Advances in research on IL-18 in the pathogenesis of allergic airway diseases / 西安交通大学学报(医学版)

The prevalence of allergic airway diseases has been increasing in recent years. Interleukin-18 (IL-18) plays an imperative role in allergic airway diseases by binding to IL-18Rα, thereby initiating the downstream proinflammatory pathway. IL-18 also binds to IL-18BP, thus inhibiting its binding to IL-18Rα. Therefore, further understanding of the role of IL-18 and its action mechanisms in allergic airway diseases is important for the treatment of allergic airway diseases, and for the development of IL-18-related biological agents.

Establishment of allergic rhinitis mouse model sensitized by Artemisia annua and its immunological study / 中华微生物学和免疫学杂志

Objective:To optimize the BALB/c mouse rhinitis model sensitized by Artemisia annua pollen allergen, and explore the humoral and cellular immune indicators that can be used for the evaluation of allergic reactions. Methods:Using BALB/c mice as experimental animals, using Artemisia annua pollen allergen extract as sensitizing protein, through different content of the main allergen Art a1 and different sensitization times, different immunization programs were set to immunize mice subcutaneously, One week and five weeks after the last immunization, Artemisia annua pollen allergen extract containing 50 μg/ml and 500 μg/ml Art a1 was used for nasal stimulation, once a day, for 1 week each time.Observe the allergic reaction of mice, detect the pathological changes of nasal tissues, determine the levels and dynamic changes of antigen-specific IgE, IgG1, IgG2a and other antibodies in the serum of each group of mice. and detect the changes in the number of antigen-specific IL-4, IL-5, IL-2, IFN-γ and other lymphocytes in the spleen of mice. Results:Sensitized mice showed obvious scratching and sneezing reactions after being stimulated by antigen; obvious allergic inflammation appeared in nasal tissue; The increase in serum level of Artemisia annua pollen-specific IgE antibody was significantly correlated with the challenge antigen; The antigen-specific IL-4 lymphocytes in the spleen of the sensitized mice were significantly increased, but the IFN-γ-specific lymphocytes did not change significantly. Conclusions:The successful establishment of a mouse model of Artemisia annua pollen allergen allergy is the first domestic use of ELISPOT technology to detect an increase in the number of antigen-specific IL-4 lymphocytes in Artemisia annua allergy mice, laying a foundation for the subsequent evaluation of the efficacy of preparations for desensitization treatment basis.

Chinese Society of Allergy Guidelines for Diagnosis and Treatment of Allergic Rhinitis

Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.

Effects of IL-18 gene combined with diterpenoid alkaloids in inducing proliferation and promoting apoptosis of tongue squamous carcinoma cells / 西安交通大学学报(医学版)

Objective To investigate the effects of human IL-18 gene combined with diterpenoid alkaloids in inhibiting the proliferation and inducing the apoptosis of tongue squamous carcinoma cells Tscca.Methods We constructed recombinant plasmid pEGFPN3-IL-18 and tranfected it into tongue squamous carcinoma cells Tscca.The transduction efficiency of the target cells was detected by fluorescent microscopy,cytotoxic effect of IL-18 gene with diterpenoid alkaloids on Tscca was detected by MTT assay,and apoptosis was detected by flow cytometry. Western blot was employed to examine the expression level of cellular signal-regulated kinase Akt/p-Akt.Results The tongue squamous cells Tscca which transfected pEGFPN3-IL-18 had significantly increased apoptosis compared with non-transfected cells (P<0 .05 ).Tongue carcinoma squamous cells cultured with diterpenoid alkaloids at the concentrations of 0 .2 ,0 .4 and 0 .6 mg/mL had significantly increased apoptosis in a dose-dependent manner (P<0.05).Human IL-18 gene combined with diterpenoid alkaloids for 48 hours inhibited significantly Tscca in a concentration-dependent manner compared with diterpenoid alkaloids alone (P<0 .05 ).The two in combination could also decrease the protein level of p-Akt dose-dependently.Conclusion The combination of pEGFPN3-IL-18 and diterpenoid alkaloids has a synergistic effect in inhibiting the growth of tongue squamous carcinoma cells Tscca.

Effect of IL-29 on trypsin-induced protease-activated receptor expression on P815 mast cells / 中国免疫学杂志

Objective:To investigate the modulatory effect of IL-29 on trypsin-induced protease activated receptors (PARs) ex-pression on P815 mast cell.Methods:After P815 mast cells were challenged with different concentrations of IL-29 alone or combined with trypsin for 2 h, 6 h and 16 h, the challenged cells were collected and analysed by flow cytometry to detect the protein expression of PARs on P815 cells, and analysed by real time RT-PCR to detect the mRNA expression of PARs on P 815 cells.Results:Compared with the corresponding control , IL-29 induced significantly decreased expression of PAR-1 at protein and mRNA level on P815 cells, and upregulated PAR-3, PAR-4 mRNA level on P815 cells, whereas IL-29 did little effect on the expressions of PAR-2,3,4 at protein level on P815 cells accordingly.Preincubation of mast cell with IL-29 did not alter trypsin-induced PAR-1 expression on P815 cells, whereas up-regulated expression of PAR-2, 3, 4 were detected when P815 cell were pre-treated with IL-29 before being challenged with trypsin compared with the corresponding control .Conclusion: IL-29 can upregulate trypsin-induced PARs expression on mast cells through which participated in mast cell related inflammation .

Identification and purification of major allergens in Artemisia sieversiana pollen / 中国免疫学杂志

Objective:To isolate,identify and purify the Artemisia sieversiana pollen ,the mostly widespread pollen among the Artemisia pollens in China.Methods: Artemisia sieversiana extract was precipitated by saturated ammonium sulfate and then electrophoresed by SDS-PAGE.The molecular mass of each protein band was determined by gel media system.The primary allergen proteins were identified by Western blot.Allergen proteins were purified and identified by DEAE-cellulose DE-52 ion exchange chroma-tography ( IEC) and Western blot.Results: We isolated more than twenty protein bands from Artemisia sieversiana pollen extract , including the most abundant six bands whose Mr were 62 kD,57 kD,38 kD,29 kD,25 kD,14 kD espectively.The protein bands with Mr were 62 kD and 16 kD had the highest binding capacity with the specific IgE from Artemisia pollen allergic patients.The DEAE-cellulose DE-32 IEC was used to purify the primary allergen proteins with Mr 62 kD and 16 kD.Conclusion:The primary allergens of Artemisia sieversiana include the allergen proteins whose Mr are 62 kD,16 kD and the allergen of Mr 62 kD and 16 kD can be purified by chromatography.

Modulation of TNF on the expression of protease activated receptors in P815 mast mells / 中华微生物学和免疫学杂志

Objective To investigate the potential influence of TNF on the expression of protease activated receptor (PAR)-1,2,3 and 4 by using P815 mast cells. Methods After being challenged with various concentrations of TNF for 2 h, 6 h and 16 h, the P815 mast cells were treated with or without Triton X-100 and the PAR expressions were detected by flow cytometry and immunofluorescent microscopy. Results Compared with the corresponding controls, TNF concentration-dependently upregulated expressions of PAR-2 and PAR-4 both in Triton X-100-treated and the untreated groups, but had no significant effect on the expression of PAR-1 and PAR-3. Moreover, no significantly different expressions of TNF-induced PAR-1, 2, 3 were observed between Triton X-100-treated and the untreated groups, whereas Triton X-100-treated PAR-4 expressions were significantly enhanced by TNF compared with the Triton X-100-untreated ones. Conclusion TNF can up-regulate PAR-2, 4 expression of P815 mast cells but has little effect on the expression of PAR-1, 3 correspondingly. And Triton X-100 treatment had no significant effect on TNF-modulated expression of PARs in P815 mast cells.

Mutation analysis of type Ⅱ hair keratin gene in a pedigree with monilethrix / 中华皮肤科杂志

Objective To investigate the mutation of type Ⅱ human hair basic keratin (hHb) gene in a family with monilethrix.Methods Scanning electron microscopy was used to observe the structure of hair shafts.With informed consent,blood samples were drawn from affected and unaffected membets in this family,as well as from 50 healthy controls.Genomic DNA was isolated from these samples.The exon 1 and exon 7 of hHb1,hHb3 and hHb6 were amplified by polymerase chain reaction (PCR).All the PCR products were sequenced directly using ABI3730 automated sequencer.DNA sequence alignment was carried out with BLAST software.Results A typical beaded appearance was observed in affected hairs by using scanning electron microscopy.There were obvious longitudinal ridges and sulcuses in hair node.and hair cuticles were irregularly shaped.Most cortex and medullary substance were absent in affected hairs of a patient.After sequence alignment,a G1289A point mutation in exon 7 of hHb6 gene,which led to a substitution of arginine for glutamide at codon 430,was detected in affected members of this family,but not in unaffected family members or 50 unrelated human controls.No mutation was observed in exon 1 or exon 7 of hHb1 and hHb3 gene or exon 1 of hHb6 gene.Conclusion The missense mutation of R430Q is a novel mutation.which may be associated with the pathogenesis of monilethrix in this pedigree.

High levels of adiponectin in the inflammatory joint of rheumatoid arthritis contribute to elevated interleukin-6, monocyte chemoattractant proteins-1 and receptor activator for nuclear factor κB ligand expression / 中华风湿病学杂志

Objective To explore the potential role of high levels of adiponectin (AD) in the inflammatory joint of rheumatoid arthritis (RA). Methods ELISA was used to measure the levels of AD, IL-Iβ,IL-6, IL-8, TNF-α, MCP-1 and MMP-9 in the synovial fluids of RA and osteroarthritis (OA), the levels of these cytokines were tested after the synovial fibroblasts (SFLs) were stimulated with AD. Doublelabeling immunohistochemistry was used to analyze the expression of AD in RA synovium. Cytokines were measured by ELISA after SFLs were stimulated with AD. The expression of RANKL was detected by real-time PCR after MH7A were treated with AD and IL-6 ANOVA, Student's t-test, Mann-Whitney U-tese, Spearman's-test were used for statistical analysis. Results High levels of AD in RA synovial fluids were correlated with IL-6 levels. Double labeling immunohistochemistry showed that AD was localized in fibroblasts. MCP-1 and IL-6 were dramatically increased in human synovial fibroblasts following incubation with recombinant AD for 24 h. RANKL mRNA was significantly increased in MH7A after treated with AD and IL-6. Conclusion High levels of AD in the inflammatory joints of RA are likely to contribute to the high expression of IL-6, MCP-1 and RANKL, which may play an important role in the chronic inflammation, osteoclasts activation and bone erosion in RA.

The expression of adiponectin and its receptors in the inflammatory joint of rheumatoid arthritis patients / 中华风湿病学杂志

Objective To study the expression of Adiponectin (AD) and its receptors Adiponectin receptor 1 (Adipo R1) and Adipo R2 in the synovial fluids and the synovium of rheumatoid arthritis (RA). Methods ELISA was used to determine the levels of AD in 23 RA and 23 osteoarthritis (OA) patients. Real-time PCR and Western blot techniques were employed to study the expression of AD, AdipoR1 and AdipoR2 in the synovium of 10 RA and OA patients. Results It was observed that approximately twice more adiponeetin in the synovial fluids of patients with RA than with OA. Adiponectin and AdipoR1, but not AdipoR2 mRNA, were significantly expressed in synovium of RA patients in comparison with OA. Adiponectin and AdipoR1 protein were wuch more expressed in synovium from RA than those from OA. Conclusion High expression of Adiponectin and AdipoRl is likely to contribute to the formation of inflammatory joints in RA.

Expression of protease activated receptors in mast cells / 临床检验杂志

Objective To investigate the expression of protease activated receptors(PARs)in mast cells.Methods Reverse transcription polymerase chain reaction(RT-PCR),flowcytometry and immunofluorescent cell staining were used to detect the expression of PARs in mast cell lines P815 and MC/9 at the levels of protein and mRNA.Results Both the P815 and MC/9 of mast cell lines expressed PAR-1,PAR-2,PAR-3 and PAR-4 at either protein or mRNA level.Conclusion The expression of all the four PARs in mast cells were detectable,which may be of significance for the further study on the function of PARs in mast cells.

How to Search and Get the Medical Information by Internet / 中华医学教育探索杂志

There is a rapid increase in medical information in the internet,so it is necessary for medical workers to know how to search and get the useful medical information. The medical information can be obtained by some search engines and internet address.

Protease-activated receptor 1 challenges human lung epithelial cells to produce MCP-1 / 第三军医大学学报

Objective To investigate the actions of PAR1 agonists and thrombin on the secretion of monocyte chemoattractant protein (MCP)-1 from human lung epithelial cells. Methods A549 cells were cultured in a 12-well culture plate. The challenge was performed by addition of various concentrations of PAR1 agonist peptides SFLLR and its reverse peptides RLLFS, thrombin or thrombin inhibitor named hirudin into each well, respectively. After 2 h or 16 h, the reactions were terminated by removal of the supernatant from each well. A sandwich ELISA was used to determine the levels of MCP-1 in supernatants. Results Following 16 h incubation, SFLLR could induce concentration-dependent secretion of MCP-1. The maximum release of MCP-1 was nearly 12-fold more than baseline release. The reverse PAR1 agonists had little effects on MCP-1 release. Thrombin could induce concentration-dependent secretion of MCP-1. As low as 3 000 U/L thrombin could induce MCP-1 release from epithelial cells, and the maximum of accumulated release of MCP-1 was observed with 10 000 U/L thrombin, which was 5-fold more than baseline release. Thrombin inhibitor hirudin could inhibit thrombin induced secretion of MCP-1. The time course showed that the actions of PAR1 agonist peptides SFLLR and thrombin initiated at 2 h and reached their peak at 16 h. Conclusion PAR1 agonist peptides and thrombin are potent secretogogue of MCP-1 release from cultured human lung epithelial cells, and PAR1 antagonists and thrombin inhibitor may possess the ability to inhibit airway inflammation.

Effect of chymase inhibitors on histamine release from human colon mast cells / 中国药理学通报

Aim To investigate the ability of chymase inhibitors o n histamine release from human colon mast cells. Methods Human ma st cells were dispersed from colon tissue with collagenase and hyaluronidase, an d were challenged with stimulus for 15 min at 37℃.A glass fibre-based fluorome tric assay was used to measure histamine in the supernatants of dispersed mast c ells.Results chymase inhibitors ZIGPFM, TPCK and ? 1-antitry psin failed to induce significant histamine release from colon mast cells. All t he chymase inhibitors were able to inhibit anti-IgE induced histamine release i n a concentration dependent manner with a maximum of 37%, 26% and 36.8% inhibit ion being achieved with 1 mmol?L -1 of ZIGPFM, 80 mmol?L -1 of TPCK , 30 mmol?L -1 of ? 1-antitrypsin, respectively. Preincubation of inhib itors of ZIGPFM and TPCK with cells for 20 min at 37℃ before challenging with a nti-IgE was able to slightly enhance their inhibitory actions. All the chymase inhibitors were able to inhibit calcium ionophore induced histamine release, th e maximum inhibition was 23.6%～35%.And the extent of inhibition by TPCK was in creased when colon mast cells were preincubated for 20 min before calcium ionoph ore being added. However, the same treament failed to improve the action of ZIGP FM. Conclusion In the current study, we found that inhibitors o f chymase were able to inhibit anti-IgE and calcium ionophore induced histamine release from human colon mast cells, which may indicate a potential of a novel therapy for the treatment of inflammatory bowel disease or other mast cell relat ed diseases.

Detection of tryptase and its clinical implication / 医学研究生学报

Tryptase,a multifunctional inflammatory mediator,is mainly secreted by activated mast cells and can initiate the development of many diseases.Accordingly the plasma tryptase level is increased with the degranulation of the mast cells,and the detection of the changes in its serum level may provide valuable evidence for the clinical diagnosis and treatment of related diseases.

Effect of chymase inhibitors on tryptase release from human colon mast cells / 中国免疫学杂志

Objective:To investigate the ability of chymase inhibitors on tryptase release from human colon mast cells.Methods:Human mast cells were dispersed from colon tissue with collagenase and hyaluronidase,and were challenged with stimulus for 15 min at 37℃.Tryptase assay performed following previous procedures.In brief,a 96-well microtitre plate was coated with antiserum to human tryptase.The tryptase levels in the samples were detected with a monoclonal antibody specific to tryptase and the reaction was visualized by addition of OPD.Results:At 15 min and 35 min following incubation,anti-IgE and calcium ionophore were able to provoke significant tryptase release from human colon mast cells.Chymase inhibitors ZIGPFM,TPCK and ?1-antitrypsin had no stimulatory effect on colon mast cells at both 15 min and 35 min incubation periods.All the chymase inhibitors were able to inhibit anti-IgE induced tryptase release in a concentration dependent manner with a maximum of 37%,40% and 36.6% inhibition being achieved with 1 ?mol/mL of ZIGPFM,80 ?mol/mL of TPCK,30 ?mol/mL of ?1-antitrypsin,respectively.Preincubation of inhibitors of ZIGPFM and TPCK with cells for 20 min at 37℃ before challenging with anti-IgE was able to slightly enhance their inhibitory actions.Amastatin,a specific inhibitor of aminopeptidase,had no effect on anti-IgE induced tryptase release.All the chymase inhibitors were able to inhibit calcium ionophore induced tryptase release,the maximum inhibition were 23%-35.3%.And the extent of inhibition by ZIGPFM was increased when colon mast cells were preincubated for 20 min before calcium ionophore being added.However,the same treament failed to improve the action of TPCK.Conclusion:We found for the first time that inhibitors of chymase were able to inhibit anti-IgE and calcium ionophore induced tryptase release from human colon mast cells,which may indicated a potential of a novel therapy for the treatment of inflammatory bowel disease or other mast cell related diseases.

Levels of IL-18, IL-16, IL-8, eotaxin and the chymase activity in the sputum of asthmatics / 中国病理生理杂志

AIM: To investigate the levels of IL-18, IL-16, IL-8, eotaxin and the chymase activity in the sputum of asthmatics. METHODS: IL-18, IL-16, IL-8 and eotaxin levels were detected with sandwich ELISA procedures and chymase activity was determined spectrophotometrically (410 nm) by the rate of hydrolysis of N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide (SAAPP). RESULTS: The specific chymase activities in the severe and moderate asthmatics were higher than that in controls. Native protease inhibitors ?_1-antitrypsin (?_1-AT) and soybean trypsin inhibitor (SBTI) inhibited 71.9% and 72.1% enzymatic chymase activity, respectively. The levels of IL-18, IL-16, IL-8 and eotaxin were significantly elevated in the sputum of patients with acute asthma. There were correlations between the levels of IL-8 and IL-16 (r=0.55, P

Effect of leupeptin, TLCK and lactoferrin on tryptase release from human colon mast cells / 中国病理生理杂志

AIM: To investigate the ability of leupeptin, TLCK and lactoferrin on tryptase release from human colon mast cells. METHODS: Human mast cells were dispersed from colon tissue with collagenase and hyaluronidase, and were challenged with stimulus for 15 min at 37 ℃. Tryptase concentrations were measured with a sandwich ELISA procedure. RESULTS: Leupeptin, TLCK and lactoferrin were not able to provoke significant tryptase release from human colon mast cells. But they were able to inhibit anti-IgE and calcium ionophore (CI)-induced tryptase release in a concentration-dependent manner. Preincubation of inhibitors of leupeptin and TLCK with cells for 20 min before challenging with anti-IgE failed to enhance their inhibitory actions. The extent of inhibition by leupeptin was increased when colon mast cells were preincubated for 20 min before CI being added. However, the same treatment failed to improve the action of TLCK. CONCLUSION: We found for the first time that leupeptin, TLCK and lactoferrin were able to inhibit anti-IgE and calcium ionophore-induced tryptase release from human colon mast cells, which may indicate a potential of a novel therapy for the treatment of inflammatory bowel disease or other mast cells-related diseases.

The roles of IL-17,18,23,25 in the allergic diseases / 中国病理生理杂志

Many newly discovered interleukins have been implicated to play an important role in the pathogenesis of the allergic diseases, for example, human interleukin (IL)-17, a T-cell derived cytokine; interleukin (IL)-18, an interferon (IFN)-gamma-inducing cytokine; interleukin (IL)-23, produced by activated dendritic cells, and interleukin (IL)-25, a recently described T helper 2 (Th2) cell-derived cytokine. Understanding their characteristic and roles in diseases may help us to know better the mechanism of the allergic diseases and develop the strategy for treating the disease. [

Role of transforming growth factor-? in the airway inflammation and remodeling in asthma / 中国病理生理杂志

Transforming growth factor-?(TGF-?)was reported to be increased in asthma in some studies. Accumulation of TGF-? in airway promotes smooth muscle cell mitogenesis and hyperplasia, and induces fibroblast and myofibroblast and smooth muscle proliferation as well as increase in protein synthesis in connective tissue(such as collagen deposition on the reticular basement membrane). The autocrine induction of collagen expression by smooth muscle may contribute to the thickening of the reticular basement membrane, irreversible fibrosis and remodeling seen in the airways in some asthmatics. TGF-? is considered to be a major fibrogenic cytokine. It can increase smooth muscle mass and lead to severe bronchial obstruction in an asthma attack.