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51.
Quantitative evaluation of liver-specific promoters from retroviral vectors after in vivo transduction of hepatocytes 总被引:3,自引:1,他引:3
Hepatic gene therapy could be used to treat a number of inherited blood diseases such as hemophilia or thrombophilia. Although liver-directed retroviral transduction can result in long-term gene expression in vivo, the low level of protein production has limited its clinical application. We reasoned that the insertion of liver-specific promoters into retroviral vectors would increase gene expression in vivo. The 347- bp human alpha 1-antitrypsin (hAAT), the 810-bp murine albumin (mAIb), the 490-bp rat phosphoenolpyruvate carboxykinase (rPECK), and the 596- bp rat liver fatty acid binding protein promoters were inserted into a Moloney murine leukemia retroviral backbone containing the hAAT reporter gene. Vectors that produced appropriately sized RNA and hAAT protein in vitro were tested in vivo by transducing regenerating rat livers. Long-term serum expression of the hAAT reporter gene was normalized to retroviral transduction efficiency as determined by using a polymerase chain reaction-based assay of genomic DNA from transduced rat livers. The hAAT, mAIb, and rPEPCK promoters were, respectively, 35- , 8-, and 0.02-fold as strong as the previously studied constitutive Pol-II promoter. We conclude that the hAAT promoter resulted in the highest expression from a retroviral vector and may result in therapeutically significant expression of other clinically significant blood proteins. 相似文献
52.
P. Osewe D. G. Addiss K. A. Blair A. Hightower M. L. Kamb J. P. Davis 《Epidemiology and infection》1996,117(2):297-304
During March-April 1993, an estimated 403000 residents of the 5-county greater Milwaukee, Wisconsin area developed cryptosporidiosis after drinking contaminated municipal water. Although the number of cases dropped precipitously after the implicated water plant closed on 9 April, cases continued to occur. To investigate risk factors for post-outbreak cryptosporidiosis, 33 Milwaukee-area residents who had laboratory-confirmed Cryptosporidium infection with onset of diarrhoea between 1 May and 27 June 1993 were interviewed by telephone. Of these, 28 (85%) had onset of diarrhoea during May, 12 (36%) had watery diarrhoea during the outbreak, and 5(15%) were HIV-infected. In a neighbourhood-matched case-control study, immunosuppression (matched odds ratio (MOR) not calculable, 95% confidence interval (CI) 3.0, infinity) and having a child less than 5 years old in the household (MOR = 17.0, CI 2.0, 395.0) were independently associated with infection. When persons who had diarrhoea during the outbreak were excluded, immunosuppression remained significantly associated with illness (MOR not calculable, CI 1.6, infinity). Cryptosporidium transmission continued after this massive waterborne outbreak but decreased rapidly within 2 months. 相似文献
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55.
Linkage of a familial platelet disorder with a propensity to develop myeloid malignancies to human chromosome 21q22.1-22.2 总被引:2,自引:4,他引:2
Ho CY; Otterud B; Legare RD; Varvil T; Saxena R; DeHart DB; Kohler SE; Aster JC; Dowton SB; Li FP; Leppert M; Gilliland DG 《Blood》1996,87(12):5218-5224
Linkage analysis was performed on a large pedigree with an autosomal dominant platelet disorder and a striking propensity in affected family members to develop hematologic malignancy, predominantly acute myelogenous leukemia. We report the linkage of the autosomal dominant platelet disorder to markers on chromosome 21q22. Four genetic markers completely cosegregate with the trait and yield maximum logarithm of difference scores ranging from 4.9 to 10.5 (theta = .001). Two flanking markers, D21S1265 and D21S167, define a critical region for the disease locus of 15.2 centimorgan. Further analysis of this locus may identify a gene product that affects platelet production and function and contributes to the molecular evolution of hematologic malignancy. 相似文献
56.
Elevation of serum cystathionine levels in patients with cobalamin and folate deficiency 总被引:14,自引:0,他引:14
Homocysteine can be methylated to form methionine by the cobalamin- (Cbl) and folate-dependent enzyme, methionine synthase; serum levels of total homocysteine are elevated in greater than 95% of patients with either Cbl or folate deficiency. Homocysteine can also condense with serine to form cystathionine in a pyridoxal phosphate-dependent reaction catalyzed by cystathionine beta-synthase. Cystathionine is subsequently cleaved to cysteine and alpha-ketobutyrate by the pyridoxal phosphate-dependent enzyme gamma-cystathionase. To assess levels of cystathionine in Cbl and folate deficiency, we developed a new capillary gas chromatographic-mass spectrometric assay and measured cystathionine in the serum of normal subjects and patients with clinically confirmed deficiencies of these vitamins. The normal range for serum cystathionine was 65 to 301 nmol/L (median = 126 nmol/L) for 50 normal blood donors. In 30 patients with clinically confirmed Cbl deficiency, values for cystathionine ranged from 208 nmol/L to 2,920 nmol/L (median = 816 nmol/L) and 26 (87%) had levels above the normal range. In 20 patients with clinically confirmed folate deficiency, cystathionine concentrations ranged from 138 nmol/L to 4,150 nmol/L (median = 1,560 nmol/L) and 19 (95%) had values above the normal range. Five homozygotes for cystathionine beta-synthase deficiency had high values for serum-total homocysteine and low or low-normal values for serum cystathionine that ranged from 30 nmol/L to 114 nmol/L even though they were on treatment with pyridoxine and had partially responded. One patient with a defect in the synthesis of 5-CH3- tetrahydrofolate and five patients with defects in the synthesis of CH3- Cbl had high values for serum-total homocysteine and high values for cystathionine that ranged from 311 nmol/L to 1,500 nmol/L even though they were on treatment with folic acid and Cbl, respectively, and had partially responded. We conclude that levels of cystathionine are evaluated in the serum of most patients with Cbl and folate deficiency and that they are useful in the differential diagnosis of an elevated serum-total homocysteine level. 相似文献
57.
58.
Congenital heart lesions involving the right ventricular outflow tract (RVOT) are a common problem in paediatric cardiology. These patients need multiple surgical interventions in the form of valved conduits over a lifetime. Surgical re-valvulation was the standard treatment option until the introduction of percutaneous pulmonary valves over a decade ago. These valves can be used to prolong the lifespan of conduits and reduce the number of re-operations. The Melody® valve (Medtronic, Minneapolis, MN, USA) was introduced as the first dedicated percutaneous pulmonary valve. Percutaneous pulmonary valves can be implanted successfully and have the advantage of short hospitalisations. We describe the first three Melody® valve implantations in Africa. 相似文献
59.
Baird JB Charles JL Streit TG Roberts JM Addiss DG Lammie PJ 《The American journal of tropical medicine and hygiene》2002,66(2):163-169
Both secondary infections and antifilarial immunity are thought to play roles in the development and progression of lymphedema. To investigate this issue, immune responses to a panel of bacterial, fungal, and parasite antigens were examined for women with lymphedema and elephantiasis (n = 28) and for women with no clinical evidence of lymphatic dysfunction who were either microfilaremic (Mf+, n = 23) or microfilaria- and filarial antigen-negative (Ag-, n = 24). The prevalence and intensity of delayed-type hypersensitivity (DTH) responses was similar for most recall antigens; for individual antigens, lymphedema patients were significantly more likely to be reactive only to Proteus. Lymphedema patients with a history of three or more attacks of adenolymphangitis in the last 18 months showed increased DTH reactivity to Trichophyton. Proliferative responses to fungal and bacterial antigens were similar for all three groups; however, antigen-negative women, independent of disease status, mounted greater responses to filarial antigen. In contrast, lymphedema patients had higher levels of antifilarial specific IgG1, IgG2, and IgG3 and higher IgG responses to streptolysin O than either Ag- or Mf+ women. In persons with lymphatic filariasis, immune reactivity is influenced by disease status as well as infection status. 相似文献
60.
Critchley J Addiss D Ejere H Gamble C Garner P Gelband H;International Filariasis Review Group 《Tropical medicine & international health : TM & IH》2005,10(9):818-825
OBJECTIVES: The Global Programme to Eliminate Lymphatic Filariasis recommends albendazole in combination with other antifilarial drugs. This systematic review examines albendazole in treatment and control of lymphatic filariasis. DATASOURCES: The Cochrane Controlled Trials Register, MEDLINE and EMBASE to April 2005; contacting experts, international organisations and drug manufacturers. METHODS: Randomised or quasi-randomised controlled trials included; two reviewers independently assessed eligibility, quality, and extracted data. We calculated the relative risk of microfilaraemia (mf) prevalence using fixed effect, or random effects model in case of heterogeneity. RESULTS: Six trials met inclusion criteria. Three trials compared albendazole with placebo: no effect was demonstrated on mf prevalence, but density was lower in one of the three studies at 6 months. Three trials added albendazole to ivermectin, with no demonstrable effect; prevalence tended to be lower at 4--6 months but not at 12 months (4--6 months; RR 0.49, 95% CI 0.18 to 1.39, n=255, 2 trials; 12 months: RR 1.00, 95% CI 0.88 to 1.13, n=348, 2 trials). Mf density was significantly lower in two of the three trials; one of two trials measuring density at 12 months showed a difference. Three trials added albendazole to diethylcarbamazine; two were small trials with no difference demonstrated; the third study tended to favour combination at 6 months (RR=0.62, 95% CI 0.32 to 1.21, n=491), with a significant difference for density. CONCLUSIONS: The effect of albendazole against adult and larval filarial parasites, alone and in combination with other antifilarial drugs, deserves further rigorous research. 相似文献