Objective:To characterize the group-29 allergens from Dermatophagoides(D.)pteronyssinus and investigate their ability to cross-react with other group-29 allergens from D.pteronyssinus as well as those from D.farinae a...Objective:To characterize the group-29 allergens from Dermatophagoides(D.)pteronyssinus and investigate their ability to cross-react with other group-29 allergens from D.pteronyssinus as well as those from D.farinae and Tyrophagus putrescentiae.Methods:Der p 29,Der f 29,and Tyr p 29 cDNA sequences were amplified from total RNA isolated from D.pteronyssinus,D.farinae and Tyrophagus putrescentiae,respectively.Then they were cloned into the pET28a vector,expressed in Rosetta2(DE3)plysS,and purified using anion exchange chromatography.The IgE-binding rates of rDer p 29 were assessed by IgE Western blotting.The four epitopes of rDer p 29 were predicted,synthesized,and detected by IgE-ELISA.The cross-reactivity among the recombinant proteins rDer p 29,rDer f 29,and rTyr p 29 was investigated using dot blot and IgE-ELISA inhibition experiments.The allergens’physiochemical properties,amino acid sequences,and tertiary structures were also compared.Results:Der p 29 was successfully expressed in Rosetta2(DE3)plysS as a single,393-bp open reading frame.Western blotting showed that the purified rDer p 29 protein exhibited an IgE-binding rate of 100%when tested on patient sera.The following four Der p 29 epitopes were predicted and synthesized:37-45(EP1),57-69(EP2),75-80(EP3),and 104-117(EP4).IgE-ELISA tests on 20 D.pteronyssinus-positive sera yielded IgE-binding rates of 85%(rDer p 29),80%(EP1),55%(EP2),40%(EP3),and 55%(EP4),respectively.The dot blot experiments further confirmed cross-reactivity among the three group-29 proteins.When used as an inhibitor,rDer p 29 demonstrated an average cross-reactive inhibition rate of 49.7%against rDer f 29 and 54.4%against rTyr p 29.When rTyr p 29 was used as an inhibitor,it showed an average cross-reactive inhibition rate of 56.3%against rDer f 29.Conclusions:A recombinant protein,rDer p 29 with strong allergenicity was produced.Moreover,it was found that rDer p 29 cross-reacted with rDer f 29 and rTyr p 29,due to their highly homologous sequences and structures.These findings highlight the importance of considering inter-species epitope cross-reactivity when diagnosing and treating allergic diseases.展开更多
Objective:To examined the immediate and 24 hours post-irradiation germicidal effects of UV-C lamp on.eggs and adults of house dust mites Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermatophagoides farinae(D.fa...Objective:To examined the immediate and 24 hours post-irradiation germicidal effects of UV-C lamp on.eggs and adults of house dust mites Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermatophagoides farinae(D.farinae).Methods:This study investigated the immediate and24 hours post irradiation mortalities of adult mites exposed to UV-C at different exposure times(5mins,10 mins,15 mins,20 mins,30 mins and 60 mins)and distances(10 cm,25 cm,33 cm,45 cm and 55 cm).Fresh eggs of the 2 dust mites were also irradiated at 10,35 and 55 cm for 0.5,1,2,3,and 5 minutes,and observed daily post-irradiation for up to 7 days.Results:Highest immediate mortality of 100%occurred with direct irradiation al 10 cm distance from UV-C lamp and for 60mins,for both species of mites.The post 24 hours mean mortality rates were(58.4±17.4)%for D.pteronyssinus and(27.7±9.7)%for D.fitrinae when irradiated for 1 hour at 55 cm distance under UV-C lamp.When mites were irradiated in the presence of culture media,the highest mortality rates were lower compared to the direct irradiation;at 10 cm distance and 60 mins exposure,the mean mortality was(74.0±6.8)%for D.pteronyssinus and(70.3±6.7)%for D.farinae.Egg hatchability for both species of mites was also notably reduced by greater than 50%following irradiation.Conclusions:Ultraviolet C irradiation is lethal to an array of organisms by damaging their nucleic acids(DNA and RNA).This study demonstrates the increasing mite mortalities with increasing exposure times and decreasing distances.展开更多
Objective:To investigate the short and long tenn efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermalophagoides farinae(D.farinae)mites.Methods:The effect of a comm...Objective:To investigate the short and long tenn efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermalophagoides farinae(D.farinae)mites.Methods:The effect of a commercial ionizer on D.pteronyssinus and D.farinae was evaluated in the laboratory,using a specially designed test.Mortality was assessed after 6,16and 24 hours for direct exposure and after 24,36,48,60 and 72 hours for exposure in simulated mattress.New batches of mites were used for each exposure time.Results:LT_(50)for direct exposure of ionizer was 10 hours for D.pteronyssinus and 18 hours for D.farinae.The LT_(50)for exposure in simulated mattress was 132 hours or 5.5 days for D.pteronyssinus and 72 hours or 3days for D.farinae.LT_(95)for direct exposure of ionizer was 36 hours for D.pteronyssinus and D.farinae.Meanwhile,the LT_(95)for exposure in simulated mattress was 956 hours or 39.8 days for D.pteronyssinus and 403 hours or 16.8 days for D.farinae.Conclusions:This study demonstrates the increasing mite mortalities with increasing exposure time of a commercial ionizer and suggests that negative ions produced by an ionizer kill dust mites and can be used to reduce natural mile populations on exposed surfaces such as floors,clothes,curtains,etc.However,there is reduced efficacy on mites inside stuffed materials as in mattresses and furniture.展开更多
Objective: To investigate the effects of dermatophagoides farinae drops combined with pidotimod on inflammatory response and immune response in children with allergic rhinitis combined with asthma. Methods: A total of...Objective: To investigate the effects of dermatophagoides farinae drops combined with pidotimod on inflammatory response and immune response in children with allergic rhinitis combined with asthma. Methods: A total of 118 children with allergic rhinitis combined with asthma who were treated in our hospital between December 2014 and May 2017 were selected as the research subjects and divided into the control group (n=59) and the dermatophagoides farinae drops group (n=59) by random number table method. Control group received both conventional therapy and pidotimod therapy, and dermatophagoides farinae drops group received both conventional therapy and dermatophagoides farinae drops combined with pidotimod therapy. The differences in inflammatory factor, cellular immunity index and humoral immunity index levels were compared between the two groups before treatment (T0), after 3 months of treatment (T1), after 6 months of treatment (T2) and after 12 months of treatment (T3). Results: At T0, inflammatory factor, cellular immunity index and humoral immunity index levels were not significantly different between the two groups. At T1, T2 and T3, serum inflammatory factor IL-2 contents of dermatophagoides farinae drops group were higher than those of control group at the corresponding time points whereas IL-4, IL-5, IL-9 and IL-18 contents were lower than those of control group at the corresponding time points;peripheral blood cellular immunity indexes CD3+, CD4+, CD8+ and CD4+/CD8+levels were higher than those of control group at the corresponding time points;serum humoral immunity indexes IgA, IgM and IgG contents were higher than those of control group at the corresponding time points whereas IgE contents were lower than those of control group at the corresponding time points. Conclusion: Dermatophagoides farinae drops combined with pidotimod therapy can effectively reduce the inflammatory response degree and optimize the cellular and humoral immune function of the children with allergic rhinitis combined with asthma.展开更多
Objective:To study the ventilation function after sublingual dermatophagoides farinae drops combined with loratadine treatment of children with asthma and allergic rhinitis and the influence on serum biochemical index...Objective:To study the ventilation function after sublingual dermatophagoides farinae drops combined with loratadine treatment of children with asthma and allergic rhinitis and the influence on serum biochemical indexes.Methods: A total of 40 children with asthma and allergic rhinitis treated in our hospital between September 2013 and March 2015 were collected and divided into the control group (n=22) who accepted loratadine therapy alone and the observation group (n=18) who accepted sublingual dermatophagoides farinae drops combined with loratadine therapy after the treatment was reviewed. Before treatment and after 6 months and 1 year of treatment, spirometer was used to test ventilation function indexes;ELISA method was used to determine the contents of inflammatory mediators;RIA method was used to determine the contents of airway remodeling indicators.Results:Before treatment, differences in ventilation function index levels as well as inflammatory mediator and airway remodeling index contents were not statistically significant between two groups of children. After 6 months and 1 year of treatment, FEV1, FVC and PEF levels of observation group were higher than those of control group;serum IL-2 content was higher than that of control group while IL-5, IL-17 and IL-33 contents were lower than those of control group;serum PDGF-BB, TGF-β1 and NF-κB contents were lower than those of control group.Conclusion:Sublingual dermatophagoides farinae drops combined with loratadine therapy can optimize the ventilation function, reduce the systemic inflammatory response and inhibit airway remodeling in children with asthma and allergic rhinitis.展开更多
Background The dust mites, which are mostly represented by Dermatophagoides spp. （Acari： Pyroglyphidae）, are the major sources of indoor allergens. Identification and characterization of these mite allergen molecul...Background The dust mites, which are mostly represented by Dermatophagoides spp. （Acari： Pyroglyphidae）, are the major sources of indoor allergens. Identification and characterization of these mite allergen molecules are an important step in the development of new effective diagnostic procedures and possible therapeutic strategies for allergic disorders associated with dust mites. Methods Total RNA was extracted from Dermatophagoides farinae. The gene coding for Der f 3 was amplified by RT-PCR with the primers designed based on previous sequence published in GenBank. The target gene was cloned intermediately into pMD19-T plasmid and finally into plasmid pET28a （＋）, expressed in E. coil BL21 at the aid of the inducer isopropyI-D-thiogalactopyranoside （IPTG）. The physicochemical properties, spatial structure of the allergen were analyzed with bioinformatics software. Results The cDNA coding for group 3 allergen of Dermatophagoides farinae from China was cloned and expressed successfully. Sequencing analysis showed that there were nineteen mismatched nucleotides in five Der f3 cDNA clones in comparison with the reference （GenBank Accession No. AY283291）, which resulted in deduced amino acid sequence incompatibility in eleven residues. Bioinformatics analysis revealed that the Der f 3 pro-protein was an extracellular hydrophobic protein, consisting of 259 amino acids with a 16 amino acid signal peptide. The protein was deduced to have three chymotrypsin active sites （53-68 AA, 108-122 AA and 205-217 AA）, one N-glycosylation site, one cAMP- and cGMP-dependent protein kinase phosphorylation site, four protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, and five N-myristoylation sites. Conclusions Der f3 is an extracellular hydrophobic protein which possesses multiple activation and phosphorylation sites. Polymorphism may exist in the Der f3 gene but this needs to be further confirmed in the future.展开更多
Derf2 gene for one of mite allergens in Dermatophagoides farinae has been cloned and expressed under regulation of 35S promoter in transgenic tobacco. The tran-scriptional analysis showed that this mite complete gene ...Derf2 gene for one of mite allergens in Dermatophagoides farinae has been cloned and expressed under regulation of 35S promoter in transgenic tobacco. The tran-scriptional analysis showed that this mite complete gene structure in genomic sequence could be spliced at prediction site. Allergenicity assay with immunological sera indicated that the extracts from the transgenic tobacco gave obvious positive IgE binding reaction with specific serum pool. This work would be of potential use in allergenicity assessment of genetically modified food.展开更多
基金supported by Jiangsu Maternal and child health research project(no.F202068)the Taihu Lake talent plan(Top-Level,no.2020THRC-GD-7)The Project of Wuxi Health Commission(no.201949).
文摘Objective:To characterize the group-29 allergens from Dermatophagoides(D.)pteronyssinus and investigate their ability to cross-react with other group-29 allergens from D.pteronyssinus as well as those from D.farinae and Tyrophagus putrescentiae.Methods:Der p 29,Der f 29,and Tyr p 29 cDNA sequences were amplified from total RNA isolated from D.pteronyssinus,D.farinae and Tyrophagus putrescentiae,respectively.Then they were cloned into the pET28a vector,expressed in Rosetta2(DE3)plysS,and purified using anion exchange chromatography.The IgE-binding rates of rDer p 29 were assessed by IgE Western blotting.The four epitopes of rDer p 29 were predicted,synthesized,and detected by IgE-ELISA.The cross-reactivity among the recombinant proteins rDer p 29,rDer f 29,and rTyr p 29 was investigated using dot blot and IgE-ELISA inhibition experiments.The allergens’physiochemical properties,amino acid sequences,and tertiary structures were also compared.Results:Der p 29 was successfully expressed in Rosetta2(DE3)plysS as a single,393-bp open reading frame.Western blotting showed that the purified rDer p 29 protein exhibited an IgE-binding rate of 100%when tested on patient sera.The following four Der p 29 epitopes were predicted and synthesized:37-45(EP1),57-69(EP2),75-80(EP3),and 104-117(EP4).IgE-ELISA tests on 20 D.pteronyssinus-positive sera yielded IgE-binding rates of 85%(rDer p 29),80%(EP1),55%(EP2),40%(EP3),and 55%(EP4),respectively.The dot blot experiments further confirmed cross-reactivity among the three group-29 proteins.When used as an inhibitor,rDer p 29 demonstrated an average cross-reactive inhibition rate of 49.7%against rDer f 29 and 54.4%against rTyr p 29.When rTyr p 29 was used as an inhibitor,it showed an average cross-reactive inhibition rate of 56.3%against rDer f 29.Conclusions:A recombinant protein,rDer p 29 with strong allergenicity was produced.Moreover,it was found that rDer p 29 cross-reacted with rDer f 29 and rTyr p 29,due to their highly homologous sequences and structures.These findings highlight the importance of considering inter-species epitope cross-reactivity when diagnosing and treating allergic diseases.
文摘Objective:To examined the immediate and 24 hours post-irradiation germicidal effects of UV-C lamp on.eggs and adults of house dust mites Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermatophagoides farinae(D.farinae).Methods:This study investigated the immediate and24 hours post irradiation mortalities of adult mites exposed to UV-C at different exposure times(5mins,10 mins,15 mins,20 mins,30 mins and 60 mins)and distances(10 cm,25 cm,33 cm,45 cm and 55 cm).Fresh eggs of the 2 dust mites were also irradiated at 10,35 and 55 cm for 0.5,1,2,3,and 5 minutes,and observed daily post-irradiation for up to 7 days.Results:Highest immediate mortality of 100%occurred with direct irradiation al 10 cm distance from UV-C lamp and for 60mins,for both species of mites.The post 24 hours mean mortality rates were(58.4±17.4)%for D.pteronyssinus and(27.7±9.7)%for D.fitrinae when irradiated for 1 hour at 55 cm distance under UV-C lamp.When mites were irradiated in the presence of culture media,the highest mortality rates were lower compared to the direct irradiation;at 10 cm distance and 60 mins exposure,the mean mortality was(74.0±6.8)%for D.pteronyssinus and(70.3±6.7)%for D.farinae.Egg hatchability for both species of mites was also notably reduced by greater than 50%following irradiation.Conclusions:Ultraviolet C irradiation is lethal to an array of organisms by damaging their nucleic acids(DNA and RNA).This study demonstrates the increasing mite mortalities with increasing exposure times and decreasing distances.
文摘Objective:To investigate the short and long tenn efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermalophagoides farinae(D.farinae)mites.Methods:The effect of a commercial ionizer on D.pteronyssinus and D.farinae was evaluated in the laboratory,using a specially designed test.Mortality was assessed after 6,16and 24 hours for direct exposure and after 24,36,48,60 and 72 hours for exposure in simulated mattress.New batches of mites were used for each exposure time.Results:LT_(50)for direct exposure of ionizer was 10 hours for D.pteronyssinus and 18 hours for D.farinae.The LT_(50)for exposure in simulated mattress was 132 hours or 5.5 days for D.pteronyssinus and 72 hours or 3days for D.farinae.LT_(95)for direct exposure of ionizer was 36 hours for D.pteronyssinus and D.farinae.Meanwhile,the LT_(95)for exposure in simulated mattress was 956 hours or 39.8 days for D.pteronyssinus and 403 hours or 16.8 days for D.farinae.Conclusions:This study demonstrates the increasing mite mortalities with increasing exposure time of a commercial ionizer and suggests that negative ions produced by an ionizer kill dust mites and can be used to reduce natural mile populations on exposed surfaces such as floors,clothes,curtains,etc.However,there is reduced efficacy on mites inside stuffed materials as in mattresses and furniture.
文摘Objective: To investigate the effects of dermatophagoides farinae drops combined with pidotimod on inflammatory response and immune response in children with allergic rhinitis combined with asthma. Methods: A total of 118 children with allergic rhinitis combined with asthma who were treated in our hospital between December 2014 and May 2017 were selected as the research subjects and divided into the control group (n=59) and the dermatophagoides farinae drops group (n=59) by random number table method. Control group received both conventional therapy and pidotimod therapy, and dermatophagoides farinae drops group received both conventional therapy and dermatophagoides farinae drops combined with pidotimod therapy. The differences in inflammatory factor, cellular immunity index and humoral immunity index levels were compared between the two groups before treatment (T0), after 3 months of treatment (T1), after 6 months of treatment (T2) and after 12 months of treatment (T3). Results: At T0, inflammatory factor, cellular immunity index and humoral immunity index levels were not significantly different between the two groups. At T1, T2 and T3, serum inflammatory factor IL-2 contents of dermatophagoides farinae drops group were higher than those of control group at the corresponding time points whereas IL-4, IL-5, IL-9 and IL-18 contents were lower than those of control group at the corresponding time points;peripheral blood cellular immunity indexes CD3+, CD4+, CD8+ and CD4+/CD8+levels were higher than those of control group at the corresponding time points;serum humoral immunity indexes IgA, IgM and IgG contents were higher than those of control group at the corresponding time points whereas IgE contents were lower than those of control group at the corresponding time points. Conclusion: Dermatophagoides farinae drops combined with pidotimod therapy can effectively reduce the inflammatory response degree and optimize the cellular and humoral immune function of the children with allergic rhinitis combined with asthma.
文摘Objective:To study the ventilation function after sublingual dermatophagoides farinae drops combined with loratadine treatment of children with asthma and allergic rhinitis and the influence on serum biochemical indexes.Methods: A total of 40 children with asthma and allergic rhinitis treated in our hospital between September 2013 and March 2015 were collected and divided into the control group (n=22) who accepted loratadine therapy alone and the observation group (n=18) who accepted sublingual dermatophagoides farinae drops combined with loratadine therapy after the treatment was reviewed. Before treatment and after 6 months and 1 year of treatment, spirometer was used to test ventilation function indexes;ELISA method was used to determine the contents of inflammatory mediators;RIA method was used to determine the contents of airway remodeling indicators.Results:Before treatment, differences in ventilation function index levels as well as inflammatory mediator and airway remodeling index contents were not statistically significant between two groups of children. After 6 months and 1 year of treatment, FEV1, FVC and PEF levels of observation group were higher than those of control group;serum IL-2 content was higher than that of control group while IL-5, IL-17 and IL-33 contents were lower than those of control group;serum PDGF-BB, TGF-β1 and NF-κB contents were lower than those of control group.Conclusion:Sublingual dermatophagoides farinae drops combined with loratadine therapy can optimize the ventilation function, reduce the systemic inflammatory response and inhibit airway remodeling in children with asthma and allergic rhinitis.
基金This work was supported by grants from National Science Foundation of China （No. 30060166 and 30671939）.
文摘Background The dust mites, which are mostly represented by Dermatophagoides spp. （Acari： Pyroglyphidae）, are the major sources of indoor allergens. Identification and characterization of these mite allergen molecules are an important step in the development of new effective diagnostic procedures and possible therapeutic strategies for allergic disorders associated with dust mites. Methods Total RNA was extracted from Dermatophagoides farinae. The gene coding for Der f 3 was amplified by RT-PCR with the primers designed based on previous sequence published in GenBank. The target gene was cloned intermediately into pMD19-T plasmid and finally into plasmid pET28a （＋）, expressed in E. coil BL21 at the aid of the inducer isopropyI-D-thiogalactopyranoside （IPTG）. The physicochemical properties, spatial structure of the allergen were analyzed with bioinformatics software. Results The cDNA coding for group 3 allergen of Dermatophagoides farinae from China was cloned and expressed successfully. Sequencing analysis showed that there were nineteen mismatched nucleotides in five Der f3 cDNA clones in comparison with the reference （GenBank Accession No. AY283291）, which resulted in deduced amino acid sequence incompatibility in eleven residues. Bioinformatics analysis revealed that the Der f 3 pro-protein was an extracellular hydrophobic protein, consisting of 259 amino acids with a 16 amino acid signal peptide. The protein was deduced to have three chymotrypsin active sites （53-68 AA, 108-122 AA and 205-217 AA）, one N-glycosylation site, one cAMP- and cGMP-dependent protein kinase phosphorylation site, four protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, and five N-myristoylation sites. Conclusions Der f3 is an extracellular hydrophobic protein which possesses multiple activation and phosphorylation sites. Polymorphism may exist in the Der f3 gene but this needs to be further confirmed in the future.
文摘Derf2 gene for one of mite allergens in Dermatophagoides farinae has been cloned and expressed under regulation of 35S promoter in transgenic tobacco. The tran-scriptional analysis showed that this mite complete gene structure in genomic sequence could be spliced at prediction site. Allergenicity assay with immunological sera indicated that the extracts from the transgenic tobacco gave obvious positive IgE binding reaction with specific serum pool. This work would be of potential use in allergenicity assessment of genetically modified food.
