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401.
New methods of structure refinement for macromolecular structure
determination by NMR 总被引:8,自引:0,他引:8
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G. Marius Clore Angela M. Gronenborn 《Proceedings of the National Academy of Sciences of the United States of America》1998,95(11):5891-5898
Recent advances in multidimensional NMR methodology have permitted solution structures of proteins in excess of 250 residues to be solved. In this paper, we discuss several methods of structure refinement that promise to increase the accuracy of macromolecular structures determined by NMR. These methods include the use of a conformational database potential and direct refinement against three-bond coupling constants, secondary 13C shifts, 1H shifts, T1/T2 ratios, and residual dipolar couplings. The latter two measurements provide long range restraints that are not accessible by other solution NMR parameters. 相似文献
402.
Nonalcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities 总被引:101,自引:0,他引:101
Sanyal AJ Campbell-Sargent C Mirshahi F Rizzo WB Contos MJ Sterling RK Luketic VA Shiffman ML Clore JN 《Gastroenterology》2001,120(5):1183-1192
BACKGROUND AND AIMS: The pathogenesis of nonalcoholic steatohepatitis (NASH) is unknown. We tested the hypothesis that NASH is associated with 2 defects: (1) peripheral insulin resistance, which increases lipolysis, delivery of free fatty acids (FFA) to the liver, and hepatic fatty acid beta oxidation, thereby creating oxidative stress; and (2) an abnormality within the hepatocytes that might render them more susceptible to injury from oxidative stress. METHODS: The hypothesis was tested by evaluation of (1) insulin resistance by a 2-step hyperinsulinemic (10 and 40 mU. m(-2). min(-1)) euglycemic clamp; (2) insulin effects on lipolysis by enrichment of [U-(13)C]glycerol; (3) frequency and severity of structural defects in hepatocyte mitochondria in vivo; (4) fatty acid beta oxidation from serum [beta-OH butyrate], release of water-soluble radioactivity from (3)H-palmitate by cultured fibroblasts and urinary dicarboxylic acid excretion; and (5) hepatic lipid peroxidation by immunohistochemical staining for 3-nitrotyrosine (3-NT). Subjects with NASH (n = 6-10 for different studies) were compared with those with fatty liver (n = 6) or normal controls (n = 6). RESULTS: NASH and fatty liver were both associated with insulin resistance, with mean glucose infusion rates (normal/fatty liver/NASH) of step 1, 4.5/1.6/0.9; step 2, 9.5/7.7/4.5 (P < 0.03 for both steps). Although baseline rates of glycerol appearance were higher in those with NASH than in those with fatty liver (means, 14.6 vs. 21.6 micromol. kg(-1). min(-1); P < 0.05), neither group significantly suppressed glycerol appearance at insulin infusion rates of 10 mU. m(-2). min(-1). NASH was associated with loss of mitochondrial cristae and paracrystalline inclusions in 9 of 10 subjects, compared with 0 of 6 subjects with fatty liver. However, no evidence of a generalized defect in fatty acid beta oxidation was noted in any group. Also, mean [beta-OH butyrate] was highest in those with NASH (means, 90 vs. 110 vs. 160 micromol/L; P < 0.04). Increased staining for 3-NT was present in fatty liver, and even greater staining was seen in NASH. CONCLUSIONS: These data indicate that peripheral insulin resistance, increased fatty acid beta oxidation, and hepatic oxidative stress are present in both fatty liver and NASH, but NASH alone is associated with mitochondrial structural defects. 相似文献
403.
404.
Serial observations of blast cell colony development from spleen cells of mice treated with 5-fluorouracil (5-FU) four days earlier revealed that either form of human interleukin-1 (IL-1 alpha or IL-1 beta) hastens the emergence of interleukin-3 (IL-3)-dependent blast cell colonies. This activity was essentially indistinguishable from the effect of interleukin-6 (IL-6) or granulocyte colony-stimulating factor (G-CSF) in the same system, an effect that we have ascribed previously to a shortening of the G0 period of the dormant stem cells. We also analyzed the time courses of colony formation from cultures of day-2 post-5-FU marrow cells supported by IL-1 alpha, IL-6, or G-CSF alone or in combination with IL-3. In the presence of IL-3, G-CSF and IL-6 but not IL-1 alpha hastened the development of colonies and increased the numbers of multilineage colonies relative to cultures of IL-3 alone. This observation, together with our previous data from the human system, suggests that the synergistic effect of IL-1 is likely due to induction of secondary growth factors, including IL-6 and G-CSF, by accessory cells in culture. The effect of IL-6 on G0 was confirmed by analysis of the cycling status of progenitor cells in short-term culture. While neither IL-3 nor IL-6 alone had any effect on the cycling status, the combination of factors resulted in a rapid recruitment of quiescent cells into cell cycle (within 48 hours) as represented by a twofold increase in the numbers of multipotential progenitors and a significant increase in the sensitivity of these cells to 3H-thymidine with high specific activity. Combinational testing of all of these synergistic factors revealed that the target cell populations for the IL-1, IL-6, and G-CSF overlap considerably, suggesting that they all may act through a common mechanism. This is further supported by our finding that cells from blast cell colonies grown in the presence of a combination of any one of the synergistic factors with IL-3 replate with higher efficiency and yield more multilineage secondary colonies than those from colonies grown in IL-3 alone. These findings provide further evidence that IL-1, IL-6, and G- CSF serve to integrate the immediate host responses to infection through augmentation of effector cells and antibody production as well as the longer term host responses by recruitment of dormant hemopoietic stem cells into active cell cycling. 相似文献
405.
It is important to evaluate not only comprehensive treatment packages, but also specific procedures to determine their contribution
to the change process. The current study examined the efficacy of cognitive modification (CM) techniques compared to supportive
therapy (ST) for university students reporting significant internalizing distress. CM consisted of three sessions of Fluency
Training (FT) to increase positive self-statements and three sessions of Thought Record (TR) training in challenging negative
thoughts. Participants (N = 57) were randomized to CM or ST and assessed at pre-, mid-, post-treatment, 1- and 3-month follow-ups.
Intent-to-treat analyses suggested that CM produced greater changes in internalizing distress and positive thinking, which
was especially apparent at midtreatment with small-to-medium differences consistently favoring CM at post-treatment and follow-up.
These data suggest that change in CM was beyond that attributable to time spent with an engaged therapist who provided a treatment
rationale, subsequent intervention and homework assignment. 相似文献
406.
407.
408.
目的:观察血小板反应素4基因G29926C(A387P)多态性与中国苏皖地区汉族人群不稳定型心绞痛的可能关系。方法:选择2004-11/2006-05在南京医科大学第一附属医院和江苏大学附属武进医院住院不稳定型心绞痛患者110例,病例均符合2002年AHA/ACC关于不稳定型心绞痛诊断指南的诊断标准,同期选择337例非冠心病者为对照。酚-氯仿法提取白细胞DNA,应用聚合酶链反应-限制性片段长度多态性方法分析血小板反应素4A387P多态性,取部分PCR扩增产物进一步测序鉴定,比较两组间多态性频率分布差异,探讨血小板反应素4基因多态性与不稳定型心绞痛发病的可能关系。结果:447例均进入结果分析。GC基因型在不稳定型心绞痛组和对照组的分布无统计学差异(5.5%,7.1%,P=0.54),未检测到CC纯合子。C等位基因频率在不稳定型心绞痛组和对照组分别为2.7%,3.6%(P=0.55)。GC基因型与不稳定型心绞痛无显著性关联(OR=0.75,95%CI:0.30 ̄1.89,P=0.54)。结论:血小板反应素4基因387A→P不是中国苏皖地区汉族人群常见的多态位点,且与不稳定型心绞痛的发病无显著相关性。 相似文献