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Objective:To study the effect of resveratrol (RES) on interleukin-1β (IL-1β)-induced chondrocytes and its pathways of action.Methods:Wistar mammary rat chondrocytes were extracted and divided into 5 groups: control group, IL-1β group, RES+IL-1β group, RES+IL-1β+EX-527 [silent information regulator 1 (SIRT1) inhibitor] group and RES+IL-1β+AS [frame transcription factor O1 (FOXO1) inhibitor] group. Quantitative real time polymerase chain reaction (qRT-PCR) was used to detect SIRT1, forkhead FOXO1 and matrix metalloproteinase 3 (MMP-3) mRNA expression. Protein expression of chondrocyte type Ⅱ collagen (Col-Ⅱ) detected by immunofluorescence, and the expression of chondrocyte SIRT1 and p-FOXO1/FOXO1 was measured by Western blot. The expression of chondrocyte inflammatory factors IL-6 and TNF-α was measured by enzyme-linked immunosorbent assay. One-way analysis of variance (ANOVA) was performed and two-way comparisons between groups were made using the least significant difference (LSD) method. P< 0.001 was statistically significant. Results:Compared to normal chondrocytes, the mRNA and protein expressions of Col-Ⅱ, SIRT1, FOXO1 and p-FOXO1/FOXO1 in chondrocytes induced by IL-1β was significantly decreased ( P<0.001). The secretion of tumor necrosis factor (TNF)-α [(24.70±2.84), t=19.24, P<0.001] and IL-6 [(3.35±0.28), t=12.97, P<0.001] was significantly increased, and the mRNA expression of MMP-3 [(2.46± 0.23), t=12.61, P<0.001] was significantly increased. The mRNA and protein expressions of Col-Ⅱ, SIRT1, FOXO1 and p-FOXO1/FOXO1 were significantly increased. The secretion of TNF-α [(12.60±1.05), t=10.14, P<0.001] and IL-6 [(2.00±0.15), t=9.89, P<0.001] was significantly reduced by RES treated IL-1β-induced chondrocytes. mRNA expression of MMP-3 [(1.30±0.14), t=10.460, P<0.001] was decreased. After adding SIRT1 inhibitor EX-527 or FOXO1 inhibitor AS, RES significantly reduced the mRNA and protein expression of Col-Ⅱ, SIRT1, FOXO1 and p-FOXO1/FOXO1 in IL-1β-induced chondrocytes ( P<0.001). The secretion of TNFα and IL-6 was significantly decreased ( P<0.001), and the mRNA expression of MMP-3 was significantly decreased ( P<0.001). Conclusion:RES significantly ameliorates IL-1β-induced cartilage extracellular matrix egradation and inflammatory responses via the SIRT1/FOXO1 pathway.
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Objective:To explore the effects of long non-coding RNA (lncRNA) FAM224A on the proliferation and migration of ovarian cancer cells by regulating the expression of miRNA-590-3p (miR-590-3p).Methods:Human ovarian cancer cell lines OC3, SKOV-3, HO-8910, A2780 and human normal ovarian epithelial cell line IOSE80 were selected, and the relative expression of FAM224A in each cell line was detected by real-time quantitative polymerase chain reaction (qRT-PCR). The cell line with the lowest relative expression level of FAM224A was screened for follow-up experiment. The cells were divided into FAM224A group (transfected with FAM224A mimic gene) and control group (transfected with control mimic gene). CCK-8 method and cell scratch test were used to detect the cell proliferation and migration ability of the two groups. The bioinformatics website LncBase v.2 predicted that the target gene that FAM224A might complementarily bind to was miR-590-3p. qRT-PCR was used to detect the relative expression levels of miR-590-3p and forkhead box protein A2 (FOXA2) mRNA, and the expressions of related proteins were detected by Western blot.Results:The relative expression levels of FAM224A in ovarian cancer cell lines OC3, SKOV-3, HO-8910, A2780 and normal ovarian epithelial cell line IOSE80 were 0.23±0.04, 0.65±0.05, 0.45±0.03, 0.63±0.08 and 1.02±0.11, respectively, and the difference was statistically significant ( F = 14.78, P < 0.01), and the cell line with the lowest relative expression level of FAM224A was OC3. The results of CCK-8 method showed that the proliferation ability of OC3 cells in the FAM224A group was lower than that in the control group on the 2nd, 3rd, 4th and 5th day of culture (all P < 0.05). The scratch healing rates of OC3 cells in the FAM224A group and the control group were (18.6±2.3)% and (71.7±7.2)%, respectively, and the difference was statistically significant ( t = 6.99, P < 0.01). The relative expression levels of FAM224A in OC3 cells in the FAM224A group and the control group was 12.36±1.45 and 1.14±0.24, respectively ( t = 13.08, P < 0.01); the relative expression levels of miR-590-3p were 0.19±0.06 and 1.04±0.20, respectively ( t = 4.01, P < 0.01); the relative expression levels of FOXA2 mRNA were 6.37±1.37 and 1.05±0.08, respectively ( t = 3.86, P < 0.01). Compared with the control group, the expression of FOXA2 protein in OC3 cells in the FAM224A group was increased, the expressions of cell proliferation protein cyclin-dependent kinase 2 (CDK2) and cyclin D3 were decreased, and the expression of cell migration protein Snail was decreased. Conclusions:FAM224A is low expressed in ovarian cancer cell lines. FAM224A reduces the proliferation and migration ability of ovarian cancer OC3 cells by inhibiting the expression of miR-590-3p.
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Objective:To observe the expression of miR-142-5p and forkhead transcription protein O subgroup 3 (FOXO3) in CD4 + T cells of experimental autoimmune uveitis (EAU) model rats, and preliminarily explore the targeting relationship between the two and the effect on EAU impact. Methods:Ten Lewis rats were randomly divided into model group and control group. Rats in the model group wree induced an EAU animal model by adoptive immunization. Twenty days after immunization, CD4 + T cells were extracted from the eyeballs and draining lymph nodes of rats in the control group and model group, and divided into control group, model group, mimic-negative control (NC) group, miR-142-5p-mimic group, and small interference (si)-NC group, si-FOXO3 group for in vitro experiments. The miR-142-5p-mimic group and si-FOXO3 group were transfected with miR-142-5p-mimic and si-FOXO3, respectively. Twenty-five Lewis rats were randomly divided into model group, mimic-NC transfected group, miR-142-5p-mimic transfected group, si-NC transfected group, and si-FOXO3 transfected group. The above-mentioned in vitro experimental groups were injected with cells respectively. Slit lamp microscopy and EAU score were performed on 4, 8, 12, 16, 20 days after immunization; on 20 days after immunization, hematoxylin-eosin staining was performed for histopathological grading. Real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression of miR-142- 5p and FOXO3 mRNA in CD4 + T cells and eye tissues of rats in each group, and helper T cell 17 (Th17) marker interleukin (IL)-17, IL-22, retinoic acid-related orphan receptor gamma (ROR gamma) relative expression level in the supernatant. Bioinformatics website and dual luciferase was used to predict the targeting relationship between miR-142-5p and FOXO3. One-way analysis of variance or t test was used for comparison between groups. Results:All rats in the model group showed symptoms of EAU to varying degrees, and the symptoms became worse with time. Compared with the control group, the relative expression of miR-142-5p mRNA in CD4 + T cells of the model group increased, and the relative expression of FOXO3 mRNA decreased. The differences were statistically significant ( t=7.374, 10.423; P=0.002, 0.001). Compared with the mimic-NC group, the relative expression of miR-142-5p mRNA in the CD4 + T cells of the miR-142-5p-mimic group increased, and the difference was statistically significant ( t=6.540, P=0.003). Compared with the model group, mimic-NC group, and si-NC group, the relative expression of IL-17, IL-22, and RORγ mRNA in CD4 + T cells in the miR- 142-5p-mimic group and si-FOCO3 group increased significantly. The difference was statistically significant ( F=26.110, 6.292, 5.269, 55.660, 10.490, 11.430; P<0.05). Compared with the mimic-NC transfected group, the relative expression of miR-142-5p mRNA in the ocular tissues of the miR-142-5p-mimic transfected rats increased significantly, and the difference was statistically significant ( t=6.690, P<0.05). Compared with the transfected si-NC group, the relative expression of FOXO3 mRNA in the eye tissue of the transfected si-FOXO3 group was significantly decreased, and the difference was statistically significant ( t=17.751, P<0.05). Rats in the mimic-NC transfected group, miR-142-5p-mimic transfected group, si-NC transfected group, and si-FOXO3 transfected group prolonged with time after immunization, and the EAU scores showed an upward trend. The EAU score and histopathological grade of rats in the miR-142-5p-mimic transfected group were higher than those in the mimic-NC transfected group, and the difference was statistically significant ( t=5.633, 6.286; P<0.05). The EAU score and histopathological grade of the rats in the transfected si-FOXO3 group were higher than those in the transfected si-NC group, and the difference was statistically significant ( t=6.852, 6.635; P<0.05). FOXO3 has a targeting relationship with miR-142-5p. Conclusions:In EAU rat CD4 + T cells, the expression of miR-142-5p is up-regulated, while the expression of FOXO3 is down-regulated. miR-142-5p targets the expression of FOXO3 to promote the development of Th17 cell-related inflammatory factors.
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Objective:To investigate the correlation of the expression of forkhead transcription factor O1 (FOXO1) with clinicopathological features and the prognosis in patients with diffuse large B-cell lymphoma (DLBCL).Methods:The data of 42 patients newly diagnosed with DLBCL in Hebei General Hospital admitted from June 2012 to January 2020 were collected. The expressions of FOXO1, phosphorylated FOXO1 (p-FOXO1) in DLBCL tissues were detected by using immunohistochemistry. The association of FOXO1 expression with clinicopathological features and the prognosis in DLBCL patients was retrospectively analyzed.Results:The positive rate of FOXO1 was 42.9% (18/42) and the positive rate of p-FOXO1 was 28.6% (12/42) in DLBCL tissues. There were no statistically significant differences in the positive rates of FOXO1 and p-FOXO1 among patients stratified by gender, age, Ann Arbor staging, immunophenotype, Eastern Cooperative Oncology Group score, lactate dehydrogenase, international prognostic index, β 2-microglobulin (β 2-MG) and primary sites (all P > 0.05). The positive rate of FOXO1 in patients with non-B symptoms was higher than that in those with B symptoms [53.6% (15/28) vs. 21.4% (3/14), χ2=3.938, P=0.047], and there was no statistically significant difference in the positive rate of p-FOXO1 among patients with or without B symptoms ( P > 0.05). The 2-year overall survival (OS) rate in FOXO1 positive group was higher than that in FOXO1 negative group (90.9% vs. 66.7%), the 2-year OS rate in p-FOXO1 positive group was lower than that in p-FOXO1 negative group (50.0% vs. 85.0%), and the differences were not statistically significant (all P > 0.05). Among patients without B symptoms, the 2-year OS rate in FOXO1 positive group was higher than that in FOXO1 negative group (100.0% vs. 50.0%, χ2=5.486, P=0.019). Among patients with primary lymph node, elevated β 2-MG and non-B symptoms, the 2-year OS rate in p-FOXO1 negative expression group was higher than that in p-FOXO1 positive group (100.0% vs. 50.0%, 100.0% vs. 25.0%, 91.7% vs. 33.3%), and the differences were statistically significant (all P < 0.05). Conclusions:FOXO1 may be involved in the development and progression of DLBCL, and FOXO1 positive expression may indicate the good prognosis of patients. These results suggest that p-FOXO1 positive expression may be related with poor prognosis.
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ObjectiveTo investigate the expression levels of forkhead box A2 (FOXA2) and forkhead box J2 (FOXJ2) in hepatocellular carcinoma (HCC) tissue and the association of FOXA2 and FOXJ2 with HCC. MethodsClinical data and pathological tissue samples were collected from 54 patients with pathologically confirmed HCC in The First Affiliated Hospital of Henan University of Traditional Chinese Medicine from January 2014 to July 2019. The immunohistochemical SP method was used to measure the protein expression levels of FOXA2 and FOXJ2 in HCC tissue, and their association with HCC-related clinicopathological features and patient prognosis was analyzed. The chi-square test and the adjusted chi-square test were used for comparison of categorical data; a Spearman correlation analysis was performed to investigate the correlation between the expression of FOXA2 and FOXJ2; the Kaplan-Meier method was used for survival analysis; Image-Pro Plus was used to perform the semi-quantitative analysis of the expression of FOXA2 and FOXJ2; the Wilcoxon rank-sum test was used for comparison between groups. ResultsThe positive rates of the protein expression of FOXA2 and FOXJ2 in HCC tissue were 70.37% (38/54) and 75.92% (41/54), respectively, and there was a significant positive correlation between the expression levels of FOXA2 and FOXJ2 (rs=0.648, P＜0.001). In both negative and positive groups, the expression level of FOXA2 was associated with tumor diameter, degree of tumor differentiation, number of tumors, and alpha-fetoprotein (χ2=5.440, 4.113, 4.352, and 3.865, P=0.020, 0.043, 0037, and 0.049), and the expression level of FOXJ2 was associated with the degree of tumor differentiation (χ2=9.267, P=0.002). The group with positive expression of FOXA2 and FOXJ2 had a significantly higher cumulative survival rate than the group with negative expression of FOXA2 and FOXJ2 (P＜0.01). ConclusionThe expression levels of FOXA2 and FOXJ2 are associated with the development, progression, and prognosis of HCC, and they have a synergistic effect in the development and progression of HCC.
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Hepatocellular carcinoma (HCC) is one of the most common malignant cancers and has high incidence and mortality rates and poor prognosis. Forkhead box (FOX) transcription factor family can regulate cell growth, differentiation, and tissue development and plays an important role in tumor. This article reviews the association of the molecular expression of the FOX family with the development, progression, and prognosis of HCC and analyzes the mechanism of action of FOX in the progression of HCC. It is pointed out that FOX family is expected to become a new target for HCC treatment.
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Objective@#To explore the prognostic value of human mitochondrial transcription termination factor 3 (hMTERF3) and forkhead box protein 3 (Foxp3) in non-small cell lung cancer (NSCLC).@*Methods@#The clinical data of 88 patients with NSCLC who were admitted to the Third Medical Center of PLA General Hospital from March 2017 to March 2018 were retrospectively analyzed. All patients were diagnosed by pathological puncture. The patients were followed-up by telephone for 12 months, and according to the prognosis, the patients were divided into good prognosis group and poor prognosis group. The pathological tissues were taken from all patients, and the expressions of hMTERF3 and Foxp3 proteins were detected by immunohistochemistry. The expressions of hMTERF3 and Foxp3 in the good prognosis group and the poor prognosis group were compared. Logistic regression model was used to analyze the risk factors of poor prognosis in patients with NSCLC.@*Results@#Of 88 patients, 61 patients (69.3%) had good prognosis and 27 patients (30.7%) had poor prognosis. The positive expression rate of hMTERF3 in the good prognosis group was 57.4% (35/61), which was significantly lower than that in the poor prognosis group (81.5%, 22/27) (χ 2= 4.766, P= 0.029). The positive expression rate of Foxp3 in the good prognosis group was 55.7% (34/61), which was significantly lower than that in the poor prognosis group (85.2%, 23/27) (χ 2= 7.113, P= 0.008). The proportions of patients with medium and high differentiation or stage Ⅰ- Ⅱ in the good prognosis group were 82.0% (50/61) and 68.8% (42/61), respectively, which were significantly higher than those in the poor prognosis group [48.15% (13/27) and 25.93% (7/27)] (both P < 0.05). Logistic regression analysis showed that the poor differentiation, stage Ⅲ-Ⅳ, hMTERF3-positive and Foxp3-positive were the risk factors for poor prognosis in NSCLC patients (all P < 0.05).@*Conclusions@#The positive expression rates of hMTERF3 and Foxp3 in patients with good prognosis are lower. The hMTERF3-positive and Foxp3-positive are risk factors for poor prognosis in NSCLC patients.
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Objective The aim of this study was to evaluate the number of Treg cells in preeclamp-sia (PE) patients, to explore the expression levels of microRNA-210 (microRNA-210) and forkhead box p3 (Foxp3) genes in preeclampsia, and to reveal the regulatory mechanism of microRNA-210 and Foxp3 in preeclampsia. Methods Serum levels of cytokines [ interleukin ( IL)-6, IL-10, IL-17, and transforming growth factor-beta 1 (TGF-β1)] were detected with enzyme-linked immunosorbent assay (ELISA). 29 pa-tients with late-onset preeclampsia (≥36 weeks of gestation) , 27 pregnant women with normal uncomplicat-ed pregnancies (≥36 weeks of gestation) and 20 healthy non-pregnant women were enrolled in the study. Reverse-transcription polymerase chain reaction ( qRT-PCR) was performed to detect mRNA expression for maternal placenta retinoic acid-related orphan receptor C (RORc), Foxp3, and miR-210. Foxp3 protein expression was evaluated by Western blot. Results ⑴The serum levels of IL-6, IL-17 and TGF-beta 1 in preeclampsia patients were significantly higher than those in normal pregnant women, and the level of Treg cytokine IL-10 was lower than that in normal pregnant women ( P <0. 05 ) . ⑵ The percentage of CD4 +CD25 +CD127 - /CD4 +T cells in peripheral blood of preeclampsia patients was significantly lower than that of normal pregnancy group and healthy non-pregnant women ( P <0. 001 ) . ⑶ The mRNA expression of Foxp3 in placenta of preeclampsia patients was significantly lower than that of normal pregnant women, RORc in preeclampsia patients was significantly higher than that of normal pregnant women, and the expres-sion of microRNA210 in preeclampsia patients was enhanced ( P<0. 01 ) . ⑷ Consistent with mRNA ex-pression results, lower protein expression levels of Foxp3 was observed in patients with PE compared with normal pregnant subjects. Conclusions Treg cells decreased in preeclampsia patients and Treg/Th17 im-balance existed in preeclampsia patients, which regulate maternal immune tolerance to fetuses. The expres-sion of Foxp3 in placenta of preeclampsia patients was significantly decreased, which was correlated with the expression of microRNA-210.
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Hepatic fibrosis refers to the pathological process of abnormal proliferation of connective tissue in the liver caused by various pathogenic factors. Hepatic fibrosis is observed in the whole process of liver injury repair and healing, and long-term chronic injury factors may induce the progression of hepatic fibrosis into liver cirrhosis. Epidemiological data show that there are 1.1805 million new cases of hepatitis B and 243 thousand cases of hepatitis C reported in 2017. Chronic viral hepatitis has brought heavy social and economic burden. Forkhead transcription factor (FOXO) belongs to the forkhead family and plays an important role in various cell life activities. Studies have shown that FOXO1/3 can regulate hepatic stellate cell activity through the TGFβ pathway and thus play an important role in hepatic fibrosis. This article reviews the research advances in the pathogenesis of hepatic fibrosis, the mechanism of action of FOXO1/3 in the regulation of hepatic fibrosis, and targeted therapy for FOXO1/3 (one of the sites for hepatic fibrosis).
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Objective To observe the level of CD4+CD25+ regulatory T cells (CD4+CD25+ Treg cells) with positive fork head transcription factor 3 (Foxp3) and changes of T-box transcription factor TBX1 (TBX1) and myocyte specific enhancer 2D (MEF2D) expression in peripheral blood of rats with acute rejection after renal transplantation,and to investigate its regulatory mechanisms by combined with renal function,plasma interleukin-10 (IL-10),interferon-γ (IFN-γ) and renal histopathological changes.Methods Rat renal transplantation model was established and divided into two groups:acute rejection group (AR group) and non-acute rejection group (non-AR group).Their renal function including serum creatinine (Scr) and blood urea nitrogen (BUN) in plasma was measured.The renal histopathology was observed by HE staining.Levels of IL-10 and IFN-γ in plasma were detected by ELISA.The proportion of CD4+CD25+ Treg cells was measured by flow cytometry.The mRNA expressions of Foxp3,TBX1 and MEF2D in CD4+CD4+Treg cells were detected by real-time PCR,and their protein expressions were tested by Western blotting.Results Compared with these in the non-AR group,the levels of BUN,Scr and IFN-γ significantly increased in AR group (all P ＜ 0.05),while IL-10 decreased (P ＜ 0.05).Renal histopathology in the acute rejection group showed glomerular hypertrophy and mesangial cell proliferation,capillary proliferation and neutrophil infiltration;renal interstitial edema and tubular necrosis,accompanied by lymphocytes,plasma cells and neutrophils infiltration.Compared with that in the non-AR group,the percentage of CD4+CD25+ Treg cells in peripheral blood was notably lowered in AR group (4.50％±0.50％ vs 5.74％±1.96％,P ＜ 0.05).The mRNA and protein expressions of Foxp3 and MEF2D were lower in AR group than those in non-AR group,while the expressions of TBX1 was elevated (all P ＜ 0.05).Conclusions In rats with acute renal allograft rejection,the percentage of CD4+CD25+ Treg cells and expressions of Foxp3,MEF2D and IL-10 decrease,while the expressions of TBX1 and IFN-γ enhance.These participate in the development of acute rejection after renal transplantation,and aggravate the renal damage.
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Objective To investigate the expressions of forkhead transcription factor O1 (FOXO1) and Kruppel like transcription factor 9 (KLF9) in non-small cell lung cancer (NSCLC) and their clinical significances. Methods A total of 102 patients with NSCLC who underwent surgical resection in the First Hospital of Yulin from September 2012 to June 2015 were selected as NSCLC group, and the paracancerous tissues more than 5 cm far from the edge of the cancer tissues were selected as the control group. The relative expressions of FOXO1 and KLF9 mRNA in NSCLC tissues and paracancerous tissues were detected by real-time quantitative polymerase chain reaction (qRT-PCR), and the immunohistochemistry was used to detect the expressions of FOXO1 and KLF9 in NSCLC tissues and paracancerous tissues. According to the mean mRNA expression in NSCLC tissues, FOXO1 and KLF9 were divided into high expression group and low expression group, their relationships with clinicopathological parameters were observed. The correlation of FOXO1 and KLF9 expressions was analyzed by Pearson method, and the survival of patients was analyzed by Kaplan-Meier method, logistic regression analysis was performed on the related factors affecting the prognosis of NSCLC. Results Compared with the control group, the expression level of FOXO1 in NSCLC group increased (1.28±0.47 vs. 2.82±0.15, t = 31.525, P < 0.05), while the expression level of KLF9 decreased (1.04±0.11 vs. 0.34± 0.07, t = 54.222, P < 0.05). The expressions of FOXO1 were lower in NSCLC tissues of patients with lymph node metastasis, high differentiation, and TNM stage Ⅲ-Ⅳ than those in NSCLC tissues of patients without lymph node metastasis, middle and low differentiation, and TNM stage Ⅰ-Ⅱ, and the differences were statistically significant (all P < 0.05). The expressions of KLF9 were higher in NSCLC tissues of patients with lymph node metastasis, high differentiation, and TNM stage Ⅲ-Ⅳ than those in NSCLC tissues of patients without lymph node metastasis, middle and low differentiation and TNM stage Ⅰ-Ⅱ, and the differences were statistically significant (all P < 0.05). Pearson correlation analysis showed that there was no significant correlation between FOXO1 and KLF9 expression (r = 0.154, P = 0.156). Kaplan-Meier analysis showed that the 3-year overall survival rate of FOXO1 high expression group was lower than that of FOXO1 low expression group (10.81％ vs. 60.71％, χ 2 = 27.341, P < 0.01). The 3-year overall survival rate of KLF9 low expression group was lower than that of KLF9 high expression group (26.32％ vs. 61.54％, χ 2 = 8.526, P = 0.003). Logistic regression analysis showed that FOXO1 was a risk factor for the prognosis of NSCLC (OR = 2.682, P = 0.003), and KLF9 was a protective factor for the prognosis of NSCLC (OR = 0.446, P = 0.012). Conclusions The high expression of FOXO1 and the low expression of KLF9 in NSCLC tissues can be used as effective indexes to predict the progression of NSCLC. The expressions of FOXO1 and KLF9 are closely related to the poor prognosis of patients.
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OBJECTIVE To invest igate the therapeutic effect and mechanism of PPAR gamma agonist on allergic rhinitis(AR) in mice. METHODS AR murine model was established by OVA sensitization and challenge. The behavior observation was used to understand the improvement effect of PIO on AR symptoms. The morphological characteristics of nasal tissues were observed by HE staining. The total RNA was extracted to investigate the level of mRNA expression of Foxp3, T-bet and GATA-3. The changes of CD4+Foxp3+T cells in spleen of mice were analyzed by flow cytometry. RESULTS BALB/c mice received OVA sensitization followed by OVA intranasal challenge, the frequencies of sneezing and nose-scratching increased signif icantly in AR group compared with control group. The frequencies decreased significantly in PIO group, compared with AR group. The continuity of nasal mucosa ciliated columnar epithelium in AR group was destroyed and appeared to be repaired in PIO group. Inflammatory cells infiltration was also markedly decreased by PIO treatment. PIO significantly increased the expression of Foxp3 mRNA(P ＜0.001) compared with AR and control group. There was no significant difference in T-bet between PIO group and AR group, but the expression of GATA-3 mRA in PIO group was significantly lower than AR group. The proportion of CD4+Foxp3+T cells in AR group (4.43％±0.25％) decreased compared with control group (5.19％±0.39％) (P ＜0.001). PIO treatment induced production of Tregs (6.35％±0.37％) compaered with control group(P ＜0.001). CONCLUSION PPAR-gamma agonist can effectively alleviate allergic symptoms of mice and regulate the balance of Th1/Th2. The role of PPAR gamma agonist in the treatment of AR may be the amplification of Tregs by promoting Foxp3 expression.
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Objective To evaluate the role of sirtuin 3 ( SIRT3)∕forkhead box O3α ( FOXO3α) signaling pathway in dexmedetomidine-induced reduction of hepatic ischemia-reperfusion ( I∕R) injury in mice. Methods Forty clean-grade C57BL∕6 mice of both sexes, aged 2 weeks, weighing 6-8 g, were di-vided into 4 groups (n＝10 each) using a random number table method: sham operation group (group S), hepatic I∕R group ( group I∕R), dexmedetomidine group ( group D) and SIRT3 inhibitor 3-TYP plus dexmedetomidine group (group T+D). Portal vein and hepatic artery supplying left and middle lobes of the liver and biliary tract were clamped resulting in ischemia of 70% of the liver in anesthetized rats. Normal sa-line 0. 25 ml was intraperitoneally injected at 1 h before establishing model, and 30 min later dexmedetomi-dine 50 μg∕kg (diluted to 0. 25 ml in normal saline) was intraperitoneally injected in group D. In group T+D, 3-TYP 5 mg∕kg (diluted to 0. 25 ml in normal saline) was intraperitoneally injected at 1 h before estab-lishing model, and 30 min later dexmedetomidine 50 μg∕kg (diluted to 0. 25 ml in normal saline) was in-traperitoneally injected. Mice were selected at 6 h after reperfusion, blood samples were obtained through eyeball, and the mice were then sacrificed and kidneys were removed for determination of the serum concen-trations of creatinine (Cr) and blood urea nitrogen (BUN), cell apoptosis (by TUNEL), malondialdehyde (MDA) content (using thiobarbituric acid method), superoxide dismutase (SOD) activity (by xanthine oxidase method), and acetylation of FOXO3α in renal tissues (by using immunoprecipitation) and for ex-amination of the pathologic changes. The damage to renal tubules was scored. Apoptosis index ( AI) was calculated. Results Compared with group S, the renal tubular damage score and AI were significantly in-creased, serum concentrations of Cr and BUN were increased, the content of MDA was increased, the ac-tivity of SOD was decreased, and the acetylation of FOXO3α was decreased in I∕R, D and T+D groups ( P＜0. 05). Compared with group I∕R, the renal tubular damage score and AI were significantly decreased, serum concentrations of Cr and BUN were decreased, the content of MDA was decreased, the activity of SOD was increased, and the acetylation of FOXO3α was decreased in group D (P＜0. 05), and no signifi-cant change was found in the parameters mentioned above in group T+D (P＞0. 05). Compared with group D, the renal tubular damage score and AI were significantly increased, serum concentrations of Cr and BUN were increased, the content of MDA was increased, the activity of SOD was decreased, and the acetylation of FOXO3α was decreased in group T+D ( P＜0. 05). Conclusion Activation of SIRT3∕FOXO3α signaling pathway is involved in dexmedetomidine-induced reduction of hepatic I∕R injury in mice.
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Objective To research the obesity-related gene (FTO) and forkhead transcription factors O1 (FoxO1) protein expression level in the livers of non-alcoholic fatty liver disease(NAFLD) rat model. Methods The animal model of NAFLD in rats was prepared by feeding high energy and high fat feed. Then the rat blood and liver tissue were collected for detecting the liver index and blood biochemical indexes, including triglycerides (TG), total cholesterol (TC), alanine aminotransferase (AST), aspartate aminotransferase(ALT) ,alkaline phosphatase(ALP),high-density lipoprotein (HDL) and low density lipoprotein(LDL) ;the liver pathological examination was performed;FTO protein and Fox O1 protein expression levels in rat liver were detected by using the immunohistochemical assay. Results The rat liver weight,body weight and liver index after 8 weeks in the model group were higher than those in the control group,the difference was statistically significant (P<0.05);the levels of AST, ALT, LDL, ALP, TG and TC in the model group were higher than those in the control group,the difference was statistically significant (P<0. 05),the HDL level in the model group was lower than that in the control group, the difference was statistically significant (P<0.05) ;the model group produced steatosis and inflammation in hepatic lobule part,while the control group had no these lesions;the FTO prot ein and FoxO1 protein expression levels in liver of the model group were higher than those in the control group,the difference was statistically significant (P<0.05). Conclusion FTO and FoxO1 interaction may disturb the normal energy and fat metabolism.
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Objective To explore the expression of forkhead box protein 3 (FOXP3) in gastric cancer cells,and its relationship with clinicpathological characteristics and prognosis of gastric cancer patients.Methods The expression profile of FOXP3 in 111 gastric adenocarcinoma tissues and 20 normal gastric mucosa were detected with immunohistochemistry,statistical analysis combined with clinical data of patients with gastric cancer.Results Positive staining of FOXP3 was mainly located in the nucleus of gastric adenocarcinoma cells with the positive rate of 42.3％ (47/112).In the normal gastric tissue,the positive rate of FOXP3 was 0％ (0/20).There were significant difference (x2 =13.207,P ＜0.05) when comparison was made between gastric adenocarcinoma and normal gastric tissue groups.Comparison of FOXP3 positive rate between gastric adenocarcinoma patients and its histological subtype (tumor size,depth of invasion,and clinical stage) showed significant difference (P ＜0.05),but (sex,age,and staging of tumor node metastasis (TNM),lympho node metastasis and differentiation) showed no significant difference (P ＞ 0.05).In the survival analysis,there was no significance in survival rate between the FOXP3 positive group and FOXP3 negative group either (P ＞ 0.05).Conclusions The FOXP3 were expressed in the human gastric cancer specimens.To a certain extent,FOXP3 was involved in the development of gastric adenocarcinoma and related to the degree of malignancy,but is not related to the prognosis of gastric cancer.
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Objective To investigate the expression of forkhead box C2 (FOXC2),Vimentin and E-cadherin protein in colorectal adenocarcinoma and the relationship of FOXC2,Vimentin and E-cadherin with the clinicopathological parameters,and the role of epithelial-mesenchymal transition (EMT) in colorec tal adenocarcinoma and the correlation between FOXC2 and EMT.Methods The expressions of FOXC2,Vimentin and E-cadherin in 102 cases of colorectal adenocarcinoma tissues and 30 cases of adjacent noncancerous tissues were detected by immunohistochemical method,and the correlation between FOXC2 and EMT and the clinicopathological parameters of colorectal adenocarcinoma were analyzed.Results The positive rates of FOXC2,Vimentin and E-cadherin in colorectal adenocarcinoma tissues were 53.9％,40.2％ and 26.5％,respectively.The positive rates of FOXC2,Vimentin and E-cadherin in adjacent noncancerous tissues were 6.7％,0 and 63.3％.Compared to paracancerous tissue,the expression of FOXC2 and Vimentin in colorectal adenocarcinoma was significantly increased,while the expression of E-cadherin was significantly decreased.The positive rates of FOXC2 and Vimentin in lymph node metastasis group were 79.1％ and 67.4％,respectively,which were significantly higher than those without lymph node metastasis (35.6％,P ＜0.01;20.3％,P ＜0.01).The positive rate of E-cadherin in lymph node metastasis group was 16.3％,which was significantly lower than that without lymph node metastasis (33.9％,P ＜ 0.01).The positive rates of FOXC2 and Vimentin in T1 +T2 phase were 41.7％ and 25.0％,respectively,which were significantly lower than those in T3 + T4 stage (83.3％,P ＜0.01;76.7％,P ＜0.01).The positive rate of Ecadherin in T1 + T2 stage group was 33.3％,which was significantly higher than that in T3 + T4 stage (10.0％,P ＜ 0.01).There was a positive correlation between FOXC2 and Vimentin in colorectal adenocarcinoma (P ＜0.01),and negatively correlated with E-cadherin expression (P ＜ 0.01).Conclusions EMT may promote the development and metastasis of colorectal adenocarcinoma,FOXC2 may be involved in colorectal cancer EMT and through the EMT to promote the malignant process of colorectal adenocarcinoma.
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Objective@#To investigate the relationship between expression of FoxM1 and BCRP in invasive breast carcinoma of no special type (IBC-NST) tissues and the clinical pathological characteristics and prognosis of the patients.@*Methods@#Seventy-eight cases of IBC-NST with excision were included. The expression of FoxM1 and BCRP was assessed by immunohistochemistry and its relationship with the clinical pathological characteristics and prognosis was evaluated.@*Results@#FoxM1 was expressed in 71.8%(56/78) of IBC-NST, and the expression was related to tumor diameter, TNM staging, ER, PR and HER2. BCRP was expressed in 53.8% (42/78) of IBC-NST, and the expression was related to age, tumor diameter, lymph node metastasis, ER and HER2. Kaplan-Meier survival analysis showed the survival time was related to tumor diameter, TNM staging, lymph node metastasis and the expression of FoxM1, BCRP, ER, PR and HER2. Cox multivariate analysis showed that TNM staging, FoxM1, BCRP, HER2 were determinants of patient survival time.@*Conclusions@#The expression of FoxM1 is associated with tumor diameter, TNM staging, ER, PR and HER2 while BCRP is associated with age, tumor diameter, lymph node metastasis, ER and HER2. Both FoxM1 and BCRP have prognostic significance in IBC-NST patients.
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Type 2 diabetes is a chronic metabolic disorder caused by a deficiency of beta cell func-tion leading to varying degrees of insulin deficiency and insulin resistance. The decline of beta cell function is the central part in the development of type 2 diabetes. The dedifferentiation of pancreatic beta cells is one of the important mechanisms of islet dysfunction. In recent years, it has been shown that the decrease of the activity of the transcription factor forkhead box O1 (FoxO1) is closely related to the dedifferentiation of beta cells. The study of the specific mechanism of FoxO1 involved in the dedifferentiation of beta cells and the redifferentiation of the differentiated cells will provide new ideas for the prevention and treatment of type 2 diabetes.
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Objective To explore the role of forkhead box protein O3a (FOXO3a) in inhibiting the proliferation and metastasis of breast cancer cells by downregulation of vascular endothelial growth factorA (VEGF-A).Methods Cell proliferation bioassay and plate clone formation assay were used to compare the changes of proliferation after siRNA interference.Wound healing and Transwell were used to compare the changes of invasion metastasis after interference.Meanwhile,the changes of VEGF-A expression in the cells before and after interference were compared by real time polymerase chain reaction (RT-PCR) and Western blot.Furthermore,the expressions of mRNA FOXO3a and VEGF-A were detected in 20 cases of breast cancer patients in breast cancer tissues and adjacent normal breast tissues.Results The MCF-7 and MDA-MB-231 cells were increased in cell proliferation and invasion after interference.Further studies found that mRNA and protein expression of VEGF-A were up-regulated in MCF-7 and MDA-MB-231cells after interference.In vivo the expression of FOXO3a was lower in 15 cases of cancer compared to normal tissues,and the expression of VEGF-A was high in 15 cases of cancer (75％).FOXO3a and VEGF-A expression was highly negatively correlated.Conclusions This study showed that FOXO3a could inhibit the proliferation and invasion of breast cancer cells,which might be regulated by VEGF-A.It provides an important theoretical evidence for targeted inhibition of breast cancer metastasis.
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Background Studies show that regulatory T cells (Treg) are a kind of T cell subsets to negatively regulate immune response,and play an important role in maintaining immune homeostasis and immune tolerance.Autoimmune uveitis is an autoimmune disease,the regulation of Treg cells in pathogenesis and progression of autoimmune uveitis is not fully unelucidated.Objective This study was to observe the dynamic changes of Treg in experimental autoimmune uveitis (EAU) rats and explore the role of Treg cells in the pathological process of EAUrats.Methods Eighty four 6-8 week-old SPF Lewis rats were randomly divided into model group and control group.The mixed emulsifier of interphotoreceptor retinoid-binding protein (IRBP)1177-1191,tuberculin (TB),complete Freund adjuvant (CFA) and PBS (300 μl) was subcutaneously injected in double rear foot pad,abdominal side and back,and only equal amount of TB,CFA and PBS emulsifier was used in the same way in the control group.Ocular inflammation symptoms was examined at 9,13,18,23,28,35 and 48 days after modeling and scored based on the severity of the inflammatory.Six rats of each group were sacrificed in above time points respectively for the histopathological examination of iris,ciliary body and retinas by haematoxylin-eosin staining.The lymphocytes were isolated and cultured from rat spleens,and the proportion of Foxp3-labelled cells,a specific marker of Treg cells,was assayed by flow cytometry.The relative expression level of Foxp3 mRNA in the lymphocytes detected by using realtime quantitative PCR (RT-PCR).The use and care of the rats complied with the ARVO Statement.Results Eye inflammatory response appeared at 8 days after immunization,showing vasodilation and hyperemia of rat iris in the model group,and the response peaked at 13 days,with exudation and hypopyon in the anterior chamber.The highest inflammatory scores were 3.75±0.42 at day 13,and the ocular inflammation reaction was gradually relieved after that and disappeared at 23 days after immunity.A significant difference in ocular inflammatory scores of model rats was found among different time points (F =81.709,P ＜ 0.001);while no inflammatory symptom was observed in the control group.Histopathology examination showed obvious infiltration of inflammatory cells in the iris,ciliary body and retinas in model rats,including neutrophils,lymphocytes and mononuclear cells.The proportion of Foxp3-labelled cells in spleen lymphocytes was (5.50 ± 0.64)％,(13.36 ± 0.98)％,(10.34 ± 0.79)％,(9.58 ± 1.02)％,(6.73 ±0.81)％ and (5.58 ± 0.47) ％ in the model group on day 13,18,23,28,35,48 respectively,with statistically significant differences in comparison with (2.80 ± 0.38) ％,(3.36 ± 0.53) ％,(3.65 ± 0.57) ％,(3.37 ± 0.43) ％,(3.33±0.50)％ and (3.13±0.61)％ in the control group (t=-6.272,-15.556,-11.910,-9.753,-6.154,-5.491,all at P＜0.01).The change trend of Foxp3 mRNA expression was consistent to the dynamic change of the proportion of Foxp3-labelled cells.Conclusions The pathogenesis and development is closely associated with the dynamic changes of CD4+ CD25 + Foxp3+ Treg cells in EAU rats.