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91.
Purification and properties of bacterially synthesized human granulocyte-macrophage colony stimulating factor 总被引:16,自引:0,他引:16
Burgess AW; Begley CG; Johnson GR; Lopez AF; Williamson DJ; Mermod JJ; Simpson RJ; Schmitz A; DeLamarter JF 《Blood》1987,69(1):43-51
Human granulocyte-macrophage colony stimulating factor (GM-CSF) has been synthesized in high yield using a temperature inducible plasmid in Escherichia coli. The human GM-CSF is readily isolated from the bacterial proteins because of its differential solubility and chromatographic properties. The bacterially synthesized form of the human GM-CSF contains an extra methionine residue at position 1, but otherwise it is identical to the polypeptide predicted from the cDNA sequence. The specific activity of 2.9 X 10(7) units/mg of protein for purified bacterially synthesized human GM-CSF indicates that despite the lack of glycosylation, the molecule is substantially in its native conformation. This molecule stimulated the same number and type of both seven- and 14-day human bone marrow colonies as the CSF alpha preparation from human placental conditioned medium. Human GM-CSF had no activity on murine bone marrow or murine leukemic cells. There was no detectable, direct stimulation of adult human erythroid burst forming units (BFU-E) by the bacterially synthesized human GM-CSF. Although impure preparations containing native human GM-CSF (eg, human placental conditioned medium) stimulated the formation of mixed colonies, even in the presence of erythropoietin, the bacterially synthesized human GM-CSF failed to stimulate the formation of mixed colonies from adult human bone marrow cells. The bacterially synthesized human GM-CSF increased N-formyl-methionyl-leucyl- phenylalanine (FMLP)-induced superoxide production and lysozyme secretion. Antibody-dependent cytotoxicity and phagocytosis by human neutrophils was stimulated by the bacterially synthesized human GM-CSF and eosinophils were also activated in the antibody-dependent cytotoxicity assay. 相似文献
92.
KA Jackman AA Miller TM De Silva PJ Crack GR Drummond CG Sobey 《British journal of pharmacology》2009,156(4):680-688
Background and purpose:
Reactive oxygen species (ROS) derived from Nox2-containing reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity is reportedly detrimental in cerebrovascular disease. However, ROS generation by other Nox isoforms may have a physiological role. No Nox2-selective inhibitors have yet been identified, and thus it is unclear whether isoform non-selective Nox inhibitors would necessarily improve outcome after stroke. We assessed the effect of apocynin on cerebrovascular ROS production and also on outcome following cerebral ischaemia when administered either before ischaemia or after cerebral reperfusion. The involvement of Nox2-containing NADPH oxidase in the effects of apocynin was assessed using Nox2−/− mice.Experimental approach:
Transient cerebral ischaemia was induced by 0.5 h middle cerebral artery occlusion followed by 23.5 h reperfusion. Mice received apocynin (2.5 mg·kg−1, i.p.) either 0.5 h before ischaemia or 1 h after reperfusion. In situ superoxide production after cerebral ischaemia-reperfusion was measured in brain sections of wild-type mice at 24 h using dihydroethidium fluorescence.Key results:
Treatment with apocynin 0.5 h before ischaemia reduced total infarct volume, neurological impairment and mortality in wild-type but not Nox2−/− mice. Conversely, treatment with apocynin 1 h after initiation of reperfusion had no protective effect. Cerebral ischaemia and reperfusion increased superoxide production in the brain at 24 h, and pretreatment but not posttreatment with apocynin reduced superoxide levels.Conclusions and implications:
Apocynin improves outcome following stroke when administered before ischaemia in wild-type but not Nox2−/− mice. 相似文献93.
Bisphosphonate associated osteonecrosis of the jaws (ONJ) usually commences at the alveolus. Comparison is made between the structure and function of long bones and alveolar bone and the differing susceptibilities of the bisphosphonates at these different sites are explored. Current concepts of the causation of ONJ are discussed. The clinical implications of these findings to dentists managing periodontal conditions are presented. 相似文献
94.
95.
96.
Rat ferritin-H: cDNA cloning, differential expression and localization during hepatocarcinogenesis 总被引:2,自引:0,他引:2
Wu CG; Groenink M; Bosma A; Reitsma PH; van Deventer SJ; Chamuleau RA 《Carcinogenesis》1997,18(1):47-52
Elevated serum ferritin levels, especially of the H subunit, accompany many
clinical malignancies. By using the subtraction-enhanced display technique,
we have recently isolated several cDNA clones which are over- expressed in
rat hepatocellular carcinoma induced by diethylnitrosamine. One
830-base-pair clone was 88% similar to human ferritin-H cDNA and encoded a
182 amino acid protein which is 97% homologous to human ferritin-H chain.
Hepatic mRNA levels of ferritin-H were increased markedly at the early
stage of diethylnitrosamine- induced hepatocarcinogenesis in the rat (6
weeks) and appeared more than 10-fold overexpressed as the tumour
progressed. In contrast, hepatic ferritin-H mRNA remained constant during
liver regeneration after a 70% partial hepatectomy. In situ hybridization
showed that over- expression of ferritin-H was exclusively localized to
preneoplastic foci, to tumour nodules and to tumour cells invading blood
vessels. These findings suggest that ferritin-H is a highly conserved
protein, its over-expression during tumour development is phenotypically
correlated with tumour initiation and/or progression, and it is useful as
an early marker for hepatocellular carcinoma.
相似文献
97.
Genomic screening for beta-sarcoglycan gene mutations: missense mutations may cause severe limb-girdle muscular dystrophy type 2E (LGMD 2E) 总被引:7,自引:3,他引:7
Bonnemann CG; Passos-Bueno MR; McNally EM; Vainzof M; de Sa Moreira E; Marie SK; Pavanello RC; Noguchi S; Ozawa E; Zatz M; Kunkel LM 《Human molecular genetics》1996,5(12):1953-1961
Autosomal recessive limb-girdle muscular dystrophies (LGMDs) are
genetically heterogeneous. A subgroup of these disorders is caused by
mutations in the dystrophin-associated sarcoglycan complex. Truncating
mutations in the 43 kDa beta-sarcoglycan gene (LGMD 2E) were originally
identified in a sporadic case of Duchenne-like muscular dystrophy, and a
common missense mutation (T151R) was identified independently in Indiana
Amish pedigrees with a milder form of LGMD. To facilitate mutational
analysis of larger numbers of patients directly from genomic DNA, as
opposed to reverse transcribed RNA from muscle biopsies, we have determined
the genomic structure of the beta-sarcoglycan gene. The open reading frame
of the beta-sarcoglycan coding region extends over six exons. Primers were
designed for PCR amplification of single exons from genomic DNA and
subsequent single strand conformation polymorphism (SSCP) analysis. We
screened 15 patients from the Brazilian LGMD patient population, 13 of whom
followed a severe course. Most of the patients had been assessed previously
for deficiency of alpha- sarcoglycan immunofluorescence on muscle biopsy
sections as a marker for disease of the sarcoglycan complex. Novel
mutations in two familial and two sporadic cases of severe childhood-onset
LGMD were identified. Only one of these patients carried a truncating
mutation (homozygous 2 bp deletion, FS164TER), while the other three
carried missense mutations (homozygous R91P, homozygous M100K, heterozygous
recessive L108R; only one allele could be identified in this family). All
three missense mutations occurred in exon 3, coding for the immediate
extracellular domain. Complete absence for all three of the known
sarcoglycans was noted by immunohistochemistry on muscle biopsy sections of
the patients.
相似文献
98.
Granell Gil M Aguar Olba F Arnau Obrer A Grau Real F Cantó Armengod A Palanca Sanfrancisco JM 《Revista espa?ola de anestesiología y reanimación》2000,47(7):293-298
OBJECTIVES: To evaluate the effects on postoperative pulmonary function and quality of analgesia of two protocols for epidural infusion of alfentanil after lung resection. PATIENTS AND METHODS: After informed consent, 30 ASA I-IV patients undergoing chest surgery (lobectomy or pneumonectomy) were randomly assigned to two groups of 15. A catheter was inserted into the epidural space at T5-7 (group T) or L2-3 (group L). After a test dose, an initial bolus of alfentanil (10 micrograms/kg) was administered. After anesthetic induction, epidural analgesia was performed with an infusion of 400 micrograms/h of alfentanil (group L) during and after surgery. Endovenous patent-controlled anesthesia (PCA) was provided with morphine. During the first 24 h after surgery, the following variables were recorded: arterial blood gas concentrations, spirometric parameters, pain on a visual analog scale (VAS) and side effects. ANOVA and Scheffé and chi-square tests were used to analyze the results (p < or = 0.05). RESULTS: In group T, PaO2 was significantly higher at 6 and 18 h (p < or = 0.05), while FEV1 and FVC were significantly higher at 12 and 18 h. Pain assessed by VAS and PCA need for morphine was significantly less in group T. CONCLUSIONS: Thoracic epidural analgesia with alfentanil and lidocaine improves postoperative lung function and reduces the need for top-up analgesia in comparison with lumbar epidural infusion of alfentanil. 相似文献
99.
100.
Summary of mutations underlying autosomal recessive congenital ichthyoses (ARCI) in Arabs with four novel mutations in ARCI‐related genes from the United Arab Emirates
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