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Dual expression of the HA protein of H5N2 avian influenza virus in a baculovirus system 总被引:3,自引:0,他引:3
Hu YC Luo YL Ji WT Chulu JL Chang PC Shieh H Wang CY Liu HJ 《Journal of virological methods》2006,135(1):43-48
Baculovirus/insect cell system is used widely for recombinant protein production. The hemagglutinin (HA) gene of H5N2 avian influenza virus (AIV) 1209 strain and the enhanced green fluorescent protein (EGFP) gene were cloned into pFastBac DUAL vector that has two promoters and cloning sites, allowing simultaneous expression of these two genes. The HA protein of AIV was fused with a hexahistidine (His6) tag for purification. The coexpression of EGFP allowed identification of the recombinant baculoviruses in Sf-9 insect cells, eliminating cumbersome and time-consuming assays. A recombinant baculovirus, Bac-HA, was generated by transfecting pBac-HA to bacmid inside DH10B(AC)Escherichia coli by site-specific transposition, followed by transfection into the Sf-9 cells. Fluorescence in the insect cells was observed from 3 days post-infection. The expressed HA protein was confirmed by Western blot using an anti-HA monoclonal antibody. Also, different detergents and incubation times on ice were tested. The two-stage extraction with Triton X-100 or Tween 20 and incubation on ice for 2h exhibited high efficiency. Since purification of HA with ConA resin resulted in low protein recovery, lentil lectin affinity column was used and was useful for HA purification. 相似文献
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Jean-Louis Bayart Jonathan Degosserie Julien Favresse Constant Gillot Marie Didembourg Happy Phanio Djokoto Valrie Verbelen Gatien Roussel Cline Maschietto Franois Mullier Jean-Michel Dogn Jonathan Douxfils 《Viruses》2022,14(4)
Rapid antigen detection (RAD) tests are commonly used for the diagnosis of SARS-CoV-2 infections. However, with the continuous emergence of new variants of concern (VOC), presenting various mutations potentially affecting the nucleocapsid protein, the analytical performances of these assays should be frequently reevaluated. One hundred and twenty samples were selected and tested with both RT-qPCR and six commercial RAD tests that are commonly sold in Belgian pharmacies. Of these, direct whole-genome sequencing identified the strains present in 116 samples, of which 70 were Delta and 46 were Omicron (BA.1 and BA.1.1 sub-lineages, respectively). The sensitivity across a wide range of Ct values (13.5 to 35.7; median = 21.3) ranged from 70.0% to 92.9% for Delta strains and from 69.6% to 78.3% for Omicron strains. When taking swabs with a low viral load (Ct > 25, corresponding to <4.9 log10 copies/mL), only the Roche RAD test showed acceptable performances for the Delta strains (80.0%), while poor performances were observed for the other RAD tests (20.0% to 40.0%). All the tested devices had poor performances for the Omicron samples with a low viral load (0.0% to 23.1%). The poor performances observed with low viral loads, particularly for the Omicron strain, is an important limitation of RAD tests, which is not sufficiently highlighted in the instructions for use of these devices. 相似文献
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Rajaraman DuraiAuthor Vitae Philip C.H. NgAuthor Vitae Happy HoqueAuthor Vitae 《AORN journal》2010,91(5):599-609
Methicillin-resistant Staphylococcus aureus (MRSA) is a serious threat to patients in health care facilities and the community. A MRSA infection can be much more severe than other bacterial infections and can be life-threatening. Resistance to common antibiotics makes treating MRSA costly and difficult. Prolonged hospitalization requiring specialized IV antibiotics also has cost implications. Treatment of MRSA can include use of antibiotics; topical therapies such as honey, topical silver, and gentian violet; and bacteriophages. Research is being conducted on new antibiotics and a MRSA vaccine. 相似文献
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Vincent Chin-Hung Chen Andrew T. A. Cheng Happy K. L. Tan Chung-Ying Chen Tony H. H. Chen Robert Stewart Martin Prince 《Social psychiatry and psychiatric epidemiology》2009,44(12):1005-1011
Objective
To investigate the case fatality proportion and associated factors in those carrying out suicide acts in Nantou, Taiwan. 相似文献9.
Chen CH Tan HK Liao LR Chen HH Chan CC Cheng JJ Chen CY Wang TN Lu ML 《Comprehensive psychiatry》2007,48(3):269-275
OBJECTIVE: Severe natural disasters can cause long-term psychological impact on the survivors. This study aimed to examine the prevalence and risk factors of posttraumatic stress symptoms and psychiatric morbidity among survivors of the severe earthquake that occurred in Chi-Chi, Taiwan, in September 21, 1999. METHODS: A total of 6412 earthquake survivors whose houses were destroyed by earthquake were recruited about 2 years after the disaster. They completed a self-report questionnaire assessing posttraumatic stress symptoms, psychiatric morbidity, and information of demographics, trauma exposure, and current living status. RESULTS: The estimated rates of posttraumatic stress disorder caseness and psychiatric morbidity were 20.9% and 39.8%, respectively. Psychiatric morbidity occurred mainly in survivors who were female, older, with low education level, and currently living in a prefabricated house. The risk factors for posttraumatic stress disorder caseness were female sex, currently living in a prefabricated house, low education level, and experienced complete destruction of property. CONCLUSION: These results showed that severe earthquake can cause long-term psychological impact in the survivors. The findings of risk factors suggest avenues for targeting postdisaster interventions. 相似文献
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Hui-Ling Lee Happy Kuy-Lok Tan Hui-Ing Ma Chia-Yu Tsai Yung-Kung Liu 《The American journal of occupational therapy》2006,60(4):435-441
OBJECTIVE: The purpose of this study was to examine the effect of a work-related stress management program on perceived work-related stress in patients with chronic schizophrenia. METHOD: A single-blind, randomized crossover design was used. Twenty-nine patients undergoing vocational training while working at paid part-time jobs in a psychiatric center were randomly assigned to receive 12 weeks of a work-related stress management program followed by 12 weeks of no treatment, or the reverse. The Work-Related Stress Questionnaire for Chronic Psychiatric Patients (WSQP) was developed for this study and used to examine the treatment effect. RESULTS: The decrease in stress was significantly greater when the participants were undergoing the program compared to the decrease of stress when they were not (t = 2.93, p = .0034, r = .49). CONCLUSION: The results show that the work-related stress management program had large short-term positive effects on the patients' perceived work-related stress. These findings support providing this type of program to employed patients with chronic schizophrenia. 相似文献