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1.
A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2–4F, 7–3G, and 11–11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC50) and percentage of cross-reactivity, respectively. MAb 2–4F exhibited the highest sensitivity, with an IC50 of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross-reactivity to other unrelated antibiotics. When MAb 2–4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%–118% for an intra-assay and 96%–113% for an inter-assay. The coefficients of variation of the assays were 3.9%–13.9% and 5.5%–14.9%, respectively.  相似文献   

2.
A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC50) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%–84.60%, with coefficients of variation of less than 12.2%.  相似文献   

3.
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4.
Objective: To evaluate the interaction between serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) and Helicobacter pylori (H. pylori) infection in patients with chronic gastritis and peptic ulcer. Methods: The serum levels of sICAM-1 in 205 patients with chronic gastric diseases were detected by ELISA method and the status of H. pylori was determined by histologic examination, RUT, 14C - UBT, and serology. The sera obtained from 18 healthy volunteers served as controls. Results: The serum levels of sICAM-1 were significantly higher in patients with H. pylori positive than those of H. pylori negative (889.43±32.52 ng/ml vs. 747.07±30.45 ng/ml, P<0.05). The serum levels of sICAM-1 in patients with mild, moderate and severe infection of H. pylori were 841.68±72.36 ng/ml, 905.43±37.59 ng/ml and 1012.54±49.34 ng/ml,respectively (P<0.05). The serum levels of sICAM-1 proved to be significantly correlated with the density of H. pylori colonization in gastric mucosa (rs =0.316, P<0.001). The serum levels of sICAM-1 in patients with chronic gastritis and peptic ulcer were significantly higher than those in healthy controls (P<0.05). Conclusions: These results indicated that H. pylori infection up-regulates the expression of sICAM-1.  相似文献   

5.
Modified 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) method was employed to synthesize the artificial antigen of norfloxacin (NOR), and New Zealand rabbits were used to produce anti-NOR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (icELISA) standard curve was established. This assay was sensitive and had a working range from 0.12 to 68.40 ng/ml, with the half maximal inhibitory concentration (IC50) and limit of detection (LOD) values of 2.7 ng/ml and 0.06 ng/ml, respectively. The produced pAb exhibited high cross-reactivity to fluoroquinolones (FQs) tested, and the IC50 values to enoxacin, ciprofloxacin, and pefloxacin were 3.1, 3.4, and 4.1 ng/ml, respectively. It also indicated that the concentrations of NaOH and methanol in assay buffer should not be higher than 10% and 30%. When spiked in milk at 5, 20, and 50 ng/ml, the recoveries for NOR, enoxacin, ciprofloxacin, and pefloxacin ranged 90.5%–98.0%, 84.0%–95.2%, 94.0%–106.0%, and 89.5%–100.0%, respectively. The results suggest that this class-specific pAb-based icELISA could be utilized for the primary screening of FQ residues in animal-original products.  相似文献   

6.
Objective: To evaluate the interaction between serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) andHelicobacter pylori (H. pylori) infection in patients with chronic gastritis and peptic ulcer. Methods: The serum levels of sICAM-1 in 205 patients with chronic gastric diseases were detected by ELISA method and the status ofH. pylori was determined by histologic examination, RUT,14C-UBT, and serology. The sera obtained from 18 healthy volunteers served as controls. Results: The serum levels of sICAM-1 were significantly higher in patients withH. pylori positive than those of H. pylori negative (889.43±32.52 ng/ml vs. 747.07±30.45 ng/ml,P<0.05). The serum levels of sICAM-1 in patients with mild, moderate and severe infection ofH. pylori were 841.68±72.36 ng/ml, 905.43±37.59 ng/ml and 1012.54±49.34 ng/ml, respectively (P<0.05). The serum levels of sICAM-1, proved to be significantly correlated with the density ofH. pylori colonization in gastric mucosa (r s=0.316,P<0.001). The serum levels of sICAM-1 in patients with chronic gastritis and peptic ulcer were significantly higher than those in healthy controls (P<0.05). Conclusions: These results indicated thatH. pylori infection up-regulates the expression of sICAM-1.  相似文献   

7.
Development and evaluation of immunoassay for zeranol in bovine urine   总被引：1，自引：0，他引：1  
A high affinity polyclonal antibody-based enzyme linked immunosorbent assay （ELISA） was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1：5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml, and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation （CVs） were in the range of 82%-127% and 3.5%-8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol （ranging from 0.2 to 10 μg/ml） were detected by the ELISA and liquid chromatography （LC） method, and good correlations were obtained between the two methods （R^2=0.9643）. We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an altemative for the conventional LC method for zeranol in bovine urine.  相似文献   

8.
This study presents a rapid, specific and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) assay for determination of risperidone (RIS) in human serum using paroxetine as an internal standard (IS). An Alltima-C 18separation. The analysis was performed by selected reaction monitoring (SRM) method, and the peak area of the m/z 411.3→ 191.1 transition for RIS was measured versus that of the m/z 330.1→192.1 transition for IS to generate the standard curves. The assay linearity of RIS was confirmed over the range 0.25～50.00 ng/ml and the limit of quantitation was 0.05 ng/ml. The linear range corresponds well with the serum concentrations of the analytes obtained in clinical pharmacokinetic studies. Intraday and interday relative standard deviations were 1.85%～9.09% and 1.56%～4.38%, respectively. The recovery of RIS from serum was in the range of 70.20%～84.50%. The method was successfully applied to investigate the bioequivalence between two kinds of tablets (test versus reference products) in 18 healthy male Chinese volunteers. The result suggests that two formulations are bioequivalent.  相似文献   

9.
The enantioselective assay for S( )- and R(-)-propafenone (PPF) in human urine that developed in this work involves extraction of propafenone from human urine and using S( )-propafenone as internal standard, chiral derivatization with 2,3,4,6-tetra-O-β-D-glucopranosyl isothiocyanate, and quantitation by an RP-HPLC system with UV detection (λ=220nm). A baseline separation ofpropafenone enantiomers was achieved on a 5-μm reverse phase ODS column, with a mixture of methanol:water:glacial acetic acid (25:12:0.02,v/v) as mobile phase. There was good linear relationship from 24.9 ng/ml to 1875.0 ng/ml for both of enantiomers. The regression equations of the standard curves based on Cs-PPF (or CR-PPF ) versus ratio of As-PPF/As (or AR-PPF/As ) were y=0.0032x-0.081, (r=0.999) for S-PPF and y=0.0033x 0.0039, (r=0.998) for R-PPF,respectively. The method's limit of detection was 12.5 ng/ml for both enantiomers, and the method's limit ofquantitation was 28.2±0.52 ng/ml for S-PPF, 30.4±0.53 ng/ml for R-PPF (RSD＜8%, n=5). The analytical method yielded average recovery of 98.9% and 100.4% for S-PPF and R-PPF, respectively. The relative standard deviation was no more than 6.11% and 6.22% for S-PPF and R-PPF, respectively. The method enabled study of metabolism of S( )- and R(-)-propafenone in human urine. The results from 7 volunteers administered 150 mg racemic propafenone indicated that propafenone enantiomers undergo stereoselective metabolism and that in the human body, S( )-propafenone is metabolized more extensively than R(-)propafenone.  相似文献   

10.
超薄液晶显示器(LCD)模块在经历了热压键合后,由于驱动芯片和玻璃基板之间的温度差异较大,会引起一定程度上的翘曲,并最终导致连接点界面分层现象的发生,影响器件连接可靠性。用实验的方法对比进行研究比较困难,我们用有限元数值模拟的方法并选择通用软件ANSYS对这一现象进行了分析;并详细分析了键合压力、温度、基板温度和IC/玻璃厚度对翘曲的影响。从分析中得知,导致玻璃翘曲的主要原因是键合温度。  相似文献   

11.
Objective To investigate the relationship between serum resistin level and acute coronary syndrome (ACS) or stable angina pectoris (SAP). Methods Sixty-five patients, with coronary artery disease, were enrolled and divided into three subgroupsacute myocardial infarction (AMI), unstable angina pectoris (UAP) and SAP, and 26 healthy people were recruited as controls in the cross-sectional study. Serum resistin levels were determined by ELISA (enzyme-linked immunosorbent assay), and WBC (white blood cell count), hsCRP (high sensitive C-reaction protein), CKmax (maximum ofcreatinkinase), CK-MBmax (maximum of isozyme of creatinkinase) and cTnImax (maximum oftroponin) were measured by standard laboratory methods. Results The serum resistin levels were 4 folds higher in AMI patients, 2.43 folds in UAP patients and 1.12 folds in SAP patients than in the healthy controls (P＜0.05). The resistin levels were also significantly different between AMI [(8.16±0.79) ng/ml], UAP [(5.59±0.75) ng/ml]and SAP [(3.45±0.56) ng/ml] groups (P＜0.01); WBC, hsCRP, CKmax, CK-MBmax and cTnImax were significantly increased in AMI patients over UAP and SAP patients. Spearman analysis showed that serum resistin levels were positively correlated with WBC (r=0.412, P=0.046), hsCRP (r=0.427, P=0.037), CKmax, CK-MBmax and cTnImax (r=0.731, 0.678, 0.656; P＜0.01). ConclusionSerum resistin levels increased with inflammatory factors and myocardial impairment. The results suggest that human resistin might play an important role in the pathogenesis of atherosclerosis and AMI as an inflammatory factor.  相似文献   

12.
Polychlorinated dibenzo-p-dioxins and furalls(PCDD/Fs)emissions in flue gas from two types of municipal solid waste incinerators(MSWIs)most commonly used in China were investigated in this study.The selected incinerators include two grate-type MSWIs:MSWl-A(350 t/d)and MSWI-B(150 t/d),and two fluidized bed MSWIs:MSWI-C(400 t/d) and MSWI-D (400 t/d),which are all equipped with semi-dry lime scrubber and bag filter except MSWI-D equipped with cyclone and wet scrubber(WS)as air pollutant conffol device(APCD).Results indicated that the emission concentration and the international toxic equivalents(I-TEQs)of the PCDD/Fs from the stacks were in the range of 1.210～10.273 ng/Nm3 and 0.019～0.201 ngI-TEQ/Nm3,respectively.They were greatly lower than the emission regulation standard of PCDD/FS in China(1.0 ng I-TEQ/Nm3).However,only the PCDD/Fs emission Ievel from MSWI-C Was below 0.1 ng I-TEQ/Nm3.Although the homologue profiles were distinct,the contributions of the 2,3,7,8-subsituted congenem to the total I-TEQ were similar among all the investigated MSWIs.Two major 2,3,7,8-substituted congeners,2,3,4,7,8-PeCDF and 1,2,3,7,8-PeCDD,account for 47% and 9%(average values)of the total I-TEQ values,respectively.The correlation between PCDD/Fs levels and composition of flue gas Was also discussed.  相似文献   

13.
目的：观察急性肾功能衰竭（ARF）家兔心肌匀浆自由基、一氧化氮的变化，探讨心肌损伤的体液机制。方法：60只家兔均分为四组（n=15）。ARF模型1组：皮下注射1％HgCl2（1．3ml／kg．bw）；ARF模型2组：肌肉注射50％甘油（10ml／kg．bw）；以等量生理盐水代替HgCl2和甘油作为对照1、2组。24h后，所有动物颈总动脉放血备检，并选择固定位置。制备10％心肌匀浆。经Aeroset型全自动生化分析仪测定血清反映肾功能的生化指标。检测心肌匀浆丙二醛（MDA）、超氧化物歧化酶（SOD）、一氧化氮（NO）及其合酶（NOS）的变化。结果：与相应对照组比较，ARF模型1、2组心匀浆SOD活性下降、MDA含量升高（P〈0．05），ARF模型1、2组心匀浆NO含量增强、NOS及iNOS活性增强（P〈0．05～0．01）。结论：ARF家兔心肌损伤的机制与自由基损伤及NO升高有关。  相似文献   

14.
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15.
本文报道丹贝中生物活性物质大豆甙元同辣根过氧化物酶偶联及其酶活性、免疫活性和光谱分析鉴定。大豆甙元—辣根过氧化物酶标记物中两者摩尔比11：1，标记物工作浓度50ng／ml，3．5ng／ml大豆甙元能与氧化酶标记物发生50％左右竞争。  相似文献   

16.
Objective:To investigate the relationship between the fatty acid-binding protein 4(FABP4)and colorectal cancer(CRC).Methods:Using an enzyme-linked immunosorbent assay(ELISA),we measured the expression of FABP4 in plasma of50 patients who underwent surgery for CRC from October 2017 to May 2018 and 50 healthy controls.The content of the visceral fat area(VFA)as seen with abdominal computed tomography(CT)scanning was measured by ImageJ software.The expression levels of FABP4,E-cadherin,and Snail proteins in CRC and adjacent tissues were determined by immunohistochemistry.Results:The mean concentration of plasma FABP4 of CRC patients was higher than that of the control group(22.46 vs.9.82 ng/m L;P<0.05).The concentration of plasma FABP4 was related to the tumor,node,metastatis(TNM)stage and lymph node metastasis and was independent of age,body mass index(BMI),tumor size and location,and the degree of differentiation of CRC.The concentration of plasma FABP4 was positively correlated with high VFA and lipoprotein-a(LPA)(P<0.05);but it was not correlated with total cholesterol(TG),total triglyceride(TC),low-density lipoprotein(LDL),high-density lipoprotein(HDL),or apolipoprotein AI(Apo-AI).The expression of FABP4 protein in CRC tissues was positively correlated with the degree of CRC differentiation,tumor stage,and lymph node metastasis.The level of FABP4 protein was negatively correlated with E-cadherin protein(r=-0.3292,P=0.0196)and positively correlated with Snail protein(r=0.5856,P<0.0001).Conclusions:High LPA and VFA were risk factors for increased plasma FABP4 in CRC patients.FABP4 protein was highly expressed in CRC tissues and associated with TNM stage,differentiation,and lymph node metastasis of CRC.The level of FABP4 in CRC tissue was correlated with E-cadherin and Snail expression,suggesting that FABP4 may promote CRC progression related to epithelial-mesenchymal transition(EMT).  相似文献   

17.
Ethanol extracts of brown seaweeds from Pakistan and China were isolated and compared for their antiallergenic activities.They included Sargassum tennerimum (ST) and Sargassum cervicorne (SC) from Pakistan,and Sargassum graminifolium turn (SG),Sargassum thunbergii (STH),and Laminariajaponica (LJ) from China.The ethanol extracts of these brown seaweeds were optimized at 85% (v/v) ethanol for the maximum yield of phlorotannin,an inhibitor against hyaluronidase.Total phlorotannins contained in the crude extracts were measured as 1.71% (SG),0.74% (STH),0.97% (LJ),3.30% (SC),and 5.06% (ST).The 50% inhibitory concentrations (IC50) of Pakistani SC and ST were 109.5 and 21 μg/ml,respectively,lower than those of Chinese SG,STH,and LJ (134,269,and 148 μg/ml,respectively).An antiallergic drug,disodium cromoglycate (DSCG),had an IC50=39 μg/ml,and a natural inhibitor of hyaluronidase,catechin,had an IC50=20 μg/ml.The IC50 of ST extract was found similar to that of catechin (21 vs 20 μg/ml) and lower than that of DSCG (21 vs 39 μg/ml).This suggests that ST is a potent inhibitor of hyaluronidase,indicating a promising future development of natural antiallergic medicines or functional foods.  相似文献   

18.
Although several antiviral drugs and vaccines are available for use against hepatitis B virus (HBV), hepatitis caused by HBV remains a major public health problem worldwide, which has not yet been resolved, and new anti-HBV drugs are in great demand. The present study was performed to investigate the anti-HBV activity of epigallocatechin-3-gallate (EGCG), a natural-origin compound, in HepG2 2.2.15 cells. The antiviral activity of EGCG was examined by detecting the levels of HBsAg and HBeAg in the supernatant and extracellular HBV DNA. EGCG effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose- and time-dependent manner, and it showed stronger effects at the level of 0.11–0.44 μmol/ml (50–200 μg/ml) than lamivudine (3TC) at 0.87 μmol/ml (200 μg/ml). EGCG also suppressed the amount of extracellular HBV DNA. The data indicated that EGCG possessed anti-HBV activity and suggested the potential of EGCG as an effective anti-HBV agent with low toxicity.  相似文献   

19.
Polychlorinated dibenzo-p-dioxins and furans （PCDD/Fs） emissions in flue gas from two types of municipal solid waste incinerators （MSWIs） most commonly used in China were investigated in this study. The selected incinerators include two grate-type MSWIs： MSWI-A （350 t/d） and MSWI-B 050 t/d）, and two fluidized bed MSWIs： MSWI-C （400 t/d） and MSWI-D （400 t/d）, which are all equipped with semi-dry lime scrubber and bag filter except MSWI-D equipped with cyclone and wet scrubber （WS） as air pollutant control device （APCD）. Results indicated that the emission concentration and the international toxic equivalents （I-TEQs） of the PCDD/Fs from the stacks were in the range of 1.210-10.273 ng/Nm^3 and 0.019-0.201 ng I-TEQ/Nm^3, respectively. They were greatly lower than the emission regulation standard of PCDD/Fs in China （1.0 ng I-TEQ/Nm^3）. However, only the PCDD/Fs emission level from MSWI-C was below 0.1 ng I-TEQ/Nm^3. Although the homologue profiles were distinct, the contributions of the 2,3,7,8-subsituted congeners to the total I-TEQ were similar among all the investigated MSWIs. Two major 2,3,7,8-substituted congeners, 2,3,4,7,8-PeCDF and 1,2,3,7,8-PeCDD, account for 47% and 9% （average values） of the total I-TEQ values, respectively. The correlation between PCDD/Fs levels and composition of flue gas was also discussed.  相似文献   

20.
To detect gatifloxacin (GAT) residue in swine urine, an electrochemical immunoassay was established. An indirect competitive immunoassay was developed, in which the coating antigen is immobilized in an enzyme-linked immunosorbent assay (ELISA) plate and GAT residue from the sample competes with the limited binding sites in added anti-GAT antibody. Horseradish peroxidase (HRP) conjugated to goat anti-rabbit IgG was used as the enzymatic label. A carbon fiber working electrode was constructed and current signals were detected by using hydrogen peroxide as a substrate and hydroquinone as an electrochemical mediator. The electrochemical immunoassay was evaluated by analysis of GAT in buffer or swine urine and an average value of half inhibition concentration (IC50) of 8.9 ng/ml was obtained. Excellent specificity of the antibody was achieved with little cross-reaction with lomefloxacin (3.0%), ciprofloxacin (3.0%), and ofloxacin (1.9%) among commonly used (fluoro)quinolones. In conclusion, the immunoassay system developed in this research can be used as a rapid, powerful and on-site analytical tool to detect GAT residue in foods and food products.  相似文献   

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