Hui-ying Wang

Neuroprotective effects of icaritin against beta amyloid-induced neurotoxicity in primary cultured rat neuronal cells via estrogen-dependent pathway

Beta-amyloid protein (Abeta) is the hallmark of pathogenic neurotoxins which contribute greatly to Alzheimers disease (AD)-associated cascade including severe neuronal loss. In present study, icaritin, an active natural ingredient from a Chinese plant, Epimedium sagittatum maxim, was investigated to assess its neuroprotective effect against the toxicity induced with Abeta(25-35) in primary cultured rat cortical neuronal cells as well as the underlying mechanisms. Abeta(25-35) induced neuronal toxicity, characterized by decreased cell viability, lactate dehydrogenase (LDH) release, and neuronal DNA condensation, which is associated with both the loss of membrane potential and the alteration of the expression of Bcl-2 family proteins. The phenotype alternation induced by Abeta(25-35) could be reversed by icaritin. Furthermore, the neuroprotective effects of icaritin mentioned above were estrogen receptor dependent due to the blocking action induced by estrogen receptor antagonist ICI 182,780 and well matched binding affinity with estrogen receptor by a receptor-ligand docking experiment. mitogen-activated protein kinase/extracellular signal-regulated kinase kinase inhibitor PD98059 weakened the protective effects, which implied mitogen-activated protein kinase/extracellular signal-regulated kinase pathway may also be involved in and partly contributed to the neuroprotective effects of icaritin.

Evaluation of the Role of IgE Responses to Der p 1 and Der p 2 in Chinese House Dust Mite-Allergic Patients

Background: The role of specific IgE (sIgE) against Der p 1 and Der p 2 in Chinese patients with house dust mite (HDM) allergy has not yet been well investigated. Methods: One hundred patients were enrolled, based on sensitization and doctor-diagnosed allergy to HDM. Questionnaires were administered to document demographic and clinical characteristics. Serum IgE reactivity to Dermatophagoides pteronyssinus (Dp) extract, Der p 1, Der p 2 and Der p 10 was measured by ImmunoCAP. Results: Almost all patients were sensitized to Der p 1 (95%) and Der p 2 (93%), with both allergens together being largely responsible for the total anti-HDM IgE response. No evidence for a significant role of Der p 10 was found. Overall, IgE responses to HDM and its 2 major allergens were higher in children than in adults in this cross-sectional study. With increasing age, IgE responses to Der p 2 become more important. A positive correlation was observed between the reaction of sIgE against Dp, Der p 1 and Der p 2 and the number of organs (including the eyes, nose, lungs and skin) that were affected in patients. Conclusions: In China, Der p 1 and Der p 2 are the dominant allergens in patients with HDM allergy. The relative importance of Der p 1 and Der p 2 changes with age, in favor of Der p 2. Overall, sIgE titers were positively associated with the number of organs affected.

Anaphylaxis and generalized urticaria from eating Chinese bayberry fruit

Chinese bayberry Myrica rubra is a very popular fruit in southeastern China. In spite of its wide consumption, no allergies to this fruit have been reported previously. Here we report on a 40-year-old woman suffering from anaphylaxis to Chinese bayberry fruit. Prick-prick skin tests revealed strong reactions to fresh Chinese bayberry fruits as well as to peach, and weaker reactions to some other fruits including apple, melon, and banana. ImmunoCAP analysis revealed identical titers of specific IgE (4.3 kUA/L) to peach extract and its lipid transfer protein (LTP, rPru p 3), which was confirmed by detection of a 9 kD band following immunoblotting. Immunoblot analysis with Chinese bayberry extract gave bands of 22, 45, and 90 kD, but no 9 kD band was recognized. There was also no evidence of LTP recognition for loquat (36 kD) or melon (24 kD). This first report of a severe allergic reaction to Chinese bayberry fruit in a patient with LTP-mediated peach allergy indicates that other as yet unidentified non-pollen related fruit allergens are involved in this new severe fruit allergy.

Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA

Fruit is one of the most commonly reported food allergy sources in China, and peach lipid transfer protein (Pru p 3) has been identified as the major allergen inducing systematic symptoms. Crude allergen extracts and single component allergens have been used in food allergy diagnosis and immunotherapy. Reliable and sensitive analytical methods to quantify Pru p 3 content in fruit will help to identify low-allergenic cultivars among the abundant peach genetic resources in China. In this study, we developed a sensitive sandwich ELISA method to measure Pru p 3 in peach fruit based on two monoclonal antibodies with high sensitivity and linearity. Significant variability was observed in peach cultivars, with a much higher Pru p 3 level in peach peel than in pulp. This method will be very useful to select peach varieties with low Pru p 3 content and as a diagnostic product to detect hidden allergens in processed food.

Anti-CD69 monoclonal antibody treatment inhibits airway inflammation in a mouse model of asthma

ObjectiveAirway inflammation and airway hyper-responsiveness (AHR) are principle pathological manifestations of asthma. Cluster of differentiation 69 (CD69) is a well-known co-stimulatory factor associated with the activation, proliferation as well as apoptosis of immune cells. This study aims to examine the effect of anti-CD69 monoclonal antibody (mAb) on the pathophysiology of a mouse model of asthma.MethodsA murine model of ovalbumin (OVA)-induced allergic airway inflammation was used in this study. Briefly, mice were injected with 20 μg chicken OVA intraperitoneally on Days 0 and 14, followed by aerosol provocation with 1% (0.01 g/ml) OVA on Days 24, 25, and 26. Anti-CD69 mAb or isotype IgG was injected intraperitoneally after OVA challenge; dexamethasone (DXM) was administrated either before or after OVA challenge. AHR, mucus production, and eosinophil infiltration in the peribronchial area were examined. The levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-5 (IL-5) in bronchoalveolar lavage fluid (BALF) were also assayed as indices of airway inflammation on Day 28 following OVA injection.ResultsPretreatment with DXM together with anti-CD69 mAb treatment after OVA provocation completely inhibited AHR, eosinophil infiltration and mucus overproduction, and significantly reduced BALF IL-5. However, treatment with DXM alone after OVA challenge only partially inhibited AHR, eosinophil infiltration and mucus overproduction, and did not diminish BALF IL-5. Treatment with either DXM or anti-CD69 mAb did not alter the concentration of BALF GM-CSF.ConclusionsAnti-CD69 mAb treatment inhibits established airway inflammation as effectively as DXM pretreatment. This study provides a potential alternative therapeutic opportunity for the clinical management of asthma and its exacerbation.總結目 的气道慢性炎症和高反应性是哮喘最主要的病理生理表现,CD69 是一种与免疫细胞活化、增殖及凋亡密切相关的共刺激分子,本研究旨在观察抗 CD69 单克隆抗体在哮喘小鼠模型中抑制气道炎症和高反应性的作用及机制探讨。创新点(1)与大多数关于哮喘的药物研究多为造模前提前用药以阻断炎症的进展相比,本研究将抗 CD69 单抗作用于气道已经形成的炎症,与临床上哮喘的治疗更接近;(2)抗CD69 单抗的独特作用在于它特异性地作用于气道活化的炎症细胞,与已有的哮喘药物以糖皮质激素相比,可能的全身反应少。方 法鸡卵白蛋白致敏激发制备哮喘模型(图1),Buxco 系统测试哮喘小鼠的气道高反应性(图6)。结 论抗CD69 单克隆抗体可抑制哮喘小鼠已经形成的气道炎症,其作用效果与地塞米松预处理相当。本研究为临床哮喘的急性发作治疗和管理提供了新的可能靶点。

IgE‐binding potencies of three peach Pru p 1 isoforms

SCOPE Pru p 1, the Bet v 1 homologue from peach, has been identified as a clinically relevant allergen. Three isoforms have been described, two in peach fruit (Pru p 1.0101 and Pru p 1.0201) and one in pollen (Pru p 1.0301). The present study aimed to compare their IgE-binding potencies. METHODS AND RESULTS Three Pru p 1 isoforms were cloned and expressed as soluble proteins with His-tags in Escherichia coli. Protein identity was confirmed by MS, circular dichroism, and RNAse activity. IgE-binding capacity using ELISA and ImmunoCAP was compared. Three Pru p 1 isoforms had quite similar IgE-binding potencies for 60% of the sera, but more than twofold between any two isoforms among 40% of the 47 sera. The mean IgE binding of Pru p 1.0201 was slightly higher than other two isoforms. In a sera pool, homologous ImmunoCAP inhibition was higher than other two heterologous isoforms. Individual serum with diverse IgE values of three isoforms demonstrated the higher IgE inhibition of specific isoform with higher IgE value. CONCLUSION A similar and variable pattern of IgE recognition was observed among three Pru p 1 isoforms. The two new isoforms can be used as more accurate diagnostic reagents.

Diagnosis and treatment of allergic diseases in Zhejiang Province: A cross-sectional survey

BackgroundThe specialty of allergy developed quickly in western countries because of the rapid increase of allergic diseases, whereas it developed relatively slowly in China. The prevalence of allergen sensitization and allergic diseases in Zhejiang Province of China is high and improving the medical services for these diseases is critically needed.ObjectiveTo investigate the working status of the diagnosis and treatment of allergic diseases, including doctor resources, diagnostic methods, and allergen-specific immunotherapy in patients of Zhejiang Province, and to provide instructions for the strategic development of subspecialties of allergic diseases.MethodsFirst we defined the doctors who treat allergic diseases, and designed a comprehensive questionnaire to collect personal and hospital information for these doctors. The questionnaires were distributed to hospitals with different ranks and from different areas in the province. The general condition of doctor’s resources, carryout of diagnostic methods, and allergen-specific immunotherapy were described and variations in the different specialties, hospitals, and areas were further analyzed.ResultsDoctors in their thirties with bachelor’s degrees were the mainstream for diagnosing and treating allergic diseases. The main specialties of the doctor resources were the specialties of Ear, Nose and Throat (ENT), Respirology, Pediatrics, and Dermatology. The Pediatrics specialty had a more reasonable infrastructure of doctor resources with more young doctors working in this subspecialty. The development of allergy subspecialty varied within hospitals at different levels or from different areas. The carryout of the skin prick test (SPT), serum specific IgE (ssIgE), and subcutaneous immunotherapy (SCIT) was best performed in provincial hospitals, while sublingual immunotherapy (SLIT) was prescribed most commonly in municipal hospitals. The performance of SPT and ssIgE in Hangzhou, Jiaxing, and Wenzhou areas was much better than that in other places. The performance of SCIT and SLIT was best in Wenzhou.ConclusionsOur survey revealed a very initial and unbalanced development for the allergy subspecialty in Zhejiang Province. Doctor resources for allergic diseases were mainly from the specialties of ENT, Respirology, and Pediatrics, and the performance of diagnosis and treatment was mainly focused on provincial and municipal hospitals. Continuous education of allergies could be extended to primary healthcare centers and more efforts should be directed to those areas with poor medical resources.總結目 的通过问卷调查浙江省过敏性疾病的诊断和治疗状况,包括治疗疾病的医生资源、诊断手段和特异性免疫治疗的开展情况,为将来过敏学科的发展提供真实可靠的信息。创新点国内首次通过问卷调查,明确浙江省过敏性疾病的诊治现状,以期为未来发展学科的政策提供信息。方 法设计调查问卷,定义诊治过敏性疾病的医生。问卷按照不同的行政地区划分,分发到不同级别的医院,收集诊治过敏性疾病的医生和医院的全面信息,然后进行分析。结 论调查显示浙江省的过敏学科的发展还处于起始阶段,且存在地区间的发展不平衡。诊治过敏性疾病的医生主要来自五官科、呼吸科和小儿科,过敏原的诊断和特异性免疫治疗主要集中在省市级医院。今后过敏学科的继续教育可扩展至初级医疗保健机构,并对缺乏医学资源的地区加大资源投入。

The PpLTP1 Primary Allergen Gene is Highly Conserved in Peach and Has Small Variations in Other Prunus Species

Peach lipid transfer protein (LTP1), Pru p 3.01, is a major allergen causing severe systemic reactions in peach allergic patients in Mediterranean countries and China. Significant expression variability has been reported among peach cultivars at both transcript and protein level. In this study, the allele diversity of the LTP1-encoding gene was assessed in a large set of peach cultivars by direct sequencing of the gene and its upstream region. Evolution of the LTP1-encoding genes in peach (Prunus persica) and three other Prunus species (Prunus kansuensis, Prunus mira, and Prunus davidiana) were also inferred. Sequence analysis revealed that LTP1-encoding genes are highly conserved among peach cultivars and wild peach. Three different allele sequences were obtained from 50 Prunus accessions on the basis of the upstream region of the LTP1-encoding gene and three allele specific markers were derived according to the polymorphic sites. These markers were used to test 316 peach cultivars, most of which clustered within the three main subpopulations of peach, ‘Yu Lu’, ‘Hakuho’ and landraces. The genotypic frequencies and allele frequencies in the oriental peach cultivars in these subpopulations except for ‘Yu Lu’ were found to be in Hardy-Weinberg equilibrium (P < 0.05). The dominant alleles were upLTP1-a in the ‘Yu Lu’ subpopulation and upLTP1-c in the ‘Hakuho’ subpopulation, allele upLTP1-b and allele upLTP1-c had the highest rates in the landraces, and the dominant allele in the three peach wild relatives was allele upLTP1-b. Furthermore, we found many light-responsive elements in the upstream region. Most of the polymorphic sites in Prunus species are located in the intron region. Phylogenetic analysis suggests that Prunus kansuensis KC311794 and Prunus mira KC311791 are related more closely to Prunus persica KC311795 than to Prunus davidiana KC311792 and KC311793. The allele sequences we derived on the basis of this variability were distributed unevenly, indicating the need to study allergenicity in different subpopulations and the association between allele sequences and allergenicity.

Artemisia pollen allergy in China: component-resolved diagnosis reveals allergic asthma patients have significant multiple allergen sensitization

Artemisia pollen allergy is a major cause of asthma in Northern China. Possible associations between IgE responses to Artemisia allergen components and clinical phenotypes have not yet been evaluated. This study was to establish sensitization patterns of four Artemisia allergens and possible associations with demographic characteristics and clinical phenotypes in three areas of China.

Identification of a 62-kDa major allergen from Artemisia pollen as a putative galactose oxidase

Around 20 years ago, a 60‐ to 70‐kDa protein was reported as a major allergen of mugwort (Artemisia vulgaris) pollen. This study was to identify and characterize its molecular properties.