Small interfering RNA-mediated knockdown of Notch1 in lung

Background The immunologic response to allergens mediated by T lymphocytes is an incipient key element in the pathogenesis of asthma, and Thl/Th2 balance is regarded as the core of asthma pathogenesis. Notch is a single-pass transmembrane receptor protein that regulates differentiation, proliferation and apoptosis in a broad range of cells. It is considered that the Notch signal pathway works in every stage of T cell development and differentiation. Whether the pathway of asthma pathogenesis is related to Notch1 remains unknown. This study is aimed to investigate whether the pathway of asthma pathogenesis is related to Notch1 by examining the effect of knockdown of the Notch1 gene by small interfering RNA on T cell differentiation. Methods An OVA-induced asthma mouse model was established. The expression of Notch1 in the tissue and T cells of the lung from asthmatic mice was detected by RT-PCR and Western blotting. The expression of Notch1 and cytokine interleukin （IL）-4 and interferon （IFN）-γ in activated lung T cells was detected by RT-PCR and enzyme-linked immunosorbent assay after blocking Notch1 by small interfering RNA. Results The mRNA and protein expression of Notch1 increased significantly both in the lung tissue and lung T cells of asthmatic mice （both P 〈0.05）. IL-4 decreased and IFN-y increased significantly in active lung T cells after Notch1 was blocked by Notchl-specific small interfering RNA （IL-4： （2.51±0.51） pg/ml vs 0.64±0.27） pg/ml protein; IFN-γ： （21.72±4.24） pg/ml vs （39.79±4.09） pg/ml protein, P 〈0.05）. Conclusion This study demonstrated that the Notch1 signal might play a role in the pathogenesis of asthma by its involvement in Thl/Th2 differentiation.     

IL-24 Expression at Maternal-fetal Interface and Its Roles in Trophoblast Invasion

In this study, the expression of IL-24 at maternal-fetal interface and the roles in extravillous trophoblast （the TEV-1 cell line） invasion were examined. Immunohistochemistry was used to detect the expression of IL-24 in villi and decidual tissue. The proliferation of TEV-1 cells under the effect of IL-24 was measured by MTT assay. The invasiveness of TEV-1 cells under the effect of recombinant IL-24 （rhIL-24） was examined by transwell system. Immunohistochemical detection showed that IL-24 was expressed in the villi and decidual tissue, and distributed in villous column, trophoblasts, stroma and blood vessels. The proliferation of TEV-1 cells was not inhibited by rhIL-24 of various concentrations. The examination of invasion in vitro showed that rhIL-24 could inhibit the invasion of TEV-1 cells in a concentration-dependent manner. The results suggested IL-24 could inhibit the invasion of TEV-1 cells. Therefore, IL-24 produced by maternal-fetal interface in human first trimester pregnancy may influence the invasion of trophoblasts and is involved in normal pregnancy.     

Heparin inhibits burn-induced spleen cell apoptosis by suppressing interleukin-1 expression

Background Epidermal burn injury may trigger significant apoptosis of the spleen cells,which might be caused by a burninduced systemic inflammatory reaction.Heparin has been shown to possess anti-inflammatory properties.Interleukin 1 （IL-1） is centrally important among pro-inflammatory cytokines.We hypothesized that heparin might inhibit burn-induced apoptosis in the spleen via suppression of the IL-1 pathway.Methods Burn injury was performed on IL-1 R＋/＋ （IL-1 receptor wild-type mouse） and IL-1 R-/-（IL-1 receptor knockout mouse） mice,and they were then treated with heparin,saline or IL-1 receptor antagonist IL-Ra.Apoptosis,IL-1α and IL-1β expression were assessed in the spleens and serum.Survival curve analysis was further applied to elucidate the mechanism of heparin＇s protective properties.Results Burn induced significant apoptosis （sham：3.6％±2.1％ vs.burn：28.8％±5.9％; P ＜0.001）and remarkable expression o IL-1α and IL-1β in the mouse spleens and serum.Heparin reduced the burn-induced apoptosis in the spleens （heparin treated：8.6％±3.4％,P ＜0.005）,which could be blocked by IL-1Ra.Heparin markedly decreased both IL-1α and IL-1β expression in the spleens and serum of burned mica.IL-1 R-/-mice demonstrated considerably less apoptosis in the spleens and had a higher survival rate after burns.Heparin did not significantly decrease apoptosis in the spleen and the mortality rate in IL-1 R-/-mice after burns.Conclusion Heparin inhibits burn-induced apoptosis of the spleen cells by suppressing IL-1 expression in mice.     

Interleukin- 1 gene polymorphism disease activity and bone mineral metabolism in rheumatoid arthritis

Objective To determine whether interleukin- 1α and 1β gene polymorphism is associated w ith rheumatoid arthritis disease activity and bone mineral metabolism, and whet her there is any relationship between IL- 1β and rheumatoid arthritis (RA) moti f gene. Methods IL- 1 gene polymorphisms were analyzed in 65 RA patients who met American College of Radiology (ACR) criteria and 60 controls. From genomic DNA, 2 polymo rphisms in each gene for IL1α- 889 and IL- 1β+3953 were typed by PCR- RFLP and HLA- DRB1 allele typing was also undertaken by PCR- SSOP. Some clinical and l aboratory parameters were collected. The allelic frequencies and carriage rate s were compared between RA patients and controls and between patients with acti ve and quiescent disease. Comparison was also made between IL- 1 polymorphism a nd parameters of bone mineral metabolism and between patients with the HLA- DRB1 RA motif plus IL- 1β2 and patients without the two alleles. Fisher test an d the analysis of variance was used to analyze the data. Results There was no significant difference in the frequency and carriage rate of IL- 1 α polymorphisms between RA patients and the controls. The β2/2 genotype of IL - 1β was more common in female RA patients compared with controls (P=0. 001 ). A lower carriage rate of IL- 1β2 occurred in male RA patients (P=0. 0 01). A higher carriage rate of IL- 1α2 is associated with a higher ESR (P =0. 008), HAQ score (P=0. 03), and vit- D(3) (P<0. 001), but conversely a lower SJC (p=0. 002), a lower RF (P=0. 002) and a lower BMD at the l um bar spine (P=0. 001). A higher frequency of IL- 1α1 is associated with a l ow er CRP value (P=0. 009). An increased IL- 1β2 carriage is associated with active rheumatoid disease as indicated by a higher CRP (P<0. 001), ESR ( P<0. 001) and pain score (P=0. 001) and a higher BMD at the lumbar spin e (P=0. 007), lower vit- D(3) and. Udpd/Crea level The presence of the HLA DR B1 RA motif and IL- 1β allele 2 at same time did not contribute to disease acti vity. Conclution Polymorphisms of the IL- β gene may affect the RA occurrence. Carriage of IL- 1β2 polymorphisms is associated with more active disease in RA and the presenc e of both the IL- 1α2 and the IL- 1β1 allele in RA influences bone resorption .     

Penetrability of interleukin-1β and its effect on the concentration of tumor necrosis factor-α and interleukin-6 in the aqueous humor of rabbits treated with interleukin-1β

Background Interleukin （IL）-1β may effectively decrease introcular pressure （IOP） when administered by subconjunctival injection in normal rabbit. However, IL-1β is a large molecular agent and an inflammation factor. The aim of this study was to evaluate the penetrability of IL-1β, and the concentrations of both tumor necrosis factor （TNF）-α and IL-6 in the aqueous humor of normal rabbits treated with IL-1β. Methods A total of 170 rabbits were used in the study and were assigned to several different treatment groups as follows： 125 of the rabbits were assigned to two groups. In one group, 33 rabbits were injected subconjunctivally with IL-1β and 39 were injected with saline alone. In the other group, 27 rabbits were given eye drops containing IL-1β （400 ng/ml） and 26 were given saline alone. Aqueous humor （AH） was drawn and the concentration of IL-1β within the fluid measured. The lOP was measured in another six rabbits after administration of eye drops containing IL-1β （400 ng/ml）. A further 20 rabbits were assigned to 3 groups as follows： eight untreated normal controls; six injected subconjunctivally with IL-1β; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of TNF-a in the fluid was measured. Another 19 rabbits were assigned to 3 groups as follows： seven untreated normal controls; and six injected subconjunctivally with IL-1β; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of IL-6 in the fluid measured. Measurement of cytokine concentration was by radio-immunoassay in all cases. Results The IL-1β concentration in the AH was higher in those animals in which it had been administered subconjunctivally （P 〈0.01）. The IL-1β concentration in the AH of the animals given eye drops was almost the same as that in the controls （P 〉0.05）. The administration of IL-1β in the form of eye drops had little effect upon lOP reduction. Lower TNF-α concentrations were seen in the AH after the subconjunctival administration of IL-1β, but the concentration of IL-6 was the same as in the normal controls. Conclusions IL-1β shows good corneal penetrability after subconjunctival injection into normal rabbit eyes. The lOP reduction induced by IL-1β is unlikely be associated with an inflammatory response.     

The Effectof BCG-PSN on T-cell Subsets and Cytokines in Vernal Conjunctiviti

The effects of BCG-PSN on T-cell subsets and cytokines in vernal conjunctivitis were observed. The level of total IgE was quantitatively determined before and after treatment with BCGPSN by allergen diagnostic instrument in vitro. The content of T-cell subsets of peripheral blood and cytokine were determined by using indirect immune fluorescence method, and IL-4 and INF-γ were quantified by ELISA. The results showed that the level of total IgE was substantially reduced (P＜0.01) after treatment in the BCG-PSN group. Meanwhile, CD8 was decreased, CD4 and CD4 /CD8 ratio elevated with significant differences (P＜0. 05) as compared with pre-treatment results. The changes in total IgE, CD 8 ,CD4 and CD4 /CD 8 ratio after treatment also presented significant differences (P＜0. 05) between BCG-PSN group and routine treatment group. The level of IL-4 in serum declined (P＜0. 05) after treatment in the BCG-PSN group, and INF-γ went up (P＜0.05). IL-4and INF-γ in serum showed significant differences (P＜0. 05) between two groups after treatment.It is concluded that BCG-PSN has a bi-directional immunoregulating effect. It can bring CD4 and CD 8- into homeostasis, thereby preventing the occurrence of anaphylaxis. At the same time, BCGPSN can restrain Th2, decrease the synthesis of IL-4, switch the balance of Th1/Th2 to Th1 side,boost up the predominance of Th1 relatively, which is propitious to perennial stabilization and recov cry of vernal conjunctivitis.     

Study on the Expression of IL-18, AcPL and IL-18BP mRNA in the Psoriatic Lesions

Interleukine- 1 8(IL- 1 8) isa newly identified cy-tokine that plays an important role in the T- helper(Th1 ) response,primarily via its ability to induceIFN- γ production in T cells and NK cells.IL- 1 8isrelated to the IL- 1 family in terms ofstructureand interms of function,and induction of IFN-γ by IL- 1 8is similar to that by IL- 1 2 in thatthey are a sole cy-tokine,and there is synergism between IL- 1 8andIL - 1 2 for IFN-γ production[1— 4 ] .In addition,mono-cytes and macro…     

The Expression of Interleukin-23 （p19/p40） and Inteleukin-12 （p35/p40） in Psoriasis Skin

Interleukin-12 (IL-12), which is composed of p35 and p40 subunit, is a proinflammatory Th1-inducing cy-tokine. Besides IL-12, p40 can be covalently linked to a p35-related protein p19. This heterodimer is known as IL-23[1]. IL-12 promotes the development of naive T cells, whereas IL-23 mediates late-stage inflammation. In this study, the expression of IL-12p35/p40 mRNA and IL-23p19/p40 mRNA were analyzed by using reverse translation-polymerase chain reaction (RT-PCR) in order to pro…     

Expression of CC Chemokine Ligand 20 and CC Chemokine Receptor 6 mRNA in Patients with Psoriasis Vuigaris

Itiswell knownthatimmune inflammatorymechanism playsanimportantroleinpsoriasis .Therelationshipbetweenchemokine ,receptorsandpsori asishasbeenconfirmed[1] .CCchemokineligand 2 0(CCL2 0 )isanewmemberinthefamilyof β chemokineandalsotheonlyligandofCCchemoki…     

IL-15在寻常型银屑病患者皮损中的表达

目的从mRNA及蛋白质水平研究白介素15（IL-15）在寻常型银屑病患者皮损及非皮损皮肤中的表达,探讨其在银屑病发病中的作用。方法采用RT-PCR和免疫组化法对28例寻常型银屑病患者皮损及非皮损皮肤中IL-15的表达情况进行了检测,同时以10例正常人皮肤作为对照。结果2种IL-15前体蛋白的mRNA在所有皮肤标本中都有表达,银屑病皮损中IL-15 mRNA的表达明显高于非皮损处皮肤（P〈0．001）和正常人皮肤（P〈0．001）,非皮损皮肤的表达与正常人皮肤相比差异无显著性（P〉0．05）。免疫组化结果显示：IL-15低表达于非皮损和正常皮肤的基底细胞层,而在银屑病皮损处高表达,且表达范围可达表皮全层及真皮乳头层。皮损处IL-15的表达明显高于非皮损（P〈0．001）和正常皮肤（P〈0．001）,非皮损与正常皮肤之间的表达无显著性差异（P〉0．05）。结论寻常型银屑病的皮损中高表达IL-15,在疾病的发生中可能起一定的作用。     

TIG2在寻常型银屑病中的表达变化及意义

目的 研究TIG2在寻常型银屑病中的作用机制.方法 用免疫组化和原位杂交的方法检测正常组织,未受累组织和银屑病皮损中TIG2蛋白和mRNA的表达变化.结果 在正常组织及未受累组织中,TIG2表达于表皮全层,在银屑病周围皮损中,可见TIG2阳性染色于棘层上部,在棘层下部及基底层中表达较少,而在银屑病中间皮损中无表达.TIG2在正常和未受累皮肤基底层中的表达高于其在银屑病边缘皮损基底层中的表达(p<0.01);TIG2在正常皮肤和未受累皮肤中的表达高于其在银屑病中间皮损中的表达(P<0.01);银屑病边缘皮损基底上层中的表达高于其在银屑病中间皮损中的表达(P<0.01).结论 TIG2可保持表皮正常分化功能,TIG2的降低可能参与银屑病的发生和发展.     

The Expression of Interleukin-17, Interferon-gamma, and Macrophage Inflammatory Protein-3 Alpha mRNA in Patients with Psoriasis Vulgaris

Interleukin 17(IL 17)isanewlyidentifiedpro inflammatorycytokinethatisproducedbyactivatedTcells.IL 17hastheabilitytoinducestromacellstoproduceproinflammatorycytokinesandhematopoieticcytokines .Soitmayparticipateinsomeinflammatoryreactionsandsomeimmunoloreg…     

Expression of LL-37, Human beta Defensin-2, and CCR6 mRNA in Patients with Psoriasis Vuigaris

Theinnateimmunesystemofhumanskincon tainsantimicrobial peptidesknownascathelicidins(LL 37)andhumanbeta defensins (hBD) [1] .Innor malskinthesepeptidesarenegligible ,buttheexpres sionofthemwillbetriggeredbyinjuryorinflamma tionoftheskin .Andtheirexpression…     

Expression of Plasmacytoid Dendritic Cells, IRF-7, IFN-α mRNA in the Lesions of Psoriasis Vulgaris

Psoriasis is a common and chronic inflammatory skin disease associated with both genetic and environ-mental risk factors. Recent researches have focused pri-marily on the role of innate immunity in psoriasis. Type Ⅰ interferons (IFNs), as represented by IFN-α, are promptly secreted upon the interaction between micro-bial pathogens and the immune system, and activate a variety of immune effector cells that belong to the innate as well as the adaptive immune system. Plasmacytoid dendritic ce…     

成纤维细胞生长因子10 mRNA在寻常型银屑病皮损中的表达

目的:研究成纤维细胞生长因子10 mRNA在寻常型银屑病皮损中的表达,探讨其在银屑病发病中的作用机制.方法:应用原位杂交法检测FGF10 mRNA在22例正常皮肤组织、银屑病皮损和未受累皮肤中的表达和分布.结果:原位杂交法检测发现,在22例正常皮肤组织、银屑病皮损和未受累皮肤表皮的全层或表皮中下层可见FGF10 mRNA表达(95%);在正常皮肤表皮的基底层或少量细胞内可见FGF10 mRNA的表达(5%);在寻常型银屑病非皮损表皮的中下层或个别细胞内可见FGF10 mRNA的表达(82%).寻常型银屑病皮损表皮中FGF10 mRNA的表达明显高于正常对照组(P＜0.05).结论:FGF10 mRNA在银屑病发病早期阶段的表达增高可能与银屑病的早期发病有关.     

miR-21及其靶蛋白RASA1在寻常型银屑病患者皮肤组织中的表达及意义

目的 探讨miR-21在寻常型银屑病患者皮损组织、皮损旁组织以及正常健康人群皮肤组织中的表达水平,同时研究其下游靶蛋白RASA1在寻常型银屑病患者皮损组织和正常人群皮肤组织中的表达水平及意义。 方法 采用实时荧光定量PCR法检测miR-21在12例寻常型银屑病患者皮损组织、皮损旁组织以及15例正常人群皮肤组织中的表达水平,差别倍数用2-ΔΔCt值表示。利用免疫组织化学Envision法检测45例寻常型银屑病患者与20例正常人群皮肤组织中RASA1蛋白的表达情况,RASA1阳性显色主要为细胞质显示棕黄色或棕褐色颗粒。 结果 经过实时荧光定量PCR检测后分析显示:寻常型银屑病患者皮损组织中miR-21的表达水平显著高于正常皮肤组织,差异具有统计学意义(P<0.01)。免疫组化结果表明:RASA1蛋白在正常皮肤组织中的阳性表达率为60.00%,显著高于寻常型银屑病患者皮损中的阳性表达率(10.00%),差异具有统计学意义(χ2=14.009,P<0.001)。 结论 miR-21在寻常型银屑病皮损组织中的表达高于正常组织,其靶蛋白RASA1在寻常型银屑病患者皮肤组织中的阳性表达显著低于正常皮肤组织;可能是由于miR-21的差异表达调控RASA1基因,导致RASA1蛋白表达受到抑制,从而参与指向寻常型银屑病的发病过程。