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41.
The color complementation assay (CCA) is a method of allele-specific DNA amplification by which competitive priming and extension of fluorescently labeled oligonucleotide primers determine the color of DNA amplification product. This diagnostic method precludes the need for radioisotopes, electrophoresis, and multiple high-stringency reaction conditions. The multiplicity of mutant globin genes present in Southeast Asians complicates clinical diagnosis and underscores the importance of DNA-based diagnostic methods. We have applied CCA to distinguish beta A and beta E alleles. Competing 15mer primers were a fluorescein-labeled complement to beta A and a rhodamine-labeled complement to beta E, identical except for their central nucleotides. A common unlabeled primer was used to amplify DNA product, the color of which was determined by the perfectly complementary primer. Color photography and spectrofluorometry, as well as a method of black-white photography that we developed to distinguish fluorescein- and rhodamine- labeled DNA, were used to record results. We applied CCA to define the complex genotype of a Thai woman with thalassemia intermedia, 96% HbE, and 4% HbF whose possible genotypes included several permutations of alpha-thalassemia, beta-thalassemia, and beta E genes. zeta-Globin gene mapping of DNA doubly digested with Bg/II and Asp 718 showed the -alpha 3.7/--SEA genotype, and CCA confirmed homozygous beta E/beta E. The CCA is useful for diagnosing the compound hemoglobin genotypes of Southeast Asians and could be applied also to prenatal diagnosis in this population. 相似文献
42.
Diagnostic role of an immunoassay-detected polymorphism of factor IX for potential carriers of hemophilia B 总被引:3,自引:1,他引:3
In hemophilia B, assays based on a monoclonal antifactor IX specific for the Thr-148 variant of an exonic polymorphism have diagnosed carriers in selected families by either establishing linkage or by indicating the presence or absence of a given normal factor IX. The sensitivity of the immunoassays for detecting heterozygous women was explored by comparing results from immunoassays with solid-phase polyclonal v the monoclonal antifactor IXs. Factor IX with the normal Ala-148 variant gave a flat dilution curve, qualitatively distinct from factor IX with the Thr-148 variant in the monoclonal assay. The two were indistinguishable in the polyclonal assay. Mixtures of equal amounts of the two types gave an intermediate result, about half as reactive in the monoclonal as compared with the polyclonal assay system. Whereas mixtures with 10% Ala-148 and 90% Thr-148 factor IXs could not readily be distinguished from Thr-148 factor IX plasma, as little as 1% of the Thr-148 protein was detected in Ala-148 factor IX plasma. The frequency of the Ala-148 variant varied in individuals with different ethnic backgrounds; it was found in 29% of white, 12% of black, and none of Asian blood donors' factor IX genes in Seattle. Only 4% of samples from South African black men were nonreactive (ie, Ala- 148). The Thr/Ala-148 dimorphism is in strong linkage disequilibrium with Taql restriction fragment length polymorphisms (RFLPs). Three recombinations were noted in normal white genes and one in a normal black factor IX gene (less than 2% of those examined). In 34 white families with at least one woman being a possible carrier, genetically, the immunoassay results were informative in 18. RFLP analyses were informative in eight of the 15 families tested. In five families each, assignment of carrier status was made to a woman by only DNA or only immunoassay results, whereas the other approach was noninformative. The immunoassays provide a rapid, inexpensive screening test and complement DNA analysis in white women who are potential carriers of hemophilia B. 相似文献
43.
Aim: Findings of hypoxia prior to death and involvement of a dysregulation of the serotonergic network in sudden infant death syndrome (SIDS) may indicate that brain‐derived neutrophic factor (BDNF) also is of importance with regard to sudden unexpected infant death. Based on this, the purpose of this study was to investigate the BDNF val66met polymorphism in SIDS cases, cases of infectious death and controls. Methods: The polymorphism was investigated in 163 SIDS cases, 34 cases of infectious death and 121 controls, using real‐time PCR and fluorescence melting curve analysis. Results: There were no differences in val66met genotype distribution between neither the SIDS cases nor the cases of infectious death and controls (p = 0.95 and p = 0.52, respectively). Conclusion: The study indicates that the val66met polymorphism is not important for sudden unexpected infant death. However, several other SNPs in the BDNF gene, as well as in other genes involved in this pathway, including G‐protein, have to be investigated to fully exclude any involvement of BDNF in SIDS. 相似文献
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Objective : Understanding patients' and their parents' perceptions towards asthma medication is important in developing strategies to ensure patient compliance. In this study, parents' perceptions towards their children's use of inhaled medication for asthma treatment were evaluated.
Methodology : A questionnaire was administered by interviewing parents of 210 asthmatic children attending the Department of Paediatrics, National University Hospital.
Results : Our results showed that 76/210 (36%) of parents either felt opposed to inhaler therapy and/or preferred oral medications. The main reasons for their reluctance to use inhalers were related to fear of dependence, side effects and overdosage, and the child's dislike for inhalers. A third of these parents felt that inhalers were only indicated for very severe asthma.
Conclusions : We conclude that a significant proportion of parents have reservations regarding the use of inhalers for the treatment of asthma. These factors should be taken into consideration when planning an effective asthma education programme. 相似文献
Methodology : A questionnaire was administered by interviewing parents of 210 asthmatic children attending the Department of Paediatrics, National University Hospital.
Results : Our results showed that 76/210 (36%) of parents either felt opposed to inhaler therapy and/or preferred oral medications. The main reasons for their reluctance to use inhalers were related to fear of dependence, side effects and overdosage, and the child's dislike for inhalers. A third of these parents felt that inhalers were only indicated for very severe asthma.
Conclusions : We conclude that a significant proportion of parents have reservations regarding the use of inhalers for the treatment of asthma. These factors should be taken into consideration when planning an effective asthma education programme. 相似文献
46.
Objective : To assess the relationship between the subtypes of hypertension in pregnancy and subsequent neonatal haematology.
Methodology : Retrospective review of the haematology of newborns of hypertensive mothers at a tertiary neonatal unit
Results : Over a 2 year period. 249 infants had full blood examinations. Nineteen (7.6%) were neutropenic and 35 (14.1%) thrombocytopenic, including 11 (4.4%) who were both neutropenic and thrombocytopenic. Neutropenia occurred only in infants whose mothers had severe pre-eclampsia and eclampsia or pre-eclampsia with pre-existing hypertension, whereas thrombocytopenia complicated all maternal hypertension subtypes. Two (10%) of the neutropenic infants developed nosocomial infection while seven (20%) of the thrombocytopenic infants bled. Thirteen (68%) of the neutropenic infants compared with 15 (43%) of the thrombocytopenic infants developed their haematological abnormality within 24 h of birth. All but two infants developed the haematological abnormality by the 5th day of life.
Conclusions : Although haematological abnormalities in infants born to hypertensive mothers are uncommon, serious neonatal complications can occur and therefore early haematological screening of these infants is recommended. 相似文献
Methodology : Retrospective review of the haematology of newborns of hypertensive mothers at a tertiary neonatal unit
Results : Over a 2 year period. 249 infants had full blood examinations. Nineteen (7.6%) were neutropenic and 35 (14.1%) thrombocytopenic, including 11 (4.4%) who were both neutropenic and thrombocytopenic. Neutropenia occurred only in infants whose mothers had severe pre-eclampsia and eclampsia or pre-eclampsia with pre-existing hypertension, whereas thrombocytopenia complicated all maternal hypertension subtypes. Two (10%) of the neutropenic infants developed nosocomial infection while seven (20%) of the thrombocytopenic infants bled. Thirteen (68%) of the neutropenic infants compared with 15 (43%) of the thrombocytopenic infants developed their haematological abnormality within 24 h of birth. All but two infants developed the haematological abnormality by the 5th day of life.
Conclusions : Although haematological abnormalities in infants born to hypertensive mothers are uncommon, serious neonatal complications can occur and therefore early haematological screening of these infants is recommended. 相似文献
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据文献记载和作者的调查,我国产麻黄属(Ephedra)植物共有13种3变种1变型。本文报道我国北方生产的7种麻黄生药形态组织学比较研究结果。7种麻黄是:草麻黄Ephedra sinica Stapf、木贼麻黄E.equisetina Bunge、中麻黄E.intermedia Schrenk ex Mey.、膜果麻黄E.przewalskii Stapf,单子麻黄E.monosperma Gmel.ex Mey.、雌雄麻黄E.fedtschenkoae Pauls和细子麻黄E.reaeliana Florin。文中附有生药性状与组织构造特征比较表及生药组织图 相似文献
50.