miR-142-3p alleviates house dust mite induced airway inflammation among children / 中国学校卫生

Objective@#To investigate the role of miR-142-3p in alleviation of house dust mite induced allergic airway inflammation among children, so as to provide insights into unraveling the pathogenesis of allergic airway inflammation.@*Methods@#Serum samples were collected from 15 patients with house dust mite induced allergic asthma and 15 healthy children in Jiangnan University Medical Center from September to November 2022, and serum miR-142-3p expression was quantified using a fluorescent quantitative real time PCR (qPCR) assay. The levels of interleukin 6 (IL 6) and tumor necrosis factor α (TNF α) were measured in the cell culture supernatant using enzyme linked immunosorbent assay (ELISA), and the expression of high mobility group box 1 (HMGB1) was detected at transcriptional and translational lvels using qPCR and Western blotting assays. The negative regulation of the HMGB1 gene by miR 142 3p was identified using a dual luciferase gene reporter assay, and the expression of downstream regulatory proteins was determined in human normal lung epithelial cells (BEAS 2B) cells transfected with miR 142 3p using Western blotting. In addition, female C57BL/6 mice at ages of 6-8 weeks were randomly assigned to the phosphate buffer saline (PBS) group, house dust mite sensitized airway inflammation group and house dust mite sensitized airway inflammation + miR 142 3p intervention group. Mouse airway inflammation was evaluated using hematoxylin eosin staining, and the expression of inflammatory cells and inflammatory cytokines were detected in mouse bronchoalveolar lavage fluid (BALF) using Giemsa staining and ELISA.@*Results@#Lower serum miR-142-3p expression was quantified among children with house dust mite induced allergic asthma than among healthy controls (1.33±0.21 vs. 4.74±0.62, t=5.22, P <0.05). Stimulation with dermatophagoides farinae extract (DFE) resulted in a reduction in miR-142-3p expression in BEAS-2B cells (0.82±0.25), while transfection with miR-142-3p mimics resulted in a rise in miR-142-3p expression in BEAS-2B cells (0.55±0.14)( t=3.31, 3.94, P <0.05). Pre treatment with miR-142-3p reduced the expression of IL 6(2.25±0.46)and TNF α(6.58±1.95) ( t=4.86, 3.38, P <0.05) in BEAS 2B cells stimulated with DFE, and treatment with miR-142-3p mimics resulted in a reduction in TLR4 and NF-κB expression in BEAS-2B cells via negative regulation of the HMGB1 expression. In addition, treatment with miR-142-3p was found to alleviate inflammatory cell infiltration in lung tissues of house dust mite sensitized mice, and results in a reduction in interleukin 4 (IL-4)[(107.60±10.43)pg/mL], interleukin 5 (IL 5)[(95.78±13.14)pg/mL] and HMGB1[(2.52±0.87)pg/mL] expression in BALF ( t=10.32, 7.29, 2.90, P <0.05).@*Conclusion@#miR-142-3p alleviates house dust mite induced allergic airway inflammation among children via negative regulation of the HMGB1/TLR4/NF-κB pathway.

Dermatophagoides farinae induces conjunctival epithelial cell damage to promote neutrophil migration and neutrophil extracellular traps formation / 中国血吸虫病防治杂志

Objective To investigate the mechanisms underlying allergic conjunctivitis caused by conjunctival epithelial cell damage, neutrophil migration and neutrophil extracellular traps (NETs) formation induced by crude extracts of Dermatophagoides farinae mite (CDM). Methods Human conjunctival epithelial cells were stimulated with 500, 1 000, 2 000, 4 000 ng/mL, and the expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-8 were detected using quantitative real-time PCR (qPCR) assay and enzyme-linked immunosorbent assay (ELISA). The culture supernatant of human conjunctival epithelial cells was collected and co-cultured with neutrophils. Neutrophil migration was measured using Transwell migration assay, and the expression of NETs markers myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) was quantified using immunofluorescence staining. Neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and then NETs were collected for treatment of human conjunctival epithelial cells. Cell apoptosis was detected using flow cytometry, and the levels of IL-6, TNF-α, IFN-γ and IL-8 were measured in the cell culture supernatant using ELISA. Results Treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL up-regulated IL-6, TNF-α, IFN-γ and IL-8 expression in human conjunctival epithelial cells. Following treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL, the culture supernatant of human conjunctival epithelial cells promoted neutrophil migration and induced increases in the staining intensity of MPO and CitH3. In addition, increased NETs triggered the apoptosis of human conjunctival epithelial cells and IL-6, TNF-α, IFN-γ and IL-8 secretion in the culture supernatant of human conjunctival epithelial cells. Conclusions CDM induces human conjunctival epithelial cell damages, thereby promoting neutrophil migration and NETs formation, while the release of NETs further aggravates human conjunctival epithelial cell damages.

Bacterial community diversity in Dermatophagoides farinae using high-throughput sequencing / 中国血吸虫病防治杂志

Objective To investigate the bacterial community diversity in Dermatophagoides farinae. Methods Laboratory-cultured D. farinae was collected, and the composition of microbial communities was determined by sequence analyses of the V4 region in the bacterial 16S ribosomal RNA (16S rRNA) gene on an Illumina PE250 high-throughput sequencing platform. Following quality control and filtering of the raw sequence files, valid reads were obtained and subjected to operational taxonomic units (OTU) clustering and analysis of the composition of microbial communities and alpha diversity index using the Usearch software, Silva database, and Mothur software. Results A total of 187 616 valid reads were obtained, and 469 OTUs were clustered based on a sequence similarity of more than 97%. OTU annotation showed that the bacteria in D. farinae belonged to 26 phyla, 43 classes, 100 orders, 167 families and 284 genera. The bacteria in D. farinae were mainly annotated to five phyla of Proteobacteria, Firmicutes, Bacteroidota, Actinobacteriota, and Acidobacteriota, with Proteobacteria as the dominant phylum, and mainly annotated to five dominant genera of Ralstonia, norank-f-Mitochondria, Staphylococcus and Sphingomonas, with Wolbachia identified in the non-dominant genus. Conclusions A high diversity is identified in the composition of the bacterial community in D. farinae, and there are differences in bacterial community diversity and abundance among D. farinae.

Establishment and performance evaluation of Der p 1/Der p 2 specific immunoglobulin E detection for the components of dust mite allergens based on chemiluminescence immunoassay / 中华核医学与分子影像杂志

Objective:To establish a quantitative detection method for the main components of dust mite allergens Der p 1, Der p 2 specific immunoglobulin E (sIgE) by using the nano-magnetic particle chemiluminescence immunoassay.Methods:The performance indexes of the established method were evaluated after setting up and optimizing the chemiluminescence detection system and immune reaction conditions of sIgE for dust mite allergen. Serum sIgE levels of 50 suspected allergic patients with dust mite were determined by this chemiluminescence method. At the same time, this method was compared with the Phadia kit and the consistency was analyzed by Kappa test. Results:The optimal amount of magnetic beads was 25 μg, the optimal reaction buffer (pH=7.4) contained 0.1 mol/L Tris-HCl and 0.25%( W/ W) casein, the optimal coating solution contatined 20 mmol/L phosphate buffer (PB) and 1%( W/ W) bovine serum albumin (BSA), and the luminescence enhancement solution contained 0.05%( V/ V) Triton X-100. The two-step immunoreaction was adopted, and the detection could be completed with 20 μl sample at the optimal reaction temperature of 37℃. The limit of detection (LOD) of the established nano-magnetic particle chemiluminescence system in detecting Der p 1 and Der p 2 sIgE antibodies were both less than 0.01 kU/L, with the linear range of 0.2-100.0 kU/L, the precision of less than 7%, and the cross contamination rate of 0.19% and 0.21%. Compared with the Phadia system, the positive and negative coincidence rate of Der p 1 were 78.0%(32/41) and 9/9 with good consistency ( Kappa=0.65, P=0.008), and the positive and negative coincidence rate of Der P 2 were 93.3%(28/30) and 85.0%(17/20) with good consistency ( Kappa=0.79, P=0.003). Conclusion:The nano-magnetic particle chemiluminescence immunoassay is successfully established for detecting dust mite allergen sIgE, which has good detection performance and good consistency with Phadia system.

Scanning electron microscopic observation of the external morphology of Dermatophagoides farinae at different developmental stages / 中国血吸虫病防治杂志

Objective To investigate the morphological characteristics of Dermatophagoides farinae at different developmental stages. Methods The cultured D. farinae was isolated, and the external morphological features of mites at various developmental stages were observed using scanning electron microscopy (SEM), including egg, larva, nymph and adult stages. Results The D. farinae egg appeared a long oval shape, and the larval mites had three pairs of legs. The nymph had four pairs of legs and underdeveloped genital pores containing genital setae and anal setae, and adult mites appeared long and oval in shape, with decorative patterns on epidermis, and had four pairs of legs. In male adult mites, remarkable thickening of the leg I and thicker and longer leg III than the leg IV were seen, and ventral genital regions were found between the basal segments of legs III and IV; the anus was surrounded by a circular peri-anal ring, with a pair of anal suckers and anal setae within the ring. In the female adult mites, slender legs III and IV with an equal length were seen, and a “λ-shape” genital hole was observed on the ventral surface, with a crescent-like genital plate in the anterior part, and the anus appeared a longitudinal slit. Conclusions An SEM observation of the external morphology of D. farinae provides understandings of the morphological characteristics of D. farinae, which is of great significance for the classification and identification.

Inmunoterapia con ácaros Valergen por vía sublingual y subcutánea en pacientes alérgicos. Cabaiguán 2010-2019 / Sublingual and subcutaneous Valergen mite immunotherapy in allergic patients in Cabaiguán 2010-2019

RESUMEN Fundamento: La inmunoterapia consiste en la administración de dosis crecientes del alérgeno para controlar la sensibilidad hacia este. Objetivo: Describir el comportamiento de la inmunoterapia por vía subcutánea o sublingual a ácaros. Metodología: Estudio retrospectivo realizado en Cabaiguán, periodo de 2010 a 2019, el universo de 267 pacientes con pruebas positivas a ácaros y la muestra de 53 pacientes. Los datos se obtuvieron del registro de datos. Resultados: Usó la vía sublingual el 60.4 %, predominó el grupo de edad entre 5-18 años (41.5 %), la vía subcutánea se utilizó en un 39.6 %, fue más frecuente en el sexo masculino (58.5 %). La vía subcutánea se indicó mayoritariamente en la rinitis alérgica (20.7 %) y la sublingual en el asma (32.1 %); para ambas, fue el Dermatofagoide pteronyssinus el ácaro de mayor sensibilidad y utilización (43.4 %), el tiempo de administración dominante fue de 3 años (67.9 %). El control de los síntomas se alcanzó con la vía sublingual (37.7 %). En la vía sublingual no se presentaron eventos adversos en el 43.3 %, con la vía subcutánea 11.3 % de los eventos adversos fueron locales y 7.6 % sistémicos leves. Conclusiones: La vía sublingual fue la más utilizada, la de mayor control de los síntomas y menos eventos adversos en el estudio.

ABSTRACT Background: Immunotherapy consists of the administration of the allergen increasing doses to control sensitivity towards it. Objective: To describe the behavior of subcutaneous or sublingual immunotherapy to mites. Methodology: Retrospective study carried out in Cabaiguán, from 2010 to 2019, the universe of 267 patients with positive tests to mites and the sample 53 patients. Data were obtained from the data record. Results: 60.4 % used sublingual, the age group between 5-18 years prevailed (41.5 %), the subcutaneous was used in 39.6 %, and it was more frequent in males (58.5 %). The subcutaneous was indicated mainly in allergic rhinitis (20.7 %) and sublingual in asthma (32.1 %); for both, Dermatofagoide pteronyssinus was the highest sensitivity and use mite (43.4 %) the dominant administration time was 3 years (67.9 %). Symptom control was achieved by sublingual (37.7 %). In sublingual, there were no adverse events in 43.3 %, with the subcutaneous, 11.3 % of the adverse events were local and 7.6 % were mild systemic. Conclusions: Sublingual was the most used, the one with the highest control of symptoms and the fewest adverse events in the study.

IgE AND IgG antibody responses to Dermatophagoides pteronyssinus in dogs with demodicosis and atopic dermatitis / Respostas de anticorpos IgE E IgG específicos à Dermatophagoides pteronyssinus em soros de cães com demodicose e dermatite atópica

Canine demodicosis is a common inflammatory parasitic skin disease caused by Demodex mites. House dust mites, such as Dermatophagoides spp., play an important role in the pathogenesis of canine atopic dermatitis (AD). The goal of this experimental work was to investigate whether demodectic dogs could be previously exposed/sensitized to house dust mites' antigens. First the prevalence of demodicosis in a southeastern region of Brazil was investigated by analyzing clinical files of dogs that were admitted to a Veterinary Hospital. Subsequently, the IgG responses to Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df) and IgE to D.pteronyssinus (Dp) were evaluatedin two groups, AD or demodicosis dogs. Additionally, the major IgE-binding Dp proteins that are recognized by sera from dogs with demodicosis and AD were evaluated. A total of 2,599 clinical files were analyzed to identify the major parasitic skin diseases in dogs from this region, considering the age, sex and breed of the animals. The epidemiological study identified 111 animals with skin diseases; from these 20.7% presented demodicosis. Afterwards, serum samples were obtained from another groups of demodicosis, AD, and healthy dogs, and analyzed for Dp and Df-specific IgG, and IgE antibody levels, Dp IgG avidity by ELISA and IgE-binding Dp-specific proteins by immunoblot. IgG and IgE antibodies to Dp were detected in sera from additional groups of dogs with AD, demodicosis or healthy, with higher IgE levels to Dp in AD than demodectic or healthy dogs. IgG to Df was detected, despite with smaller levels compared to Dp in sera from demodectic dogs, and also in healthy dogs. Immunoblot showed IgE-binding to Dp proteins in sera of dogs with demodicosis and AD; with strong reactivity for the 72 and 116 kDa antigens detected by sera from demodicosis dogs. However, sera from healthy dogs >12 months old also presented reactivity to these bands. In conclusion, the detection of Dp-IgG and IgE antibodies in sera from demodectic dogs indicates previous exposure and sensitization to the house dust mite, respectively, more than cross-reactivity between demodex mites and Dp antigens detected by canine antibodies. Additionally, higher Dp-specific IgE levels were found in dogs with AD compared with those with demodicosis or healthy, suggesting that Dp-specific IgE could better discriminate dogs with AD from healthy ones or even those with demodicosis.

Demodicose canina é uma doença inflamatória comum da pele causada por ácaros do gênero Demodex. Ácaros da poeira doméstica como Dermatophagoides spp. desempenham papel importante na patogênese da dermatite atópica canina (DA). O objetivo desse trabalho experimental foi investigar se cães com demodicose poderiam ser previamente expostos/sensibilizados com antígenos de ácaros da poeira doméstica. A princípio, investigou-se a prevalência de demodicose em uma região sudeste do Brasil, analisando-se prontuários clínicos de cães admitidos em um Hospital Veterinário. Posteriormente, as respostas de IgG a Dermatophagoides pteronyssinus (Dp) e D. farinae (Df) e IgE a D. pteronyssinus (Dp) foram avaliadas em dois grupos, DA ou demodicose. Também foram avaliadas as principais proteínas Dp reconhecidas por anticorpo IgE presente em soros de cães com demodicose e DA. Um total de 2.599 prontuários clínicos foram analisados para identificar as principais doenças parasitárias da pele em cães dessa região, considerando a idade, sexo e raça dos animais. O estudo epidemiológico detectou 111 animais com doenças de pele e destes, 20,7% apresentavam demodicose. Posteriormente, amostras de soro foram obtidas de outros grupos de cães com demodicose, DA ou saudáveis, e analisadas quanto aos níveis de IgG e IgE específicos para Dp e Df, avidez de IgG a Dp por ELISA e proteínas específicas de Dp reconhecidas por IgE por immunoblot. Anticorpos IgG e IgE para Dp foram detectados em soros de grupos adicionais de cães com DA, demodicose ou saudáveis, com níveis mais altos de IgE para Dp na DA do que no soro de animais saudáveis. Níveis de IgG específicos para Df foram detectados, apesar serem menores em comparação com os detectados para Dp em soros de cães demodéticos, e também em cães saudáveis. A análise de immunoblot demonstrou detecção de IgE para proteinas de Dp em soros de cães com demodicose e DA; com forte reatividade para os antígenos de 72 e 116 kDa detectados por soros de cães com demodicose. No entanto, soros de cães saudáveis > 12 meses de idade também apresentaram reatividade a essas bandas. Em conclusão, a detecção de anticorpos Dp-IgG e IgE específicos em soros de cães demodéticos indica exposição prévia e sensibilização aos ácaros, respectivamente, mais do que reatividade cruzada entre ácaros Demodex e antígenos Dp detectados por anticorpos caninos. Além disso, níveis de Dp-IgE específicos mais elevados encontrados em cães com DA, sugerem que esses anticorpos poderiam discriminar melhor cães com DA daqueles saudáveis ou mesmo demodéticos.

Cloning and sequence analysis of leptin receptor overlapping transcript-like 1 gene from Dermatophagoides farinae / 中国血吸虫病防治杂志

Objective To obtain the leptin receptor overlapping transcript-like 1 encoding gene (LepROTL1 gene) from Dermatophagoides farina, investigate the molecular characteristics of the gene and construct a prokaryotic expression vector to express this gene. Methods The LepROTL1 gene-encoding sequence fragments were captured based on the transcriptome sequencing results, and the full-length gene fragments were amplified from total RNA of D. farinae using a RT-PCR assay, and used to construct the expression plasmid pET28a(+)-LepROTL1, followed by sequencing. The plasmid was transformed into E. coli BL21 (DE3) T1R for the induction of IPTG expression. The expression product was characterized by SDS-PAGE and Western blotting. Bioinformatics analyses were performed to analyze the sequence and the molecular characteristics of its encoded protein. Results The amplification products of the RT-PCR assay showed a clear band on agarose gel electrophoresis, and sequencing analysis of the pET28a(+)-LepROTL1 plasmid showed 417 bp in length of the coding gene from the start codon ATG to the termination codon TAA. Following the plasmid transformation into E. coli and induction with IPTG, a specific band was seen on SDS-PAGE, indicating successful expression. Bioinformatics analysis showed that the LepROTL1 gene-encoded protein was composed of 134 amino acids, and had a relative molecular weight of 14 378.13 Da, a hydrophilicity index of 1.149, and certain hydrophobicity. The secondary structure was composed of alpha-helix (19 aa, 14.18%), extended strand (48 aa, 35.82%) and random coil (67 aa, 50.00%). The deduced amino acid sequence was used to obtain homologous genes by BLAST, and the phylogenetic tree showed that D. farinae was clustered with D. pteronyssinus. Conclusion The full-length sequences and expression plasmid of the LepROTL1 gene are obtained, and the molecular features of the gene are demonstrated using bioinformatics analyses, which provide insights into further studies on the gene.

Investigation of temporal and spatial characteristics of allergic rhinitis epidemics by Baidu Index in China / 中华耳鼻咽喉头颈外科杂志

Objective@#To explore the temporal and spatial characteristics of search volume (SV) of allergic rhinitis (AR) and related search terms on Baidu Index, as well as the correlation between SV from Baidu Index and epidemiology of AR.@*Methods@#SV of AR and related search terms in mainland area of China from January 2012 to December 2016 were extracted from Baidu Index. Firstly, we investigated the temporal and spatial characteristics of SV of AR and related search terms ("pollen allergy" and" dust mite allergy", etc.), and explored the correlation between SV of AR and SV of related search terms. Secondly, the SV of specific search terms in Beijing and Guangzhou were compared with the actual pollen count in Beijing from April to September in 2016 and the monthly outpatient volume of AR patients in the Otorhinolaryngology Hospital, The First Affiliated Hospital of Sun Yat-sen University from 2014 to 2016 respectively to analyze the consistency between SV from Baidu Index and the real-world data. Spearman correlation analysis was used to explore the correlation between different search terms according to the normality and homogeneity of variance of the data. P<0.05 was considered statistically significant.@*Results@#SV of AR showed repetitively seasonal pattern, with the first peak in May and the second peak from August to September. SV of AR in northeast China, north China and northwest China were correlated with both SV of "pollen allergy" and "dust mite allergy" (r value was 0.533, 0.638, 0.609, 0.791, 0.517, 0.682, respectively, all P<0.05). In east China, central China, south China and southwest China, SV of AR showed significant correlation only with SV of "dust mite allergy" (r value was 0.897, 0.884, 0.799, 0.937, respectively, all P<0.05), but not with SV of "pollen allergy" (r value was 0.110, 0.171, 0.020, 0.265, respectively, all P>0.05). The correlation between pollen count and the SV of AR on the day and SV of "pollen allergy" on the next day was the highest (r value was 0.692 and 0.713, respectively, all P<0.05). The SV of AR was correlated with the monthly outpatient volume of AR patients one month and two months later (r value was 0.523 and 0.503, respectively, all P<0.05).@*Conclusion@#The SV of AR and related search terms show different trends in different times and regions which are consistent with the AR epidemiological survey, and significantly correlate with pollen count and AR outpatient volume.

Effects of Vacuuming Mattresses on Allergic Rhinitis Symptoms in Children

PURPOSE: To evaluate the effects of daily vacuuming of mattresses on the concentration of house dust mite (HDM) allergens and on allergic rhinitis (AR) symptoms in children sensitized to HDM. METHODS: Forty children between the ages of 6 and 12 years with mild persistent AR and sensitized only to HDM were enrolled and randomly allocated to 2 groups. Caregivers of children in the experimental group cleaned the children's rooms and vacuumed their mattresses daily for 2 weeks. Caregivers of children in the control group cleaned the children's rooms without vacuuming mattresses. Symptoms of AR were checked weekly and dust samples were collected from the mattresses before and after the study. RESULTS: Demographics at the beginning of the study were not significantly different between the 2 groups. In the experimental group, symptoms of AR and dust weight were significantly decreased after 2 weeks (total symptoms of AR, P <0.001; sneezing, P < 0.001; rhinorrhea, P <0.001; nasal obstruction, P < 0.001; itching, P <0.001; and dust weight, P = 0.006). The concentrations of HDM allergens were not changed significantly (Der p1, P = 0.333; Der f1, P = 0.841). In the control group, there were no significant changes in symptoms of AR, dust weight, or the concentration of HDM allergens. CONCLUSIONS: Our findings showed that daily vacuuming of mattresses reduced dust weight and symptoms of AR. However, the concentration of HDM allergens did not significantly decrease.

Performance of magnetic nanoparticle chemiluminescence immunoassay in detection of allergen-specific immunoglobulin E / 第二军医大学学报

Objective To evaluate the performance of magnetic nanoparticle chemiluminescence immunoassay (NM-CLIA) in detection of allergen-specific immunoglobulin E (sIgE) antibodies to Dermatophagoides pteronyssinus (International Allergen Code D1) and Dermatophagoides farinae (International Allergen Code D2). Methods A total of 489 serum samples from the patients with suspected allergic disease (244 cases caused by D1, and 245 caused by D2), who were treated at Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine, were detected by NM-CLIA and immunofluorescence assay, respectively. χ2 test and Kappa test were used to evaluate the correlation between the two methods in detection of D1 and D2 sIgE antibodies. The limit of detection (LoD), linear range and precision of NM-CLIA in detection of D1 and D2 sIgE antibodies were verified by the standard method of American Clinical Laboratory Standardization Association. Results The LoDs of NM-CLIA in detecting D1 and D2 sIgE antibodies were both less than 0.01 U/mL, the linearity ranged from 0.1 to 100 U/mL, the within-run precision was less than 5%, and the between-run precision was less than 8%. Methodological comparison results showed that NM-CLIA and immunofluorescence assay had good consistency in detecting D1 and D2 sIgE antibodies. For D1, the positive coincidence rate and negative coincidence rate were 95% and 92%, respectively (χ2＝174.45, P0.001, Kappa＝0.843), and the ±1 class agreement was 95.6%; for D2, the positive coincidence rate and negative coincidence rate were 91% and 97%, respectively (χ2＝154.263，P0.001，Kappa＝0.787), and the ±1 class agreement was 94.2%. Conclusion NM-CLIA has good correlation with immunofluorescence assay in detecting D1 and D2 sIgE antibodies, and has good LoD, linear range and precision, suggesting that it can be recommended for clinical testing of D1 and D2 sIgE antibodies.

Experimental study on dendritic cells modified by dust mite allergen gene in the treatment of allergic rhinitis in mouse / 中华耳鼻咽喉头颈外科杂志

Objective@#To explore the therapeutic effects of dendritic cells (DC) modified by the dust-mite-allergen(Der p1) gene on mouse model of allergic rhinitis (AR).@*Methods@#DC modified by the Der p1 gene (Der p1-DC) were prepared.Using random number table, 24 Balb\c mice were divided into four groups: immature DC (imDC)/AR group, dexamethasone/AR group, Der p1-DC/AR group and control group, with 6 mice in each group.AR mouse model was built with Der p1 and the mouse model of AR was established.The AR mice were respectively given by abdominal injection of Der p1-DC, imDC and dexamethasone.Normal control mice were treated with physiologic saline.ELISA method was used for determining the content of IgE, IgG1and histamine in blood.The relative expression of mRNA of IL-4 and IL-13 on nasal mucosa with protein was analyzed by RT-PCR and Westen blot methods.All the data were statistically analyzed by SPSS 19.0 statistical software, and the variance analysis was used in multiple groups of average samples.@*Results@#The contents of IgE, IgG1 and histamine in the mice of Der p1-DC/AR group were lower than those in imDC/AR group ((0.560±0.110) OD 450 nm vs (1.150±0.280) OD 450 nm, (0.690±0.054) OD 450 nm vs (0.920±0.125) OD 450 nm, (4 145±670) pg/ml vs (7 685±669) pg/ml, t value was 4.80, 4.14, 9.16, respectively, all P<0.05), and the expression of IL-4 and IL-13 on nasal mucosa in Der p1-DC/AR group was remarkedly lower than those in imDC/AR group (0.41±0.25 vs 1.59±1.02, 0.26±0.01 vs 1.10±0.09, t value was 2.75, 22.72, respectively, all P<0.05). There was no statistically significant difference between the mice treated with Der p1-DC and dexamethasone group.@*Conclusions@#The results showed that Der p1-DC could reduce inflammation in AR mice and decrease the expression of IL-4 and IL-13. It suggested that Der p1-DC can be used in the immunotherapy of AR mouse.

Evaluation of efficacy of 1 year after completing the 2-year specific sublingual immunotherapy treatment course in allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

Objective@#To evaluate the efficacy of treatment at 1 year after completing the 2-year specific sublingual immunotherapy in patients with allergic rhinitis (AR).@*Methods@#A retrospective analysis of 124 patients with AR who were allergic to dust mite and treated in the First Affiliated Hospital of Wenzhou Medical University since 2012 was performed. All patients achieved sublingual immunotherapy (SLIT) for 2 years and had a regular follow-up over 1 year. Efficacy evaluation index such as VAS score, total nasal symptom score and total medication score were compared between the time of SLIT for 2 years, and 1 year after termination of SLIT. Paired t test was used for the comparison of scores.@*Results@#Compared with those before treatment, the VAS score, total nasal symptom score and total medication score at the time of SLIT for 2 years were significantly decreased in all patients with AR [(9.40±5.96) vs (24.78±6.36), (4.53±2.06) vs (9.51±2.02), (0.42±0.87) vs (3.02±0.41), t value was 17.627, 24.600, 5.331, respectively, all P<0.01]. Compared with 2 years SLIT treatment, VAS score and total nasal symptom score were significantly increased than 1 year after termination of immunotherapy [(12.52±6.92) vs (9.40±5.96), (5.30±2.36) vs (4.53±2.06), t=5.199, 3.744, respectively, all P<0.01], but the total medication score showed no significant difference [(0.34±0.84) vs (0.42±0.87), t=-1.043, P>0.05].@*Conclusions@#The symptoms after 2 years SLIT in patients with AR have been well controlled. One year after the termination of SLIT, there is a certain rebound in symptoms, 2 years SLIT may not be the ideal treatment cycle for AR.

Usefulness of House Dust Mite Nasal Provocation Test in Asthma

PURPOSE: We previously reported that the skin prick test was sensitive and the serum specific immunoglobulin E test was specific for predicting positive airway responses to house dust mites (HDMs) in patients with asthma. Because the nose and bronchus are one airway, the nasal provocation test would be more specific for predicting the bronchial responses to HDM than the skin test. METHODS: The allergy skin prick test and nasal and bronchial provocation tests using HDM (Dermatophagoides farinae) were performed in 41 young men (age, 19–28 years) who wanted military certification for asthma. The nasal responses to HDM was scored according to the severity of rhinorrhea, sneezing, and nose itching. RESULTS: The prevalence of a positive skin prick test to HDM did not significantly differ between patients with (n=24) and without (n=17) an early airway reaction (EAR; 79.2% vs 70.6%, P=0.534). However, the prevalence of a positive nasal test was significantly higher in the airway responders than in the others (37.5% vs 0%, P=0.005). The concordance of a positive response to the nasal test (κ=0.332, P=0.004) but not to the skin prick test (κ=0.091, P=0.529) was significant with an EAR. The diagnostic sensitivity of the nasal test (37.5%) was lower than that of the skin prick test (79.2%), but the specificity was higher (100% vs 29.4%). CONCLUSIONS: The skin prick test is more sensitive, whereas the nasal test is more specific and accurate, for predicting an EAR to HDM in patients with asthma.

Usefulness of Allerkin House Dust Mite Extract for Nasal Provocation Testing

OBJECTIVES: We evaluated the clinical usefulness of Allerkin (Lofarma) for nasal provocation testing (NPT) in patients with rhinitis symptoms, by examining changes in nasal symptoms and acoustic parameters after exposure to house dust mite (HDM) extract. METHODS: Twenty patients (16 males and 4 females, mean age: 29.6±14.6 years) were enrolled. We performed skin prick test (SPT) before and 15 and 30 minutes after intranasal challenge with Allerkin HDM extract, and we evaluated symptom changes (nasal obstruction, rhinorrhea, sneezing, and itching) using a visual analogue scale. We also evaluated changes in acoustic parameters such as total nasal volume (TNV) and minimal cross-sectional area (MCA) before and after challenge. RESULTS: Group A (the nonallergic group, n=8) showed negative results for all tested aeroallergens in SPT and nonprovocative results ( 29% decrease in TNV/MCA from the baseline value). Patients in group C showed significant aggravation of nasal obstruction compared to those in group A (P < 0.05). Thirty minutes after HDM challenge, patients in groups B and C showed significantly greater decreases in MCA compared to those in group A (P < 0.01). CONCLUSION: Allerkin HDM extract can be a useful provocative agent in NPT for diagnosing allergic rhinitis and local allergic rhinitis.

House Dust Mite Sensitization Is Inversely Associated with Plasma 25-Hydroxyvitamin D3 Levels in Patients with Severe Atopic Dermatitis

BACKGROUND: The relationship between atopic dermatitis (AD) and low vitamin D levels has been studied. Emerging evidence has implicated vitamin D as a critical regulator of immunity, playing a role in both the innate and cell-mediated immune systems. However, the effect of vitamin D on house dust mite (HDM) sensitization in patients with AD has not been established. OBJECTIVE: We investigated the association between vitamin D levels and HDM sensitization according to AD severity. METHODS: In total, 80 patients (43 men and 37 women) with AD were included. We classified AD severity using Rajka and Langeland scores. Laboratory tests included serum 25-hydroxyvitamin D3, total immunoglobulin E (IgE), and specific IgE antibody titer against Dermatophagoides farinae and D. pteronyssinus. RESULTS: There were no differences in vitamin D levels between the mild or moderate AD and severe AD groups. In the severe AD group, high HDM sensitization group had lower serum vitamin D levels compared to low HDM sensitization group with statistical significance. In addition, a significant negative correlation was found between vitamin D levels and HDM sensitization in the severe AD group. CONCLUSION: Our results demonstrate that low vitamin D levels may link to high HDM sensitization in patients with the severe AD. Further elucidation of the role of vitamin D in HDM sensitization may hold profound implications for the prevention and treatment of AD.

Therapeutic effects of orally administered CJLP55 for atopic dermatitis via the regulation of immune response

Atopic dermatitis (AD) is an inflammatory skin condition accompanied by symptoms such as edema and hemorrhage. Kimchi is a traditional fermented Korean dish consisting of various probiotics. In this study, the therapeutic effect of Lactobacillus plantarum CJLP55 isolated from Kimchi was studied in AD-induced mice. Orally administered Lactobacillus strain, CJLP55, suppressed AD symptoms and high serum IgE levels. CJLP55 administration reduced the thickness of the epidermis, infiltration of mast cells and eosinophils into the skin lesion, enlargement of axillary lymph nodes, and increase in cell population in axillary lymph nodes. CJLP55 treatment decreased the production of type 2 cytokines, such as interleukin (IL)-4, IL-5, IL-10, IL-12, interferon (IFN)-γ, and IL-6,which were stimulated by house dust mite extracts, in the axillary lymph node cells. Orally administered CJLP55 exhibited a therapeutic effect on house dust mite-induced AD in NC/Nga mice after onset of the disease by altering immune cell activation. The Lactobacillus strain, CJLP55, isolated from Kimchi, suppressed AD. Our results suggest its possible use as a potential candidate for management of AD.

A specific immune therapeutic effect of Der p2 T cell epitope vaccine on asthma mice / 中国血吸虫病防治杂志

Objective To investigate the specific immune therapeutic effect of the T cell fusion peptide vaccine from group II allergens from Dermatophagoides pteronyssinus(Der p2). Methods Thirty mice were randomly divided into three groups, namely a negative control group(a PBS group),positive control group(an asthma group)and protein Der p2 T cell fusion epit?ope for specific immunotherapy(SIT)group(a Der p2 T group). The extract of house dust mites(HDM)was used to establish the asthmatic models in BALB/c mice,and the PBS group was always used with PBS buffer. Thirty minutes before spray inhala?tion from 25 to 27 days,the mice of the Der p2 T group were respectively injected subcutaneously with the therapeutic proteins for SIT,then the serum and bronchoalveolar lavage fluid(BALF)were collected. ELISA was used to assay the levels of IFN?γ, IL?4,and IL?13 in BALF,as well as serum levels of specific IgE and IgG2a. The lung tissue sections were stained with haematox?ylin and eosin(H&E)for pathological examinations. Results The ELISA detection revealed that the number of eosinophil in BALF of the asthmatic mice was(5.57 ± 0.64)× 105/ml,which was significantly higher than that in the PBS group[(0.50 ± 0.30)× 105/ml,P < 0.01],the number of eosinophil in the Der p2 T immunotherapy group decreased significantly[(3.45 ± 0.36)×105/ml,P<0.01]. The content of IFN?γin the PBS group,asthma group and Der p2 T group were(267.00 ± 21.98), (155.80 ± 20.53)pg/ml and(234.40 ± 24.46)pg/ml respectively. Compared with the asthma group,the mice with Der p2 T vac?cine specific immune treatment produced a high level of IFN?γ(P<0.01). The content of IL?4 in the PBS group,asthma group and Der p2 T group were(23.40 ± 5.96),(53.28 ± 8.26)pg/ml and(30.00 ± 5.50)pg/ml respectively. Compared with the asth?ma group,the content of IL?4 in the mice of the Der p2 T treatment group was significantly lower(P<0.01). Compared with the asthma group[(308.10 ± 28.32)pg/ml],the content of IL?13 in BALF of the mice in the Der p2 T treatment group was signifi?cantly decreased,which was[(174.50 ± 25.99)pg/ml,P<0.01]. The content of IL?13 in the PBS group was(95.99 ± 31.14) pg/ml. The lung tissue sections showed that the lung inflammation in the p2 T Der group was significantly less than that in the asthma group,and the inflammatory cell infiltration was significantly decreased,and airway epithelial construction remodeled. Conclusion The Der p2 T cell fusion epitope,which is as vaccines for specific immunotherapy with asthma models,can allevi?ate effectively allergic inflammation of airway and lung in the mice,and it may be used as a candidate vaccine for asthma.

Efficacy evaluation of specific sublingual immunotherapy with standardized dust mites drops on different age patients with allergic rhinitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To evaluate the difference of efficacy of sublingual immunotherapy on different age patients with allergic rhinitis. METHODS 229 patients with dermatophagoides farina drops allergic rhinitis who have finished SLIT at least 2 years were enrolled and analyzed by retrospective analysis. All patients were divided into different age groups: pre-adolescence(n=107), adolescence(n=53), adulthood(n=69). Indexes for therapeutic effects of SLIT visual analogue scale(VAS) score, rhinitis symptom scores, medication scoreintra- and inter groups were assessed during different stage of treatment(3 months, 1 year and 2 years). Data were analyzed with SPSS 19.0 software. RESULTS Compared with prior treatment, after the SLIT for 3 months, 1 year, 2 years, VAS scores, rhinitis symptom scores, medication scoresof all the patients with allergic rhinitis are significantly decreased; Differences of rhinitis symptom scores(H=0.844, 4.153, 2.669, P>0.05), VAS score(H=1.356, 3.720, 0.313, P >0.05), medication scores(H=1.044, 5.841, 3.399, P>0.05) between groups had no statistical significance at different stage after treatment(3 months, 1 year and 2 years); The differences between 2 years and 1 year of SLIT treatment showed statistical significance in the VAS scores(Z=1.635, P =1.635). CONCLUSION SLIT with dust mites drops had good curative effect and security in patients with allergic rhinitis, which hadno obvious difference between different age groups.

Short-term clinical efficacy of sublingual immunotherapy in children with allergic rhinitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the short-term clinical efficacy of sublingual immunotherapy in children with allergic rhinitis. METHODS Forty-four patients, from 3-14 years of age, with mite-sensitive allergic rhinitis were treated sublingually with standardized dermatophagoidesfarinae extract. After treatment of 3 or 6 months using the dermatophagoidesfarinae extract, the total nasal symptoms score(including nasal congestion, nasal itching, rhinorrhea and sneezing scores), total rhinitis medicine scores, visual analog scale scores and the percentage of blood eosinophilic granulocyte(Eos%) before and after treatment were evaluated. SPSS 20.0 software was used to analyze the data. RESULTS After 3 or 6 months' treatment,thenasal symptoms score(including nasal congestion, nasal itching, rhinorrhea and sneezing scores), total rhinitis medicine scores and visual analog scale scoreshad significantly reduced,comparing with that before treatment(all P0.05), while nasal itching scores had significantly reduced(Z=-4.185, P<0.05). After 6 months' treatment, the percentage of Eos% had significantly reduced compared with that before treatment(Z=-4.544, P<0.05). CONCLUSION SLIT could be an effective treatment in patients with allergic rhinitis sensitive to house dust mite. It would have effect after3 months treatment and Eos%reduced significantly after 6 months treatment.