Preparation of Cathepsin L1 Monoclonal Antibody Against Fasclala gigantica and Establishment of Double Antibody Sandwich ELISA

【Objective】 This study was intend to obtain cathepsin L1(rFgCat L1) specific monoclonal antibody and construct the double antibody sandwich ELISA.【Method】 Five BALB/c mice were immunized with 1 mg/mL rFgCat L1 protein for four times.Mouse splenocytes were isolated and fused with SP2/0 cells to construct hybridoma cells.Strong positive hybridoma cell lines were screened, 1×10⁶ cells were injected intraperitoneally per mouse to prepare monoclonal antibodies.Antibody titer and antigenic epitope were detected using ELISA method, antibody subtype and specificity were identified using Western blotting method.The double antibody sandwich ELISA was constructed by combining the anti-rFgCat L1 polyclonal antibody, and its sensitivity and specificity were tested.The positive and negative critical value was screened by 20 negative sera with positive control, and the constructed double antibody sandwich ELISA was verified by 47 goat positive sera and 47 dairy cow positive sera.【Result】 After immunization, the antibody titers in serum of 4 mice were all more than 10⁴.After isolated mouse with the highest immune response spleen cells were fused with SP2/0 cells total of 8 of them were positive cell lines were obtained after selective culture.5D5 and 7G6 were identified as strong positive strains with stable antibody secretion.After multiple subcloning screens and subcultures, the antibodies secreted in the cell supernatant were stable, with titers of 2⁹ and 2¹⁰ respectively, with ascites titers of 10⁷ and 10⁸.Western blotting and antibody subtype identification kits identified that the two antibodies were IgG1 type and the light chain was kappa type, both of which could specifically bind FgESP.According to the same antigen site was recognized by the two kinds of antibodies, the antigen titer of the two monoclonal antibodies were comparied, 7G6 was used as the coating antibody, and anti-rFgCat L1 was used as the enzyme-labeled secondary antibody.The optimized condition of method was that 7G6 was coated at a concentration of 2 μg/mL, the dilution concentration of anti-rFgCat L1 polyclonal antibody was 25 μg/mL, the dilution of Don-HRP-conjugated was 1∶4 000, 5% skimmed milk powder was selected as the blocking solution and the color development time was 25 min.The method was proved that could recognize the lowest antigen concentration of 0.625 μg/mL, also could specifically recognize antigen of Fasciola fasciatus.The constructed sandwich ELISA method was used for antigen detection of 47 dairy cow positive serum and 47 goat positive serum infective samples kept in the laboratory and the positive antigen rate were 72.3% and 78.7%, respectively.【Conclusion】 Anti-rFgCat L1 monoclonal antibody was successfully prepared and the double-sheet sandwich ELISA method for fascioliasis was constructed, which provided a good theoretical basis and material basis for the development of low-cost and rapid diagnostic kits.     

装饰单板贴面胶合板与杉木复合制造地板技术

为了降低实木复合地板生产的材料消耗和成本,直接采用装饰单板贴面胶合板与杉木组合制造实木复合地板.结果表明,按改进工艺制造的复合地板不仅具有国标所要求的物理力学性能,而且可降低材料成本30%左右.     

建宁县杂交水稻制种的几种栽培模式

福建省建宁县是国家级杂交水稻种子生产基地,该文总结了其单季制种模式、早制—晚制连作模式和春制—稻栽培模式及关键技术。     

松香酯羟丙基季铵盐表面活性剂的合成、表征及性能

以脱氢枞酸、丙烯海松酸、马来海松酸为原料分别与2,3-环氧丙基三甲基氯化铵反应,得到单、双、三季铵盐Ⅰ、Ⅱ及Ⅲ3种松香酯羟丙基季铵盐表面活性剂。采用IR,1H NMR,13C NMR对产物结构进行了表征,并对产物的临界胶束浓度(Ccmc)、表面张力(γcmc)、乳化性能、泡沫性能及抑菌性能进行了研究。结果表明,3种松香酯羟丙基季铵盐表面活性剂的Ccmc值分别为3.58×10-4、2.53×10-4和3.25×10-4mol/L,对应的γcmc分别为38.9、28.9、33.5 m N/m;乳化时间分别为31、33、38 min;表面活性剂Ⅰ的起泡性较好,起泡高度达到115 mm、5 min后泡沫高度为85 mm;随着季铵盐数量的增加,亲水亲油平衡值(HLB)增大,表面活性剂的亲水性增强;季铵盐Ⅰ对金黄色葡萄球菌具有良好的抑制活性,最小抑菌浓度为4 mg/L,抑菌效果优于市售的新洁尔灭,与氨苄青霉素钠相当。     

沙发椅背倾斜角度对坐姿舒适度影响的研究

采用肌电测试方法以及主观评价与客观测试相结合的方式,分析了沙发椅背倾角与坐姿舒适度之间的关系,为从人体工程学的角度优化沙发椅背设计以及对沙发舒适性的评价提供依据.实验研究表明,当沙发坐垫硬度为89N,沙发椅背硬度为32N,沙发椅背高度为600mm,椅背倾角分别为100°、110°、120°时,使用倾角为110°的椅背总体舒适感最好,其次为使用倾角120°的椅背,使用倾角为100°的椅背总体舒适感最差     

新干县生态环境问题及林业可持续发展对策

对江西省新干县生态环境的基本情况进行了分析,在自然地理分异形成的自然区域的基础上,分析了区域性生态基础、区域性经济技术环境、区域性综合社会特征以及林业比较效益的弱质性与林业可持续发展的关系,并对林业可持续发展的道路进行了一定的探索。     

Cx43 is involved in electrical remodeling of atrial myocytes through regulating L-type calcium current

AIM: To investigate whether the association of connexin 43(Cx43) and L-type calcium channel involved in the pathogenesis of atrial fibrillation (AF). METHODS: The biochemical assays and whole-cell patch-clamp technique were used to study the expression of Cx43 in human atrial tissue. The co-localization of Cx43 and L -type calcium channel, and the regulation of L-type calcium current in atrial myocytes were investigated. RESULTS: The expression of Cx43 at mRNA and protein levels was decreased in human atrial tissues of AF patients. In cultured atrium-derived myocytes (HL-1 cells), knockdown of Cx43 significantly inhibited the mRNA expression of L-type calcium channel α1c subunit, as well as L-type calcium current. Co-localization of Cx43 with L-type calcium channel α1c subunit in mouse atrial myocytes was observed. CONCLUSION: The decrease in Cx43 is involved in the pathogenesis of AF, probably through reducing the L-type calcium current in atrial myoctyes by co-localization with L-type calcium channel, thus representing the potential pathogenesis in atrial fibrillation.     

甲基红褪色光度法测定数量溴酸根离子

研究了盐酸介质中 ,在氯化纳的催化作用下 ,溴酸根离子氧化甲基红褪色的最佳条件 .吸收波长为 5 18nm .表观摩尔吸光系数为 7.2×10 4 L· m ol- 1 · cm- 1 ,溴酸根离子在 3.0～ 18μg/2 5 m L范围内符合比尔定律 .方法用于化学试剂中微量溴酸根的测定 ,结果令人满意 .     

TLC法测定虎杖中白藜芦醇的含量

本文建立了虎杖白藜芦醇含量测定的薄层扫描分析方法：以硅胶G为薄层吸附荆，氯仿：丙酮：乙酸：水(4：4：0．5：0．2)为展开剂，在紫外灯(365nm)下观察荧光斑点，在365n／n处并进行扫描测定，线性方程为：Y=(13．826S’10^-6)-0．8698(Y为白藜芦醇浓度ug／ml，S为峰面积)。并对虎杖茎、根、叶进行了含量测定。结果表明：用TIC法测定虎杖中白藜芦醇含量方法简便、快捷、准确度高、重复性好。虎杖鲜根茎中含量达0．548％。