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 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 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 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
Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samp...Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samples were collected from nine cities. Major allergens Der p 1 and Der f I from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.Results HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from 〈0.006 to 9.2 μg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.Conclusion HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.展开更多
Objective:To examine the acaricidal effects of the essential oil of Cymbopogon citratits leaf extract(lemongrass)and ethanolic Azadirachta indica leaf extract(neem)against house dust mites Dermatophagoides farinae(D.f...Objective:To examine the acaricidal effects of the essential oil of Cymbopogon citratits leaf extract(lemongrass)and ethanolic Azadirachta indica leaf extract(neem)against house dust mites Dermatophagoides farinae(D.farinae)and Dermalophagoides pteronyssinus(D.pteronyssinus).Methods:Twenty-five adults mites were placed onto treated filter paper that is soaked with plant extract and been tested at different concentrations(50.00%,25.00%,12.50%,6.25%and 3.13%)and exposure times(24hrs,48hrs,72hrs and 96 hrs).All treatments were replicated 7 times,and the experiment repeated once.The topical and contact activities of the two herbs were investigated.Results:Mortalities from lemongrass extract were higher than neem for both topical and contact activities.At 50%concentration,both 24 hrs topical and contact exposures to lemongrass resulted in more than 91%mortalities for both species of inites.At the same concentration and exposure time,neem resulted in topical mortalities of 40.3%and 15.7%against D.pteronyssinus and D.farinae respectively;contact mortalities were 8.0%and 8.9%against the 2 mites,respectively.There was no difference in topical mortalities of D.pteronyssinus from exposure to concentrations oflemongrass and neem up to 12.50%;lemongrass was more effective than neem at the higher concentrations.Conclusions:Generally,topical mortalities of D.farinae due to lemongrass are higher than that due to neem.Contact mortalities of lemongrass are always higher that neem against both species of mites.展开更多
The house dust mites (Dermatophagoides farinae, Derf) are the major source of aeroaller gens implicated in the expression of atopic disorders, including asthma, allergic rhinitis and atopic dermatitis. In particular, ...The house dust mites (Dermatophagoides farinae, Derf) are the major source of aeroaller gens implicated in the expression of atopic disorders, including asthma, allergic rhinitis and atopic dermatitis. In particular, strong circumstantial evidence suggests that house dust mite antigens are important precipitating factors of asthma. Many house dust mite allergens are proteases that can elicit airway inflammation by stimulating the release of cytokines from bronchial epithelial cells. To investigate whether Der f allergen proteases induced cytokine production from the epithelial cell line BEAS-2B, BEAS-2B cells were cultured with 4 different concentrations of Derf (0.02, 0.2, 2, 20μg/ml) for 24-96 h, after which supernatants were assayed for interleukin (IL)-6 and IL-8 with ELISA. Reverse transcription-PCR was also performed. The cell sheets were intact throughout the observation in control group without any exposure to Der f antigen. In the experimental groups cells treated with Der f allergen showed changes in the anchorage status of the monolayer. There was a significant increase in the level of cytokine production compared with the untreated sample. The release of IL-6 and IL-8 increased in a concentration-dependent manner (P <0.05, respectively) with the addition of increasing dosage of Der f to the cell sheets. Levels of IL-6 and IL-8 began to rise at 24 h and 48 h after allergen exposure, and they increased significantly in the supematants at 72 h and 96 h. At the same time the concentration dependence of induction of IL-6 and IL-8 expression as well as an increase in the expression of IL-6 and IL-8 mRNA manifested evidently. HDM-induced airway inflammation may include Der f-mediated release of inflammatory mediators, and the proteolytic activity of an allergen may stimulate the release of proinflammatory cytokines from human bronchial epithelium. It is suggested that IL-6 and IL-8 production by bronchial epithelial cells may play a role in the pathogenesis of allergic asthma.展开更多
Dermatophagoides farinae group 2 (Der f2) is a highly polymorphic allergen that shows a distinct pattern of sequence divergence. The effect of the variants on antibody and T cell responses has not been compared. The a...Dermatophagoides farinae group 2 (Der f2) is a highly polymorphic allergen that shows a distinct pattern of sequence divergence. The effect of the variants on antibody and T cell responses has not been compared. The aim of the present study was to evaluate IgE binding, transcription and translations for IL-5, IFN-γ and TGF-β induced by mite allergens. Sera from 24 HDM-allergic patients and 20 non-allergic subjects were measured for IgE reactivity by ELISA. PBMC was cultured with mite allergens (Df, rDerf2) and mitogen (PHA). The supernatants and cell pellet obtained were evaluated for cytokine production by ELISA and cytokine gene expression by RT-PCR, respectively. Four patients showed IgE reactivity to both allergens. Five patients showed IgE reactivity to Df. Other allergic patients and all non-allergic subjects did not show IgE reactivity to mite allergens. Both allergens showed similar levels of IL-5 and IFN-γ transcriptions in allergic patients and non-allergic subjects. The rDer f2 induced IL-5 protein from allergic patients higher than non-allergic subjects, while Df showed IL-5 protein from allergic patients similar to non-allergic subjects. Df induced IFN-protein from allergic patients higher than non-allergic subjects whereas rDer f2 induced IFN-protein from allergic patients similar to non-allergic subjects. The ratio of IFN-γ to IL-5 production after stimulation with rDer f2 was higher in non-allergic subjects than in allergic patients. Our data demonstrated that the changes in the sequence of rDer f2 compared with native Df had effect on cytokine production in both allergic patients and non-allergic subjects.展开更多
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by elevated immunoglobulin E (IgE), mast cell infiltration and skin lesions including pruritus, erythema and eczema. Cudrania tricuspidata ex...Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by elevated immunoglobulin E (IgE), mast cell infiltration and skin lesions including pruritus, erythema and eczema. Cudrania tricuspidata extracts have been clinically administered for a long time in the East Asia including Korean and China as a home-remedy to diminish the inflammation of gastritis and hepatitis. To examine whether it works on AD or not, an AD-like animal model was experimented in this study. AD was induced by applying Dermatophagoides farinae (D. farinae) extract to the backs of 9-week old NC/Nga mice for 21 days. Following this, an ethanol extract of C. tricuspidata stems (EECT) was applied topically for 14 days to the sensitized skin, while distilled water was used as a control (EECT0 mice). Anti-AD effects of EECT were evaluated using scores for AD-like skin lesions, serum IgE levels and mast cell counts in the skin dermal layers to assess inflammation. Topically applied ethanol extract of Cudrania tricuspidata stems (EECT 7.5, 25 and 75 mg/mL) markedly reduced AD-like skin lesions after 4 days (by 30.1%, 31.4% and 38.5%, respectively) and also after 14 days (by 63.6%, 66.1% and 49.6%, respectively), while distilled water improved AD by 17.8% and 38.7%, respectively (p D. farinae extract (p = 0.003) and EECT attenuated the mast cell overproduction, and reduced mast cell degranulation markedly. Attenuation was most obvious in the early stage of EECT treatment when the AD was most acute.展开更多
Background Allergen micro-arrays are powerful tools for screening of serum IgE-reactivity. In this study allergen micro-arrays were used to identify dominating IgE-binding allergens and cross-reactivity patterns among...Background Allergen micro-arrays are powerful tools for screening of serum IgE-reactivity. In this study allergen micro-arrays were used to identify dominating IgE-binding allergens and cross-reactivity patterns among selected Chinese allergy patients. Methods The study was conducted using patient sera from the cities of Guangzhou, Nanjing, Chengdu and Shenyang. In total 100 sera with Dermatophagoides pteronyssinus （Der p） specific IgE-levels higher than 50 kU/L were selected for testing against 103 individual allergens. Results Among 100 selected patients, 95% showed IgE-reactivity towards house-dust mite allergens Dermatophagoides farinae （Der f） 1, Der f 2 and Der p 2 and 94% were IgE positive against Der p 1, and 60% of sera contained IgE reacting against allergen Euroglyphus maynei （Eur m） 2. IgE against cat allergen, Felisdomesticus （Fel d） 1, was seen in 20%. Only 2% showed specific IgE-reactivity to Der p 10, a panallergen belonging to the tropomyosin family. Serum IgE-reactivity towards other allergens was in general low. IgE-reactivity against pollen allergens showed geographic differences. Conclusions This study clearly confirms that group 1 and group 2 are major allergens of house dust mites. These selected house-dust mite allergy patients are close to being mono-sensitized. Der p 10 is not an important allergen for cross-reactivity. Specific IgE-sensitization towards pollen allergens is low in southern China compared to other regions. The prevalence of food and stinging insect allergens known to give rise to IgE-mediated cross-reactivity is 2% or less.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 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 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 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Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samples were collected from nine cities. Major allergens Der p 1 and Der f I from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.Results HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from 〈0.006 to 9.2 μg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.Conclusion HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.
文摘Objective:To examine the acaricidal effects of the essential oil of Cymbopogon citratits leaf extract(lemongrass)and ethanolic Azadirachta indica leaf extract(neem)against house dust mites Dermatophagoides farinae(D.farinae)and Dermalophagoides pteronyssinus(D.pteronyssinus).Methods:Twenty-five adults mites were placed onto treated filter paper that is soaked with plant extract and been tested at different concentrations(50.00%,25.00%,12.50%,6.25%and 3.13%)and exposure times(24hrs,48hrs,72hrs and 96 hrs).All treatments were replicated 7 times,and the experiment repeated once.The topical and contact activities of the two herbs were investigated.Results:Mortalities from lemongrass extract were higher than neem for both topical and contact activities.At 50%concentration,both 24 hrs topical and contact exposures to lemongrass resulted in more than 91%mortalities for both species of inites.At the same concentration and exposure time,neem resulted in topical mortalities of 40.3%and 15.7%against D.pteronyssinus and D.farinae respectively;contact mortalities were 8.0%and 8.9%against the 2 mites,respectively.There was no difference in topical mortalities of D.pteronyssinus from exposure to concentrations oflemongrass and neem up to 12.50%;lemongrass was more effective than neem at the higher concentrations.Conclusions:Generally,topical mortalities of D.farinae due to lemongrass are higher than that due to neem.Contact mortalities of lemongrass are always higher that neem against both species of mites.
文摘The house dust mites (Dermatophagoides farinae, Derf) are the major source of aeroaller gens implicated in the expression of atopic disorders, including asthma, allergic rhinitis and atopic dermatitis. In particular, strong circumstantial evidence suggests that house dust mite antigens are important precipitating factors of asthma. Many house dust mite allergens are proteases that can elicit airway inflammation by stimulating the release of cytokines from bronchial epithelial cells. To investigate whether Der f allergen proteases induced cytokine production from the epithelial cell line BEAS-2B, BEAS-2B cells were cultured with 4 different concentrations of Derf (0.02, 0.2, 2, 20μg/ml) for 24-96 h, after which supernatants were assayed for interleukin (IL)-6 and IL-8 with ELISA. Reverse transcription-PCR was also performed. The cell sheets were intact throughout the observation in control group without any exposure to Der f antigen. In the experimental groups cells treated with Der f allergen showed changes in the anchorage status of the monolayer. There was a significant increase in the level of cytokine production compared with the untreated sample. The release of IL-6 and IL-8 increased in a concentration-dependent manner (P <0.05, respectively) with the addition of increasing dosage of Der f to the cell sheets. Levels of IL-6 and IL-8 began to rise at 24 h and 48 h after allergen exposure, and they increased significantly in the supematants at 72 h and 96 h. At the same time the concentration dependence of induction of IL-6 and IL-8 expression as well as an increase in the expression of IL-6 and IL-8 mRNA manifested evidently. HDM-induced airway inflammation may include Der f-mediated release of inflammatory mediators, and the proteolytic activity of an allergen may stimulate the release of proinflammatory cytokines from human bronchial epithelium. It is suggested that IL-6 and IL-8 production by bronchial epithelial cells may play a role in the pathogenesis of allergic asthma.
文摘Dermatophagoides farinae group 2 (Der f2) is a highly polymorphic allergen that shows a distinct pattern of sequence divergence. The effect of the variants on antibody and T cell responses has not been compared. The aim of the present study was to evaluate IgE binding, transcription and translations for IL-5, IFN-γ and TGF-β induced by mite allergens. Sera from 24 HDM-allergic patients and 20 non-allergic subjects were measured for IgE reactivity by ELISA. PBMC was cultured with mite allergens (Df, rDerf2) and mitogen (PHA). The supernatants and cell pellet obtained were evaluated for cytokine production by ELISA and cytokine gene expression by RT-PCR, respectively. Four patients showed IgE reactivity to both allergens. Five patients showed IgE reactivity to Df. Other allergic patients and all non-allergic subjects did not show IgE reactivity to mite allergens. Both allergens showed similar levels of IL-5 and IFN-γ transcriptions in allergic patients and non-allergic subjects. The rDer f2 induced IL-5 protein from allergic patients higher than non-allergic subjects, while Df showed IL-5 protein from allergic patients similar to non-allergic subjects. Df induced IFN-protein from allergic patients higher than non-allergic subjects whereas rDer f2 induced IFN-protein from allergic patients similar to non-allergic subjects. The ratio of IFN-γ to IL-5 production after stimulation with rDer f2 was higher in non-allergic subjects than in allergic patients. Our data demonstrated that the changes in the sequence of rDer f2 compared with native Df had effect on cytokine production in both allergic patients and non-allergic subjects.
文摘Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by elevated immunoglobulin E (IgE), mast cell infiltration and skin lesions including pruritus, erythema and eczema. Cudrania tricuspidata extracts have been clinically administered for a long time in the East Asia including Korean and China as a home-remedy to diminish the inflammation of gastritis and hepatitis. To examine whether it works on AD or not, an AD-like animal model was experimented in this study. AD was induced by applying Dermatophagoides farinae (D. farinae) extract to the backs of 9-week old NC/Nga mice for 21 days. Following this, an ethanol extract of C. tricuspidata stems (EECT) was applied topically for 14 days to the sensitized skin, while distilled water was used as a control (EECT0 mice). Anti-AD effects of EECT were evaluated using scores for AD-like skin lesions, serum IgE levels and mast cell counts in the skin dermal layers to assess inflammation. Topically applied ethanol extract of Cudrania tricuspidata stems (EECT 7.5, 25 and 75 mg/mL) markedly reduced AD-like skin lesions after 4 days (by 30.1%, 31.4% and 38.5%, respectively) and also after 14 days (by 63.6%, 66.1% and 49.6%, respectively), while distilled water improved AD by 17.8% and 38.7%, respectively (p D. farinae extract (p = 0.003) and EECT attenuated the mast cell overproduction, and reduced mast cell degranulation markedly. Attenuation was most obvious in the early stage of EECT treatment when the AD was most acute.
文摘Background Allergen micro-arrays are powerful tools for screening of serum IgE-reactivity. In this study allergen micro-arrays were used to identify dominating IgE-binding allergens and cross-reactivity patterns among selected Chinese allergy patients. Methods The study was conducted using patient sera from the cities of Guangzhou, Nanjing, Chengdu and Shenyang. In total 100 sera with Dermatophagoides pteronyssinus （Der p） specific IgE-levels higher than 50 kU/L were selected for testing against 103 individual allergens. Results Among 100 selected patients, 95% showed IgE-reactivity towards house-dust mite allergens Dermatophagoides farinae （Der f） 1, Der f 2 and Der p 2 and 94% were IgE positive against Der p 1, and 60% of sera contained IgE reacting against allergen Euroglyphus maynei （Eur m） 2. IgE against cat allergen, Felisdomesticus （Fel d） 1, was seen in 20%. Only 2% showed specific IgE-reactivity to Der p 10, a panallergen belonging to the tropomyosin family. Serum IgE-reactivity towards other allergens was in general low. IgE-reactivity against pollen allergens showed geographic differences. Conclusions This study clearly confirms that group 1 and group 2 are major allergens of house dust mites. These selected house-dust mite allergy patients are close to being mono-sensitized. Der p 10 is not an important allergen for cross-reactivity. Specific IgE-sensitization towards pollen allergens is low in southern China compared to other regions. The prevalence of food and stinging insect allergens known to give rise to IgE-mediated cross-reactivity is 2% or less.
