Huaiqihuang Granules Affect Differentiation of Th17 Cells in IgA Vasculitis Nephritis Mice by Regulating AMPK/ACC Pathway / 中国实验方剂学杂志

ObjectiveTo observe the intervention effect of Huaiqihuang granules （HQH） on immunoglobulin A vasculitis nephritis （IgAVN） mice and explore the underlying therapeutic mechanism. MethodFifty SPF-grade male Kunming mice were randomly divided into a normal group， an IgAVN model group， a dexamethasone group （2.5 mg·kg-1·d-1）， a low-dose HQH group （4 g·kg-1·d-1）， and a high-dose HQH group （8 g·kg-1·d-1）. The mouse model was established using oral administration of gliadin combined with intravenous injection of India ink. After successful modeling， the mice were euthanized after 4 weeks of gastric gavage according to groups. The 24 h urinary total protein （24 h UTP）， urine β2-microglobulin （β2-MG）， serum total protein， albumin， IgA， etc. were detected in each group. Flow cytometry was used to determine the proportion of T helper 17 （Th17） cells in spleen cell suspension. Western blot was employed to detect the expression of adenosine 5'-monophosphate-activated protein kinase α （AMPKα）， phosphorylated AMPKα （p-AMPKα）， acetyl-CoA carboxylase 1 （ACC1）， and phosphorylated ACC1 （p-ACC1） in Th17 cells. Pathological changes in the spleen and kidneys were observed. ResultCompared with the normal group， the IgAVN model group showed significant increases in 24 h UTP， urine β2-MG， total cholesterol （P<0.05）， serum interleukin-17 （IL-17）， IgA， Th17 proportion in the spleen cell suspension， and IL-17 expression in the spleen tissue （P<0.01）， and significantly decreased serum total protein， albumin， p-AMPKα/AMPKα， and p-ACC1/ACC1 expression of Th17 cells （P<0.01）. Compared with the IgAVN model group， in the 4th week， the 24 h UTP， urine β2-MG， serum IL-17， IgA levels， and renal IgA deposition were significantly reduced in each treatment group （P<0.01）， and the Th17 proportion and IL-17 expression in spleen tissue were significantly decreased （P<0.05， P<0.01）. Serum albumin levels significantly increased （P<0.05）. Compared with the IgAVN model group， the dexamethasone group and the high-dose HQH group showed increases in serum total protein （P<0.01）， p-AMPKα/AMPKα， and p-ACC1/ACC1 expression of Th17 cells （P<0.05， P<0.01）. The high-dose HQH group showed a significant decrease in total cholesterol level （P<0.05）. Various treatment groups showed different degrees of improvement in spleen and kidney pathological changes. ConclusionHQH may affect Th17 cell differentiation by regulating the AMPK/ACC pathway， correcting immune inflammatory disorders， and exerting therapeutic effects on IgAVN.

Effects of allergens on the expression of IL-18, IL-18-binding protein a and IL-18 receptor α by peripheral blood CD4 + Th17 cells of patients with allergic rhinitis / 中华微生物学和免疫学杂志

Objective:To investigate the expression of IL-18, IL-18-binding protein a(IL-18BPa) and IL-18 receptor α(IL-18Rα) by peripheral blood CD4 + Th17 cells of patients with allergic rhinitis (AR) and the effects of allergens on their expression. Methods:This study enrolled 45 outpatients with AR and 23 healthy control subjects receiving physical examination in the First Affiliated Hospital of Jinzhou Medical University from October 2019 to September 2020. According to the results of skin prick test, the 45 patients were divided into two groups: AR group with positive results (24 cases) and nAR group with negative results (21 cases). Blood samples of them were collected. Flow cytometry was used to analyze the effects of allergens on the expression of IL-18, IL-18BPa and IL-18Rα at protein level by peripheral blood CD4 + Th17 cells. The level of IL-17A in plasma was measured by Bioplex system, and its correlation with the percentage of IL-18 + Th17 cells was analyzed. Results:Compared with the healthy control group, the AR group showed increased ratios of CD4 + Th17 and IL-18 + Th17 cells ( P<0.01), decreased ratio of IL-18BPa + Th17 cells ( P<0.01), enhanced mean fluorescence intensity (MFI) of IL-18BPa ( P<0.01) and reduced MFI of IL-18Rα ( P<0.01); the nAR group showed enhanced MFI of IL-18BPa ( P<0.000 1) and reduced MFI of IL-18Rα ( P<0.000 1). The ratio of IL-18 + Th17 cells and the MFI of IL-18Rα in the AR group were higher than those in the nAR group ( P<0.05, P<0.01). House dust mite extract and Platanus pollen extract induced the expression of IL-18 and IL-18BPa by CD4 + Th17 cells of AR patients ( P<0.05). Moreover, house dust mite extract directly induced the CD4 + Th17 cells isolated from the healthy control subjects to express IL-18 and IL-18R ( P<0.05). Compared with healthy control subjects, AR patients had higher level of IL-17A in plasma and it was moderately correlated with the ratio of IL-18 + Th17 cells ( P<0.05). Conclusions:Allergens may be involved in the pathogenesis of AR by inducing blood CD4 + Th17 cells to express IL-18 and IL-18Rα.

To Explore the Effect And Mechanism of Traditional Chinese Medicine on Chronic Urticaria Based on Th17/Treg Balance Axis / 世界科学技术-中医药现代化

Immune imbalance is believed to play a dominant role in the pathogenesis of chronic urticaria.While Th1/Th2 imbalance used to be considered as the main contributing factor of the development of chronic urticaria.Recently,Th17/Treg imbalance is found to be an important immune mechanism leading to the development of chronic urticaria.To be more specific,according to traditional Chinese medicine(TCM)'s comprehensive understanding of the etiology of chronic urticaria.it is generally believed that the pathogenesis of chronic urticaria is due to a lack of innate endowment,a lack of solidity of the body surface,and repeated exposure to six pathogenic factors.Another possible reason lies with dietary disorders that generate heat and wind or chronic illness and weakness and loss of nourishment of qi and blood.Therefore,in terms of the treatment,from the perspective of sthenia syndrome,it is advisable to remove the wind and disperse the pathogenic factors and clear the dampness and heat.From the perspective of asthenia syndrome,it is advisable to nourish the qi and blood and support the righteousness.As for mixed excess and deficiency,both support the healthy atmosphere and dispel the pathogenic factors are important.Regarding the effects of TCM on the balance of Th17/Treg in chronic urticaria and immune diseases,it mainly involved herbal compounding,herbal active ingredients and single herbs.However,the research attention has been drawn to investigating the role of TCM in the treatment of chronic urticaria and various immune diseases based on the research outcomes in modern pharmacological research.This can not only provide scientific evidence for TCM treatment of chronic urticaria,but also bring benefits to more patients with immune diseases.Therefore,the author reviews the recent research progress of TCM on the effects of Th17/Treg immune imbalance in chronic urticaria and other immune diseases by explaining the effects of Th17 and Treg cells in chronic urticaria.

Study on the mechanism of Wenyang huazhuo tongluo formula contained serum inhibiting the proliferation of Th17 cells in scleroderma patients / 中国药房

OBJECTIVE To study the effects of Wenyang huazhuo tongluo formula conta ined serum on the expression and methylation of retinoic acid related nuclear orphan receptor （RORγt），and to explore the mechanism of its regulation of Th 17 cell proliferation in the treatment of scleroderma. METHODS Wistar rats were given 15，30，60 g/（kg·d）Wenyang huazhuo tongluo formula intragastrically to prepare different doses of drug-contained serum ，and peripheral blood of scleroderma patients were collected to sort Th 17 cells. Using blank serum as blank control ，methyltransferase inhibitor decitabine （DCA）as positive control ， Th17 cells were treated with different concentrations of drug-contained serum ，and then the proliferation of Th 17 cells was detected by CCK- 8；mRNA expressions of RORγt and IL-17 were detected by qRT-PCR ，and the protein expression of RORγt was detected by Western blot assay ；the protein expression of IL- 17 in the cell supernatant was detected by ELISA ，and the methylation level of RORγt gene promoter was detected by methylation-specific PCR. The transcriptional activity of RORγt gene promoter was detected by dual-luciferase assay. RESULTS Compared with blank control ，Wenyang huazhuo tongluo formula contained serum and DCA could inhibit the proliferation of Th 17 cells（P＜0.05），reduced the mRNA and protein expressions of RORγt and IL-17，enhanced the methylation level of RORγt gene promoter and attenuated ， the transcription activity of RORγt gene promoter. Except for mRNA expression of Th 17 and the methylation level of RORγt gene promoter in drug-contained serum low-dose group ，there were statistical significance in above indexes of other groups（P＜0.05）. CONCLUSIONS Wenyang huazhuo tongluo formula can promote the methylation lev el of RORγt gene，and inhibit the expression of RORγt and the secretion of IL-17，so as to inhibit the proliferation of Th 17 cells.

Effect of Asiaticoside on Expression of Th17/Treg Cells in DBA/1 Mice with Collagen-Induced Arthritis / 中国实验方剂学杂志

ObjectiveTo observe the effect of asiaticoside （AC） on the expression of T helper 17 （Th17） cells and regulatory T （Treg） cells in DBA/1 mice with collagen-induced arthritis （CIA）. MethodMale SPF DBA/1 mice were randomized into six groups according to body weight： control group， CIA group， methotrexate group （MTX group， ip， 0.5 mg·kg-1）， and AC low-， medium-， and high-dose groups （ig， 5， 15， 45 mg·kg-1， respectively）. Modeling was performed in rats other than the control group. To be specific， they were immunized with bovine type Ⅱ collagen and complete Freund's adjuvant on the first day and with bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21st day. Administration began on the day of the second immunization， once a day for 28 days. On the 49th day， related tissues were collected. Then， hematoxylin-eosin （HE） staining was performed to observe the pathological changes of the joints. Immunohistochemical method was used to detect the expression of interleukin-17 （IL-17） and forkhead box protein-3 （FoxP3）， the markers of Th17 and Treg cells， respectively， immunofluorescence double staining the expression of IL-17 and FoxP3 in CD4+T cells of mouse joint tissue， and flow cytometry the proportions of Th17 and Treg cells in mouse lymph nodes. ResultCompared with the control group， CIA group demonstrated joint disorder， damage of articular cartilage and bone， severe bone erosion （P<0.01）， increase in stained CD4 and IL-17 and the integral absorbance （IA） （P<0.01）， decrease in stained FoxP3 and the IA （P<0.01）， rise of Th17/Treg ratio （P<0.01）， elevation of Th17 expression in mouse lymph nodes （P<0.01）， and reduction in Treg expression （P<0.01）. Compared with CIA group， MTX group and three AC groups showed normal joints， alleviated bone erosion and damage， intact and smooth joint surface， and decrease in stained IL-17 and IA （P<0.05， P<0.01）， and MTX group and AC medium-dose and high-dose groups registered decrease in stained CD4 and IA （P<0.01） and reduction in Th17/Treg ratio （P<0.05， P<0.01）. Moreover， AC medium-dose and high-dose groups showed rise in stained FoxP3 and IA （P<0.05， P<0.01）. In the lymph nodes of mice， decrease in expression of Th17 cells （P<0.05， P<0.01） and the increase in expression of Treg cells （P<0.05， P<0.01） were observed in all the three AC group. ConclusionAC can regulate Th17/Treg balance by inhibiting the expression of Th17 cells and promoting the expression of Treg cells in CIA mice.

Aging weakens Th17 cell pathogenicity and ameliorates experimental autoimmune uveitis in mice

Aging-induced changes in the immune system are associated with a higher incidence of infection and vaccination failure. Lymph nodes, which filter the lymph to identify and fight infections, play a central role in this process. However, careful characterization of the impact of aging on lymph nodes and associated autoimmune diseases is lacking. We combined single-cell RNA sequencing (scRNA-seq) with flow cytometry to delineate the immune cell atlas of cervical draining lymph nodes (CDLNs) of both young and old mice with or without experimental autoimmune uveitis (EAU). We found extensive and complicated changes in the cellular constituents of CDLNs during aging. When confronted with autoimmune challenges, old mice developed milder EAU compared to young mice. Within this EAU process, we highlighted that the pathogenicity of T helper 17 cells (Th17) was dampened, as shown by reduced GM-CSF secretion in old mice. The mitigated secretion of GM-CSF contributed to alleviation of IL-23 secretion by antigen-presenting cells (APCs) and may, in turn, weaken APCs' effects on facilitating the pathogenicity of Th17 cells. Meanwhile, our study further unveiled that aging downregulated GM-CSF secretion through reducing both the transcript and protein levels of IL-23R in Th17 cells from CDLNs. Overall, aging altered immune cell responses, especially through toning down Th17 cells, counteracting EAU challenge in old mice.

Effect of mechanical stimulation by rubbing abdomen by interfering with Th17 cell function on intestinal homeostasis in IBS-C model mice / 国际生物医学工程杂志

Objective:To investigate the possible mechanism of rubbing abdomen to regulate intestinal homeostasis in irritable bowel syndrome with constipation (IBS-C) mouse models.Methods:IBS-C mouse models of intestinal immune dysfunction were established using trinitrobenzene sulfonic acid (TNBS)-induced C57BL/J6 male mice. Thirty C57BL/J6 mice were randomly divided into three groups: the control group, the model group, and the mogul group. After 7 days of modeling, mice in the mogul group were given a mogul mechanical stimulation intervention once per day for 2 weeks, while mice in the control and model groups were not given any intervention. Changes in serum levels of interleukin-6 (IL-6) and IL-17A were detected by enzyme-linked immune sorbent assay (ELISA) and immunohistochemistry. The gene expression and protein levels of IL-17A and IL-23 were detected by quantitative real-time fluorescence PCR (qRT-PCR) and Western Blot, respectively. The morphological changes were observed by HE staining. The CD44 and CD62L expression changes were observed by immunofluorescence staining.Results:Compared with the model group, the levels of IL-6 and IL-17A in the serum of mice in the mogul group were decreased, and the expression of IL-6 and IL-7A in the tissues was down-regulated (all P<0.001). In addition, the gene expression and protein expression levels of IL-17A and IL-13 in the tissues of mice in the mogul group were decreased (all P<0.001). HE staining results showed that the mogul mechanical stimulation intervention could repair colonic tissues and reduce the inflammatory response. Immunofluorescence staining results showed that mogul mechanical stimulation intervention could downregulate the expression of CD44 but had no modulating effect on the expression of CD62L. Conclusions:Rubbing abdomen can improve intestinal homeostasis in IBS-C model mice by regulating changes in Th17 cell function.

Advances in mechanisms for psoriasis and drug regulation / 药学学报

Psoriasis, bringing great trouble to patients' life, is a common chronic immune skin disease which is relapsed and difficult to cure. Further understanding of the pathogenesis of psoriasis will be benefit for potential drug targets and the development of therapeutic drugs. In recent years, "interleukin-23/T helper17 (IL-23/Th17) cell axis" has attracted more and more attention in the pathogenesis of psoriasis. Some drugs targeting the cell axis have also been successfully listed in the market to improve psoriasis and achieved good results. This review focuses on the "IL-23/Th17 cell axis" system to elaborate the role of a variety of immune cells involved in the pathogenesis of psoriasis, such as dendritic cells, macrophages, neutrophils and T cells, and summarizes multiple therapeutic antibodies targeting this cell axis in the treatment of psoriasis. In addition, this review also summarizes the current small molecule drugs for the treatment of psoriasis.

Primary study on the effect of glucose on mouse CD4+ T cell differentiation and its mechanism / 上海交通大学学报（医学版）

Objective :To study the effect of glucose on mouse CD4+ T cell differentiation. Methods :Mouse naïve CD4+ T cells cultured in the regulatory T cell (Treg), Th1, Th17 or Th2 differention condition were treated with different concentrations of glucose for 5 days. Treg, Th1, Th17 or Th2 percentages were measured by flow cytometry. Quantitative real-time PCR was used to detect the gene expressions of related cytokines and transcriptional factors. Results :The proportions of Treg and Th2 as well as the gene expressions of transforming growth factor-β, interleukin-4 (IL-4) and IL-13, and transcriptional factors, Foxp3 (forkhead box P3) and Gata3 (GATA binding protein 3), were increased significantly with the treatment of increasing concentration of glucose. On the contrary, with the glucose treatment, the percentages of Th1 and Th17 were reduced, and the gene expressions of the related cytokines and cytokine receptors, such as interferon-γ, IL-17A, IL-17F, IL-22 and IL-23R, and the related transcriptional factors, Tbx21 (T-box transcription factor 21) and RORC (RAR related orphan receptor C), were decreased consistently. Conclusion :Glucose promotes Treg and Th2 differentiation while inhibits Th1 and Th17 differentiation in vitro.

Clinical Efficacy of Taohua Tang and Buzhong Yiqi Tang in Treatment of Deficiency Syndrome of Crohn's Disease / 中国实验方剂学杂志

Objective:To observe clinical efficacy of Taohua Tang and Buzhong Yiqi Tang on Crohn's disease (CD) at active phase (deficiency-cold in spleen and stomach), in order to observed its effect on Th1 and Th17 cytokines. Method:According to random number table, 86 patients with CD were divided into control group (42 cases) and observation group (44 cases). The control group (mild) was given SASP, 3-4 g·d-1, Po, tid. The control group (moderate or poor efficacy of SASP) was given prednisone acetate, 0.75 mg·kg-1·d-1, Po, tid. Observation group was given Taohua Tang and Buzhong Yiqi Tang in addition to therapy of the control group, 1 dose·d-1. The course of treatment was 12 weeks. Before and after treatment, Best CDAI, SES-CD, IBDQ and deficiency syndrome were scored, and levels of CRP, ESR, ALB, HB, PLT, IFN-γ, TNF-α, IL-2 and IL-17 were measured before and after treatment. Result:After treatment, the effect of traditional Chinese medicine(TCM) syndromes in the observation group was better than that in the control group (Z=2.058, PPPPZ=2.112, PZ=2.288, PPPγ, TNF-α, IL-2 and IL-17 levels in the observation group were lower than those in the control group (PConclusion:In addition to the therapy of conventional western medicine, Taohua Tang and Buzhong Yiqi Tang in treatment of deficiency syndrome of Crohn's disease (CD) can control the activity degree of the disease, reduce the degree of illness and inflammation, and improve the remission rate and the quality of life, with a better clinical efficacy than the pure western medicine therapy.

Clinical Efficacy of Shensuyin in Treating Adult Viral Myocarditis with Syndrome of Insufficiency of Lung-Qi / 中国实验方剂学杂志

Objective: To investigate the efficacy of Shensuyin in treating viralmyocarditis (VMC) with syndrome of insufficiency of lung-qi and the effect on levels of Th17 and Treg cells and relevant factors. Method: One hundred-four VMC cases were regaded as object of study and randomly divided into control group and observation group, with 52 cases in each group. Control group was treated with routine therapy by reference to ‘Chinese Guidelines for Diagnosis and Treatment of Heart Failure 2014’. In addition to the therapy of control group, observation group on the basis of treatment in the control group with Shensuyin, 1 dose/d, bid. One course of treatment was 8 weeks for both groups. Scores of Shensuyin, serum levels of Troponin I (cTnI) and cardiac free fatty acid binding protein (H-FABP), heart function and total efficacy were compared for both groups. Flow cytometry was used to detect peripheral blood levels of Th17 and Treg cells for the two groups. Serum levels of interleukin (IL) -17, IL-21, IL-10 were detected in both groups. Result: After treatment, scores of syndrome of Yin and Yang deficiency (shortness of breath, fatigue, dull chest pain, bad breath, cough) of observation group were obviously lower than those of control group (PPPPPConclusion: In addition to the routine therapy of VMC, the efficacy of Shensuyin has a significant effect in treating VMC with syndrome of lung qi deficiency, and the regulatory effect on levels of Th17 and Treg cells and relevant factors may be one of the effective ways.

Expressions of Treg/Th17 cells in post liver transplantation acute rejection in pediatric recipients diagnosed with biliary atresia / 中华肝胆外科杂志

Objective To study the expressions of regulatory T cell (Treg) and T helper cell 17 (Thl7) in post liver transplantation (LT) acute rejection in pediatric recipients diagnosed with biliary atresia.Methods 12 pediatric recipients with post-LT acute rejection in Tianjin First Center Hospital from July 2016 to April 2017 were included in the rejection group,and 22 patients with normal graft functions for more than 1 year post-LT were included in the stable group.The pre-LT primary disease in all the recipients was biliary atresia with cholestatic cirrhosis.Ten healthy children were included in the control group.Peripheral blood samples were taken before and at 1 week after anti-rejection treatment in the rejection group.Blood samples were taken during follow-up in the stable group and in the normal control.The proportions of Treg cell and Th17 cell were detected by flow cytometry.Results The proportions of Treg cells in the rejection group and the stable group were both significantly lower than the control group (P＜0.05).There was no significant difference between the rejection group and the stable group (P＞0.05).The proportion of Th17 cells in the rejection group was significantly higher than the stable group (P＜0.05) and the control group (P＜ 0.05),respectively;and the proportion in the stable group was significantly higher than the control group (P＜0.05).Meanwhile,there was also significant increase in the proportion of CD3+CD8+CD28+T in the rejection group (P＜0.05).After anti-rejection therapy,the proportions of Treg cell and Th17 cell became significantly lower than before therapy (P＜0.05).Conclusions The balance in Treg/Th17 cells occurred in the acute phase of rejection post-LT in pediatric recipients with biliary atresia,which persisted into the early period after anti-rejection therapy.

Mannan-binding lectin regulates Treg/Th17 cell balance in mice with Candida albicans infection / 中华微生物学和免疫学杂志

Objective To investigate the regulation of Treg/Th17 axis by mannan-binding lectin (MBL) in mice with Candida albicans ( C. albicans ) infection. -ethods MBL gene-knockdown (MBL-/-) C57BL/6 mice and wild-type (WT) C57BL/6 mice were divided into four groups. C. albicans strains (2×107 CFU) were injected intraperitoneally into the mice of infection groups. Paraffin sections of mouse liver and kidney tissues were prepared on 3 d. Histopathological changes were observed with hematox-ylin and eosin ( HE) and Periodic acid-Schiff ( PAS) staining. Flow cytometry was performed to analyze Th17 and Treg cells in mice on 7 d. The levels of IL-10 and IL-17A were measured by ELISA. CD4+T cells were obtained from spleen cells by magnetic sorting. Expression of Foxp3 and RORγt at mRNA and protein levels were detected by qRT-PCR and Western blot, respectively. Results The mouse model of C. albicans infection was established successfully. After infection, the MBL-/- mice had higher percentages of Th17 cells, but lower percentages of Treg cells than the WT mice. ELISA results showed that compared with the WT mice with C. albicans infection, the MBL-/- mice had significantly increased IL-17A and decreased IL-10 after infection. Moreover, the expression of RORγt at both mRNA and protein levels was up-regula-ted, while that of Foxp3 was down-regulated in the MBL-/- mice than in the WT mice following infection. Conclusions MBL regulates the immune balance of Treg/Th17 cells in mice infected with C. albicans through promoting the differentiation of CD4+ T cells into Treg cell subsets and inhibiting the differentiation into Th17 cell subsets.

The role of intrahepatic regulatory T cells/helper thymphorytes 17 balance in patients with different phases of chronic hepatitis B virus infection / 中华传染病杂志

Objective To analyze the change of intrahepatic regulatory T cells (Treg )/helper thymphorytes (Th)17 balance in patients with different phases of chronic hepatitis B virus (HBV ) infection ,and to explore the role of Treg/Th17 balance in maintaining immune tolerance and inducing immune clearance ,and its influence on disease progression .Methods Sixty-eight patients with chronic HBV infection who underwent liver biopsy in Tianjin Second People′s Hospital were included .The 68 patients included 20 cases in immune tolerant (IT) phase ,36 cases in immune clearance (IC) phase and 12 cases in inactive phase .Eight healthy liver transplant donors were collected as healthy controls .The intrahepatic Treg/Th17 levels were detected by immuno-histochemical method . The changes of Treg/Th17 balance in patients with different phases of chronic HBV infection ,and the relationship between Treg/Th17 balance and the decreases of hepatitis B surface antigen (HBsAg ) , hepatitis B antigen (HBeAg) and HBV DNA levels in the peripheral blood were analyzed in patients with IC phase at two weeks of admission .Results The intrahepatic Treg and Th17 levels in IC phase group were the highest , then and they were higher in inactive phase group were higher than those in IT phase group ,And they were the lowest in control group .The Treg level in IC phase group increased significantly compared with the other three groups (all P< 0 .01) ,and there were no significant differences among the other three groups (all P> 0 .05) .The Th17 level between IT phase group and inactive phase group was not significantly different (P> 0 .05) ,while the differences were not significant in other groups (all P>0 .05) .Treg/Th17 ratio of IT phase group was the highest ,then the ratio of control group was higher than that of inactive phase group ,and IC phase group was the lowest ratio .The differences between IC phase group ,control group and IT phase group were significant (all P< 0 .05) ,and the difference between inactive phase group and IT phase group was also significant (all P<0 .05);and there was no significant difference among other groups (all P>0 .05) .The decreases of HBsAg ,HBeAg and HBV DNA levels in the peripheral blood at two weeks admission were negatively correlated with the intrahepatic Treg cell level in patients in IC phase of chronic HBV infection ( r= -0 .941 ,-0 .869 ,and -0 .883 ,respectively ,both P<0 .01) .The Treg ,Th17 levels and their ratio in IC phase group with different degree of inflammation and fibrosis had significant differences :G4 group > G3 group > G2 group ,S3 group > S2 group > S1 group (all P<0 .05) .Conclusions There is no change of the Treg/Th17 balance in IT phase ,and Treg has no influence on maintaining immune tolerance in chronic HBV infection .T he imbalance of Treg/Th17 is observed in IC phase .Th17 may actively participate in the immune-mediated liver injury and the development of hepatic fibrosis in CHB patients .Treg may inhibit inflammation and reduce liver injury via the negative feedback regulation mechanism ,and may impede the eradication of HBV simultaneously .

Exclusive enteral nutrition's moderating effect on Th17 cell of juvenile rat with inflammatory bowel disease / 国际儿科学杂志

Objective Through the TNBS-induced inflammatory bowel disease model in young rats,to observe the level of Th17 cell related cytokine IL-17A on the intestinal tissue of IBD young rats model after feeding EEN,Peptisorb,and investigate the enteral nutrition's regulative effects on Thl7 cells.Methods Thirty six four-week healthy male Sprague-Dswley rats were selected and divided into 4 groups,blank control group and EEN control group with 8 rats in each group,IBD group and IBD-EEN group with 10 rats in each group:blank control group and EEN control group were given normal saline enema,then fed with normal diet and enteral nutrition separately;IBD group and IBD-EEN group were given 5％ TNBS enema,establish the IBD model,then fed with normal diet and enteral nutrition separately.The weights,defecate characters and occult blood situations of all rats were determined.Also,we carried out the activity index score of disease according to the Cooper score method.On the seventh day,all rats were sacrificed and the colon specimens were collected.We observed mucosal injury by HE staining and expression level of Thl7 cell related cytokine IL-17A that were detected by immunohistochemical staining.Results Compared with IBD group,IBD-EEN group was improved significantly in terms of defecate character and occult blood situation.Both IBD group and IBD-EEN group gained their body weights than before,(39.95 ± 2.929) g and (38.40 ± 4.545) g respectively after 7 days (P≥0.05).In IBD group,gross tissue and pathological section displayed significant intestinal inflammation change,showing typical inflammatory bowel disease change;under the microscope,intestinal mucosa showed a large amount of inflammatory cell infiltration,a part of section displayed inflammatory granuloma formation,normal glandular structure was damaged.After EEN treatment,DAI scores werr decreased significantly;intestinal inflammation changes were relieved;intestinal mucosa inflammatory cell infiltration were reduced.Immunohistochemical study showed that the expression of IL-17A cytokine in the intestinal mucosa of IBD group was higher than that of IBD-EEN group (P ＜ 0.05).Conclusion The exclusive enteral nutrition treatment possessed the effect of reducing IBD young rat's intestinal mucosal inflammation,the inner mechanism may be related with the changing expression of Th17-cell cytokines,IL-17A.

Role of Notch-Dll4 signaling pathway in autoimmune damage of Hashimoto thyroiditis / 中华内分泌代谢杂志

Forty patients with Hashimoto thyroiditis ( HT) and 20 healthy subjects with matched age-and sex-features ( NC) were selected. The patients with HT were further divided into normal thyroid function ( HT-A) and hypothyroidism ( HT-B) groups. Real-time PCR was performed to evaluate the expressions of Notch1, Dll4, and retinoid-related orphan receptor ( ROR )-γt mRNA. Flow-cytometry was used to detect the percentage of Th17 cells. Thyroid function, thyroid peroxidase antibody ( TPOAb) , and thyroglobulin antibody ( TgAb) were detected by electrochemiluminescence immunoassaies. The results showed that the Notch1, Dll4, ROR-γt mRNA levels and Th17 cell percentage were significantly increased in HT group compared with NC group (all P<0.01), especially in HT-B group. In HT patients, Notch1 and Dll4 mRNA expression levels were positively correlated with Th17 cell percentage and its transcription factor ROR-γt ( all P<0.01) . Besides, there were significantly positive correlations of Notch1 and Dll4 mRNA expressions with TPOAb and TgAb titers (P<0.05 or P<0.01). These results suggest that Notch1-Dll4 signaling pathway might be involved in the pathogenesis of thyroid-specific autoimmune damage by regulating Th17 cells in HT patients.

Expression difference of Treg, Th17 cells and related cytokines between pre and post laparoscopic treatment of ovarian endometrioma / 中华内分泌外科杂志

Objective To investigate the changes and clinical significance of Treg and Th17 cells in patients with ovarian endometrioma (OEM) pre-and post-laparoscopy.Methods 36 patients with OEM undergoing laparoscopic surgery and confirmed by the pathology were enrolled as the experiment group,and 25 patients with fallopian tube obstruction who received laparoscopic examination were enrolled as the control group.The peripheral blood samples were collected from the control group and the experiment group before operation as well as from the experiment group 3 months after operation.Intraoperative peritoneal fluid were also collected from both groups.The levels of IL-17,IL-22,IL-23,IL-10 and TGF-β in serum and peritoneal fluid were detected by ELISA.The proportion of Treg and Th17 cells in peripheral blood was detected by flow cytometry.Results Compared with the control group,the levels of IL-17,IL-22 and IL-23 in the peritoneal fluid and serum of the experiment group increased significantly,while the levels of IL-10 and TGF-β were significantly decreased (P＜0.05).The proportion of Th17 cells in the peripheral blood of the experiment group increased significantly,and the proportion of Treg cells decreased significantly (P＜0.05).Compared with pre-laparoscopic treatment,the serum IL-17,IL-22 and IL-23,and the proportion of Th17 cells in peripheral blood were significantly decreased in the experiment group,and the levels of IL-10 and TGF-β in serum increased significantly (P＜0.05).Conclusions In patients with OEM,Treg and Th17 cell imbalance and the expression of related cytokines in disorder may be an important factor in the occurrence and development of disease.Detection of Treg/Th17 cells and their cytokines plays an important role in evaluating the severity of the disease and judging the efficacy of laparoscopic treatment and prognosis of OEM.

The correlation between STAT3 phosphorylation and the activity of Behcet's disease / 复旦学报（医学版）

Objective To explore the role of signal transducer and activator of transcription 3 (STAT3) phosphorylation in the pathogenesis of Behcet's disease (BD),and to investigate the association between STAT3 phosphorylation and disease activity in BD patients.Methods Peripheral blood mononuclear cell (PBMC) was isolated from 15 mL peripheral boood of 10 active BD patients (BD-A),10 BD patients in remission (BD-R) and 10 healthy controls (HC) respectively.The blockade of STAT3 phosphorylation was performed by Stattic.The PBMC was divided into Stattic subgroup (treated with 2.5 μmol/L stattic and 1 640 medium,5 mL) and blank control subgroup (treated with 5 mL 1 640 medium),respectively.The protein levels of phosphorylated STAT3 (pSTAT3) and STAT3 were examined by flow cytometry and Western blot.The protein and mRNA levels of TNF-α,IFN-γ and IL-17 were tested by RT-PCR and ELISA.Two-way ANOVA and Bonferroni post hoc test were used to analyze the results.Results Compared with HC,the BD patients showed higher protein levels of pSTAT3 and STAT3,and higher protein and mRNA levels of TNF-α,IFN-γ and IL-17;compared with blank control subgroup,the protein levels of pSTAT3 and STAT3,and protein and mRNA levels of TNF-α,IFN-γ and IL-17 decreased in Stattic subgroup.In the BD-A group,the protein level of pSTAT3,and protein and mRNA levels of TNF-α,IFN-γ and IL-17 were significantly higher than those in the BD-R group.Conclusions An increased activation of the STAT3 pathway may contribute to the pathogenesis of BD and relate to disease activity in BD patients by inducing TH1 and Th17 cells activation.

The role of interleukin-17A in acute paraquat induced lung injury in mice / 中华急诊医学杂志

Objective To investigate the role of interleukin-17A (IL-17A) in acute paraquat (PQ)-induced lung injury in mice.Methods A total of 120 healthy SPF grade ICR male mice were randomly (random number) divided into three groups (n =40 in each):normal saline control group (NS),PQ poisoning group (PQ) and antibody neutralization group (PQ + Ab).Mice of PQ group and PQ + Ab group were given 5 mg/mL PQ by one gavage in a dosage of 25 mg/kg body weight,and 5 μg IL-17A neutralizing antibody intra-peritoneally administered immediately after PQ poisoning in PQ + Ab group;Equivalent volume of normal saline instead of PQ was given to mice of NS group.Six survival mice from each group were taken for experiment at 8 h,1 d,3 d,5 d,7 d after PQ poisoning:Wet to dry ratio (W/D) of lung was determined in mice of each group.HE staining of lung tissue was used to observe the histopathological changes under the light microscope and the pathological scores were graded;Serum interleukin-17A (IL-17A),interleukin-22 (IL-22),interleukin-6 (IL-6),transforming growth factor-β (TGF-β) were detected with enzyme linked immunosorbent assay (ELISA);Expression of interleukin-23 receptor (IL-23R) in lung tissue was determined with immunohistochemical;real-time fluorescence quantification PCR (qRT-PCR) was used to detect the expression of retinoic acid related solitary nuclear receptors' mRNA in lung tissue.Results After administration of PQ,W/D ratio increased (P ＜ 0.01),lung injury was observed in mice of PQ and PQ + Ab groups,levels of cytokines (IL-17A,IL-22,IL-6 and TGF-β) in serum elevated (P ＜0.05),and the expressions of IL-23R mRNA and RORγt mRNA increased (P＜0.01).But in PQ +Ab group,W/D ratio decreased (P ＜0.05),lung injury was alleviated,the levels of cytokines (IL-17A,IL-22,IL-6 and TGF-β) decreased (P ＜ 0.05),and the expressions of IL-23R mRNA and RORγt mRNA reduced (P ＜ 0.05).Conclusions Since IL-17A involves in the lung injury of the mice induced by acute paraquat poisoning,blockade of IL-17A significantly alleviates the acute lung injury in mice.

The effects of bone marrow mesenchymal stem cells on the treatment of pulmonary fibrosis in silicosis mice / 中国职业医学

OBJECTIVE: To observe the effects of bone marrow mesenchymal stem cells( BMSCs) on the treatment of pulmonary fibrosis in silicosis mice. METHODS: Specific pathogen free healthy male C57BL/6 mice were randomly divided into control group,silicosis group and treatment group with 10 mice in each group. The mice of the control group were given one intra-tracheal injection of 20. 0 μL 0. 90% sodium chloride solution. The silicosis group and treatment group received one 20. 0 μL( mass concentration 250 g/L) of silica dust suspension. After 4 weeks,mice in treatment group were injected with 250. 0 μL of BMSCs suspension( cell density 2 × 10~9/L) by tail vein and silicosis group injected with 250. 0 μL of 0. 90% sodium chloride solution instead,once a week with continuous treatment for 4 weeks. Control group was not given any treatment. Mice were euthanized two weeks after the last treatment. Pathological sections were observed,pulmonary fibrosis score( Ashcroft scores) was marked. Lung coefficient was measured. Lung tissue hydroxyproline( HYP) level and serum transforming growth factor β1( TGF-β1) level were measured. The level of pulmonary fibrosis was scored and the percentages of T helper cell 17( Th17 cell) and regulatory T cell( Treg cell) of spleen and hilar lymph node( HLN) were measured by flow cytometry. RESULTS: The results of lung histopathological examination showed that the pulmonary fibrosis was severe in silicosis group. Massive collagen fiber accumulation and silicotic nodule were found. In treatment group,fibrosis was mild,little collagen fiber accumulation and silicotic nodule were found. The lung coefficient,Aschcroft scores,lung tissue HYP level,serum TGF-β level and the percentage of Th17 cell of spleen and HLN in silicosis group were higher than that of control group( P < 0. 05),while the above indexes of treatment group were lower than that of silicosis group( P < 0. 01). The percentage of Treg cell of spleen and HLN in silicosis group were lower than that of control group( P < 0. 05),while those indexes of treatment group were higher than that of silicosis group( P < 0. 01).CONCLUSION: BMSCs could effectively alleviate the pulmonary fibrosis in silicosis mice and correct the imbalance of Th17/Treg.