The aim of this work was to develop a liquid chromatography-tandem mass spectrometry method for the determination of milk allergen and egg allergen in food products.Signature peptides GGLEPINFQTAADQAR,VGINYWLAHK,VLVLD...The aim of this work was to develop a liquid chromatography-tandem mass spectrometry method for the determination of milk allergen and egg allergen in food products.Signature peptides GGLEPINFQTAADQAR,VGINYWLAHK,VLVLDTDYK,FFVAPFPEVFGK,and NAVPITPTLNR were confirmed and synthesized as the quantitative peptide of ovalbumin,α-lactalbumin,β-lactoglobulin,α_(S1)-casein andα_(S2)-casein,the relative isotope-labeled internal standards were used in the quantitative analysis.Linear range was in the range of0.5-5000.0 nmol/L for egg and milk allergen in bread,cake,cookie,rice crust and wheat flour samples with free from egg and milk,the limits of detection of milk allergens and egg allergen were in the range between0.94 mg/100 g and 56.71 mg/100 g,limits of quantification of milk allergens and egg allergen were in the range between 2.36 mg/100 g and 141.78 mg/100 g.The recoveries ranged from 76.7%to 122.8%,the relative standard deviations were in the range of 1.60%-15.60%.The developed method has been successfully used for the detection of egg and milk allergen in various food samples.展开更多
Vaccine is a principal and highly cost-effective medical method on controlling infectious diseases and improving population health.Various vaccines are eagerly needed in the low-income countries along the Belt&Roa...Vaccine is a principal and highly cost-effective medical method on controlling infectious diseases and improving population health.Various vaccines are eagerly needed in the low-income countries along the Belt&Road.With the good quality to reach WHO prequalification standard,and the abundant capacity to fulfill the demand from market abroad,upon the platform of China-proposed"the Belt&Road Initiative",the vaccines manufactured in China are exporting to countries worldwide.The independent innovative vaccines'R&D system,which fruited a series of innovative infectious diseases vaccines(EV71 vaccine,sIPV,HEV,Ebola vaccine,etc.)to be launched in Chinese market,indicates that China has developed rapidly from"a great vaccine-production country"to"a powerful vaccine-innovation country".The implementation of National Innovation-driven Development Strategy would further push forward the developm ent and internationalization process of Chinese innovative vaccines.Therefore,the China-invented vaccines will make an important role in the prevention and control of infectious disease in various countries and become one of the most powerful weapons in fighting the global epidemic event in the future.展开更多
目的探讨粉尘螨变应原Derf1mRNA疫苗对哮喘小鼠的特异性免疫治疗效果。方法 50只BALB/c小鼠随机分为5组(10只/组),分别为PBS组、Derf1变应原致敏组、Derf1变应原免疫治疗组、β-actin mRNA免疫治疗组和Derf1 mRNA免疫治疗组。分别于第0...目的探讨粉尘螨变应原Derf1mRNA疫苗对哮喘小鼠的特异性免疫治疗效果。方法 50只BALB/c小鼠随机分为5组(10只/组),分别为PBS组、Derf1变应原致敏组、Derf1变应原免疫治疗组、β-actin mRNA免疫治疗组和Derf1 mRNA免疫治疗组。分别于第0、第7和第14天,PBS组小鼠腹腔注射PBS,其余4组小鼠则腹腔注射10μg Der f 1进行致敏,建立小鼠哮喘模型。自第21天起,除PBS组小鼠雾化吸入PBS,其他4组小鼠均雾化吸入100μg/ml Der f 1变应原,30 min/次,1次/d,连续7 d,观察并记录小鼠哮喘发作情况。5组小鼠于最后1次雾化吸入致敏2周后,背部皮下分别注射100μl PBS、1μg Der f1(维持致敏)、10μg Derf1(免疫治疗)、2μgβ-actin mRNA和2μg Derf1mRNA,1次/周,连续3周。最后1次皮下注射后2周处死小鼠。收集各组小鼠的支气管肺泡灌洗液(BALF),ELISA法检测γ干扰素(IFN-γ)和白介素13(IL-13)水平,并计数嗜酸粒细胞(EOS);收集各组小鼠脾组织,分离脾细胞,除PBS组外,其他4组均加入10μg/ml Der f 1培养72 h,ELISA法检测脾细胞培养上清液中IFN-γ和IL-13水平;取各组小鼠眼球血,ELISA法检测血清中总IgE以及变应原特异性IgE(sIgE)、IgG1(sIgG1)和IgG2a(sIgG2a)抗体水平。HE染色观察各组小鼠肺组织切片。结果除PBS组外,其他4组小鼠雾化吸入致敏后,均出现急性哮喘发作症状。小鼠BALF中,Der f 1 mRNA免疫治疗组和Der f 1免疫治疗组的IFN-γ水平分别为(897.56±105.73)和(864.48±70.62)pg/ml,均明显高于Derf1变应原致敏组[(209.05±52.28)pg/ml]和β-actin mRNA免疫治疗组[(219.47±64.72)pg/ml](P<0.01);两者IL-13水平分别为(241.64±31.41)和(321.94±41.07)pg/ml,则明显低于Derf1变应原致敏组[(520.62±43.77)pg/ml]和β-actin mRNA免疫治疗组[(507.22±42.26)pg/ml](P<0.01);两者EOS数量分别为(1.33±0.44)×105和(1.48±0.39)×105个/ml,均明显低于Derf1变应原致敏组[(3.54±0.52)×105个/ml]和β-actin mRNA免疫治疗组[(2.98±0.53)×105个/ml](P<0.01)。脾细胞培养上清的ELISA检测结果显示,Der f 1 mRNA免疫治疗组和Der f 1免疫治疗组的IFN-γ水平分别为(420.91±69.92)和(334.92±43.72)pg/ml,明显高于Der f 1变应原致敏组[(123.75±15.48)pg/ml]和β-actin mRNA免疫治疗组[(128.84±59.00)pg/ml](P<0.01);两者IL-13水平[(268.51±40.42)和[(285.26±62.21)pg/ml]则显著低于Der f 1变应原致敏组[(613.89±51.54)pg/ml]和β-actin mRNA免疫治疗组[(524.05±39.12)pg/ml](P<0.01)。血清中抗体水平的ELISA检测结果显示,Der f 1 mRNA免疫治疗组的总IgE、sIgE和sIgG1抗体水平分别为(33.72±9.78)、(22.76±8.09)和(17.87±7.59)ng/ml,均显著低于Der f 1变应原致敏组[(94.34±11.66)、(65.67±9.47)和(75.18±9.52)ng/ml]和β-actin mRNA免疫治疗组[(86.48±10.26)、(62.36±8.35)和(69.51±8.98)ng/ml](P<0.01);其sIgG2a抗体水平为(7.74±0.88)ng/ml,则明显高于Der f 1变应原致敏组[(2.81±1.17)ng/ml]和β-actin mRNA免疫治疗组[(1.06±0.11)ng/ml](P<0.01)。肺组织切片HE染色镜检结果显示,与Der f 1变应原致敏组相比,Der f 1 mRNA免疫治疗组小鼠的气道上皮和肺泡上皮细胞结构基本完整,炎症细胞浸润明显减少。结论 Derf1mRNA疫苗可有效纠正Th1/Th2失衡。展开更多
基金supported by National Key Research and Development Program of China(2019YFC1606400)Science and Technology Project of State Administration for Market Regulation(2021MK023)+1 种基金Hebei Province High-level Talent Funding Program(A201901008)Research Project of Hebei Administration for Market Regulation(2020ZD12)。
文摘The aim of this work was to develop a liquid chromatography-tandem mass spectrometry method for the determination of milk allergen and egg allergen in food products.Signature peptides GGLEPINFQTAADQAR,VGINYWLAHK,VLVLDTDYK,FFVAPFPEVFGK,and NAVPITPTLNR were confirmed and synthesized as the quantitative peptide of ovalbumin,α-lactalbumin,β-lactoglobulin,α_(S1)-casein andα_(S2)-casein,the relative isotope-labeled internal standards were used in the quantitative analysis.Linear range was in the range of0.5-5000.0 nmol/L for egg and milk allergen in bread,cake,cookie,rice crust and wheat flour samples with free from egg and milk,the limits of detection of milk allergens and egg allergen were in the range between0.94 mg/100 g and 56.71 mg/100 g,limits of quantification of milk allergens and egg allergen were in the range between 2.36 mg/100 g and 141.78 mg/100 g.The recoveries ranged from 76.7%to 122.8%,the relative standard deviations were in the range of 1.60%-15.60%.The developed method has been successfully used for the detection of egg and milk allergen in various food samples.
文摘Vaccine is a principal and highly cost-effective medical method on controlling infectious diseases and improving population health.Various vaccines are eagerly needed in the low-income countries along the Belt&Road.With the good quality to reach WHO prequalification standard,and the abundant capacity to fulfill the demand from market abroad,upon the platform of China-proposed"the Belt&Road Initiative",the vaccines manufactured in China are exporting to countries worldwide.The independent innovative vaccines'R&D system,which fruited a series of innovative infectious diseases vaccines(EV71 vaccine,sIPV,HEV,Ebola vaccine,etc.)to be launched in Chinese market,indicates that China has developed rapidly from"a great vaccine-production country"to"a powerful vaccine-innovation country".The implementation of National Innovation-driven Development Strategy would further push forward the developm ent and internationalization process of Chinese innovative vaccines.Therefore,the China-invented vaccines will make an important role in the prevention and control of infectious disease in various countries and become one of the most powerful weapons in fighting the global epidemic event in the future.
文摘目的探讨粉尘螨变应原Derf1mRNA疫苗对哮喘小鼠的特异性免疫治疗效果。方法 50只BALB/c小鼠随机分为5组(10只/组),分别为PBS组、Derf1变应原致敏组、Derf1变应原免疫治疗组、β-actin mRNA免疫治疗组和Derf1 mRNA免疫治疗组。分别于第0、第7和第14天,PBS组小鼠腹腔注射PBS,其余4组小鼠则腹腔注射10μg Der f 1进行致敏,建立小鼠哮喘模型。自第21天起,除PBS组小鼠雾化吸入PBS,其他4组小鼠均雾化吸入100μg/ml Der f 1变应原,30 min/次,1次/d,连续7 d,观察并记录小鼠哮喘发作情况。5组小鼠于最后1次雾化吸入致敏2周后,背部皮下分别注射100μl PBS、1μg Der f1(维持致敏)、10μg Derf1(免疫治疗)、2μgβ-actin mRNA和2μg Derf1mRNA,1次/周,连续3周。最后1次皮下注射后2周处死小鼠。收集各组小鼠的支气管肺泡灌洗液(BALF),ELISA法检测γ干扰素(IFN-γ)和白介素13(IL-13)水平,并计数嗜酸粒细胞(EOS);收集各组小鼠脾组织,分离脾细胞,除PBS组外,其他4组均加入10μg/ml Der f 1培养72 h,ELISA法检测脾细胞培养上清液中IFN-γ和IL-13水平;取各组小鼠眼球血,ELISA法检测血清中总IgE以及变应原特异性IgE(sIgE)、IgG1(sIgG1)和IgG2a(sIgG2a)抗体水平。HE染色观察各组小鼠肺组织切片。结果除PBS组外,其他4组小鼠雾化吸入致敏后,均出现急性哮喘发作症状。小鼠BALF中,Der f 1 mRNA免疫治疗组和Der f 1免疫治疗组的IFN-γ水平分别为(897.56±105.73)和(864.48±70.62)pg/ml,均明显高于Derf1变应原致敏组[(209.05±52.28)pg/ml]和β-actin mRNA免疫治疗组[(219.47±64.72)pg/ml](P<0.01);两者IL-13水平分别为(241.64±31.41)和(321.94±41.07)pg/ml,则明显低于Derf1变应原致敏组[(520.62±43.77)pg/ml]和β-actin mRNA免疫治疗组[(507.22±42.26)pg/ml](P<0.01);两者EOS数量分别为(1.33±0.44)×105和(1.48±0.39)×105个/ml,均明显低于Derf1变应原致敏组[(3.54±0.52)×105个/ml]和β-actin mRNA免疫治疗组[(2.98±0.53)×105个/ml](P<0.01)。脾细胞培养上清的ELISA检测结果显示,Der f 1 mRNA免疫治疗组和Der f 1免疫治疗组的IFN-γ水平分别为(420.91±69.92)和(334.92±43.72)pg/ml,明显高于Der f 1变应原致敏组[(123.75±15.48)pg/ml]和β-actin mRNA免疫治疗组[(128.84±59.00)pg/ml](P<0.01);两者IL-13水平[(268.51±40.42)和[(285.26±62.21)pg/ml]则显著低于Der f 1变应原致敏组[(613.89±51.54)pg/ml]和β-actin mRNA免疫治疗组[(524.05±39.12)pg/ml](P<0.01)。血清中抗体水平的ELISA检测结果显示,Der f 1 mRNA免疫治疗组的总IgE、sIgE和sIgG1抗体水平分别为(33.72±9.78)、(22.76±8.09)和(17.87±7.59)ng/ml,均显著低于Der f 1变应原致敏组[(94.34±11.66)、(65.67±9.47)和(75.18±9.52)ng/ml]和β-actin mRNA免疫治疗组[(86.48±10.26)、(62.36±8.35)和(69.51±8.98)ng/ml](P<0.01);其sIgG2a抗体水平为(7.74±0.88)ng/ml,则明显高于Der f 1变应原致敏组[(2.81±1.17)ng/ml]和β-actin mRNA免疫治疗组[(1.06±0.11)ng/ml](P<0.01)。肺组织切片HE染色镜检结果显示,与Der f 1变应原致敏组相比,Der f 1 mRNA免疫治疗组小鼠的气道上皮和肺泡上皮细胞结构基本完整,炎症细胞浸润明显减少。结论 Derf1mRNA疫苗可有效纠正Th1/Th2失衡。
