Forkhead Box f1基因与新生儿肺出血的研究进展

新生儿肺出血仍然是新生儿期主要的危重疾病及死亡原因。最新的研究提示,一种新的转录因子Forkhead Box f1(Foxf1)的表达参与新生大鼠肺出血的发病机制,Foxf1表达缺陷将导致Notch信号系统异常,并引起致死性肺出血。

FoxO4 is the main forkhead transcriptional factor localized in the gastrointestinal tracts of pigs

Forkhead box （Fox） proteins play critical roles in the regulation of differentiation, proliferation, immunity and aging of cells. Most studies on Fox proteins are limited to cultured cells and rodent. The aim of the current study is to detect by immunohistrochemistry whether FoxO1, FoxO3a and FoxO4 proteins are localized in the stomach and intestine of the pig. The results showed that FoxO4 exists in the mucosa in all parts of the stomach and intestine; FoxO3a exists mainly in the lamina propria and muscularis of some parts. However, FoxOl is not detectable in all parts of the stomach and intestine. Collectively, the results of the present study indicate that there exists a distinct expression pattern of Fox proteins, and that FoxO4 is a primary forkhead transcriptional factor localized in the gastrointestinal tracts of the pig.

Propofol postconditioning ameliorates hypoxia/reoxygenation induced H9c2 cell apoptosis and autophagy via upregulating forkhead transcription factors under hyperglycemia

Background:Administration of propofol,an intravenous anesthetic with antioxidant property,immediately at the onset of post-ischemic reperfusion(propofol postconditioning,P-PostC) has been shown to confer cardioprotection against ischemia–reperfusion(I/R) injury,while the underlying mechanism remains incompletely understood.The forkhead box O(FoxO) transcription factors are reported to play critical roles in activating cardiomyocyte survival signaling throughout the process of cellular injuries induced by oxidative stress and are also involved in hypoxic postconditioning mediated neuroprotection,however,the role of FoxO in postconditioning mediated protection in the heart and in particular in high glucose condition is unknown.Methods:Rat heart-derived H9c2 cells were exposed to high glucose(HG) for 48 h,then subjected to hypoxia/reoxygenation(H/R,composed of 8 h of hypoxia followed by 12 h of reoxygenation) in the absence or presence of postconditioning with various concentrations of propofol(P-PostC) at the onset of reoxygenation.After having identified the optical concentration of propofol,H9c2 cells were subjected to H/R and P-PostC in the absence or presence of FoxO1 or FoxO3a gene silencing to explore their roles in P-PostC mediated protection against apoptotic and autophagic cell deaths under hyperglycemia.Results:The results showed that HG with or without H/R decreased cell viability,increased lactate dehydrogenase(LDH) leakage and the production of reactive oxygen species(ROS) in H9c2 cells,all of which were significantly reversed by propofol(P-PostC),especially at the concentration of 25 μmol/L(P25)(P<0.05,NC vs.HG;HG vs.HG+HR;HG+HR+P12.5 or HG+HR+P25 or HG+HR+P50 vs.HG+HR).Moreover,we found that propofol(P25) decreased H9c2 cells apoptosis and autophagy that were concomitant with increased FoxO1 and FoxO3a expression(P<0.05,HG+HR+P25 vs.HG+HR).The protective effects of propofol(P25) against H/R injury were reversed by silencing FoxO1 or FoxO3a(P<0.05,HG+HR+P25 vs.HG+HR+P25+siRNA-1 or HG+HR+P25+siRNA-5).Conclusions:It is concluded that propofol postconditioning attenuated H9c2 cardiac cells apoptosis and autophagy induced by H/R injury through upregulating FoxO1 and FoxO3a under hyperglycemia.

MiR-19a-3p regulates the Forkhead box F2-mediated Wnt/β-catenin signaling pathway and affects the biological functions of colorectal cancer cells

BACKGROUND Colorectal cancer(CRC)is one of the most common malignancies worldwide.AIM To explore the expression of microRNA miR-19a-3p and Forkhead box F2(FOXF2)in patients with CRC and the relevant mechanisms.METHODS Sixty-two CRC patients admitted to the hospital were enrolled into the study group,and sixty healthy people from the same period were assigned to the control group.Elbow venous blood was sampled from the patients and healthy individuals,and blood serum was saved for later analysis.MiR-19a-3p mimics,miR-19a-3p inhibitor,miR-negative control,small interfering-FOXF2,and short hairpin-FOXF2 were transfected into HT29 and HCT116 cells.Then quantitative polymerase chain reaction was performed to quantify the expression of miR-19a-3p and FOXF2 in HT29 and HCT116 cells,and western blot(WB)analysis was conducted to evaluate the levels of FOXF2,glycogen synthase kinase 3 beta(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),β-catenin,p-β-catenin,α-catenin,Ncadherin,E-cadherin,and vimentin.The MTT,Transwell,and wound healing assays were applied to analyze cell proliferation,invasion,and migration,respectively,and the dual luciferase reporter assay was used to determine the correlation of miR-19a-3p with FOXF2.RESULTS The patients showed high serum levels of miR-19a-3p and low levels of FOXF2,and the area under the curves of miR-19a-3p and FOXF2 were larger than 0.8.MiR-19a-3p and FOXF2 were related to sex,tumor size,age,tumor-nodemetastasis staging,lymph node metastasis,and differentiation of CRC patients.Silencing of miR-19a-3p and overexpression of FOXF2 suppressed the epithelialmesenchymal transition,invasion,migration,and proliferation of cells.WB analysis revealed that silencing of miR-19a-3p and FOXF2 overexpression significantly suppressed the expression of p-GSK-3β,β-catenin,N-cadherin,and vimentin;and increased the levels of GSK-3β,p-β-catenin,α-catenin,and Ecadherin.The dual luciferase reporter assay confirmed that there was a targeted correlation of miR-19a-3p with FOXF2.In addition,a rescue experiment revealed that there were no differences in cell proliferation,invasion,and migration in HT29 and HCT116 cells co-transfected with miR-19a-3p-mimics+sh-FOXF2 and miR-19a-3p-inhibitor+si-FOXF2 compared to the miR-negative control group.CONCLUSION Inhibiting miR-19a-3p expression can upregulate the FOXF2-mediated Wnt/β-catenin signaling pathway,thereby affecting the epithelial-mesenchymal transition,proliferation,invasion,and migration of cells.Thus,miR-19a-3p is likely to be a therapeutic target in CRC.

Nuclear accumulation of β-catenin and forkhead box O3a in colon cancer:Dangerous liaison

The WNT/-catenin and phosphoinositide 3-kinase(PI3K/AKT) signaling cascades both have been implicated in the formation and progression of colorectal cancer.Oncogenic PI3K/AKT signaling suppresses the activity of forkhead box O3a(FOXO3a) transcription factor through phosphorylation leading to its nuclear exclusion.Inhibition of the PI3K/AKT signaling by PI3K or AKT inhibitors results in the translocation of FOXO3a to the nucleus,and is considered to be a promising therapeutic strategy for many cancers including colon cancer.Now,however,a new study in Nature Medicine has revealed a nuclear interaction of-catenin with FOXO3a as a promoter of metastatic progression in colon cancer.The work has important implications for the treatment of colon cancers,suggests a companion biomarker strategy to enable a personalized medicine approach,and offers an alternative therapeutic strategy to overcome resistance to PI3K and AKT inhibitors.

Forkhead box protein A2 and T helper type 2-mediated pulmonary inflammation

The transcription factor forkhead box protein A2(FOXA2, also known as hepatocyte nuclear factor 3β or transcription factor 3β), has been found to play pivotal roles in multiple phases of mammalian life, from the early development to the organofaction, and subsequently in homeostasis and metabolism in the adult. In the embryonic development period, FOXA2 is require d for the formation of the primitive node and notochord, and its absence results in embryonic lethality. Moreover, FOXA2 plays an important role not only in lung development, but also in T helper type 2(Th2)-mediated pulmonary inflammation and goblet cell hyperplasia. In this article, the role of FOXA2 in lung development and Th2-mediated pulmonary inflammation, as well as in goblet cell hyperplasia, is reviewed. FOXA2 deletion in airway epithelium results into Th2-mediated pulmonary inflammation and goblet cell hyperplasia in developing lung. Leukotriene pathway and signal transducers and activators of transcription 6 pathway may mediate this inflammation through recruitment and activation of denditric cell during lung developments. FOXA2 is a potential treatment target for lung diseases with Th2 inflammation and goblet cell hyperplasia, such as asthma and chronic obstructive pulmonary disease.

7-difluoromethoxyl-5,4'-di-n-octylgenistein inhibits growth of gastric cancer cells through downregulating forkhead box M1

AIM: To investigate whether the 7-difluoromethoxyl-5, 4'-di-n-octylgenistein (DFOG), a novel synthetic genistein analogue, affects the growth of gastric cancer cells and its mechanisms. METHODS: A series of genistein analogues were prepared by difluoromethylation and alkylation, and human gastric cancer cell lines AGS and SGC-7901 cultured in vitro were treated with various concentrations of genistein and genistein analogues. The cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cells were incubated by DFOG at different concentrations. The growth inhibitory effects were evaluated using MTT and clonogenic assay. The distribution of the phase in cell cycle was analyzed using flow cytometric analysis with propidium iodide staining. The expression of the transcription factor forkhead box M1 (FOXM1) was analyzed by reverse transcription-polymerase chain reaction and Western blotting. The expression levelsof CDK1, Cdc25B, cyclin B and p27KIP1 protein were detected using Western blotting. RESULTS: Nine of the genistein analogues had more effective antitumor activity than genistein. Among the tested analogues, DFOG possessed the strongest activity against AGS and SGC-7901 cells in vitro. DFOG significantly inhibited the cell viability and colony formation of AGS and SGC-7901 cells. Moreover, DFOG efficaciously arrested the cell cycle in G2/M phase. DFOG decreased the expression of FOXM1 and its downstream genes, such as CDK1, Cdc25B, cyclin B, and increased p27KIP1 at protein levels. Knockdown of FOXM1 by small interfering RNA before DFOG treatment resulted in enhanced cell growth inhibition in AGS cells. Up-regulation of FOXM1 by cDNA transfection attenuated DFOG-induced cell growth inhibition in AGS cells. CONCLUSION: DFOG inhibits the growth of human gastric cancer cells by down-regulating the FOXM1 expression.

Correlation of Forkhead Box c2 with Subtypes and Invasive Ability of Invasive Breast Cancer

Forkhead Box c2 （FOXC2） is a member of forkhead/winged-helix family of transcription factors. The relationship between FOXC2 and invasive breast cancers, including basal-like breast cancer （BLBC, a subtype of breast cancer）, remains to be elucidated. In this study, immunohistochemistry was used to detect the expression of FOXC2 in samples from 103 cases of invasive breast cancers and 15 cases of normal mammary glands. The relationship between FOXC2 and clinical parameters of invasive breast cancers such as patient＇s age, tumor size, lymph node metastasis, tumor grade, the expression of ER, PR, HER-2 and p53, and Ki-67 labeling index （LI） was evaluated. The expression of FOXC2 was detected in parent MCF7 ceils, MCF cells transfected with FOXC2 expression vectors and MDA-MB- 435 cells by immunohistochemistry and Western blotting. Transwell assay was used to determine the invasive ability of these cells. The results showed that FOXC2 was strongly expressed in basal epithelial cells in normal mammary glands and weakly expressed or even not expressed in glandular epithelial cells. The majority of invasive breast cancers （71.8%, 74/103） had negative or weak expression of FOXC2. However, FOXC2 was strongly expressed in 60.7% of BLBCs. Moreover, FOXC2 was related with tumor grade, p53 expression, ki-67 LI and lymph nodes metastasis. It was expressed in FOXC2- transfected MCF cells and MDA-MB-435 cells but not in parent MCF cells. Transwell assay revealed that MCF cells transfected with FOXC2 expression vectors were more aggressive than the parent MCF ceils, suggesting a positive correlation between FOXC2 and the invasion of breast cancer. It was con- cluded that there is a significant association between FOXC2 and the metastasis of invasive breast can- cer. FOXC2 may be used as a new marker for the diagnosis and prognosis prediction of different sub- tvpes of invasive breast cancers.

Forkhead Box q1 promotes invasion and metastasis in colorectal cancer by activating the epidermal growth factor receptor pathway

BACKGROUND Colorectal cancer(CRC)is an extremely malignant tumor with a high mortality rate.Little is known about the mechanism by which forkhead Box q1(FOXQ1)causes CRC invasion and metastasis through the epidermal growth factor receptor(EGFR)pathway.AIM To illuminate the mechanism by which FOXQ1 promotes the invasion and metastasis of CRC by activating the heparin binding epidermal growth factor(HB-EGF)/EGFR pathway.METHODS We investigated the differential expression and prognosis of FOXQ1 and HB-EGF in CRC using the Gene Expression Profiling Interactive Analysis(GEPIA)website(http://gepia.cancer-pku.cn/index.html).Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blotting were used to detect the expression of FOXQ1 and HB-EGF in cell lines and tissues,and we constructed a stable lowexpressing FOXQ1 cell line and verified it with the above method.The expression changes of membrane-bound HB-EGF(proHB-EGF)and soluble HB-EGF(sHB-EGF)in the lowexpressing FOXQ1 cell line were detected by flow cytometry and ELISA.Western blotting was used to detect changes in the expression levels of HB-EGF and EGFR pathway-related downstream genes when exogenous recombinant human HB-EGF was added to FOXQ1 knockdown cells.Proliferation experiments,transwell migration experiments,and scratch experiments were carried out to determine the mechanism by which FOXQ1 activates the EGFR signaling pathway through HB-EGF,and then to evaluate the clinical relevance of FOXQ1 and HB-EGF.RESULTS GEPIA showed that the expression of FOXQ1 in CRC tissues was relatively high and was related to a lower overall survival rate.PCR array results showed that FOXQ1 is related to the HB-EGF and EGFR pathways.Knockdown of FOXQ1 suppressed the expression of HB-EGF,and led to a decrease in EGFR and its downstream genes AKT,RAF,KRAS expression levels.After knockdown of FOXQ1 in CRC cell lines,cell proliferation,migration and invasion were attenuated.Adding HB-EGF restored the migration and invasion ability of CRC,but not the cell proliferation ability.Kaplan–Meier survival analysis results showed that the combination of FOXQ1 and HB-EGF may serve to predict CRC survival.CONCLUSION Based on these collective data,we propose that FOXQ1 promotes the invasion and metastasis of CRC via the HB-EGF/EGFR pathway.

Forkhead box P3 and indoleamine 2,3-dioxygenase co-expression in Pakistani triple negative breast cancer patients

BACKGROUND Forkhead box P3(FOXP3)is a specific marker for immunosuppressive regulatory T(T-reg)cells.T-regs and an immunosuppressive enzyme,indoleamine 2,3-dioxygenase(IDO),are associated with advanced disease in cancer.AIM To evaluate the co-expression of FOXP3 and IDO in triple negative breast cancer(TNBC)with respect to hormone-positive breast cancer patients from Pakistan.METHODS Immunohistochemistry was performed to analyze the expression of FOXP3,IDO,estrogen receptor,progesterone receptor,and human epidermal growth factor receptor on tissues of breast cancer patients(n=100):Hormone-positive breast cancer(n=51)and TNBC(n=49).A total of 100 patients were characterized as FOXP3 negative vs positive and further categorized based on low,medium,and high IDO expression score.Univariate and multivariate logistic regression models were used.RESULTS Out of 100 breast tumors,25%expressed FOXP3 positive T-regs.A significant coexpression of FOXP3 and IDO was observed among patients with TNBC(P=0.01)compared to those with hormone-positive breast cancer.Two variables were identified as significant independent risk factors for FOXP3 positive:IDO expression high(adjusted odds ratio(AOR)5.90;95%confidence interval(CI):1.22-28.64;P=0.03)and TNBC(AOR 2.80;95%CI:0.96-7.95;P=0.05).CONCLUSION Our data showed that FOXP3 positive cells might be associated with high expression of IDO in TNBC patients.FOXP3 and IDO co-expression may also suggest its involvement in disease,and evaluation of FOXP3 and IDO expression in TNBC patients may offer a new therapeutic option.

Forkhead box protein P1, a key player in neuronal development?

Forkhead box protein P1（FOXP1）is a transcription factor belonging to the forkhead box（FOX）proteins,a family of transcriptional regulators sharing a highly conserved forkhead DNA-binding domain（Bacon and Rappold,2012）.Previous reports have proposed a role for FOXP1 in functionally regulating the central nervous system（CNS）,while mutations in FOXP1 have been implicated in cognitive abnormalities（Bacon and Rappold, 2012）.

Clinical implications of forkhead box M1, cyclooxygenase-2, and glucose-regulated protein 78 in breast invasive ductal carcinoma

BACKGROUND Breast infiltrating ductal carcinoma(BIDC)represents the largest heterotypic tumor group,and an in-depth understanding of the pathogenesis of BIDC is key to improving its prognosis.AIM To analyze the expression profiles and clinical implications of forkhead box M1(FOXM1),cyclooxygenase-2(COX-2),and glucose-regulated protein 78(GRP78)in BIDC.METHODS A total of 65 BIDC patients and 70 healthy controls who presented to our hospital between August 2019 and May 2021 were selected for analysis.The peripheral blood FOXM1,COX-2,and GRP78 levels in both groups were measured and the association between their expression profiles in BIDC was examined.Additionally,we investigated the diagnostic value of FOXM1,COX-2,and GRP78 in patients with BIDC and their correlations with clinicopathological features.Furthermore,BIDC patients were followed for 1 year to identify factors influencing patient prognosis.RESULTS The levels of FOXM1,COX-2,and GRP78 were significantly higher in BIDC patients compared to healthy controls(P<0.05),and a positive correlation was observed among them(P<0.05).Receiver operating characteristic analysis demonstrated that FOXM1,COX-2,and GRP78 had excellent diagnostic value in predicting the occurrence of BIDC(P<0.05).Subsequently,we found significant differences in FOXM1,COX-2,and GRP78 levels among patients with different histological grades and metastasis statuses(with vs without)(P<0.05).Cox analysis revealed that FOXM1,COX-2,GRP78,increased histological grade,and the presence of tumor metastasis were independent risk factors for prognostic death in BIDC(P<0.001).CONCLUSION FOXM1,COX-2,and GRP78 exhibit abnormally high expression in BIDC,promoting malignant tumor development and closely correlating with prognosis.These findings hold significant research implications for the future diagnosis and treatment of BIDC.

TMED3 promotes prostate cancer via FOXO1a and FOXO3a phosphorylation

Background:Transmembrane emp24 trafficking protein 3(TMED3)is associated with the development of several tumors;however,whether TMED3 regulates the progression of prostate cancer remains unclear.Materials and Methods:Short hairpin RNA was performed to repress TMED3 in prostate cancer cells(DU145 cells)and in a prostate cancer mice model to determine its function in prostate cancer in vitro and in vivo.Results:In the present study,we found that TMED3 was highly expressed in prostate cancer cells.In vitro,shTMED3 treatment suppressed the proliferation,invasion,and migration and promoted the apoptosis of DU145 cells.Additionally,the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed a strong correlation between TMED3 and forkhead box O transcription factor(FOXO)pathway.Furthermore,TMED3 inhibition efficiently decreased FOXO1a and FOXO3a phosphorylation.In vivo,TMED3 downregulation suppressed the apoptosis,growth,and metastasis of prostate cancer cells via FOXO1a and FOXO3a.Conclusion:The present findings show that TMED3 participates in the regulation of prostate cancer progression via FOXO1a and FOXO3a phosphorylation,thereby revealing a novel mechanism underlying prostate cancer development and suggesting that TMED3 inhibition may serve as a novel strategy for prostate cancer treatment.

SF3B1/FOXM1/JUNB轴调控SOX21表达对宫颈癌细胞生物学行为的影响研究

目的分析剪接因子3B亚基1/叉头框转录因子M1/转录因子活化蛋白激酶B(SF3B1/FOXM1/JUNB)轴调控转录因子21抗体(SOX21)表达对宫颈癌细胞生物学行为的影响。方法选取2022年3月至2023年12月新疆医科大学附属肿瘤医院收治的50例宫颈癌患者的癌旁组织及癌组织作为研究对象,利用实时荧光定量PCR检测SF3B1、FOXM1、JUNB、SOX21表达;通过Transwell、细胞计数试剂8(CCK8)检测宫颈癌细胞生物学行为(增殖、迁移、侵袭);利用蛋白质印迹法测定SF3B1、FOXM1、JUNB、SOX21蛋白表达。结果与癌旁组织相比,宫颈癌组织JUNB表达低,FOXM1、SOX21、SF3B1表达高,差异有统计学意义(P<0.05);与si-NC组相比,si-SF3B1/FOXM1/JUNB组0 h OD450值高,侵袭细胞数、迁移细胞数、(24、48 h)OD450值、SF3B1、FOXM1、JUNB低,差异有统计学意义(P<0.05);与OE-NC组相比,OE-SOX21组迁移细胞数、(0、24、48 h)OD450值、SOX21、侵袭细胞数高,差异有统计学意义(P<0.05);与si-SF3B1/FOXM1/JUNB+OE-NC组相比,si-SF3B1/FOXM1/JUNB+OE-SOX21组SOX21、SF3B1、FOXM1、JUNB、(0、24、48 h)OD450值、侵袭细胞数、迁移细胞数高,差异有统计学意义(P<0.05)。结论SF3B1/FOXM1/JUNB轴通过激活SOX21表达可促进宫颈癌细胞侵袭、增殖、迁移。

Forkhead转录因子调控干细胞的命运决定

Forkhead蛋白是一类在进化上高度保守的重要转录因子家族。目前,已经发现超过100种编码Forkhead蛋白的基因,Forkhead蛋白的典型结构特征是含有Forkhead结构域,即一段由100个氨基酸组成的进化上保守的"Forkhead"DNA结合序列。Forkhead蛋白的主要功能包括调控与胚胎发育、细胞生长、增殖、分化、干细胞干性保持、应激反应、语言形成以及长寿相关基因的表达。Forkhead基因的突变或者调控异常会导致发育缺陷以及肿瘤形成。因此,对于不同Forkhead蛋白家族成员功能的深入研究,有助于更好地揭示相关疾病的发病机制,为针对这些疾病的预防和治疗提供理论依据。该文对国内外Forkhead蛋白在干细胞中的主要功能方面的研究成果进行了总结,综述了近年来的最新研究成果。

清血消脂降糖方对2型糖尿病大鼠胰岛素抵抗的影响及机制

目的探讨清血消脂降糖方对2型糖尿病(T2DM)大鼠胰岛素抵抗(IR)的改善作用及潜在机制。方法采用腹腔注射30 mg/kg链脲佐菌素联合高脂高糖饲料喂养的方法建立T2DM大鼠模型。实验设置正常对照组,模型组,清血消脂降糖方低、高剂量组(6.525、13.05 g/kg,以生药量计)和二甲双胍组(阳性对照,0.18 g/kg),每组8只。给药组大鼠灌胃相应药物,正常对照组、模型组大鼠灌胃等体积生理盐水,每日1次,持续6周。测定大鼠体重、空腹血糖(FBG)并进行口服葡萄糖耐量试验,检测大鼠血清中空腹胰岛素(FINS)水平并计算胰岛素抵抗指数(HOMA-IR)、胰岛素敏感指数(ISI),检测大鼠血清中血脂四项、肝功能、氧化应激指标及炎症因子水平,观察大鼠肝组织病理变化,检测大鼠肝组织中蛋白激酶R样内质网激酶(PERK)、叉头框蛋白O1(FOXO1)蛋白磷酸化水平。结果与模型组相比,清血消脂降糖方高剂量组和二甲双胍组大鼠体重、ISI、高密度脂蛋白胆固醇、超氧化物歧化酶水平显著升高(P<0.05),FBG、糖负荷120 min时的血糖值、葡萄糖曲线下面积、FINS、HOMA-IR、低密度脂蛋白胆固醇、总胆固醇、甘油三酯、丙氨酸转氨酶、天冬氨酸转氨酶、碱性磷酸酶、丙二醛、白细胞介素6、肿瘤坏死因子α、C反应蛋白水平均显著降低(P<0.05),肝组织病理损伤明显改善,肝组织中PERK、FOXO1蛋白磷酸化水平显著降低(P<0.05)。结论清血消脂降糖方可调节糖脂代谢、炎症因子及氧化应激水平,缓解T2DM大鼠IR,其作用机制可能与抑制PERK/FOXO1信号通路有关。

特发性间质性肺炎病人血清STAT3和FOXM1水平与病情程度及预后的关系研究

目的探讨特发性间质性肺炎(IIP)病人血清信号转导及转录激活因子3(STAT3)、叉头框转录因子M1(FOXM1)表达水平与其病情程度和预后的关系。方法选取2017年1月至2022年10月四川省建筑医院收治的298例IIP病人为IIP组,选取同时期该院收治的298例普通型间质性肺炎(UIP)病人为UIP组,另选取同时期在该院体检的280例健康人员为对照组。采用酶联免疫吸附测定(ELISA)检测血清STAT3与FOXM1表达水平,并采用全肺纤维化高分辨率CT(HRCT)评分评价IIP病情严重程度。分析三组受试者血清STAT3与FOXM1表达水平差异。对IIP组病人随访3个月,根据其生存情况分为预后良好组与预后不良组,分析预后不良与预后良好组IIP病人血清STAT3与FOXM1表达水平差异,并采用Spearman法分析IIP病人血清STAT3与FOXM1表达水平与全肺纤维化HRCT评分之间的关系。采用logistic回归分析IIP病人预后不良的影响因素,绘制受试者操作特征曲线(ROC曲线)评估血清STAT3与FOXM1表达水平对IIP病人预后不良的预测效能。结果IIP组、UIP组、对照组血清STAT3[(1.50±0.39)ng/L、(1.32±0.31)ng/L、(1.01±0.27)ng/L]、FOXM1[(34.56±5.64)ng/L、(22.69±4.11)ng/L、(15.51±3.94)ng/L]水平均依次降低(P<0.05)。IIP病人随访3个月共有41例病人死亡,预后不良发生率为13.76%(41/298);预后不良组IIP病人血清STAT3、FOXM1水平均高于预后良好组(P<0.05)。Spearman相关性分析表明,IIP病人血清STAT3、FOXM1水平均与全肺纤维化HRCT评分正相关(rs=0.65,rs=0.57;P<0.001)。另预后不良组年龄、Ⅱ型肺泡细胞表面抗原(KL-6)、白细胞计数、红细胞沉降率(ESR)、C反应蛋白(CRP)水平、全肺纤维化HRCT评分均高于预后良好组(P<0.05);第一秒用力呼气量(FEV1)、最大自主通气量(MVV)均低于预后良好组(P<0.05)。多因素logistic回归分析显示,年龄、KL-6、白细胞、ESR、CRP、STAT3、FOXM1水平、全肺纤维化HRCT评分均是导致IIP病人预后不良的危险因素(P<0.05),FEV1、MVV为保护因素(P<0.05)。ROC分析结果表明,血清STAT3、FOXM1水平联合预测IIP病人预后的灵敏度高于STAT3、FOXM1单独预测的灵敏度(χ^(2)=8.10、6.12,P=0.002、0.008);联合预测的曲线下面积(AUC)高于STAT3、FOXM1单独预测的AUC(Z=3.15、2.54,P=0.002、0.011)。结论IIP病人血清STAT3、FOXM1表达水平均上升,二者与IIP病情程度正相关,均对IIP病人预后具有良好的预测价值,且联合预测效能更高。

叉头盒E1基因与中国汉族人群非综合征型唇腭裂的关联研究

目的探究叉头盒E1(FOXE1)基因所在单倍型区域hg19 chr9:100560865-100660865附近的单核苷酸多态性(SNPs)位点与中国西部汉族人群非综合征型唇腭裂(NSCL/P)发生的相关性。方法第一阶段纳入159名NSCL/P患者,采用目标区域捕获测序的方法,拟筛查FOXE1基因所在单倍型区域附近与NSCL/P发生相关的SNPs位点。第二阶段,选择21个常见SNPs位点,在1000名非综合征型单纯唇裂(NSCLO)患者,1000名非综合征型单纯腭裂(NSCPO)患者和1000名正常对照样本中进行验证。使用PLINK软件对研究人群进行哈迪-温伯格平衡(HWE)分析,对常见变异进行关联分析,对罕见变异进行基因负荷分析并使用Mutation Taster等软件对非同义突变的SNPs进行功能预测。结果第一阶段,共筛选出126个变异位点,包括76个SNPs变异位点和50个插入/缺失。纳入的所有SNPs在研究人群中遵循HWE。对筛选出的罕见变异进行功能有害性预测和基因负荷分析,差异均无统计学意义;对常见变异的关联分析表明,FOXE1基因的rs13292899位点与NSCL/P发生显著相关(P=1.85E-27),并且该位点与NSCLO(P=6.41E-23)、NSCLP(P=2.36E-15)发生也有相关性。随后在验证阶段,发现rs79268293(P=0.013,P=0.022)、rs10983951(P=0.0092,P=0.0076)、rs117227387(P=0.0092,P=0.0076)、rs3758250(P=0.0092,P=0.0076)和rs116899397(P=0.0092,P=0.0076),这5个SNPs位点与NSCLO和NSCPO均有显著关联性;另外,rs13292899(P=0.0085)、rs74606599(P=0.0083)、rs143226042(P=0.0083)和rs117236550(P=0.01),这4个SNPs位点与NSCLO发生相关;rs12343182(P=0.0087)、rs10119760(P=0.012)、rs10113907(P=0.012)和rs13299924(P=0.012),这4个SNPs位点与NSCPO发生相关。结论本研究在FOXE1基因上发现了一个新的易感SNPs位点rs13292899与NSCL/P及NSCLO发生密切相关,以及其他13个SNPs位点与NSCLO或NSCPO发生相关。

Expression and significance of fat mass and obesity associated gene and forkhead transcription factor O1 in non-alcoholic fatty liver disease

Background Non-alcoholic fatty liver disease （NAFLD） is a complex disorder and has been closely linked to obesity.The fat mass and obesity-associated （FTO） gene is a newly discovered gene related to obesity,which enhances oxidative stress and tipogenesis in NAFLD.The forkhead transcription factor O1 （FoxO1） is another important gene involved in NAFLD,which causes lipid disorders when insulin resistance appears in the liver.However,the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated.This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.Methods This study includes two parts referred to as animal and cell experiments.Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model.Aspartate aminotransferase （AST）,alanine aminotransferase （ALT）,total triglyceride （TG）,total cholesterol （TC）,alkaline phosphatase （ALP）,high-density lipoprotein （HDL）,and low-density lipoprotein （LDL） were measured.Immunohistochemical analysis was used to detect the expression and histological localization of FTO,FoxO1,and adenosine monophosphate （AMP）-activated protein kinase （AMPK）.The L02 cells were exposed to high fat for 24,48,or 72 hours.Oil red O staining was used to detect intracellular lipid droplets.Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.Results At the end of 10 weeks,ALP,ALT,AST,and LDL were significantly increased （P ＜0.01）,while TC and TG were also significantly higher （P ＜0.05）.In addition,HDL was significantly decreased （P ＜0.05）.The FTO and FoxO1 proteins were weakly expressed in the control group,but both FTO and FoxO1 were expressed significantly higher （P ＜0.01） in the experimental group,and the expression of the two factors was significantly correlated.AMPK in the high-fat group showed a low level of correlation with FTO,but not with FoxO1.Oil Red O staining results showed that the cells cultured in 50％ fetal bovine serum for 24,48,or 72 hours exhibited steatosis.FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group （P ＜0.01）.The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours （P ＜0.05）.Conclusions A high-fat diet leads to higher expression of FTO,phosphorylation of FoxO1,and decreased phosphorylation of AMPK.These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.

Forkhead box protein 3 mRNA expression in the peripheral blood of kidney-transplant recipients with acute rejection

Background Regulatory T cells （Tregs） are immunologically and clinically interesting not least because of the important role they play in allograft rejection. Likewise, expression of the transcription factor forkhead box protein 3 （FOXP3）, detected in transplant biopsies, is also of intere.（;t because of its role in the development of regulatory T cells. In this study, we investigated the relationship between FoxP3 mRNA expression and acute organ rejection in kidney-transplant recipients. Methods In this prospective study, FoxP3 mRNA expression levels in peripheral blood samples from 10 recipients of living relative-donor kidney transplants were measured before transplantation as well as at the 14th and 90th days post-transplantation. In addition, 46 first-time kidney-transplant recipients participated in a cross-sectional study, with 28 patients classified as having acute organ rejection; whilst the remaining 18 patients had functionally stable allografts. FoxP3 mRNA expression levels in peripheral blood samples were compared between these two different groups. Results Before transplantation mean FoxP3 mRNA levels vs. GADPH mRNA levels （Ig（FoxP3 mRNA/GADPH mRNA）） in the 10 recipients were 1.11±0.67. The mean FoxP3 mRNA expression levels measured at 14th and 90th days post-transplantation were significantly higher than before transplantation （1.69±0.38, P=0.03; 1.44±0.21, P=0.04, respectively）. Additionally, the mean FoxP3 mRNA levels vs. GADPH mRNA expression levels （Ig（FoxP3 mRNA/GADPH mRNA）） were significantly higher in recipients suffering acute rejection compared with those with stable allografts （1.77±0.61 and 1.43±0.27, respectively, P=0.03）. Conclusions After kidney transplantntion, FoxP3 mRNA levels were found to increase in the peripheral blood of all recipients. Considerably higher FoxP3 rnRNA levels were observed in recipients suffering acute rejection. These results suggest that FoxP3 mRNA levels in peripheral blood samples can be used as a diagnostic tool for identifying acute rejection.