies of FePro-labeled HSC or MSC will provide an accurate representation of in vivo trafficking of unlabeled cells. 相似文献
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A method is described for the isolation of Chlamydia psittaci using cell culture treated with 5-iodo-2-deoxyuridine, and subsequent identification by direct fluorescent antibody staining. The method was applied to 110 sets of tissues from a variety of avian specimens submitted for diagnosis. Chlamydiae were isolated and identified in 24 specimens: 13 from parrots, 7 from turkeys and 4 from pigeons. 相似文献
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Asher Y. Rosinger Samantha M. Olson Sascha R. Ellington Janice Perez-Padilla Regina M. Simeone Caitlin S. Pedati Betsy A. Schroeder Gilberto A. Santiago Freddy A. Medina Jorge L. Muoz-Jordn Laura E. Adams Romeo R. Galang Miguel Valencia-Prado Sonia Bakkour Candimar Coln Mary Goodwin Dana Meaney-Delman Jennifer S. Read Lyle R. Petersen Denise J. Jamieson Carmen C. Deseda Margaret A. Honein Brenda Rivera-García Carrie K. Shapiro-Mendoza 《Emerging infectious diseases》2021,27(5):1505
We evaluated nucleic acid amplification testing (NAAT) for Zika virus on whole-blood specimens compared with NAAT on serum and urine specimens among asymptomatic pregnant women during the 2015–2016 Puerto Rico Zika outbreak. Using NAAT, more infections were detected in serum and urine than in whole blood specimens. 相似文献
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Jinhui Ma Kerry Siminoski Peiyao Wang Jacob L Jaremko Khaldoun Koujok Mary Ann Matzinger Nazih Shenouda Brian Lentle Nathalie Alos Elizabeth A Cummings Josephine Ho Kristin Houghton Paivi M Miettunen Rosie Scuccimarri Frank Rauch Leanne M Ward the Canadian STOPP Consortium 《Journal of bone and mineral research》2021,36(7):1255-1268
Vertebral fractures are clinically important sequelae of a wide array of pediatric diseases. In this study, we examined the accuracy of case-finding strategies for detecting incident vertebral fractures (IVF) over 2 years in glucocorticoid-treated children (n = 343) with leukemia, rheumatic disorders, or nephrotic syndrome. Two clinical situations were addressed: the prevalent vertebral fracture (PVF) scenario (when baseline PVF status was known), which assessed the utility of PVF and low lumbar spine bone mineral density (LS BMD; Z-score <−1.4), and the non-PVF scenario (when PVF status was unknown), which evaluated low LS BMD and back pain. LS BMD was measured by dual-energy X-ray absorptiometry, vertebral fractures were quantified on spine radiographs using the modified Genant semiquantitative method, and back pain was assessed by patient report. Forty-four patients (12.8%) had IVF. In the PVF scenario, both low LS BMD and PVF were significant predictors of IVF. Using PVF to determine which patients should have radiographs, 11% would undergo radiography (95% confidence interval [CI] 8–15) with 46% of IVF (95% CI 30–61) detected. Sensitivity would be higher with a strategy of PVF or low LS BMD at baseline (73%; 95% CI 57–85) but would require radiographs in 37% of children (95% CI 32–42). In the non-PVF scenario, the strategy of low LS BMD and back pain produced the highest specificity of any non-PVF model at 87% (95% CI 83–91), the greatest overall accuracy at 82% (95% CI 78–86), and the lowest radiography rate at 17% (95% CI 14–22). Low LS BMD or back pain in the non-PVF scenario produced the highest sensitivity at 82% (95% CI 67–92), but required radiographs in 65% (95% CI 60–70). These results provide guidance for targeting spine radiography in children at risk for IVF. © 2021 American Society for Bone and Mineral Research (ASBMR). 相似文献
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To review the characteristics of reported outbreaks of acute rheumatic fever in the United States, and to determine if there is an increase in the incidence of acute rheumatic fever in the population served by the Hospital for Sick Children, Toronto, Ontario, the authors conducted a literature search and a retrospective review of inpatients and outpatients, satisfying the revised Jones criteria for the diagnosis of acute rheumatic fever, from 1972 to 1988. Patients satisfying the revised Jones criteria for the time period 1972-88 were included in the study. There have been eight articles reporting an increase in acute rheumatic fever in the United States. In three, the majority of children were white and from middle class suburban/rural communities in different geographic locations. Mucoid strains of group A streptococci were implicated but not confirmed as being associated with the outbreaks in three. The results of the chart review at the Hospital for Sick Children revealed that 83 cases satisfied the revised Jones criteria. The number of cases per 100,000 children (aged 18 years or less) per year, decreased progressively over the study period. Polyarthritis was the most frequently seen major criterion occurring in 73% of patients (61 of 83). The most frequently affected ethnic groups were Italians 23%, Afro-Canadians 19% and Orientals 8%. The reported outbreaks in the United States are multifocal and predominantly confined to white middle class children residing in suburban/rural communities. There was no evidence of an increase in the number of cases of acute rheumatic fever seen in the population served by the Hospital for Sick Children; there was a progressive decline in number of cases over the study period. The results facilitate the characterization of acute rheumatic fever within North America into three different patterns of occurrence. 相似文献
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Our previous study demonstrated that meals, particularly when rich in fat, significantly reduced the pain induced by the cold pressor stimulus in healthy human subjects. To determine the mechanisms involved, the aim of this study was to bypass the taste and cognitive component of food and to investigate the scope of these analgesic effects with direct intragastric infusion of pure macronutrients in a group of 16 healthy human volunteers (eight male and eight female) on the response to cold-induced pain. All subjects underwent the cold pressor test (CPT) on three occasions in a counterbalanced order: before and after intragastric intubation and infusion of isoenergetic fat (10% intralipid), carbohydrate (CHO-maltodextrin), and a control infusion of isotonic saline. All solutions were of equal volume and administered at room temperature. The CPT was carried out four times on each test day, once before intubation, and 0.5, 1.5, and 2.5 h after intragastric infusion. Radial pulse and blood pressure measurements and visual analogue scales of mood/emotional state were carried out before and after each CPT. There were no significant differences in pain scores between the three test conditions, suggesting that by bypassing the cognitive and taste component of eating, the trigger for any postingestive analgesic effects of food are lost. 相似文献