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1.
《Journal of dairy science》2022,105(1):585-594
Herd-level diagnosis of paratuberculosis using a pool-milk ELISA (pool size: n ≤ 50) is a novel, economical, and convenient method to identify blood serological Mycobacterium avium ssp. paratuberculosis (MAP) antibody–positive herds. To date, the diagnostic performance of the pool-milk ELISA has been described only under laboratory conditions where herd prevalence was simulated by the preparation of milk pools consisting of milk samples of cows with a known MAP status determined by fecal culture. In our observational study, test performance under field conditions was studied using pooled milk and individual blood samples. A total of 486 herds within the MAP prevalence reduction program of Lower Saxony, from which pooled milk and individual blood ELISA results were available, were assigned to this study. Data were analyzed for the period between January 1 and December 31, 2018, the first year after herd testing became obligatory in this federal state of Germany. To evaluate whether pooled milk samples reliably distinguish between herds with a MAP-apparent blood serological within-herd prevalence (MAP-Ab-WHPapp) ≥5% and herds with a MAP-Ab-WHPapp <5%, the distribution of the MAP-Ab-WHPapp was compared between pool-positive and pool-negative herds. The MAP-Ab-WHPapp was 3.4% (median; 95% confidence interval = 0–11.4%) in pool-positive herds and 1.2% (median; 95% confidence interval = 0–6.4%) in pool-negative herds. Only 10.8% (n = 12) of the pool sample–negative herds had a MAP-Ab-WHPapp ≥5% and were therefore false negatives, given the aims of the MAP prevalence reduction program. Hence, the pool-milk sampling strategy seems well suited to distinguish between herds with a MAP-Ab-WHPapp ≥ 5% and herds with a MAP-Ab-WHPapp <5% since only 10% of serum MAP-ELISA positive herds were missed. Employing a logistic regression model, we estimated that the minimum blood serological MAP-Ab-WHPapp to detect a pool-positive herd with a probability of 95% was 8%, which fits well with the aim of the MAP prevalence reduction program to focus on herds with a MAP-Ab-WHPapp of ≥5%. Despite the limitations of the control approach, which include milk pool sample collection and a low sensitivity of the ELISA used in milk pools and serum samples, the aims of the MAP prevalence reduction program can be achieved. The results of these field data support that pool-milk sample ELISA is a useful, economical, and low labor–intensive tool to identify herds seropositive for MAP in a MAP prevalence reduction program.  相似文献   
2.
为了建立一种检测猪肉中司帕沙星(SPFX)残留的方法,并且在检测时具有特异性高,背景低和无需样品预处理的特点。该研究制备了间接竞争性酶联免疫吸附剂和特异性抗司帕沙星单克隆抗体。结果表明,该抗体与氟甲喹(11.99%)、氟罗沙星(11.14%)、恩诺沙星(4.60%)、环丙沙星(0.31%)几乎没有交叉反应。该方法测定了添加SPFX的猪肉中的药物残留,检测内变异系数小于5.68%,检测间变异系数小于5.51%。检测间和检测内的平均回收率分别为102%~106%和101%~104%。该抗体对SPFX的IC50为31μg/L,没有抗体具有四种结构相关化合物和其他化合物。高效液相色谱(HPLC)和酶联免疫吸附试验(ELISA)的结果进一步证实了icELISA快速检测猪肉中SPFX的可靠性和准确性。该研究表明icELISA法检测猪肉中SPFX残留分析是一种可靠、简便、低成本的方法。  相似文献   
3.
Acute myocardial infarction (MI) is one of the most common causes of death worldwide. Pituitary adenylate cyclase activating polypeptide (PACAP) is a cardioprotective neuropeptide expressing its receptors in the cardiovascular system. The aim of our study was to examine tissue PACAP-38 in a translational porcine MI model and plasma PACAP-38 levels in patients with ST-segment elevation myocardial infarction (STEMI). Significantly lower PACAP-38 levels were detected in the non-ischemic region of the left ventricle (LV) in MI heart compared to the ischemic region of MI-LV and also to the Sham-operated LV in porcine MI model. In STEMI patients, plasma PACAP-38 level was significantly higher before percutaneous coronary intervention (PCI) compared to controls, and decreased after PCI. Significant negative correlation was found between plasma PACAP-38 and troponin levels. Furthermore, a significant effect was revealed between plasma PACAP-38, hypertension and HbA1c levels. This was the first study showing significant changes in cardiac tissue PACAP levels in a porcine MI model and plasma PACAP levels in STEMI patients. These results suggest that PACAP, due to its cardioprotective effects, may play a regulatory role in MI and could be a potential biomarker or drug target in MI.  相似文献   
4.
采用酶标抗原建立了高效、高灵敏的三聚氰胺直接竞争的ELISA(enzyme linked immunosorbent assay,ELISA)检测方法。方法 以三聚氰胺单克隆抗体包被作为固相抗体,辣根过氧化酶(Horseradish Peroxidase,HRP)标记的抗原与标准品(或样品)中三聚氰胺竞争结合抗体,建立了直接竞争酶联免疫检测体系。结果 以三聚氰胺为标准品建立标准曲线,得到方法的IC50为8.84μg/L,灵敏度为0.65μg/L,线性范围0.9-35μg/L;检测样品的回收率为70.0%-127.9%,与三聚氰酸交叉反应率为60%,其他结构类似物基本无交叉反应。结论 本方法灵敏度高、特异性强、检测快速, 可以满足实际样品的检测需求。  相似文献   
5.
Although Canadian dairy herds have been infected with bovine leukemia virus (BLV) for years, recent research has put new emphasis on the potential negative effects of this infection. Consequently, BLV control is becoming more favorable; however, BLV control cannot be successful without identifying infected animals. Bovicheck BLV (Biovet, Saint-Hyacinthe, QC, Canada) is currently the only assay licensed by the Canadian Centre for Veterinary Biologics. The first goal of this study was, therefore, to determine the reproducibility of the Bovicheck BLV assay for serum samples derived from Canadian cattle. The second goal was to evaluate and compare 5 different ELISA and determine their test characteristics using serum samples from Canadian herds. The considered ELISA were Bovicheck BLV, ID Screen BLV Competition (IDvet, Grabels, France), Idexx Leukosis Serum X2 Ab Test (Idexx Europe B.V., Hoofddorp, the Netherlands), Svanovir BLV gp51-Ab (Svanova, Uppsala, Sweden), and the Serelisa BLV Ab Mono Indirect (Synbiotics, Lyon, France). Eighty serum samples from Canadian cattle provided by Prairie Diagnostic Services (PDS; Saskatoon, SK, Canada) and an additional 80 serum samples from Canadian dairy and beef herds were used for the study. The Bovicheck BLV assay yielded the same results for all PDS-derived samples, implying a high level of reproducibility and robustness of this assay. Additionally, the comparison of the assays' results showed high agreement between assays, with Cohen's kappa values between κ = 0.91 and κ = 1. Furthermore, using original test results of the field samples as true status, relative diagnostic sensitivity and specificity were calculated. Relative diagnostic sensitivity of all tests was 100%. False-positive results were probable; therefore, the following relative diagnostic specificities were determined: 100% for Bovicheck BLV, Idexx Leukosis Serum X2, and Svanovir BLV; 95% for ID Screen BLV; and 97% for Serelisa BLV. When considering other test characteristics, ID Screen BLV is exceptional due to considerable practical advantages.  相似文献   
6.
Mycoplasma bovis is an important pathogen causing disease and substantial economic losses in cattle. However, knowledge of the dynamics of antibody responses in individual cows in the face of an outbreak is currently extremely limited. The use of commercial antibody tests to support clinical decision-making and for surveillance purposes is therefore challenging. Our objective was to describe the dynamics of M. bovis antibody responses in 4 Danish dairy herds experiencing an acute outbreak of M. bovis-associated disease, and to compare the antibody dynamics between dairy cows with different disease manifestations. A total of 120 cows were examined using a standardized clinical protocol and categorized into 4 disease groups: “mastitis,” “systemic,” “nonspecific,” and “none.” Paired blood and milk samples were collected and tested using a commercial M. bovis antibody–detecting ELISA. Plots of raw data and generalized additive mixed models with cow and herd as random effects were used to describe serum and milk antibody dynamics relative to the estimated time of onset of clinical disease. Cows with mastitis had high optical density measurement (ODC%) of antibodies in both milk and serum at disease onset. The estimated mean ODC% in milk was below the manufacturer's cut-off for the other groups for the entire study period. The estimated mean serum ODC% in the “systemic” group was high at onset of disease and stayed above the cut-off until 65 d after disease onset. However, the lower 95% confidence interval (CI) for the mean ODC% was only above the manufacturer's cut-off between 7 and 17 d after onset of disease. The CI of the “systemic” and “none” groups did not overlap at any time between the day of disease onset and 65 d after disease onset, and the estimated mean ODC% for both the “nonspecific” and “none” groups were generally below the cut-off for the majority of the study period. In conclusion, the serum antibody responses were highly dynamic and showed a high level of variation between individual cows. This strongly suggests that serology is unlikely to be useful for individual diagnosis of M. bovis-associated disease in dairy cows. However, it might still be useful for herd- or group-level diagnosis. Antibodies in milk were only increased in cows with M. bovis mastitis, indicating that milk antibody measurements only have diagnostic utility for cows with mastitis.  相似文献   
7.
Gluten is the main family of storage proteins found in barley. During malting and brewing, some of the barley malt's proteinaceous material is hydrolysed into peptides or to amino acids. Most of the gluten proteins are removed with the spent grains and with hot‐ and cold‐breaks. However, some gluten proteins and especially gluten‐derived peptides can remain throughout the brewing process and will hamper the gluten‐free (≤20 ppm) status of the beer. In this work, three production batches (a, b and c) of 51 Belgian barley malt beers from 24 breweries were analysed with the sandwich (R7001) and competitive (R7021) Ridascreen gliadin R5‐ELISA to quantify gluten proteins and peptides. Although the majority of the beers contained low‐gluten protein concentrations of ≤20 ppm (a/45, b/47, c/48), only a minority were truly gluten‐free with ≤20 ppm gluten peptides (a/18, b/17, c/15). The grain bill had no influence on the measured gluten concentration, but the use of (combined) clarification techniques and presence of wheat malt in the grist was respectively a positive and negative influence. Ten beers, from four breweries, were gluten free in all analysed samples. These included two wheat beers, reflecting the importance of effective clarification in the management of gluten. These results explore the feasibility of the production of gluten‐free barley malt beers. Copyright © 2018 The Institute of Brewing & Distilling  相似文献   
8.
Tumor-associated (TA) autoantibodies have been identified at the early tumor stage before developing clinical symptoms, which holds hope for early cancer diagnosis. We identified a TA autoantibody from HBx-transgenic (HBx-tg) hepatocellular carcinoma (HCC) model mouse, characterized its target antigen, and examined its relationship to human HCC. The mimotopes corresponding to the antigenic epitope of TA autoantibody were screened from a random cyclic peptide library and used for the detection of serum TA autoantibody. The target antigen of the TA autoantibody was identified as an oncogenic bi-functional purine biosynthesis protein, ATIC. It was upregulated in liver cancer tissues of HBx-tg mouse as well as human HCC tissues. Over-expressed ATIC was also secreted extracellularly via the cancer-derived exosomes, which might cause auto-immune responses. The cyclic peptide mimotope with a high affinity to anti-ATIC autoantibody, CLPSWFHRC, distinguishes between serum samples from HCC patients and healthy subjects with 70.83% sensitivity, 90.68% specificity (AUC = 0.87). However, the recombinant human ATIC protein showed a low affinity to anti-ATIC autoantibody, which may be incompatible as a capture antigen for serum TA autoantibody. This study indicates that anti-ATIC autoantibody can be a potential HCC-associated serum biomarker and suggests that autoantibody biomarker’s efficiency can be improved by using antigenic mimicry to native antigens present in vivo.  相似文献   
9.
结合间接竞争反应机制,运用酶联免疫吸附技术(enzyme-linked immunosorbent assay,ELISA),从抗原(antigen,Ag)包被时间、体系反应温度、酶标二抗作用时间、显色时间等主要因素开展快速检测黄曲霉毒素改进方法的研究。研究结果表明适宜的优化条件为:采用Ag包被20 h、反应温度24℃左右、酶标二抗作用时间30 min、底物显色时间15min。通过对饲料等11种样本的检测得出:半抑制浓度IC50<0.1μg/L,添加回收率在67%~116%,灵敏度达到0.03μg/L,线性系数>0.99,板内变异小于5%。检测试验结果良好,表明该改进方法具有可行性。  相似文献   
10.
为制备土霉素(Oxytetracycline,OTC)完全抗原,获得免疫学特性良好的土霉素鼠源多抗血清并建立OTC免疫学检测方法,采用重氮化法在OTC分子上引入羧基,进而利用改进碳二亚胺法将其与载体蛋白偶联制备完全抗原,通过红外扫描、紫外扫描和凝胶电泳鉴定其偶联效果,然后将完全抗原免疫BLAB/c小鼠获得多抗血清(pAb)并对其免疫学特性进行鉴定,通过优化条件建立OTC间接竞争ELISA检测方法。结果表明,偶联效果良好,免疫的4只小鼠多抗血清效价均达到1∶12 800;4只小鼠多抗血清均有抑制效果,其中2号小鼠产生的多抗血清效果最佳,基于该多抗血清建立的OTC间接竞争ELISA检测方法半数抑制浓度(IC_(50))为32.92ng/mL,检测限(LOD)为1.3ng/mL,牛奶样品中添加回收率在84.70%~87.45%,与金霉素、四环素的交叉反应率分别为5.27%,4.10%,与其他竞争物交叉反应率0.4%。该试验成功合成了OTC完全抗原,获得了免疫学特性良好的OTC多抗血清,并建立了OTC免疫学检测方法。  相似文献   
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