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Quantification of Basal and Stimulated ROS Levels as Predictors of Islet Potency and Function 总被引:2,自引:0,他引:2
B. Armann M. S. Hanson E. Hatch A. Steffen L. A. Fernandez 《American journal of transplantation》2007,7(1):38-47
We have developed a luminol-based assay using intact islets, which allows for quantification of reactive oxygen species (ROS). In addition, an index capable of characterizing metabolic and mitochondrial integrity prior to transplantation was created based on the capacity of islets to respond to high glucose and rotenone (mitochondrial respiratory chain complex I inhibitor) by production of ROS. To validate this assay, lipid peroxidation and antioxidative defense capacity were evaluated by detection of malondialdehyde (MDA) levels and glutathione peroxidase activity (GPx), respectively. Also, flow cytometric analyses of ROS (dihydroethidine), apoptosis (Annexin V, active caspases), necrosis (Topro3), and mitochondrial membrane potential (JC-1) were done in parallel to correlate with changes in luminol-measured ROS. ATP/ADP ratios were quantified by HPLC and the predictive value of ROS measurement on islet functional potency was correlated with capacity to reverse diabetes in a streptozotocin-induced diabetic NOD.scid mouse model as well as in human transplant recipients. Our data demonstrate that levels of ROS in islets correlate with the percentage of apoptotic cells and their functional potency in vivo. The ROS indices following glucose and rotenone exposure are indicative of metabolic potency and mitochondrial integrity and can be used as surrogate markers to evaluate the quality of islets prior to transplantation. 相似文献
3.
The optimal conditions for inactivation of Bordetella pertussis organisms with glutaraldehyde for the production of a safe and potent whole cell pertussis vaccine were investigated. Two bacterial harvests from B. pertussis strain 10536 were treated with glutaraldehyde, each with 0.025, 0.05 and 0.1% concentrations of glutaraldehyde for 10, 60 and 120 min. The nine types of glutaraldehyde-inactivated pertussis vaccine (GIPV) and conventional heat-inactivated pertussis vaccine (HIPV) preparations made from two bacterial harvests were comparatively evaluated for the mouse weight gain test (MWGT), potency, and the histamine-sensitization (HS) and leucocytosis-promoting-factor (LPF) tests. The minimum period for killing the B. pertussis organisms with glutaraldehyde was>10 min for 0.025%, 10 min for 0.05% and 5 min for 0.1% concentration. The average loss in opacity varied from 5 to 10% for GIPV preparations and was 14% for HIPV preparations. The GIPV preparations except those inactivated with 0.025% glutaraldehyde for 10 min (GIPV-A) were much less toxic than the HIPV preparations in the MWGT. The GIPV-A preparations did not pass the MWGT. The GIPV preparations were also much less toxic in HS and LPF tests than the HIPV preparation. The potency of GIPV preparations inactivated with 0.05% glutaraldehyde for 10 min (GIPV-D) was similar to that of HIPV preparations. The prolonged treatments with glutaraldehyde reduced the potency. The GIPV-D preparation with good potency and less toxicity was found to be inactivated with glutaraldehyde under optimal conditions. All the preparations were innocuous in the abnormal toxicity test. 相似文献
4.
Chymases (EC 3.4.21.39) are mast cell serine proteinases that are variably expressed in different species and, in most cases, display either chymotryptic or elastolytic substrate specificity. Given that chymase inhibitors have emerged as potential therapeutic agents for treating various inflammatory, allergic, and cardiovascular disorders, it is important to understand interspecies differences of the enzymes as well as the behavior of inhibitors with them. We have expressed chymases from humans, macaques, dogs, sheep (MCP2 and MCP3), guinea pigs, and hamsters (HAM1 and HAM2) in baculovirus-infected insect cells. The enzymes were purified and characterized with kinetic constants by using chromogenic substrates. We evaluated in vitro the potency of five nonpeptide inhibitors, originally targeted against human chymase. The inhibitors exhibited remarkable cross-species variation of sensitivity, with the greatest potency observed against human and macaque chymases, with Ki values ranging from ∼0.4 to 72 nM. Compounds were 10-300-fold less potent, and in some instances ineffective, against chymases from the other species. The X-ray structure of one of the potent phosphinate inhibitors, JNJ-18054478, complexed with human chymase was solved at 1.8 Å resolution to further understand the binding mode. Subtle variations in the residues in the active site that are already known to influence chymase substrate specificity can also strongly affect the compound potency. The results are discussed in the context of selecting a suitable animal model to study compounds ultimately targeted for human chymase. 相似文献
5.
Four experiments were organized around a central question: What is the form of relationship between estimated stress level on the one hand and situation strain, personal resources and social support, on the other? The first experiment examined the form of the relationship between estimated level of stress, situation strain and personal resources. The participants were students. They integrated situation strain and personal resources information in a non‐additive way. In particular, the effect of personal resources on the estimated level of stress varied as a function of the level of situation strain considered. When the situation strain was low, the stress level related with this circumstance largely depended on the personal resources of the individual. When the situation strain was high, the stress level related with this circumstance was much less dependent on the personal resources of the individual. The second experiment replicated these results among first‐aid workers, fire‐fighters and persons that had recently been injured. The third and fourth experiments replicated these results in various conditions differing as regards the level of social support. Copyright © 2002 John Wiley & Sons, Ltd. 相似文献
6.
不同层次护生对临床教学质量的评价 总被引:3,自引:0,他引:3
目的分析不同层次护生对临床教学质量评价的差异,以便采取有针对性的教学方法.方法护生在每个科室实习结束后,采用自行设计的临床教学评价表对每名护生进行问卷调查,然后分别从中专、大专、本科护生填写的调查表中各随机抽取30%进行分析.结果对病房教学氛围和护士长教学管理的评价,本科与中专、大专护生间比较,差异有显著性意义(均P<0.05),中专与大专护生间无差异(P>0.05);对带教老师的评价,本科与中专、大专与中专护生间均差异有显著性意义(P<0.01,P<0.05),本科与大专间无差异(P>0.05).结论不同层次护生对临床教学质量评价的侧重点不同,本科生对病房教学氛围及护士长教学管理的要求较高,大专生次之.在临床教学中应根据护生的学历层次,采取有针对性的教学方法,以便不断提高临床教学质量. 相似文献
7.
M. Johnson 《Allergy》1995,50(S23):11-14
Fluticasone propionate is a new corticosteroid based on the androstane nucleus. It is more lipophilic than beclomethasone dipropionate (BDP) and budesonide, and binds more avidly to human lung tissue. It has an absolute affinity (KD ) of 0.5 nM for the glucocorticoid receptor and a relative receptor aflinity 1.5- and 3.0-times greater than that of beclomethasone-17-monopropionate (17-BMP) and budesonide, respectively. The rate of association with the receptor is faster and the rate of dissociation slower than with standard corticosteroids. As a result, the half-life of the corticosteroid-receptor complex is >10 h. Fluticasone propionate is also highly selective for the glucocorticoid receptor, with little or no activity at other steroid receptors. Pretreatment with fluticasone propionate signiflcantly inhibits the increase in mast cell numbers in the nasal mucosa of rats chronically exposed to toluene di-isocyanate (TDI), and suppresses TDI-induced mast cell degranulation. It is more potent in vitro than dexamethasone, BDP and budesonide in inhibiting anti-CD3-induced human T-lymphocyte proliferation, in attenuating tumour necrosis factor-α-induced endothelial cell adhesion molecule expression, and in increasing secretory leucocyte protease inhibitor levels in airway epithelial cells. It is also more potent and longer-acting than other corticosteroids in inhibiting oedema formation, interleukin-5 (IL-5)-induced blood eosinophilia, and IL-5- or platelet activating factor-stimulated eosinophil accumulation in the lung. Fluticasone propionate therefore has increased intrinsic glucocorticoid potency and high topical anti-inflammatory activity. 相似文献
8.
Anne‐Marie Baronet 《Family relations》2003,52(2):137-142
This study examined the impact of family support and relationship difficulties between the caregiver and the care recipient on caregivers' satisfaction received from caregiving activities and subjective burden. Ninety‐seven individuals caring for a spouse or an adult child with schizophrenia or bipolar disorder participated in the study. Using hierarchical multiple regression analyses, findings showed that relationship difficulties between the caregiver and the care recipient were associated with both satisfaction received from caregiving activities and subjective burden, after the effects of personal, contextual, and stressor variables were controlled. Family support was not associated with caregivers' appraisal of the situation. 相似文献
9.
Ljiljana Ševaljević Sanja Marinković Desanka Bogojević Svetlana Matić Bogdan Bošković 《Archives of toxicology》1989,63(5):406-411
We have studied the effect of soman intoxication on serum acute phase reactants (APR) levels, and the relationship of the APR and corticosterone concentrations and the immunosuppressive activity of the serum. One day after the injection of 1.8 LD50 soman the concentrations of 2-macroglobulin (2-MG) and 1-acid glycoprotein (AGP) in the serum of antidote protected rats increased 4- and 7-fold, respectively, whereas those of hemopexin (Hx), haptoglobin (Hp) and cysteine protease inhibitor (CPI) were two to three times higher than in the controls. A similar magnitude of increase of serum acute phase reactants levels was observed when 0.3 LD50 soman was administered at 24-h intervals over the 5-day period. The relationship of changes in the APR concentration, corticosterone level and immunosuppressive activity of the serum was also comparable to that observed in the acute phase response to tissue injury. 相似文献
10.
O'Goiman's (1979) criticisms of the orienting response (OR) significance hypothesis are not well-founded. The hypothesis is not based solely on electrodermal data—studies reporting significance effects for heart rate and pupillary OR are cited. The hypothesis does not discard novelty but suggests that an interaction between uncertainty and significance triggers OR. Differential reactivity across OR components reflects stimulus- and individual-response factors, and does not require separate “significance registers.” Since the hypothesis specifically maintains that appraisals of stimulus significance affect the OR, not the mere fact of cognitive appraisal per se, no general “cognitive appraisal effect” was ever expected. O'Gorman confuses the difficulty of estimating the subject's spontaneous judgments of significance with the relative ease of manipulating significance in the laboratory. He considers classic OR theory more “objective” only because he fails to look closely enough at the complex judgmental processes involved in stimulus intake, model building, and matching functions. O'Gorman's contentions of different initial- and test-ORs, and of a simple additive relationship between significance and OR, lack supportive data and are challenged by available evidence. Literature is cited suggesting that: a) scanning is continuously biased toward the detection of significant stimuli; b) detection of a sgnificant stimulus feature triggers increased information scanning, increasing the possibility that previously unrecognized uncertainty will be detected, and c) lowers the criterion level of the OR “threshold,” increasing the likelihood of OR if uncertainty is detected. Evidence is cited suggesting that the significance-OR, and perhaps attention in general, is mediated by neocortical-limbic interaction, the limbic' “motivational” evaluator of stimulus input steering and switching neocortical “intellectual” analyzers of stimulus pattern. 相似文献