Serum autophagy-related gene 5 level in stroke patients: correlation with CD4+ T cells and cognition impairment during a 3-year follow-up

Abstract Autophagy-related gene (ATG) 5 regulates blood lipids, chronic inflammation, CD4+ T-cell differentiation, and neuronal death and is involved in post-stroke cognitive impairment. This study aimed to explore the correlation of serum ATG5 with CD4+ T cells and cognition impairment in stroke patients. Peripheral blood was collected from 180 stroke patients for serum ATG5 and T helper (Th) 1, Th2, Th17, and regulatory T (Treg) cell detection via enzyme-linked immunosorbent assays and flow cytometry. The Mini-Mental State Examination (MMSE) scale was completed at enrollment, year (Y)1, Y2, and Y3 in stroke patients. Serum ATG5 was also measured in 50 healthy controls (HCs). Serum ATG5 was elevated in stroke patients compared to HCs (P<0.001) and was positively correlated to Th2 cells (P=0.022), Th17 cells (P<0.001), and Th17/Treg ratio (P<0.001) in stroke patients but not correlated with Th1 cells, Th1/Th2 ratio, or Treg cells (all P>0.050). Serum ATG5 (P=0.037), Th1 cells (P=0.022), Th17 cells (P=0.002), and Th17/Treg ratio (P=0.018) were elevated in stroke patients with MMSE score-identified cognition impairment vs those without cognition impairment, whereas Th2 cells, Th1/Th2 ratio, and Treg cells were not different between them (all P>0.050). Importantly, serum ATG5 was negatively linked with MMSE score at enrollment (P=0.004), Y1 (P=0.002), Y2 (P=0.014), and Y3 (P=0.001); moreover, it was positively related to 2-year (P=0.024) and 3-year (P=0.012) MMSE score decline in stroke patients. Serum ATG5 was positively correlated with Th2 and Th17 cells and estimated cognitive function decline in stroke patients.

Discussion on the effects of glycyrrhetinic acid on cough variant asthma mice by adjusting Th2 deviation based on molecular docking technique / 国际中医中药杂志

Objective:To evaluate the therapeutic effect of glycyrrhetinic acid on cough variant asthma (CVA) mice based on molecular docking technique; To explore the possibility of its treatment for cough variant asthma.Methods:The software of Autodock Vina was used for molecular docking. The mice were divided into control group, model group, prednisone acetate group, glycyrrhetinic acid high-, medium-, and low-dosage groups according to the random number table method, with 8 mice in each group. Except for the blank control group, all other groups were induced by egg protein to establish cough variant asthma models. Glycyrrhetinic acid high-, medium-, and low-dosage groups were orally administered glycyrrhetinic acid suspension at 20, 10, and 5 mg/kg, while the prednisone acetate group was orally administered prednisone acetate at 5 mg/kg. The blank control group and model group were orally administered equal volumes of physiological saline, once per day for 14 consecutive days. The animal asthma behavior was observed after drug administration. The secretion of bronchial mucus in lung tissue were observed by AB-PAS staining and the index of spleen were recorded. The protein expressions of Gata3, IL-4 and IL-13 in the spleen tissue were determined by Western blot.Results:Molecular docking results showed that glycyrrhetinic acid had good binding ability to Th2-related factors Gata3, IL-4 and IL-13. Results of animal experiment showed that compared with the model group, the mucus secretion decreased in glycyrrhetinic acid groups, the index of the spleen of mice obviously decreased, protein expression levels of IL-4 and IL-13 in the spleen tissue of mice in glycyrrhetinic acid high-, medium-, and low-dosage groups decreased ( P<0.05), and Gata3 in glycyrrhetinic acid medium- and low-dosage groups decreased ( P<0.05). Conclusion:Glycyrrhetinic acid can correct the shift of Th2 in the immune system of cough variant asthma mice and has a certain therapeutic effect.

Correlation of neutrophil extracellular traps with T cells differentiation in Hashimoto′s thyroiditis mice and vitamin D′s relief effect / 中华内分泌代谢杂志

Objective:To investigate correlation between neutrophil extracellular traps(NETs) formation and T cell subsets in mice with experimental autoimmune thyroiditis(EAT) and the impact of active vitamin D intervention.Methods:Six-week-old female BALB/c mice were randomly divided into Control group, EAT group and 1, 25 dihydroxy vitamin D 3[1, 25(OH) 2D 3] treatment group(VitD group; n=6/group). HE staining was used to observe thyroid pathology. Plasma thyroglobulin antibody(TGAb), thyroid peroxidase antibody(TPOAb), and 1, 25(OH) 2D 3 were measured by ELISA. Peripheral NETs formation, Th1, Th2, and Th17 cell ratio from spleen were measured by flow cytometry. Correlation between NETs formation rate and Th1, Th2, and Th17 cell ratio was analyzed. Results:Compared with Control group, mice in EAT group had significantly increased thyroid inflammation scores, thyroiditis morbidity, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + )and Th17 cell proportions( P were <0.001, 0.002, 0.007, <0.001, <0.001, 0.003, 0.001, and 0.002, respectively), and significant decreased 1, 25(OH) 2D 3, Th1 cell proportions, Th1/Th2(CD4 + IL-4 + ), Th1/Th2(CD4 + IL-13 + ), and Th1/Th17 ratios( P were 0.010, 0.018, 0.010, 0.005, and 0.007, respectively). Compared with the EAT group, the VitD group had lower thyroid inflammation scores, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportions( P were 0.044, 0.007, <0.001, 0.001, 0.014, 0.008, and 0.001, respectively), and significant higher Th1 cell ratio, Th1/Th2(CD4 + IL-13 + ) and Th1/Th17 ratio( P were 0.011, 0.009, and 0.003, respectively). The Th1/Th2(CD4 + IL-4 + ) was not significantly increased in VitD group compared with EAT group( P=0.174). NETs formation rate was positively correlated with Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportion( r were 0.65, 0.59, and 0.61; and P were 0.004, 0.010, and 0.007, respectively), but not with Th1 cell proportion( r=-0.47, P=0.051). Conclusion:EAT mice were more prone to NETs formation. Active vitamin D may relieve immune imbalance with increased Th2 and Th17 cell ratio and decreased Th1 cell ratio by reducing the formation of NETs in EAT mice. Vitamin D played the protective role in thyroid by reducing thyroid pathological damage and thyroid autoantibody levels, and relived overall lymphocyte imbalance.

Effect of T helper 1 to T helper 2 ratio on the prognosis of patients with multiple myeloma / 白血病·淋巴瘤

Objective:To investigate the effect of T helper 1 (Th1) to T helper 2 (Th2) ratio (Th1/Th2) on the prognosis of patients with multiple myeloma (MM).Methods:The clinical data of 168 MM patients who were newly diagnosed in the Affiliated Drum Tower Hospital of Nanjing University Medical School from January 2016 to January 2021 were retrospectively analyzed. Disease staging was defined according to the Chinese guidelines for diagnosis and treatment of MM (2020 edition). Risk stratification was based on the Mayo stratification of myeloma and risk-adapted therapy (mSMART) 3.0. The levels of Th1 and Th2 in peripheral blood of patients were detected by flow cytometry. Th1/Th2 was compared among patients with different disease staging and risk stratification. Using mSMART 3.0 risk stratification as the gold standard, the receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value of Th1/Th2 for determining high-risk MM. According to the optimal cut-off value, patients were divided into high Th1/Th2 group (≥ optimal cut-off value) and low Th1/Th2 group (< optimal cut-off value). The progression-free survival (PFS) of the two groups was analyzed by Kaplan-Meier method. Multivariate Cox proportional hazards model was used to analyze the influencing factors of PFS.Results:There were 40, 62 and 66 patients with international staging system (ISS) stages Ⅰ, Ⅱ and Ⅲ, respectively, with Th1/Th2 [ M ( IQR)] of 19.20 (18.98), 15.93 (14.40) and 14.47 (12.01), respectively ( H = 6.68, P = 0.036). There were 31,102 and 35 patients with revised international staging system (R-ISS) stages Ⅰ, Ⅱ and Ⅲ, respectively, with Th1/Th2 of 19.67 (21.92), 14.87 (11.36) and 13.50 (12.80), respectively ( H = 7.26, P = 0.027). There were 99 and 69 patients with mSMART 3.0 high-risk and standard-risk, respectively, and the Th1/Th2 of high-risk patients was lower than that of the standard-risk patients [14.70 (11.93) vs. 17.72 (16.80), U = 2 612.00, P = 0.009]. ROC curve analysis showed that the area under the curve for Th1/Th2 to determine high-risk MM was 0.618 (95% CI 0.531-0.705, P = 0.010), and the optimal cut-off value was 16.55 and there were 81 and 87 cases in the high Th1/Th2 group and low Th1/Th2 group. With a median follow-up of 28 months (1-70 months), the median PFS time for all patients was 36 months (95% CI 29-43 months); PFS in high Th1/Th2 group was better than that in low Th1/Th2 group [median PFS time: 39 months (95% CI 26-51 months) vs. 28 months (95% CI 21-34 months), P = 0.040]. Multivariate Cox regression analysis showed that renal impairment (with vs. without: HR = 2.340, 95% CI 1.350-4.053, P = 0.002) and low Th1/Th2 (high vs. low: HR = 0.551, 95% CI 0.344-0.882, P = 0.013) were independent risk factors for PFS in MM patients. Conclusions:The imbalance between Th1 and Th2 is associated with the prognosis of MM patients, and patients with low Th1/Th2 are at high risk of progression. Th1/Th2 can be used as a prognostic indicator for MM.

The relationship between C3, C4, Th1/Th2 levels and the MG-ADL score in patients with myasthenia gravis and its efficacy in predicting the transition of ocular muscle type to systemic type / 中国医师杂志

Objective:To investigate the relationship between C3, C4, Th1/Th2 levels and the Myasthenia Gravis Daily Living Scale (MG-ADL) score in patients with myasthenia gravis (MG) and its efficacy in predicting the transition of ocular muscle type to systemic type.Methods:A retrospective study of 94 patients with ophthalmic MG admitted to Haikou People's Hospital from April 2017 to April 2020 was conducted. According to whether they had converted to systemic MG within 6 months, they were divided into transformation group ( n=35) and non-transformation group ( n=59). The levels of C3, C4 and Th1/Th2, as well as the score of MG-ADL and Quantitative Myasthenia Gravis (QMG) were compared between the two groups before and 1 and 3 months after treatment. The correlation between C3, C4 and Th1/Th2 levels and MG-ADL and OMG scores, as well as the related influencing factors of the transformation from ocular muscle type to systemic type was analyzed. The efficiency of each index in predicting the transformation from ocular muscle type to systemic type was analyzed. Results:At 1 and 3 months after treatment, the C3 and C4 in both groups were significantly higher than before treatment, and Th1/Th2 was significantly lower than before treatment; the C3 and C4 in the non-transformation group were higher than that in the transformation group, while Th1/Th2 was lower than that in the transformation group (all P<0.05). The MG-ADL and QMG scores in 2 groups at 1 and 3 months after treatment were significantly lower than those before treatment, and those in the non-transformation group were lower than those in the transformation group (all P<0.05). C3 and C4 levels were negatively correlated with MG-ADL and QMG scores (all P<0.05), while Th1/Th2 levels were positively correlated with MG-ADL and QMG scores (all P<0.05). At 1 and 3 months after treatment, C3, C4 and Th1/Th2 were the influencing factors for the transformation from ocular muscle type to systemic type (all P<0.05). The area under the curve (AUC) of C3, C4 and Th1/Th2 combined to predict the transformation from ocular muscle type to systemic type at 3 months after treatment was 0.939, and the best predictive sensitivity and specificity were 91.43% and 88.14%, respectively. Conclusions:There is a good linear relationship between C3, C4, Th1/Th2 levels and MG-ADL scores in MG patients, and it has a high efficiency in predicting the transition of ocular muscle type to systemic type.

T helper type 2 immune response in alopecia areata / 中华皮肤科杂志

Alopecia areata (AA) is a common inflammatory and non-scarring hair loss condition with unknown pathogenesis, and relapses are common in some patients. Evidence has demonstrated that allergy takes part in the early onset, severe condition, recurrence, and prolonged process in AA. Allergy to dust mites may be one of the reasons for refractory severe AA, especially in childhood, possibly due to the predominance of T helper type 2 (Th2) immune response. Desensitization can suppress the Th2 immune response, alter the immune balance, and reduce disease severity during AA relapses. In addition, high IgE levels may predict favorable efficacy of dupilumab in AA patients before treatment, while high interleukin-4 levels may predict the ineffectiveness of topical immunotherapy with diphenylcyclopropenone, which works by antagonizing Th1 immune response. Therefore, serum total IgE, specific IgE to dust mites, and interleukin-4 can be considered as biomarkers, revealing the predominance of Th2 immune response in AA patients. This article focuses on the relationship between allergy and AA, as well as the role of anti-allergic reactions and desensitization in the treatment of AA, aiming to provide ideas for precise and individualized treatment of AA.

An investigation into clinical significance of serum chitinase 3-like protein 1 in pemphigus vulgaris / 中华皮肤科杂志

Objective:To detect the serum level of chitinase-3-like protein 1 (YKL-40) in patients with pemphigus vulgaris, and to analyze its correlation with the severity of pemphigus vulgaris.Methods:From January 2017 to May 2018, serum samples were collected from 38 patients with pemphigus vulgaris in Department of Dermatology, the Affiliated Hospital of Southwest Medical University, and those collected from 14 age-, gender- and body mass index-matched healthy volunteers served as controls. Serum levels of YKL-40 and Th1/Th2/Th17-related cytokines were detected by using Luminex ? 200 TM system. Mann-Whitney U test was used to compare serum levels of cytokines between the patient group and control group; binary logistic regression was used to investigate factors independently related to the severity of pemphigus vulgaris; a receiver operating characteristic (ROC) curve was drawn, and the area under the curve, sensitivity and specificity were calculated to evaluate the ability of YKL-40 to predict the severity of pemphigus vulgaris. Results:Compared with the control group, the patient group showed significantly increased serum levels of YKL-40 (expressed as median[ Q1, Q3]: 15.22 [14.19, 15.93] vs. 13.64 [13.21, 14.63]μg/L, z=-3.88, P < 0.05) , interleukin (IL) -6 (2.05 [1.49, 4.21] vs. 1.57[1.38, 1.75]ng/L, z=-2.44, P < 0.05) , IL-7 (7.45[5.63, 11.63] vs. 3.77[2.21, 5.97]ng/L, z=-3.26, P < 0.05], IL-8 (6.59[3.60, 14.73] vs. 4.36[2.96, 6.53]ng/L, z=-1.96, P < 0.05) , IL-2R-α (509.08 [386.36, 757.67] vs. 336.44[309.86, 458.71]ng/L, z=-2.35, P < 0.05) , and C5a (100.35 [78.31, 140.84] vs. 72.08 [37.23, 82.08] ng/L, z = -3.04, P < 0.05) . The concentration of serum YKL-40 gradually decreased along with the reduction of lesion areas ( r = 0.63, P < 0.001) , and YKL-40 was independently correlated with the severity of pemphigus vulgaris ( P = 0.025, OR = 46.54, 95% CI: 1.61, 1 347.19) . The area under the curve of YKL-40 was 0.783 (95% CI: 0.613, 0.953) for distinguishing between patients with severe to extremely severe pemphigus vulgaris and those with mild to moderate pemphigus vulgaris. Conclusion:The serum level of YKL-40 is strongly correlated with the severity of pemphigus vulgaris, and has a potential value in predicting the severity of this disease.

Eficácia e segurança do dupilumabe no tratamento da rinossinusite crônica com polipose nasal / Efficacy and safety of dupilumab in the treatment of chronic rhinosinusitis with nasal polyposis

A rinossinusite crônica (RSC) é uma síndrome caracterizada pela inflamação da mucosa nasal e dos seios paranasais por pelo menos 12 semanas, acometendo de 5% a 12% da população geral. A síndrome é associada a alta morbidade e considerada um grande problema de saúde pública devido a sua prevalência, seu custo para a sociedade e ao impacto que acarreta na qualidade de vida dos pacientes e em seu desempenho escolar ou profissional. Ademais, a RSC está associada a diversas comorbidades, como dermatite atópica, distúrbios respiratórios do sono, conjuntivite, otite média, asma e problemas emocionais. O dupilumabe é eficaz e seguro no tratamento da RSC com polipose nasal. A eficácia é progressiva no primeiro ano de tratamento, e a posologia de 300 mg a cada duas semanas é superior em relação à de cada quatro semanas. A interrupção do tratamento com 24 semanas acarreta a perda parcial de seus efeitos benéficos. O imunobiológico também é eficaz no controle da asma nos pacientes que apresentam essa doença como comorbidade. Alguns pacientes podem apesentar aumento transitório de eosinófilos sanguíneos, e 2,7% desenvolveram conjuntivite como reação adversa nos estudos SINUS-24 e SINUS-52. O dupilumabe é uma excelente opção terapêutica no tratamento concomitante de múltiplas doenças caracterizadas pela inflamação de tipo II.

Chronic rhinosinusitis (CRS) is a syndrome characterized by inflammation of the nasal mucosa and paranasal sinuses for at least 12 weeks, affecting 5% to 12% of the general population. The syndrome is associated with high morbidity and is considered a major public health problem because of its prevalence, its cost to society, and the impact it has on patients' quality of life and on their school or professional performance. Furthermore, CRS is associated with several comorbidities, such as atopic dermatitis, sleep-disordered breathing, conjunctivitis, otitis media, asthma, and emotional problems. Dupilumab is effective and safe in the treatment of CRS with nasal polyposis. Effectiveness is progressive in the first year of treatment, and a dosage of 300 mg every two weeks is more effective than that of every four weeks. Discontinuing treatment at 24 weeks results in partial loss of its beneficial effects. The biological drug is also effective in controlling asthma in patients who have this disease as a comorbidity. Some patients may experience a transient increase in blood eosinophils, and 2.7% developed conjunctivitis as an adverse reaction in the SINUS-24 and SINUS-52 studies. Dupilumab is an excellent therapeutic option in the concomitant treatment of multiple diseases characterized by type II inflammation.

Significance of Th1/Th2 cytokines in prognostic stratification of acute myeloid leukemia / 白血病·淋巴瘤

Objective:To investigate the significance of Th1/Th2 cytokines in prognostic stratification of acute myeloid leukemia (AML).Methods:A total of 83 patients with newly diagnosed AML from June 2017 to April 2019 in the First People's Hospital of Yunnan Province were collected. According to the Chinese guidelines for diagnosis and treatment of adult acute myeloid leukemia (non-acute promyelocytic leukemia) (2017 edition), AML patients were divided into poor prognosis group (45 cases), moderate prognosis group (19 cases), and good prognosis group (19 cases); moderate prognosis plus poor prognosis was treated as the not good prognosis. Mann-Whitney U test and Kruskal-Wallis H test were used to compare the expression differences of Th1/Th2 cytokines in peripheral blood of different prognosis groups; cytokines with statistical differences among different prognosis groups were selected, and the cut-off value of AML patients with different prognostic stratification distinguished by cytokines was determined by using receiver operating characteristic (ROC) curve. Finally, patients were divided into ≥ cut-off value group and <cut-off value group according to the cut-off value, and then the association of both groups with the prognostic stratification in guideline was also analyzed. Results:The median expression level of tumor necrosis factor (TNF)-β of patients in moderate prognosis group [3.80 pg/ml (2.75 pg/ml, 15.32 pg/ml)] was higher than that of patients in poor prognosis group [2.78 pg/ml (1.28 pg/ml, 3.36 pg/ml)] and good prognosis group [1.61 pg/ml (0.83 pg/ml, 3.04 pg/ml)] ( U=216, P=0.02; U = 312, P < 0.05); the median expression level of TNF-β in good prognosis group was lower than that in poor prognosis group ( U = 562, P = 0.048). There were no statistically significant differences in the expression levels of Th1/Th2 cytokines of AML patients with different prognostic stratification (all P>0.05).The cut-off value of TNF-β was 3.23 pg/ml in good prognosis group and moderate prognosis group, the area under the ROC curve was 0.866 (95% CI 0.753-0.978, P < 0.05); among 26 patients with TNF-β≥ 3.23 pg/ml, 25 (96.2%) patients had not good prognosis. The cut-off value was 3.62 pg/ml for distinguishing between moderate prognosis group and poor prognosis group, the area under the ROC curve was 0.747 (95% CI 0.610-0.884, P = 0.02); 18 (100%) patients with TNF-β≥ 3.62 pg/ml had not good prognosis. The cut-off value was 2.19 pg/ml for distinguishing between good prognosis group and not good prognosis group, the area under the ROC curve was 0.719 (95% CI 0.595-0.842, P = 0.04); among 53 patients with TNF-β≥2.19 pg/ml, 46 (86.8%) patients had not good prognosis. Conclusions:The high expression of TNF-β may indicate that the prognosis of AML patients is not good. When the level of TNF-β was equal or greater than 3.62 pg/ml, it may contribute to the prognostic stratification of AML patients.

Effect of epigallocatechin gallate on Th1/Th2 immune balance in patients with plaque-type psoriasis / 中华皮肤科杂志

Objective@#To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.@*Methods@#A total of 33 patients with plaque-type psoriasis vulgaris were enrolled, and peripheral blood mononuclear cells (PBMC) were isolated and cultured. The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay. PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours. Flow cytometry was performed to determine proportions of Th1 and Th2 cells, enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin[IL]-2, interferon[IFN]-γ) and Th2 cytokines (IL-4, IL-10) in the cell culture supernatant, and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor) . Statistical analysis was carried out by using t test.@*Results@#According to the MTT assay results, EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments. Compared with the control group, the EGCG group showed significantly decreased number of Th1 cells (t = 3.43, P = 0.026) , increased number of Th2 cells (t = 6.68, P = 0.026) , and decreased Th1/Th2 ratio (P < 0.05) . The levels of IL-2 and IFN-γ in the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L, 1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ± 196.45 ng/L, 3 287.63 ± 235.54 ng/L respectively) , while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L, 285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L, 165.46 ± 32.25 ng/L respectively) . Compared with the control group, the EGCG group showed significantly decreased T-bet mRNA expression (t = 11.99, P < 0.001) , but increased GATA3 mRNA expression (t = 18.62, P < 0.001) .@*Conclusion@#EGCG can reduce the number of Th1 cells, inhibit the production of Th1 cytokines and transcription factors, and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors, followed by the modulation of Th1/Th2 immune imbalance.

Research advances in interaction between il-4 and il-13, and m2 alveolar macrophages in pulmonary fibrosis / 中国药理学与毒理学杂志

Type-2 helper T cells (Th2) are a group of cells differentiated from CD4∗ T cells after antigen stimulation. lnterleukin-4(IL-4) and IL-13 secreted by Th2 cells act on macrophages and acti-vate them into M2 macrophages via "alternative activation". When lung tissue is continuously stimulated by the antigen, the alveolar and interstitial macrophages are activated by IL-4 and IL-13 secreted by Th2 cells to form M2 type alveolar macrophages, thereby participating in pulmonary fibrosis (PF). The process shows that M2 alveolar macrophages play an important role in PF. In this review, we investigated the role of cytokine IL-4 and IL-13 in PF, the anti-inflammatory and profibrotic effects of M2 alveolar macrophages, the role of IL-4 and IL-13 receptors in the treatment of PF and the dual role of M2 alveolar macrophages activated by IL-4 and IL-13 in PF. Hoping to spark novel ideas for the study of anti-PF drugs.

Inhibitory effect of astragalus polysaccharides on pulmonary inflammation in asthma model mice and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the inhibitory effects of astragalus polysaccharides (A P S) with different molecular weights on the pulmonary inflammaton in the asthmatic mice, and to elucidate their mechanisms. Methods: A total of 30 female BALB/c mice were selected. They were randomly divided into normal control group, asthma model group (model group), low molecular weight APS (APS-low) group, midde molecular wieght APS (APS-middle) and high molecular weight APS (APS-high) group, with 6 mice in each group. The mouse models of asthma in model group and APS groups were established by injecting and inhaling ovalbumin (OVA). The mice in APS-low, APS-middle and APS- high groups were separately given 0. 1 mL of 4 500, 15 000, 30 000 molecular weight APS for intrapeitoneal injecton before inhaled OVA The mice in normal control group were treated with the same amount of normal saline instead of OVA injected and inhalled. The behavioral changes of the mice were observed during the atomization with OVA, and the total number of WBC and the counts of inflammatory cells in the bronchoalveolar lavage fluid (BALF) were observed with light microscope; the pathological changes of lung tissue was observed by HE staining. Cytometric Bead Array (CBA) method was used to detect the levels of IL-4 and IFN -y in the serum as well as BALF. The spleen CD4 T cells were extracted and cultured, and the ratios of T h l, Th2, Thl7 and Treg cells were detected by flow cytometry. CBA method was used to detect the levels of IL-4, IFN-y, IL-17, and IL-10 in the culture supernatant. Results: Compared with normal control group, the mice in model group showed asthma-like symptoms such as sneezing, snouting nose and shortness of breath, inflammatory infiltration of lung tissue, airway mucosal edema, smooth muscle thickening; the IL-4 levels in serum and BALF were increased (P < 0 . 05), and the IFN-y levels in serum and BALF were significantly decreased (P < 0 . 05); IL-4 and IL-17 levels in cell culture supernatant were significantly increased (P < 0 . 05), and the IFN-y and IL-10 levels were significantly decreased (P < 0 . 05); the ratios of Th2 and Thl7 cells were significantly increased (P < 0 . 05), and the ratios of T h l and Treg cells were significantly decreased (P < 0 . 05). Compared with model group, the asthmatic symptoms of the mice in APS groups, for example, scratching nose and mouth, shortness of breath, and irritability, were relieved to varying degrees; the inflammatory cell infiltration and wall thickening were significantly improved; the levels of IL-4 in serum and BALF were significantly reduced (P < 0 . 05), and the IFN -y levels were increased (P < 0 . 05); the levels of IL-4 and IL-17 in the culture supernatant were significantly reduced (P < 0 . 05) and the levels of IFN-y and IL-10 were increased (P < 0 . 05); the ratios of Th2 and Thl7 cells were significantly decreased (P < 0 . 05), and the ratos of T h l and Treg cells were increased (P < 0 . 05). Compared between three APS groups, the asthmatic symptoms and lung tissue inflammatory infiltration were lightened obviously in APS-low group, the IL-4 levels and the ratios of Th2 and Thl7 cells in serum and BALF were significantly decreased (P < 0. 05), and the IFN-y levels in serum and BALF and the ratios of Th2 and Thl7 cells were significantly increased (P < 0 . 05). Conclusion: APS can significantly improve the situaton of airway inflammaton infiltraton and the symptoms of the asthmatic mice. APS plays a therapeutic role for asthma by regulating the Th 1 / Th 2 and Th l7 / Treg cell balance, decreasing the levels of IL-4 and IL-17 and increasing the levels of IFN -y and IL-10 level; the low molecular weight APS shows an obvious effect.

The role of Th1 and Th2 cells in the severity of acute pancreatitis / 中华急诊医学杂志

Objective To investigate the relationship between the levels of Thl and Th2 cells and their cytokines IFN-γ,TNF-αt,IL-2,IL-4,IL-5 and IL-13 in peripheral blood of patients with acute pancreatitis and the clinical severity of acute pancreatitis,and to evaluate the diagnostic value and significance for the severity of acute pancreatitis.Methods This study enrolled 72 patients with acute pancreatitis and 30 healthy controls admitted to the Emergency Surgery Department of the First Affiliated Hospital of Zengzhou University from January 2015 to June 2017.The patients were divided into the mild MAP group,moderate MSAP group,and severe SAP group according to the 2012 Atlanta classification criteria.Whole blood and serum samples were taken on the day of admission for all confirmed cases,the number of Th1 and Th2 cells was detected by flow cytometry,the expression levels of IFN-γ,TNF-α,IL-2,IL-4,IL-5 and IL-13 were determined by ELISA.The total length of hospital stay and length of hospital stay in ICU of each group were recorded.The number of Th 1 and Th2 cells and the expression of related cytokines in patients with mild,moderate and severe acute pancreatitis were compared by one-way analysis of variance.Pearson linear correlation was used to analyze the correlation between Th1and Th2 cytokines and the length of hospital stay and the length of hospital stay in ICU.A P＜0.05 was considered statistically significant.Results Compared with the healthy controls,the number of Th1 and Th2 cells,the levels of Th1 cytokine IFN-γ,TNF-α IL-2 and Th2 cytokine IL-4 were significantly increased in acute pancreatitis.Th1 cells in the SAP group were significantly higher than those in the MAP and MSAP groups (F=11.137,P＜0.01),while Th2 cells in the SAP group were significantly lower than those in the MAP and MSAP groups (F=9.493,P＜0.01),and there was no significant difference between the MAP group and MSAP group.The expression level of IFN-γ was higher in the SAP and MSAP groups than that in the MAP group (F=8.605,P=0.001).The expression level of TNF-α was higher in the SAP group than those in the MSAP and MAP groups (F=1 1.847,P＜0.01),and the IL-4 expression level was significantly lower in the SAP group than those in the MAP and MSAP groups (F=8.042,P=0.001).The expression level of IFN-γ was positively correlated with the length of hospital stay and the length of hospital stay in ICU (r=0.569,P=0.014;r=0.538,P=0.021).The expression level of TNF-α was positively correlated with the length of hospital stay (r=0.475,P=0.046).The expression level of IL-4 was negatively correlated with the length of hospital stay and the length of hospital stay in ICU (r==0.577,P=0.012;r=-0.657,P=0.003).Conclusions Th1 and Th2 cells and their associated inflammatory factors IFN-γ,TNF-α,IL-2 and IL-4 are elevated in patients with acute pancreatitis.The increase in the number of Th1 proinflammatory cytokines IFN-γ,TNF-α and Th1 cells tends to aggravate the development of acute pancreatitis,and the increase in the number of Th2 cells and the anti-inflammatory cytokine IL-4 are associated with a better prognosis.

Myeloid-derived suppressor cells aggravate Sjögren's syndrome by inhibiting the response of Th2 cells / 医学研究生学报

Objective Changes in the number and function of myeloid-derived suppressor cells (MDSC) were reported in clinical and experimental Sjögren’s syndrome (SS), whereas the underlying mechanisms of MDSCs in SS remain to be elucidated. This study was to observe the changes in the pathologic structure and function of the submandibular gland and salivary flow in SS mice after adoptive transfer or deletion of MDSCs and explore the action mechanisms of MDSCs. Methods Ten 4-week-old non-obese diabetic (NOD) mice (without SS-like symptoms) received adoptive transfer of purified MDSCs at 1×106 per mouse (the MDSC group, n = 5) or injection of PBS (the PBS group, n = 5). Another ten 10-week-old NOD mice were injected intraperitoneally with anti-Gr1 antibodies (the anti-Gr1 group, n = 5) or commensurable Rat IgG2b isotype antibodies (the Rat IgG2b group, n = 5). At 2 weeks after treatment, we determined the salivary flow rate, examined lymphocytic infiltration in the submandibular glands, and counted the MDSCs on Th2 cells in different groups of the mice. Results Compared with the PBS group, the NOD mice of the MDSC group showed significantly reduced Th2 cells in the peripheral blood ([0.67 ± 0.13] % vs [0.16 ± 0.07] %, P < 0.05) and spleen ([0.80 ± 0.13] % vs [0.37 ± 0.04] %, P < 0.05) and salivary flow ([78.70 ± 6.80] vs [33.85 ± 11.25] µL, P < 0.05), but increased numbers of MDSCs in the peripheral blood ([1.54 ± 0.14] vs [5.47 ± 1.54] ×105, P < 0.05) and spleen ([1.09 ± 0.23] vs [4.50 ± 1.04] ×105, P < 0.05). In comparison with the Rat IgG2b group, the animals of the anti-Gr1 group exhibited remarkably decreased Th2 cells in the peripheral blood ([0.55 ±0.09] % vs [0.92 ± 0.10] %, P < 0.05) and spleen ([0.63 ± 0.08] % vs [1.10 ± 0.06] %, P < 0.05) and salivary flow ([56.48 ± 14.18] vs [121.20 ± 10.34] µL, P < 0.05), as well as decreased numbers of MDSCs in the peripheral blood ([1.53 ± 0.12] vs [0.35±0.16] ×105, P < 0.05) and spleen ([2.53 ± 1.10] vs [0.91±0.07] ×105, P < 0.05). The adoptive transfer of MDSCs aggravated while the injection of anti-Gr1 antibodies attenuated lymphocytic infiltration in the submandibular gland of the mice. Conclusion MDSCs participate in the pathogenesis Sjögren’s syndrome by suppressing the response of Th2 cells, which suggests that increasing the response of Th2 cells by inhibiting MDSCs could be a novel target for the treatment of Sjögren’s syndrome.

Antiallergic Effect of Hizikia fusiformis in an Ovalbumin-Induced Allergic Rhinitis Mouse Model

OBJECTIVES: The extract of Hizikia fusiformis is known to exhibit anticancer, antiatopic and antioxidant activities. We aimed to investigate the extract of H. fusiformis on allergic rhinitis inflammation in a mouse model. METHODS: The 4-week-old BALB/c mice were randomly assigned into four groups: group A, control group (n=9); group B, allergic rhinitis group (n=10); group C (n=10) received 300 mg/kg of H. fusiformis during nasal challenging period; group D (n=10) received 600 mg/kg of H. fusiformis during general sensitization period and 300 mg/kg of H. fusiformis during nasal challenging period. Allergic inflammation was made with ovalbumin (OVA) and alum then challenged intranasally with OVA. H. fusiformis was intraperitoneally administered 3 hours before the OVA administration. Allergic symptom score and the levels of immunoglobulin G1 (IgG1), IgG2a, OVA-specific IgE antibodies, levels of cytokines in the nasal mucosa and in spleen cell culture supernatant, such as tumor necrosis factor alpha (TNF-α), interleukin 4 (IL-4), IL-5, IL-13, and IL-10 were assessed. The percentage of regulatory T cell was analyzed by flow cytometry. Eosinophilic infiltration and goblet cell hyperplasia were also evaluated. RESULTS: H. fusiformis administered groups C and D showed significant inhibitory effects on nasal symptoms, IL-13 mRNA expression and eosinophil infiltration/goblet cell hyperplasia in the nasal tissue; OVA-specific IgE production in serum (P<0.05). In group D, H. fusiformis treatment downregulated IL-4, IL-5, IL-13, TNF-α, and IL-10 cytokine expression in splenocyte culture as well as significantly decreased IgG2a, IgG1 levels in serum compared with group B (P<0.05). However, the expressions of IL-5, interferon-γ and forkhead box P3 mRNA did not change in groups C and D. CONCLUSION: H. fusiformis could induce antiallergic inflammation by suppressing the T-helper type 2 cytokine production (IL-13) locally and systemically, OVA-specific IgE formation, goblet cell hyperplasia, and eosinophilic infiltration in a mouse model of allergic rhinitis. Thus, H. fusiformis could be considered as a potential therapeutic agent in treating allergic rhinitis.

Research advances in Th1/Th2 balance and fibrotic diseases / 临床肝胆病杂志

Body immunity is closely associated with the pathogenesis of fibrotic diseases. In CD4+ T lymphocyte subsets, T helper 1 (Th1)/T helper 2 (Th2) balance plays an important regulatory role in the development and progression of many diseases. This article describes the association of Th1/Th2 balance with liver fibrosis, pulmonary fibrosis, and renal fibrosis. It is believed that in addition to the measurement of single cytokines, further studies are needed to clarify the specific association between T helper cells and other cells and related mechanisms.

Effect of epigallocatechin gallate on Th1/rh2 immune balance in patients with plaque-type psoriasis / 中华皮肤科杂志

Objective To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.Methods A total of 33 patients with plaque-type psoriasis vulgaris were enrolled,and peripheral blood mononuclear cells (PBMC) were isolated and cultured.The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay.PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours.Flow cytometry was performed to determine proportions of Th 1 and Th2 cells,enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin [IL]-2,interferon [IFN]-γ) and Th2 cytokines (IL-4,IL-10) in the cell culture supernatant,and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor).Statistical analysis was carried out by using t test.Results According to the MTT assay results,EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments.Compared with the control group,the EGCG group showed significantly decreased number of Th1 cells (t =3.43,P =0.026),increased number of Th2 cells (t =6.68,P =0.026),and decreased Th1/Th2 ratio (P ＜ 0.05).The levels of IL-2 and IFN-γin the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L,1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ±196.45 ng/L,3 287.63 ± 235.54 ng/L respectively),while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L,285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L,165.46 ± 32.25 ng/L respectively).Compared with the control group,the EGCG group showed significantly decreased T-bet mRNA expression (t =11.99,P ＜ 0.001),but increased GATA3 mRNA expression (t =18.62,P ＜ 0.001).Conclusion EGCG can reduce the number of Th1 cells,inhibit the production of Th 1 cytokines and transcription factors,and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors,followed by the modulation of Th 1/Th2 immune imbalance.

Study on the expression and significance of Th1/Th2 cells in the blood of patients with primary gouty arthritis / 中华风湿病学杂志

Objective This study is aimed to investigate the possible role of Th1/Th2 cell in the pathogenesis of primary gout arthritis. Methods The peripheral blood of 21 acute gout patients (AG), 20 intermittent gout patients (IG) and 20 healthy controls (HC) were collected. The clinical data and laboratory indicators of them were enrolled. The percentages of Th1 and Th2 cells were detected by flow cytometry (FCM). The expression of GATA-3, T-bet, IL-4 and interferon (IFN)-γ mRNA in Peripheral blood mononuclear cells (PBMCs) were measured using Real time quantitative polymerase chain reaction (PCR). The protein expression levels of IL-4 and IFN-γ in serum were detected by enzyme-linked immuno sorbent assay (ELISA). The measurement data were compared by one factor analysis of variance test. The correlation between variables was used by Spearman correlation analysis. Results The percentage of Th1 cells in peripheral blood of the AG group was [(23.2 ±8.3)％], and the IG group was [(20.5 ±9.3)％], which were significantly higher (F=6.520, P<0.05) than the percentage of HC group [(14.8±3.8)％]. There was no significant difference between AG group and IG group. The percentage of Th2 cells in peripheral blood of AG group were [(1.9 ±0.7)％], which was significantly lower (F=8.267, P<0.05) than and the percentage of HC group [(3.4±1.8)％] and IG group [(3.3± 1.2)％]. There was no significant difference between the IG group and the HC group. And the proportion of TH1/Th2 cells in the AG group was higher than that of the IG group and the HC group (F=10.406, P<0.01). The expression of T-bet mRNA and IFN-γ mRNA in the AG group and IG group were higher than that in the HC group (F=4.942, P=0.010)、(F=4.458, P=0.016). However, there was no significant difference between IG group and AG group. The expression of GATA-3 mRNA and IL-4 mRNA was significantly lower (F=3.564, P=0.035) (F=5.385, P=0.007) in the AG group and IG group when compared to the HC group. And there was no significant difference between the AG and the IG group. The IFN-γ level increased in the AG and IG group compared to the HC group (F=7.659, P=0.001). The IL-4 levels in the AG group was lower (F=7.099, P=0.002) than those of the IG and HC group. Conclusion Th1 cells in the peripheral blood of patients with GA are increased and accompanied with the decrease of Th2 cells. The results of this study suggest that the imbalance of Th1/Th2 cells in the inflammatory and immune response plays a critical role in the pathogenesis of GA.

Distinct characteristics of nasal polyps with and without eosinophilia / Diferenças nas características de pólipos nasais com e sem eosinofilia

Abstract Introduction Eosinophilic and noneosinophilic Nasal polyps (NPs) are different subtypes of NPs and require different treatment methods. Objective To compare the histologic characteristics, mRNA and protein expression between Nasal Polyps with and without eosinophilia. Methods NPs tissues were obtained from eighty-six NPs patients during surgery. Eosinophilic and noneosinophilic NPs were distinguished according to immunochemical results of the specimen. The histological, mRNA and protein expression features were compared between the two groups. Results In eosinophilic NPs, we observed a significantly higher GATA-3, IL-5, IL-4, IL-13 mRNA and protein expression. In noneosinophilic NPs, IL-17, IL-23 and RORc mRNA and protein expression were increased. Immunohistochemistry tests showed, more mast cells and less neutrophils in eosinophilic NPs compared with noneosinophilic NPs. Eosinophilic NPs patient presented more severe symptom scores when compared to noneosinophilic NPs. Conclusion We demonstrate for the first time that Th2 is the predominant reaction in eosinophilic NPs while Th17 is the predominant reaction in noneosinophilic NPs. Our study may provide new treatment strategy for NPs.

Resumo Introdução Pólipos nasais (PNs) eosinofílicos e não eosinofílicos são diferentes subtipos de PNs e requerem diferentes métodos de tratamento. Objetivo Comparar as características histológicas e a expressão de mRNAs e proteínas entre PNs com e sem eosinofilia. Método Amostras de PNs foram obtidos de 86 pacientes durante a cirurgia. PNs eosinofílicos e não eosinofílicos foram diferenciados segundo os resultados imunoistoquímicos de cada amostra. As características histológicas e de expressão de mRNAs e de proteínas foram comparadas entre os dois grupos. Resultados Em PNs eosinofílicos, observamos uma expressão significativamente maior dos mRNAs e proteínas GATA-3, IL-5, IL-4 e IL-13. Nos PNs não eosinofílicos, aumentou a expressão dos mRNAs e das proteínas IL-17, IL-23 e RORc. Nos testes imunoistoquímicos, observamos maior número de mastócitos e menor número de neutrófilos nos PNs eosinofílicos, em comparação com PNs não eosinofílicos. Os pacientes com PNs eosinofílicos obtiveram escores de sintomas mais graves vs. PNs não eosinofílicos. Conclusão Demonstramos, pela primeira vez, uma reação Th2 predominante em PNs eosinofílicos e uma reação Th17 predominante em PNs não eosinofílicos. Nosso estudo pode proporcionar novas estratégias terapêuticas para a rinossinusite crônica.

Changes of serum Th1/Th2 cytokines in peripheral blood pre-and post-liver transplantation / 天津医药

Objective To explore the trend of changes of Th1 and Th2 cytokines within 6 months after adult liver transplantation (LT). Methods Twenty-three patients from Tianjin First Center Hospital were chosen as the patient group with an average age of (52.7 ± 7.6), range from 37 to 63 years old, including 21 males and 2 females. Twenty healthy staffs from Tianjin First Center Hospital formed the control group (C) with 15 males and 2 females whose average age was (31.0 ± 6.1) ranged from 22 to 24 years old. The patient group was treated with tacrolimus after LT as main immunosuppressive drug. The peripheral blood at time points before (T0) and 1 month (T1), 3 months (T3), 6 months (T6) after LT at 9:00 AM were collected. The blood sample was also collected form control group but only one time. Levels of IL-2, IFN-γ, IL-10 and TGF-βwere detected by ELISA. Results (1) The concentration of IL-2 showed a continuous up-going trend, which was not such obvious between T1 and T0, and until T3 reached a higher concentration than T0. The concentration at T6 was higher than T0 and T1. There were no significant differences in concentrations of T0 to T3 between patient group and control group, while T6 reached a higher concentration in patient group than that of the control group. (2) The concentration of IFN-γexperienced a shortly down-going trend from T0 to T3, and started rising, reached the peak at 3 months after the operation, then started its down-going trend. There were no significant differences in the concentrations of IFN-γfrom T1 to T6, and T3 reached a higher concentration than T1 while T6 was lower than T3. Only at T3, the concentration of IFN-γwas higher in patient group than that of control group. (3) There were no significant differences in the concentrations of IL-10 at various time points in patient group, and there were no significant differences in the concentrations of IL-10 at different time points between two groups (P>0.05). (4) The concentration of TGF-βshowed a gradual decline after the operation, and reached its bottom at T6, and which was lower than T0 to T3. Compared with the control group, the down-going trend was not such obviously at T0 and T1, and the concentration was down at T3 and T6(P<0.05). Conclusion Our results suggest that there is a tendency of an increasing Th1 cytokine expression at early stage in post-transplantation, while the TGF-βof Th2 cytokine is a decreasing trend. This tendency may associate with the autoimmunity response caused by LT and the immunosuppressive drugs.