Huoxue Tongjiang decoction-resisted reflux esophagitis by activation stem cell factor/c-kit/interstitial cell of cajal pathway and regulating the T-helper 17/regulatory T-cells balance in rats

Background:Huoxue Tongjiang decoction(HXTJD)is an effective prescription for treating reflux esophagitis(RE).We investigated the effects of HXTJD on esophageal motility and mucosal inflammation in a rat RE model.Methods:Chemical composition of HXTJD was analyzed by ultrahigh-performance liquid chromatography Q-Orbitrap mass spectrometry(MS).The change rates of mean contraction tension forces,mean amplitudes,and mean frequencies for the lower esophageal sphincter(LES)were recorded using the isolated tissue bath system,mechanical tension transducer,and PowerLab physiological recorder.After weighing the stomach,the phenol red labeling method was used to measure the gastric emptying rate.The LES ultrastructure was observed through transmission electron microscopy.Immunofluorescence and western blotting were used to detect the number of interstitial cells of Cajal(ICC)and the expression levels of c-kit protein,connexin43(Cx43),and stem cell factor(SCF).Flow cytometric analysis and enzyme-linked immunosorbent assay were conducted to detect the percentages of T helper 17(Th17)cells and regulatory T(Treg)cells and the serum concentrations of interleukin 6(IL-6),interleukin 17(IL-17),and interleukin 10(IL-10)in the rats.Results:We identified 28 chemical constituents in HXTJD.Regarding esophageal motility,we revealed that HXTJD increased the mean contraction tension forces,mean amplitudes,and mean frequency change rate of LES and the gastric emptying rate;decreased stomach weight;and improved the LES ultrastructure.Additionally,HXTJD increased the number of ICC-positive cells,and c-kit,Cx43,and SCF expression levels.Regarding esophageal inflammation,HXTJD significantly decreased the percentage of Th17 cells,and IL-6 and IL-17 concentrations,and increased the percentage of Treg cells and IL-10 concentration.Conclusion:HXTJD was found to be efficacious in the rat RE model.It may promote esophageal motility and alleviate the inflammatory response by activating the SCF/c-kit/ICC pathway and regulating the Th17/Treg cell balance.

中药通过调节Th17/Treg平衡干预胃癌的研究进展

胃癌(GC)是一种较为常见的消化道恶性肿瘤。辅助性T细胞17(Th17)和调节性T细胞(Treg)是CD4+T细胞的分化亚群,Th17/Treg的失衡已被证实与GC的发病进程密切相关。中药不仅可改善GC患者的生存预后,而且在GC的术后化疗中起到了增效减毒的效果。本文系统整理了中药通过调节Th17/Treg平衡干预GC的作用规律。结果发现,中药复方可通过发挥健脾益气、健脾温阳、化瘀解毒等功效以调节GC的Th17/Treg平衡;中药及其有效成分通过调节Th17/Treg平衡来干预GC的作用机制主要表现为抑制Th17、Treg的过度分化及其转录因子、细胞因子的过度表达,逆转GC Th17/Treg平衡向Th17或Treg的过度漂移,从而恢复GC Th17/Treg的免疫平衡。

miR-142在儿童自身免疫性甲状腺疾病中表达及与Th17/Treg失衡关系的研究

目的 探究微小RNA-142 (microRNA-142,miR-142)在儿童自身免疫性甲状腺疾病(autoimmune thvroid disease,AITD)中表达及其与辅助性T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cell,Treg)失衡的关系。方法 前瞻性选取2019年1月—2022年12月住院治疗的89例AITD患儿为研究对象,包括48例Graves病患儿(Graves disease,GD;设为GD组)和41例桥本甲状腺炎患儿(Hashimoto thyroiditis,HT;设为HT组),另选取55例同期体检的健康儿童为对照组,比较各组血清miR-142、甲状腺球蛋白抗体(antithyroglobulin antibody,TG Ab)、抗甲状腺过氧化物酶抗体(antithyroperoxidase antibody,TPOAb)、Th17/Treg及白细胞介素-17 (interleukin-17,IL-17)表达差异。结果 GD组和HT组miR-142、TPOAb、TGAb、Th17、Th17/Treg、IL-17表达高于对照组,Treg水平低于对照组(P<0.05)。Pearson相关分析显示,GD组miR-142与TPOAb、TGAb、Th17、Th17/Treg、IL-17呈正相关(分别r=0.711、0.728、0.785、0.716、0.709,P<0.001),与Treg呈负相关(r=-0.725,P<0.001);HT组miR-142与TPOAb、TGAb呈正相关(分别r=0.752、0.717,P<0.001)。结论 miR-142在儿童AITD中高表达,且其表达可能与GD患儿Th17/Treg失衡有关。

Treg/Th17 cell balance and phytohaemagglutinin activation of T lymphocytes in peripheral blood of systemic sclerosis patients

AIM To investigate T-cell activation, the percentage of peripheral T regulatory cells(Tregs), Th17 cells and the circulating cytokine profile in systemic sclerosis(SSc).METHODS We enrolled a total of 24 SSc patients and 16 healthy controls in the study and divided the patients as having diffuse cutaneous SSc(dc SSc, n = 13) or limited cutaneous SSc(lc SSc, n = 11). We performed a further subdivision of the patients regarding the stage of the disease-early, intermediate or late. Peripheral venous blood samples were collected from all subjects. We performed flow cytometric analysis of the activationcapacity of T-lymphocytes upon stimulation with PHA-M and of the percentage of peripheral Tregs and Th17 cells in both patients and healthy controls. We used ELISA to quantitate serum levels of human interleukin(IL)-6, IL-10, tissue growth factor-β1(TGF-β1), and IL-17 A.RESULTS We identified a decreased percentage of CD3+CD69+ cells in PHA-stimulated samples from SSc patients in comparison with healthy controls(13.35% ± 2.90% vs 37.03% ± 2.33%, P < 0.001). However, we did not establish a correlation between the down-regulated CD3+CD69+ cells and the clinical subset, nor regarding the stage of the disease. The activated CD4+CD25+ peripheral lymphocytes were represented in decreased percentage in patients when compared to controls(6.30% ± 0.68% vs 9.36% ± 1.08%, P = 0.016). Regarding the forms of the disease, dc SSc patients demonstrated lower frequency of CD4+CD25+ T cells against healthy subjects(5.95% ± 0.89% vs 9.36% ± 1.08%, P = 0.025). With regard to Th17 cells, our patients demonstrated increased percentage in comparison with controls(18.13% ± 1.55% vs 13.73% ± 1.21%, P = 0.031). We detected up-regulated Th17 cells within the lc SSc subset against controls(20.46% ± 2.41% vs 13.73% ± 1.21%, P = 0.025), nevertheless no difference was found between dc SSc and lc SSc patients. Flow cytometric analysis revealed an increased percentage of CD4+CD25-Foxp3+ in dc SSc patients compared to controls(10.94% ± 1.65% vs 6.88% ± 0.91, P = 0.032). Regarding the peripheral cytokine profile, we detected raised levels of IL-6 [2.10(1.05-4.60) pg/m L vs 0.00 pg/m L, P < 0.001], TGF-β1(19.94 ± 3.35 ng/m L vs 10.03 ± 2.25 ng/m L, P = 0.02), IL-10(2.83 ± 0.44 pg/m L vs 0.68 ± 0.51 pg/m L, P = 0.008), and IL-17 A [6.30(2.50-15.60) pg/m L vs 0(0.00-0.05) pg/m L, P < 0.001] in patients when compared to healthy controls. Furthermore, we found increased circulating IL-10, TGF-β, IL-6 and IL-17 A in the lc SSc subset vs control subjects, as it follows: IL-10(3.32 ± 0.59 pg/m L vs 0.68 ± 0.51 pg/m L, P = 0.003), TGF-β1(22.82 ± 4.99 ng/m L vs 10.03 ± 2.25 ng/m L, P = 0.031), IL-6 [2.08(1.51-4.69) pg/m L vs 0.00 pg/m L, P < 0.001], and IL-17 A [14.50(8.55-41.65) pg/m L vs 0.00(0.00-0.05) pg/m L, P < 0.001]. Furthermore, circulating IL-17 A was higher in lc SSc as opposed to dc SSc subset(31.99 ± 13.29 pg/m L vs 7.14 ± 3.01 pg/m L, P = 0.008). Within the dc SSc subset, raised levels of IL-17 A and IL-6 were detected vs healthy controls: IL-17 A [2.60(0.45-9.80) pg/m L vs 0.00(0.00-0.05) pg/m L, P < 0.001], IL-6 [2.80(1.03-7.23) pg/m L vs 0.00 pg/m L, P < 0.001]. Regarding the stages of the disease, TGF-β1 serum levels were increased in early stage against late stage, independently from the SSc phenotype(30.03 ± 4.59 ng/m L vs 13.08 ± 4.50 ng/m L, P = 0.017).CONCLUSION It is likely that the altered percentage of Th17 and CD4+CD25-Fox P3+ cells along with the peripheral cytokine profile in patients with SSc may play a key role in the pathogenesis of the disease.

川芎-独活-牛膝配伍在骨关节炎发病中TH17/Treg细胞免疫平衡的调节作用

目的:探讨川芎-独活-牛膝配伍在骨关节炎发病中Th17/Treg细胞免疫平衡的调节作用。方法:购置SPF级无菌C57BL/6小鼠40只,随机取小鼠10只为空白对照组;剩余30只小鼠采用Hulth-Telhag方法构建膝关节炎动物模型,并随机分为模型组、异戊酸组和川芎-独活-牛膝配伍组,各10只。空白对照组和模型组腹腔灌注等体积生理盐水,异戊酸组腹腔灌胃异戊酸,川芎-独活-牛膝配伍组采用川芎-独活-牛膝水煎液灌胃,各组均持续7d干预,比较各组HE染色结果、炎性因子水平及Th17/Treg细胞免疫。结果:HE染色下异戊酸组伴有轻度关节软骨破坏,可见少许炎症细胞浸润;川芎-独活-牛膝配伍组关节软骨破坏不明显,炎症细胞亦相对较少;川芎-独活-牛膝配伍组IL-17、IL-6、IL-4低于异戊酸组(P<0.05);IL-10和TGF高于异戊酸组(P<0.05);流式细胞仪测定结果表明:川芎-独活-牛膝配伍组干预7d后Th17细胞低于异戊酸组和模型组(P<0.05);Treg细胞高于异戊酸组和模型组(P<0.05)。结论:川芎-独活-牛膝配伍用于骨关节炎中能减轻膝关节滑膜及软组织损伤,降低体内炎性因子水平,其机制可能与Th17/Treg细胞免疫平衡的调节有关。

Effect of saikosaponin A on Treg and Th17 immune balance in depressive rats

Objective:To investigate the Effect of saikosaponin A on Treg and Th17 immune balance in depressive rats.Methods:The rat depression model was established with reference to the Katz method,and the rats were randomly divided into control group,model group,western medicine group,and saikosaponin A group.The western medicine group was given 1.2 mg/kg/d of fluoxetine,and the saikosaponin A group was given 25 mg/kg/d of saikosaponin A,while the control group and model group were given the same volume of normal saline.The evaluation of depression in Rats was analyzed by Openfield-test and sugar water preference test.Flow cytometry was used to detect the expression of Th17 and Treg cells.And the expression of IL-17,IL-23,TNF-α,IL-10,TGF-βwere detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the control group,the horizontal exercise score,vertical exercise score,and sugar preference of the model group decreased significantly(P<0.05).Compared with the model group,the above indicators were significantly increased in the western medicine group and saikosaponin A group(P<0.05).Flow cytometry showed that compared with the control group,the Th17 cells,Th17/Treg cell ratio in model group increased significantly,whereas the Treg cells decreased significantly(P<0.05).Compared with the model group,The Th17 cells and Th17/Treg ratio in western medicine group and saikosaponin A group decreased,while the Treg cells increased significantly(P<0.05).ELISA showed that compared with control group,the serum levels of IL-17,IL-23 and TNF-αin model group increased,while the levels of IL-10 and TGF-βdecreased(P<0.05).Compared with model group,the levels of IL-17,IL-23 and TNF-αdecreased,while the levels of IL-10 and TGF-βincreased in western medicine group and saikosaponin A group(P<0.05).Conclusion:Saikosaponin A can reduce the degree of depression by regulating the imbalance of Th17/Treg cells and the secretion of inflammatory cytokines in depressed rats.

Detection and Clinical Significance of Th17/Treg Cell-Related Factors in Patients with Gestational Diabetes Mellitus

Objective:To investigate the detection of Th17/Treg cell-related factors in patients with gestational diabetes mellitus(GDM)and its clinical significance.Methods:In this study,a retrospective cohort research method was used to collect the clinical data of 42 patients who were hospitalized in the Affiliated Hospital of Hebei University and received the diagnosis of GDM from January 2018 to December 2022,as well as 42 patients with normal pregnancies during the same period.The Th17/Treg expression levels and metabolism-related indexes in the peripheral blood of patients were detected by radioimmunoassay.Results:The relative expression of Th17 in the serum of patients in the GDM group was significantly higher than that of the control group,and the level of Treg was significantly lower than that of the control group(P<0.05);the levels of FBG,FINS,2hBG,TC,TG and HOMA-IR of the patients in the GDM group were significantly higher than that of the control group,and the level of HOMA-βwas significantly lower than that of the control group(P<0.05).Conclusion:The imbalance of the Th17/Treg cell ratio in patients with GDM may be related to their disease progression and prognosis,providing new ideas and strategies for the clinical treatment of GDM.

Paeonol attenuated vascular fibrosis through regulating Treg/Th17 balance in gut microbiota-dependent manner

OBJECTIVE Atherosclerosis(AS)is a chronic inflammatory disease characterized by the accumulation of lipids,vascular fibrosis,and inflammation.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chinese medicine,Cortex Moutan,which exhibits anti-AS effects.Our previous work demonstrated that gut microbiota plays an important role during AS treatment as it affects the efficacy of Pae.However,the mechanism of Pae in protecting against vascular fibrosis as related to gut microbiota has yet to be elucidated.To investigate the anti-fibrosis effect of Pae on AS mice and demonstrate the underlying gut microbiota-dependent mechanism.METHODS ApoE-/-mice were fed with high-fat-diet(HFD)to replicate the AS model.HE and Masson staining were used to observe the plaque formation and collagen deposition.Gut microbiota alteration and short-chain fatty acids(SCFAs)production were analyzed through 16S rRNA sequencing and LC-MS/MS.The frequency of immune cells in spleen were phenotyped by flow cytometry.The mRNA expression of aortic inflammatory cytokines were detected by qRT-PCR.The protein expression of LOX and fibrosis related indicators were examined by Western blotting.RESULTS Pae restricted the development of AS and collagen deposition.Notably,the anti-fibrosis effect of Pae was achieved by regulating the gut microbiota.16S rRNA sequencing and LC-MS/MS data indicated that the relative abundance of SCFAs-producing bacteria and SCFAs production was increased.Additionally,Pae administration selectively up-regulated the frequency of regulatory T(Treg)cells as well as down-regulated the ratio of T helper type 17(Th17)cells in the spleen of AS mice,improving the Treg/Th17 balance.In addition,as expected,Pae intervention significantly down-regulate the mRNA expression levels of pro-inflammatory cytokines IL^(-1)β,IL-6,TNF-αand IL^(-1)7 in the aorta tissue,up-regulate the levels of anti-inflammatory factor IL^(-1)0,a marker of Treg cells.Finally,Pae′s intervention in the gut microbiota resulted in the restoration of the balance of Treg/Th17,which indirectly down-regulated the protein expression level of LOX and fibrosis-related indicators(MMP-2/9 and collagenⅠ/Ⅲ).CONCLUSION Pae attenuates vascular fibrosis in a gut microbiota-dependent manner.The underlying protective mechanism is associated with the improved Treg/Th17 balance in spleen mediated through the increased microbiota-derived SCFAs production.

Influence of budesonide and salbutamol atomization inhalation on Th17/Treg balance in patients with bronchial asthma and its correlation with airway remodeling

Objective:To study the influence of budesonide and salbutamol atomization inhalation on Th17/Treg balance in patients with bronchial asthma and its correlation with airway remodeling.Methods:A total of 90 patients with bronchial asthma who received systemic treatment in our hospital between July 2013 and April 2016 were divided into control group (n=45) and observation group (n=45) according to random number table. Patients in the control group were treated with salbutamol atomization inhalation alone while those in observation group were treated with budesonide and salbutamol atomization inhalation. The Th17/Treg ratio in peripheral blood as well as serum contents of airway remodeling-related indicators was compared between two groups before and after treatment. The correlation between Th17/Treg balance and airway remodeling in patients with bronchial asthma was detected by Pearson test. Results:Before treatment, the differences in peripheral blood Th17/Treg ratio as well as serum contents of inflammatory mediators, growth factor indexes and collagen metabolism indexes were not statistically significant between two groups of patients. After treatment, peripheral blood Th17/Treg ratio in observation group was lower than that in control group, serum CRP, IL-8, IL-13 and TNF- contents in observation group were lower than those in control group, serum PDGF-BB, b-FGF and VEGF contents in observation group were lower than those in control group, and serum HA, PCⅢ and LN contents in observation group were lower than those in control group. The Th17/Treg ratio in patients with bronchial asthma after treatment was positively correlated with the levels of airway remodeling-related indexes.Conclusion:Budesonide combined with salbutamol atomization inhalation can optimize the Th17/Treg balance in patients with bronchial asthma, and Th17/Treg is directly correlated to the degree of airway remodeling.

Correlation of serum YKL-40 content with eosinophil function as well as Th1/Th2 and Treg/Th17 immunity in patients with allergic rhinitis

Objective: To discuss the correlation of serum YKL-40 content with eosinophil function as well as Th1/Th2 and Treg/Th17 immunity in patients with allergic rhinitis. Methods: A total of 310 patients with allergic rhinitis who were treated in the hospital between January 2015 and May 2017 were selected as allergic rhinitis group and 100 healthy volunteers who received physical examination in the hospital during the same period were selected as normal control group. The differences in serum YKL-40 content, eosinophil function as well as Th1/Th2 and Treg/Th17 cytokine contents were compared between the two groups. Pearson test was used to assess the correlation between serum YKL-40 content and disease severity in patients with allergic rhinitis. Results: Serum YKL-40 content of allergic rhinitis group was higher than that of normal control group;serum eosinophil function IgE and Eotaxin-2 contents of allergic rhinitis group were higher than those of normal control group;serum Th1 cytokines IFN-γand IL-12 contents were lower than those of normal control group while Th2 cytokines IL-4 and IL-13 contents were higher than those of normal control group;serum Treg cytokine TGF-β content was lower than that of normal control group while Th17 cytokine IL-17 content was higher than that of normal control group. Pearson test showed that serum YKL-40 content in patients with allergic rhinitis was directly correlated with the eosinophil function as well as Th1/Th2 and Treg/Th17 immunity. Conclusion: Serum YKL-40 content is abnormally high in patients with allergic rhinitis, and the specific expression is positively correlated with the disease severity.

Correlation between Treg/Th17 imbalance and the liver injury caused by systemic inflammatory response and oxidative stress in children with sepsis

Objective: To study the correlation between Treg/Th17 imbalance and the liver injury caused by systemic inflammatory response and oxidative stress in children with sepsis. Methods:A total of 43 children who were diagnosed with sepsis in Shanghai Jiading District Central Hospital between February 2016 and March 2017were selected as the sepsis group of the study, and children who received physical examination in Outpatient Clinic of Shanghai Jiading District Central Hospital during the same period were selected as the control group of the study. The peripheral blood was collected to determine the contents of Treg and Th17 as well as the expression of oxidative stress molecules, and serum was collected to determine the contents of inflammatory reaction molecules, oxidative stress molecules and liver injury indexes. Results: Peripheral blood Treg and Th17 contents as well as Treg/Th17 of sepsis group were significantly higher than those of control group;serum ALT, AST, TBIL, γ-GT, MIF, sTREM-1, sVCAM-1 and PCT contents as well as peripheral blood MPO and Nrf-2 mRNA expression of sepsis group were significantly higher than those of control group and positively correlated with peripheral blood Treg/Th17 while peripheral blood Keap-1 mRNA expression as well as serum SOD and GSH contents was significantly lower than that of control group and negatively correlated with peripheral blood Treg/Th17. Conclusion: The increase of Treg/Th17 in peripheral blood of children with sepsis is closely related to the activation of inflammatory response and oxidative stress response and the injury of liver function.

电针刺激上巨虚、天枢对溃疡性结肠炎模型大鼠Th17相关特异性因子及Treg的影响

目的观察电针刺激上巨虚、天枢对溃疡性结肠炎(ulcerative colitis,UC)模型大鼠辅助性T细胞17(T helper cell 17,Th17)相关特异性因子及调节性T细胞(T regulatory cell,Treg)的影响,探讨合募配穴理论指导下电针刺激大肠腑下合穴、募穴治疗UC的作用机制,为特定穴的临床应用提供实验依据。方法50只SD大鼠随机分出10只作为空白组,剩余40只UC模型造模成功的大鼠随机分为模型组、合募配穴组、天枢组和上巨虚组,每组10只。2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)/乙醇法制备UC大鼠模型。合募配穴组予以电针刺激双侧天枢、上巨虚,天枢组予以电针刺激双侧天枢,上巨虚组予以电针刺激双侧上巨虚,每次20 min,每天1次,共10次。观察大鼠一般情况,计算大鼠疾病活动指数(disease active index,DAI);肉眼观察大鼠结肠黏膜损伤情况,评估结肠黏膜损伤指数(colonic mucosa damage index,CMDI),HE染色观察结肠形态学变化;流式细胞术检测大鼠脾脏CD4+T细胞中Treg细胞百分比,荧光定量PCR检测大鼠结肠组织中转化生长因子-β(transforming growth factor-β,TGF-β)、维甲酸相关孤儿受体γt(retinoid acid related orphan receptor gamma t,RORγt)、白细胞介素-17A(interleukin-17A,IL-17A)mRNA相对表达量。结果(1)与空白组比较,模型组一般情况不佳,肠黏膜坏死,有明显溃疡面,炎性浸润严重,DAI及CMDI评分显著升高(P<0.05),Treg细胞百分比及TGF-βmRNA相对表达量显著降低(P<0.05),RORγt、IL-17A mRNA相对表达量显著升高(P<0.05);(2)与模型组比较,合募配穴组、天枢组及上巨虚组一般情况明显好转,炎性浸润和结肠黏膜坏死情况明显减轻,DAI及CDMI评分显著降低(P<0.05),Treg细胞百分比及TGF-βmRNA相对表达量显著升高(P<0.05),RORγt、IL-17A mRNA相对表达量显著降低(P<0.05);(3)与天枢组和上巨虚组比较,合募配穴组Treg细胞百分比显著升高(P<0.05),RORγt mRNA相对表达量显著降低(P<0.05)。结论电针刺激大肠合募穴上巨虚、天枢对UC有良好的治疗作用,合募配穴疗效优于单穴疗效,其作用机制可能是通过调节Th17/Treg平衡,降低炎性细胞因子表达,以调控肠黏膜免疫失衡状态,恢复肠黏膜免疫稳态,减轻炎症反应。

YTE-17 inhibits colonic carcinogenesis by resetting antitumor immune response via Wnt5a/JNK mediated metabolic signaling

The density and composition of lymphocytes infiltrating colon tumors serve as predictive factors for the clinical outcome of colon cancer.Our previous studies highlighted the potent anti-cancer properties of the principal compounds found in Garcinia yunnanensis(YTE-17),attributing these effects to the regu-lation of multiple signaling pathways.However,knowledge regarding the mechanism and effect of YTE-17 in the prevention of colorectal cancer is limited.In this study,we conducted isobaric tags for relative and absolute quantification(iTRAQ)analysis on intestinal epithelial cells(IECs)exposed YTE-17,both in vitro and in vivo,revealing a significant inhibition of the Wnt family member 5a(Wnt5a)/c-Jun N-terminal kinase(JNK)signaling pathway.Subsequently,we elucidated the influence and mechanism of YTE-17 on the tumor microenvironment(TME),specifically focusing on macrophage-mediated T helper 17(Th17)cell induction in a colitis-associated cancer(CAC)model with Wnt5a deletion.Additionally,we performed the single-cell RNA sequencing(scRNA-seq)on the colonic tissue from the Wnt5a-deleted CAC model to characterize the composition,lineage,and functional status of immune mesenchymal cells during different stages of colorectal cancer(CRC)progression.Remarkably,our findings demon-strate a significant reduction in M2 macrophage polarization and Th17 cell phenotype upon treatment with YTE-17,leading to the restoration of regulatory T(Treg)/Th17 cell balance in azoxymethane(AOM)/dextran sodium sulfate(DSS)model.Furthermore,we also confirmed that YTE-17 effectively inhibited the glycolysis of Th17 cells in both direct and indirect co-culture systems with M2 macrophages.Notably,our study shed light on potential mechanisms linking the non-canonical Wnt5a/JNK signaling pathway and well-established canonical b-catenin oncogenic pathway in vivo.Specifically,we proposed that Wnt5a/JNK signaling activity in IECs promotes the development of cancer stem cells with b-catenin activity within the TME,involving macrophages and T cells.In summary,our study undergoes the po-tential of YTE-17 as a preventive strategy against CRC development by addressing the imbalance with the immune microenvironment,thereby mitigating the risk of malignancies.

消痈乳康汤联合中医换药对肉芽肿性乳腺炎临床疗效及Treg/Th17免疫平衡机制的影响

目的:观察消痈乳康汤联合中医换药治疗肉芽肿性乳腺炎(granulomatous mastitis,GM)的临床疗效及对调节性T细胞(regulatory T cell,Treg)/辅助性T细胞17(T helper cell 17,Th17)免疫平衡机制以及免疫相关指标的影响。方法:选取实施乳腺导管切除术后GM患者97例,按照完全随机原则分为观察组(48例)及对照组(48例)。对照组术后予西医常规治疗,观察组在对照组基础上加服消痈乳康汤治疗,共治疗4周。比较两组临床症状、疼痛评分、QLQ-C30评分,外周血Treg细胞、白细胞介素10(interleukin-10,IL-10)、转化生长因子β_(1)(trans forming growth factor-β_(1),TGF-β_(1))水平,Treg/Th17比值及免疫相关指标,并统计其临床疗效、复发率及不良反应率。结果:治疗后观察组乳房包块最大直径、乳房皮肤评分、疼痛评分及外周血Treg细胞、Treg/Th17比值、IL-10、TGF-β_(1)、IgG、IgA水平均明显低于治疗前与对照组治疗后(P<0.05);QLQ-C30总评分及各维度评分均明显高于治疗前与对照组治疗后(P<0.05)。观察组治疗总有效率[95.83%(46/49)]明显高于对照组[75.51%(37/48)](P<0.05),观察组不良反应发生率[6.25%(3/48)]低于对照组[16.33%(8/49)](P<0.05),且随访复发率低于对照组(P<0.05)。结论:消痈乳康汤联合中医换药治疗GM安全有效,能够明显缓解临床症状,加速疾病康复,改善患者生活质量,还可能是通过调控Treg/Th17免疫平衡机制治疗GM,并能够提升患者免疫功能,同时降低疾病复发风险。

Restoring the Treg cell to Th17 cell ratio may alleviate HBV-related acute-on-chronic liver failure

AIM: To investigate the role of T helper 17 cells (Th17) and regulatory T cells (Treg) in hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF).METHODS: We enrolled 79 patients with HBV infection into the study, 50 patients with HBV-related ACLF and 29 patients with chronic hepatitis B (CHB), from the First Affiliated Hospital of Medical College from January 2009 to June 2012. The ACLF patients were diagnosed according to the criteria recommended by The 19th Conference of the Asian Pacific Association for the Study of the Liver in 2009. Twenty healthy individuals with a similar gender and age structures to the two patient groups were also included as the normal controls (NC). Of the 50 ACLF patients, 28 were subsequently classified as non-survivors: 19 patients died from multiorgan failure, 3 underwent liver transplantation, and 6 discontinued therapy during follow-up because of financial reasons. The remaining 22 ACLF patients whose liver and anticoagulation function recovered to nearly normal levels within the next 6 mo were classified as survivors. The number of circulating Treg and Th17 cells was determined upon diagnosis and during the 8th week of follow-up through flow cytometry. RESULTS: The percentage of circulating Treg cells in the ACLF group was significantly higher than that in the CHB group (5.50% ± 1.15% vs 3.30% ± 1.13%, P < 0.01). The percentages of circulating Th17 cells in the ACLF and the CHB groups were significantly higher than that in the NC group (6.32% ± 2.22% vs 1.56% ± 0.44%, P < 0.01; 3.53% ± 1.65% vs 1.56% ± 0.44%, P < 0.01). No significant difference in Treg cell to Th17 cell ratio was observed between the ACLF group and the CHB group (0.98 ± 0.44 vs 1.12 ± 0.64, P = 0.991), whereas those in the two HBV infection groups were significantly lower than that in the NC group (1.85 ± 1.22; both P < 0.01). The percentage of Treg cells in the survivors during the 8th week of follow-up was significantly lower than that during peak ACLF severity [total bilirubin (TBIL) peak] (3.45% ± 0.97% vs 5.18% ± 1.02%, P < 0.01). The percentage of Th17 cells in survivors during the 8th week of follow-up was significantly lower than that during the peak TBIL (2.89% ±0.60% vs 5.24% ± 1.46%; P < 0.01). The Treg cell to Th17 cell ratio during the 8 th week of follow-up was significantly higher than that during the TBIL peak (1.22 ± 0.36 vs 1.10 ± 0.54; P < 0.05). CONCLUSION: Restoring the Treg cell to Th17 cell ratio during the follow-up phase of ACLF could maintain the immune system at a steady state, which favours good prognosis.

Effects of early enteral nutrition on Th17/Treg cells and IL-23/IL-17 in septic patients

BACKGROUND The imbalance of Th17/Treg cells and the IL-23/IL-17 axis have been confirmed to be associated with sepsis and various inflammatory diseases. Early enteral nutrition (EEN) can modulate the inflammatory response, improve immune dysfunction, and prevent enterogenic infection in critically ill patients;however, the precise mechanisms remain unclear. Considering the important roles of Th17 and Treg lymphocytes in the development of inflammatory and infectious diseases, we hypothesized that EEN could improve the immune dysfunction in sepsis by maintaining a balanced Th17/Treg cell ratio and by regulating the IL- 23/IL-17 axis. AIM To investigate the effects of EEN on the Th17/Treg cell ratios and the IL-23/IL-17 axis in septic patients. METHODS In this prospective clinical trial, patients were randomly divided into an EEN or delayed enteral nutrition (DEN) group. Enteral feeding was started within 48 h in the EEN group, whereas enteral feeding was started on the 4th day in the DEN group. The Th17 and Treg cell percentages and the interleukin levels were tested on days 1, 3, and 7 after admission. The clinical severity and outcome variables were also recorded. RESULTS Fifty-three patients were enrolled in this trial from October 2017 to June 2018. The Th17 cell percentages, Th17/Treg cell ratios, IL-17, IL-23, and IL-6 levels of the EEN group were lower than those of the DEN group on the 7th day after admission (P < 0.05). The duration of mechanical ventilation and of the intensive care unit stay of the EEN group were shorter than those of the DEN group (P <0.05). However, no difference in the 28-d mortality was found between the two groups (P = 0.728). CONCLUSION EEN could regulate the imbalance of Th17/Treg cell ratios and suppress the IL- 23/IL-17 axis during sepsis. Moreover, EEN could reduce the clinical severity of sepsis but did not reduce the 28-d mortality of septic patients.

Correlation between microRNA-21 and expression of Th17 and Treg cells in microenvironment of rats with hepatocellular carcinoma

Objective: To study the correlation between mi R-21 and Treg/Th17 ratio in the microenvironment of rats with hepatocellular carcinoma. Methods: Diethylnitrosamine was used to build the hepatocel ular carcinoma model of rats; the content of Treg cells and Th17 cells and the expression of mi R-21 in the peripheral blood of rats with hepatocellular carcinoma were detected. The statistical analysis was performed on the correlation between mi R-21 expression and Treg/Th17 ratio. Results: Hepatocellular carcinoma model of rats was successfully constructed. The proportion of Th17 cells among all CD4+T cells in the peripheral blood of rats with hepatocellular carcinoma was 5.319%, which was higher than the control group; while the proportion of Treg cells was 9.472%, which was higher than the control group. Treg/Th17 ratio in the model group was 1.781, compared with 1.478 in the control group. The expression of mi R-21 was increased in the peripheral blood of rats with hepatocellular carcinoma and it showed a positive correlation with the ratio of Treg/Th17. Conclusions: There is a positive correlation between the expression level of miR-21 and the ratio of Treg/Th17.

Mechanism of Liancao-Xieli capsule in the treatment of ulcerative colitis through the differentiation of Th17 and Treg cells

Objective:To observe the effect of Liancao-Xieli Capsule on STAT signal pathway and T cell differentiation in mouse model of ulcerative colitis;Methods:Forty BALB/c mice were randomly divided into the control group,model group,mesalazine group and Liancao-Xieli capsule group.Except the control group,the other three groups were treated with 3%dextran sodium sulfate free drinking water to construct the model of ulcerative colitis.During the modeling period,each group was given corresponding drugs for intervention,while the control group and the model group were given the same volume of distilled water by gavage as the control.After treatment,HE staining was used to observe the pathological changes of colon tissue,flow cytometry was used to detect the proportion of Th17 and Treg cells in spleen and mesenteric lymph nodes,and ELISA was used to detect TGF-β,IL-6 and IL-17A in colon tissue.Western blot was used to detect the expression levels of related proteins in STAT3/ROR-γt and STAT5/Foxp3 signaling pathways.Results:Compared with the model group,the body weight,colon length and the content of TGF-βin the colon tissue of the mice in the Liancao-Xieli capsule group increased significantly after the experiment,while the DAI score,colon histopathology score,and the contents of IL-6 and IL-17A in the colon tissue were significantly reduced,and the difference was statistically significant(P<0.01).At the same time,Liancao-Xieli capsule can reduce the ratio of Th17 cells and the ratio of Th17/Treg in the spleen and mesenteric lymph node tissues of UC mice,and increase the ratio of Treg cells,and the difference is statistically significant when compared with the model group(P<0.01).In addition,compared with the model group,the expression levels of p-STAT3 and RORγt protein in the colon tissue of the Liancao-Xieli capsule group were significantly reduced,and the expression levels of p-STAT5 and Foxp3 protein were significantly increased after treatment,and the differences are statistically significant(P<0.01),while the expression levels of STAT3 and STAT5 proteins in colon tissue did not change significantly,and the difference was not statistically significant(P>0.05);Conclusion:Liancao-Xieli Capsule can regulate the immune balance of Treg/Th17 and improve the intestinal inflammation of UC.Its mechanism of action is mainly through inhibiting STAT3/ROR-γt and promoting the activation of STAT5/Foxp3 signaling pathway.

黄芪多糖对免疫抑制模型小鼠Treg细胞及Th17细胞亚群的影响

本研究旨在探讨黄芪多糖对免疫抑制模型小鼠CD4+CD25+调节性T细胞(Treg细胞)和CD4+Th17细胞亚群功能的影响。通过小鼠腹腔注射环磷酰胺建立免疫抑制动物模型,应用黄芪多糖对免疫抑制小鼠进行治疗,采用流式细胞仪分析脾脏中CD4+T细胞、CD4+CD25+Treg细胞的比例,ELISA检测血清IL-17的水平。结果表明:与模型对照组比较,黄芪多糖高、中剂量明显降低小鼠脾脏CD4+T细胞数的比例(P<0.05),同时黄芪多糖上调小鼠脾脏CD4+CD25+Treg细胞数的比例(P<0.05),降低免疫抑制小鼠血清IL-17的水平,尤其1.0 g/(kg·d)剂量组的IL-17水平下降较为明显(P<0.01)。提示黄芪多糖具有提高免疫抑制小鼠CD4+CD25+Treg细胞数的比例、降低CD4+T细胞的数量及抑制IL-17分泌活性的作用。

多发性骨髓瘤患者骨髓中Treg/Th17细胞失衡研究

本研究通过检测多发性骨髓瘤(MM)患者骨髓微环境中调节性T细胞(Treg)和Th17细胞的比例,探讨Treg/Th17细胞失衡与多发性骨髓瘤发生发展的关系。应用流式细胞术检测37例MM患者和12例正常对照者骨髓中Treg和Th17细胞的表达量,对其中19例MM患者同时检测了外周血中Treg和Th17细胞的表达量。结果表明:MM组骨髓中Treg细胞比例高于正常对照组(P<0.05),自ISS分期的Ⅰ+Ⅱ期至Ⅲ期逐渐增高(P<0.05);进一步研究发现,MM患者骨髓中Treg细胞水平高于外周血,差异有统计学意义(P<0.05)。MM组骨髓中Th17细胞比例与正常对照组无差别(P>0.05),临床分期间无差别(P>0.05)。MM患者骨髓中Th17细胞比例与外周血无差别(P>0.05)。MM组Treg/Th17的比值高于正常对照组(P<0.05),自ISS分期的Ⅰ+Ⅱ期至Ⅲ期逐渐增高(P<0.05)。结论 MM患者骨髓微环境中存在Treg/Th17比例失衡,这种失衡可能促进了MM的病程进展。