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Huang CP Liu YT Nakatsuji T Shi Y Gallo RR Lin SB Huang CM 《Proteomics. Clinical applications》2008,2(9):1234-1245
Proteomics is a powerful tool for the identification of proteins, which provides a basis for rational vaccine design. However, it is still a highly technical and time‐consuming task to examine a protein's immunogenicity utilizing traditional approaches. Here, we present a platform for effectively evaluating protein immunogenicity and antibody detection. A tetanus toxin C fragment (Tet‐c) was used as a representative antigen to establish this platform. A cell wall‐anchoring sialidase‐like protein (SLP) of Propionibacterium acnes was utilized to assess the efficacy of this platform. We constructed an Escherichia coli vector‐based vaccine by overexpressing Tet‐c or SLP in E. coli and utilized an intact particle of E. coli itself as a vaccine (E. coli Tet‐c or SLP vector). After ultraviolet (UV) irradiation, the E. coli vector‐based vaccines were administered intranasally into imprinting control region mice without adding exogenous adjuvants. For antibody detection, we fabricated antigen microarrays by printing with purified recombinant proteins including Tet‐c and SLP. Our results demonstrated that detectable antibodies were elicited in mice 6 weeks after intranasal administration of UV‐irradiated E. coli vector‐based vaccines. The antibody production of Tet‐c and SLP was significantly elevated after boosting. Notably, the platform with main benefits of using E. coli itself as a vaccine carrier provides a critical template for applied proteomics aimed at screening novel vaccine targets. In addition, the novel immunogenic SLP potentially serves as an antigen candidate for the development of vaccines targeting P. acnes‐associated diseases. 相似文献
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结核分枝杆菌融合抗原Ag85B-ESAT6真核表达载体的构建及鉴定 总被引:1,自引:0,他引:1
以结核分枝杆菌H37Rv株的基因组作为模板,将Ag85B和ESAT6编码基因进行PCR扩增,然后采用基因剪接重叠扩增PCR法(gene SOEing)将其通过疏水甘氨酸接头(GGIGIAPG)连接融合,定向克隆至质粒Pvax1中,构建结核分枝杆菌Ag85B-ESAT6融合抗原的真核表达质粒Pvax1/AE.经单双限制性内切酶图谱、PCR产物及DNA测序分析等多种方法鉴定,证实Pvax1/AE真核表达质粒构建成功.为进一步研究其结核核酸疫苗原型的免疫保护效果奠定了基础. 相似文献
25.
Cytokine flow cytometry (CFC) is a multiparameter assay of antigen-specific T cell function, potentially useful in the monitoring of experimental vaccines and progression of infectious diseases and cancer. Automation of CFC assays would greatly facilitate their use in clinical trials and involves several components. We describe here the migration of these assays to 96-well plates, the use of sample-handling robotics, and the use of lyophilized antigen and antibody plates to help automate CFC. Together, these elements can produce an integrated system capable of walkaway automation of an entire assay, resulting in the reproducible processing of potentially hundreds of samples per day. Implementation of such systems has begun to be undertaken by our group and others. 相似文献
26.
In order to develop more effective immunological strategies to prevent vibriosis of farmed marine fish in Hong Kong and southern China, various vaccine preparations including formalin-, phenol-, chloroform- and heat-killed whole cell bacterins and subcellular lipopolysaccharides (LPS), as well as different administration routes, were investigated. Fish immunized with the subcellular LPS exhibited the best protection [Relative Percent of Survival (RPS) = 100], while fish immunized with whole cell bacterins displayed varying degrees of protection (RPS ranged from 28 to 80), in descending order: formalin-killed > phenol-killed > heat-killed > chloroform-killed bacterins. Regarding various administration routes, fish immunized with two intraperitoneal (i.p.) injections exhibited the best protection, and the RPS values were 100 or 85 upon higher or lower doses of pathogenic V. alginolyticus challenges. Both oral vaccination and a combination of injection/immersion trial were also effective, which achieved relatively high protection (the RPS values ranged from 45 to 64.3). However, two hyperosmotic immersions could not confer satisfactory protection, especially when fish were exposed to the severe pathogenic bacteria challenge. Marked elevations of serum agglutinating antibody titer were detected in all immunized fish. Macrophage phagocytosis was enhanced significantly, especially in the fish immunized by formalin- and phenol-killed bacterins through various administration routes. Both adaptive (specific antibody) and innate (phagocytic activity) immunity elicited by different immunization strategies were in parallel with the degree of protection offered by each of them. Although all vaccination trials had no significant effect on the serum hematocrit and hemoglobin levels, the circulating lymphocyte counts were significantly elevated in the fish immunized with LPS, formalin- and phenol-killed bacterins. Serum cortisol levels appeared to be reduced in all immunized fish except the trial of hyperosmotic immersion, which indicated the stressful impact on vaccinated fish. 相似文献
27.
Molecularly imprinted polymers by phase inversion technique for the selective recognition of saccharides of biomedical interest in aqueous solutions
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Niccoletta Barbani Elisabetta Rosellini Marco Donati Paolo Costantino Caterina Cristallini Gianluca Ciardelli 《Polymer International》2017,66(6):900-907
The purpose of this work was the preparation and characterization of polymeric membranes for the selective recognition of saccharides using molecular imprinting technology associated with phase inversion. A system able to bind saccharides with high selectivity is particularly important in the pharmaceutical sector, since some of these compounds are constituents of molecules which can exert serious toxic effects even at very low concentrations. Two polymeric matrices were prepared using poly(ethylene‐co‐vinyl alcohol) copolymers, with an ethylene molar content of 32% and 44%, and were imprinted with two different saccharide molecules: maltose and 2‐keto‐3‐deoxy‐d ‐manno‐octulosonate (KDO). Matrices imprinted against maltose and KDO showed an easy template extraction, high binding capability and satisfactory selectivity, particularly for the matrix with an ethylene molar content of 44%. © 2017 Society of Chemical Industry 相似文献
28.
Shin-ichiro Ohno Gregor P. C. Drummen Masahiko Kuroda 《International journal of molecular sciences》2016,17(2)
Many types of cells release phospholipid membrane vesicles thought to play key roles in cell-cell communication, antigen presentation, and the spread of infectious agents. Extracellular vesicles (EVs) carry various proteins, messenger RNAs (mRNAs), and microRNAs (miRNAs), like a “message in a bottle” to cells in remote locations. The encapsulated molecules are protected from multiple types of degradative enzymes in body fluids, making EVs ideal for delivering drugs. This review presents an overview of the potential roles of EVs as natural drugs and novel drug-delivery systems. 相似文献
29.
To increase antibody secretion and dose sparing, squalene-in-water aluminium hydrogel (alum)-stabilised emulsions (ASEs) have been developed, which offer increased surface areas and cellular interactions for higher antigen loading and enhanced immune responses. Nevertheless, the squalene (oil) in previous attempts suffered from limited oxidation resistance, thus, safety and stability were compromised. From a clinical translational perspective, it is imperative to screen the optimal oils for enhanced emulsion adjuvants. Here, because of the varying oleic to linoleic acid ratio, soybean oil, peanut oil, and olive oil were utilised as oil phases in the preparation of aluminium hydrogel-stabilised squalene-in-water emulsions, which were then screened for their stability and immunogenicity. Additionally, the underlying mechanisms of oil phases and emulsion stability were unravelled, which showed that a higher oleic to linoleic acid ratio increased anti-oxidative capabilities but reduced the long-term storage stability owing to the relatively low zeta potential of the prepared droplets. As a result, compared with squalene-in-water ASEs, soybean-in-water ASEs exhibited comparable immune responses and enhanced stability. By optimising the oil phase of the emulsion adjuvants, this work may offer an alternative strategy for safe, stable, and effective emulsion adjuvants. 相似文献
30.
目的评价卡介菌纯蛋白衍生物(purified protein derivative of BCG,BCG-PPD)的稳定性。方法将3批BCGPPD样品置2~8℃存放30个月,分别于第0、3、6、9、12、18、24、30个月进行物理外观检查、pH和效价测定,第0、12、18、24、30个月同时进行鉴别试验、无菌试验、装量差异、异常毒性及苯酚含量检测;于(25±2)℃条件下存放12个月,第0、1、2、3、6、9、12个月进行物理外观检查、pH和效价测定;于(37±2)℃条件下存放12周,在第0、1、2、3、4、8、12个月进行外观检查、pH和效价测定。结果 3批BCG-PPD于2~8℃存放不同时间样品的鉴别试验、物理外观、装量差异、无菌试验及异常毒性检测结果均合格,pH为7. 12~7. 25,苯酚含量为2. 48~2. 72 g/L,效价为0. 92~1. 13,均呈稳定状态。3批BCG-PPD样品于(25±2)和(37±2)℃条件下保存,物理外观检查均合格,pH分别为7. 12~7. 32和7. 11~7. 20,效价分别为0. 93~1. 06和0. 91~1. 02,均符合要求。结论 BCG-PPD具有良好的稳定性,可满足将其有效期延长至24个月的要求。 相似文献