Objective To investigate the expression of circular RNA(Circ RNAS)Actinin α 4(ACTN4)and platelet thrombin protein 1(THBS1)in intrahepatic cholangiocarcinoma(ICC)and their relationship with clinicopathological characteristics and prognosis,and provide reference for clinical diagnosis and treatment.Method A retrospective analysis was conducted on 84 ICC patients diagnosed and treated in the Second Affiliated Hospital of Xian Jiaotong University from May 2017 to June 2020.The expressions of CircACTN4 mRNA,THBS1 mRNA and protein were detected by real-time fluorescent quantitative PCR and immunohistochemistry.Pearson correlation analysis was used to analyze the correlation between CircACTN4 mRNA and THBS1 mRNA in ICC cancer tissue.The prognostic differences of ICC patients with different CircACTN4 mRNA and THBS1 mRNA expressions were compared by the Kaplan-Meier method(log rank test).COX regression analysis was performed to identify prognostic factors in ICC patients.The prognostic value of CircACTN4 mRNA and THBS1 mRNA in evaluating the risk of death in ICC patients was assessed by receiver operating characteristic(ROC)curve analysis.Results The expression of CircACTN4 mRNA in ICC tissues(3.14±0.42)was higher than that in adjacent tissues(0.76±0.25),with significant difference(t=44.094,P＜0.001).The positive rates of THBS1 mRNA(2.82±0.36)and protein positive rate(92.86％)in ICC tissues were higher than those in adjacent tissues(0.81±0.24,7.14％),and the differences were statistically significant(t/x2=42.068,123.429,all P＜0.001).CircACTN4 mRNA was positively correlated with THBS1 mRNA in ICC cancer tissue(r=0.669,P＜0.001).The expressions of CircACTN4 mRNA,THBS1 mRNA in ICC cancer tissues with TNM stage Ⅲ,low differentiation,and lymph node metastasis were higher than those in TNM stage Ⅰ～Ⅱ,high differentiation,and non-lymph node metastasis cancer tissues,and the differences were statistically significant(x2=7.949,9.164,12.207;23.270,18.625,19.828,all P＜0.001).The 3-year cumulative survival rates of the high expression group of CircACTN4 mRNA and THBS1 mRNA were lower than those of the low expression group of CircACTN4 mRNA(25.00％vs 56.82％)and THBS1 mRNA(19.51％vs 62.79％),and the differences were statistically significant(Log rank x2=13.601,24.310,all P＜0.001).CircACTN4 mRNA(OR=1.839,95％CI:1.228～2.753),THBS1 mRNA(OR=1.744,95％CI:1.245～2.443),lymph node metastasis(OR=1.925,95％CI:1.316～2.816),TNM staging(OR=1.613,95％CI:1.223～2.126),and tumor differentiation(OR=1.510,95％CI:1.205～1.892)were independent factors affecting the prognosis of ICC.The area under the curve of the combination detection of circACTN4 and THBS1 mRNA on the prognosis of death in patients with ICC was 0.868,which was greater than that of the single index(0.812 and 0.784),with significant differences(Z=3.348,3.847,all P＜0.001).Conclusion The expressions of CircACTN4 mRNA and THBS1 mRNA were increased in ICC,and they were associated with TNM stage,differentiation,and lymph node metastasis.These markers may serve as novel indicators to evaluate the poor prognosis of ICC patients.

Objective To analyze the expression level of YEATS2 in hepatocellular carcinoma(HCC)about its clinical prognosis and therapeutic value based on biological information from the cancer genome atlas(TCGA)and human protein atlas(HPA)databases.Methods The mRNA expression data and clinical information of HCC were downloaded from the TCGA database,the expression of YEATS2 between HCC tissues and normal tissues was analyzed by using the R software,and the protein expression differences were preliminary verified by the HPA database.The expression differences of YEATS2 between various clinical features of HCC were compared,and their effects on the survival of HCC patients by Kaplan-Meier method and COX regression analysis were then evaluated.Receiver operating characteristic(ROC)curves were plotted to evaluate their diagnostic values.The biological functions of YEATS2 in HCC were analyzed using gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis.The relationship between YEATS2 expression and tumor microenvironment(TME)was analyzed by the"ESTIMATE"algorithm,and its relationship with tumor-infiltrating immune cells(TIICs)was assessed by CIBERSORT.Analysis of YEATS2 expression levels to immune checkpoints and drug sensitivity was performed using the R package.Results The expression of YEATS2 was increased in HCC tissues(P=4.96e-21),and its expression level was correlated with age,clinical stage,pathological grade and T stage(all P＜0.05).Overall survival(OS)(P＜0.001)and progression-free survival(FPS)(P=0.016)were decreased in HCC patients with high expression of YEATS2,COX regression results showed that the expression level ofYEATS2 was associated with poor prognosis in HCC patients(OS:HR=2.167,95％CI:1.441～3.261,P=2.06e-04),and it was an independent risk factor for predicting poor prognosis in HCC patients(OS:HR=1.891,95％CI:1.243～2.877,P=0.003).The ROC curve suggested the AUCs for 1,3 and 5 years were 0.677,0.622 and 0.612,respectively,indicating good predictive ability.The TCGA database screened a total of 6 764 differential genes in the YEATS2 high and low expression groups,of which 4 094 genes were up-regulated and 2 670 genes were down-regulated in the YEATS2 high expression group.The results of GO and KEGG enrichment analyses showed that the differentially differentiated genes in the YEATS2 high expression group were mainly enriched in immunoregulation,and cell cycle regulation drug resistance pathway.The results of the TME score showed that the YEATS2 high expression group caused a decrease in immunity score(P＜0.01).The correlation between YEATS2 and TIICs showed that YEATS2 expression was positively correlated with the level of M0-type macrophage infiltration levels(r=0.48,P＜0.001)and 23 immune checkpoint genes(r=0.20～0.46,all P＜0.05),and was negatively correlated with the CD8+T-cells,plasma cells and monocyte(r=-0.26,-0.29,-0.30,P=0.021,0.011,0.008).Drug sensitivity analysis showed that the half maximal inhibitory concentration(IC50)of cabozantinib,lincitinib,doxorubicin,and cyclobenzaprine in patients with high expression of YEATS2 was higher than those in patients with low expression(all P＜0.01).Conclusion YEATS2 was highly expressed in HCC,and the expression level was associated with poor prognosis in HCC patients.YEATS2 can be used as a biomarker for the clinical early diagnosis,prognosis and immunotherapy of HCC,which may provide new ideas for clinical diagnosis and treatment.

Objective To explore the diagnostic value of serum neuronal Per-Arnt-Sim domain protein 4(NPASDP-4)and myelin basic protein(MBP)expression in patients with Parkinson's disease in relation to cognitive impairment(CI)and severity.Methods Selected and 138 Parkinson's disease patients admitted to the 909th Hospital of the Joint Logistics Support Force of the People's Liberation Army of China as the Parkinson's disease group,and 69 healthy people in the physical examination center of the hospital were in the healthy control group.Patients with Parkinson's disease were divided into normal cognitive function group(n=55),mild CI group(n=51)and dementia group(n=32)according to whether CI occurred and its severity.General data of subjects was collected,the serum levels of NPASDP-4 and MBP were detected by ELISA,correlation analysis was adopted by Spearman rank correlation or Pearson linear correlation,diagnostic value was analyzed by ROC curve,and influencing factors were analyzed by multivariate Logistic regression.Results Compared with the healthy control group,the levels of serum NPASDP-4(6.75±0.48 ng/ml vs 2.38±0.31 ng/ml)and MBP(8.34±0.65 μg/L vs 3.54±0.42 μg/L)in the Parkinson's disease group were increased with statistical significance(r=68.751,55.761,all P＜0.05).There were significant differences in H-Y stage among the normal cognitive function group,mild CI group and dementia group(x2=7.788,P＜0.05).Compared with the group with normal cognitive function(47.92±11.63 score),the mild CI group(50.78±13.69 score)and the dementia group(41.95±10.36 score)showed an increase in UPDRS-Ⅲ scores,and the differences were statistically significant(H=6.672,all P＜0.05).In normal cognitive function group,mild CI group and dementia group,the course of disease,and serum NPASDP-4(5.89±0.40,6.83±0.55,8.12±0.54 ng/ml)and MBP(6.65±0.56,8.94±0.69,10.27±0.70μg/L)levels were significantly increased(H=207.950,355.594,allP＜0.05),while MMSE score(28.47±0.94,24.51±1.35,17.09±2.57 score),MoCA score(27.45±1.03,20.18±1.92,11.75±2.53 score)and GPCOG total score(13.47±0.69,10.25±1.04,8.97±0.82 score)were significantly decreased,and the differences were statistically significant(H=515.005,775.933,327.584,all P＜0.05),respectively.The serum levels of NPASDP-4 and MBP in Parkinson's disease patients were significantly positively correlated with the course of disease(r=0.316,0.358),H-Y stage(r=0.345,0.384)and UPDRS-Ⅲ score(r=0.371,0.396),and significantly negatively correlated with MMSE score(r=-0.468,-0.517),MoCA score(r=-0.504,-0.569)and GPCOG total score(r=-0.527,-0.538)(all P＜0.05),respectivey.The areas under the curve(AUC)of the serum levels of NPASDP-4,MBP and their combination in diagnosing of Parkinson's disease were 0.850,0.930 and 0.960,respectively.The AUC of the serum levels of NPASDP-4 and MBP and their combination in diagnosing the severity of CI in patients with Parkinson's disease were 0.866,0.803 and 0.933,respectively.H-Y stage metaphase[OR(95％CI):4.725(1.742～12.814)],H-Y stage advanced[OR(95％CI):5.083(1.919～13.464)],UPDRS-Ⅲ score[OR(95％CI):3.257(1.464～7.246)],NPASDP-4[OR(95％CI):5.324(1.516～18.701)]and MBP[OR(95％CI):5.769(2.459～13.533)]were the influential factors for CI in patients with Parkinson's disease(all P＜0.05).NPASDP-4[OR(95％CI):4.768(2.382～9.543)]and MBP[OR(95％CI);5.846(3.141～10.882)]were the influential factors for the severity of CI in patients with Parkinson's disease(all P＜0.05).Conclusion The serum levels of NPASDP-4 and MBP in patients with Parkinson's disease were high,and they were closely related to CI and its severity,which may have certain clinical diagnostic value.

Objective To explore the expression of serum Hepcidin,long non-coding RNA myocardial infarction-associated transcript(LncRNA MIAT)and creatinine(CRE)in patients with bipolar disorder(BPD)and their relationship with the severity of the disease.Method In this study,150 patients with BPD who were admitted to Baoji Central Hospital from March 2021 to March 2023 were selected as the study subjects and were included in the BPD group.Another 150 healthy subjects were selected as the control group.Hamilton Depression Scale(HAMD)was used to evaluate depression.Young mania ratings scale(YMRS)was used to evaluate the degree of mania.All subjects were tested for serum Hepcidin,LncRNA MIAT and CRE levels.The levels of serum Hepcidin,LncRNA MIAT and CRE were compared between control group and BPD group.The levels of serum Hepcidin,LncRNA MIAT and CRE in BPD groups with different depression/mania degrees were compared.Receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of serum Hepcidin,LncRNA MIAT and CRE in BPD.The relationship between serum Hepcidin,LncRNA MIAT and CRE and the severity of BPD was analyzed.Results The levels of serum Hepcidin(46.17±9.17 ng/L),LncRNA MIAT(11.92±2.65)and CRE(74.02±11.89 μ mol/L)in BPD group were higher than those in control group(22.16±5.23 ng/L,4.11±0.95,57.16±8.61 μ mol/L),with significant differences(t=-27.856,-43.079,-14.066,all P＜0.05).The levels of serum Hepcidin,LncRNA MIAT and CRE increased with the increase of depression/mania in BPD patients(F=22.036,25.463,17.499;19.552,15.791,18.335,all P＜0.05).The combined detection of serum Hepcidin,LncRNA MIAT and CRE(95％CI:0.866～0.949)for BPD diagnosis had an area under the ROC curve of 0.914,with high specificity and sensitivity,which was significantly higher than that of serum Hepcidin alone(95％CI:0.693～0.816),LncRNAMIAT(95％CI:0.696～0.819)alone and CRE(95％CI:0.587～0.701)detection alone(AUC=0.759,0.762,0.614,all P＜0.05).Serum Hepcidin,LncRNA MIAT and CRE were positively correlated with depression severity in BPD patients(r=0.784,0.771,0.826,all P＜0.05).Serum Hepcidin,LncRNA MIAT and CRE were positively correlated with mania severity in BPD patients(r=0.806,0.793,0.831,all P＜0.05).Conclusion The levels of serum Hepcidin,LncRNA MIAT and CRE in BPD patients were higher than those in the normal population,and they were positively correlated with the severity of the disease.Combined detection may have a high diagnostic value.

Objective To explore the expression of mocro RNA(miR)-142-5p and suppressor of cytokine signaling 1(SOCS1)mRNA in peripheral blood mononuclear cells(PBMC)of mice and clinical patients with ankylosing spondylitis(AS)and their impact on immune function.Methods The mRNA levels of miR-142-5p and SOCS1 in PBMC of 30 patients with AS(Patient group)and 30 healthy controls(Health group)treated in the First People's Hospital of Shangqiu from January 2022 to March 2023 were measured by quantitative real time PCR(qRT-PCR).AS mice models were induced by bovine proteoglycan combined with complete Freund's adjuvant,and these mice were divided into control group,model group,NC group and antagomir group.Normal saline was injected into tail vein in control group and model group,and NC-antagomir and miR-142-5p-antagomir were injected into tail vein in NC group and antagomir group,respectively.After 2 weeks of treatment,the arthritis symptom scores of mice in each group were evaluated.The morphology of ankle joint was evaluated by hematoxylin eosin(HE)staining.The levels of Th1 cytokine interferon-γ(IFN-γ),Th2 cytokine interleukin-4(IL-4),Th17 cytokine interleukin-17(IL-17)and Treg cytokine forkhead box protein P3(FOXP3)in PBMC of mice were detected by ELISA method.The mRNA and protein expression of miR-142-5p,SOCS1,IFN-γ,IL-4,IL-17 and FOXP3 in PBMC and ankle joints were detected by qRT-PCR and Western blot.Results Compared with Health group,the level of miR-142-5p in PBMC of patient group was increased(1.00±0.21 vs 3.03±0.99,t=10.997,P＜0.001),while the level of SOCS1 mRNA was decreased(1.00±0.18 vs 0.41±0.09,t=15.956,P＜0.001).Compared with control group,miR-142-5p level(1.00±0.04 vs 4.00±0.52)and the mRNA and protein levels of IFN-γ and IL-17 in ankle joint tissue of model group were increased,while the mRNA and protein levels of SOCS1,IL-4 and FOXP3 were decreased,with significant differences(t=23.356,31.420,48.056,47.224,38.035,29.007,54.183,28.123,55.155,26.758,45.346,all P＜0.05).The arthritis symptom score was increased(7.83±0.94 vs 0.00±0.00,t=22.212,P＜0.05),and the ankle joint structure was damaged.Serum IFN-γ,IL-17 levels,IFN-γ/IL-4 ratio(0.81±0.08 vs 2.08±0.33)and IL-17/FOXP3 ratio(0.41±0.03 vs 1.27±0.10)were increased,and the differences were statistically significant(t=15.382,35.779,8.934,35.130,all P＜0.05).Compared with NC group,miR-142-5p level(3.89±0.33 vs 1.47±0.10),the mRNA and protein levels of IFN-γ and IL-17 in ankle joint tissue of antagomir group were decreased,while the mRNA and protein levels of SOCS1,IL-4 and FOXP3 were increased,and the differences were statistically significant(t=18.846,22.969,43.454,32.617,23.259,20.881,41.832,11.994,32.977,15.190,35.834,all P＜0.05).The arthritis symptom score was decreased(7.42±1.24 vs 2.75±0.75,t=13.233,P＜0.05),and the shape of the ankle joint of the rats was improved.Serum IFN-γ,IL-17 levels,IFN-γ/IL-4 ratio(1.22±0.11 vs 1.91±0.19)and IL-17/FOXP3 ratio(0.69±0.05 vs 1.23±0.12)were decreased,and the differences were statistically significant(t=8.688,22.972,3.785,22.007,all P＜0.05).Conclusion MiR-142-5p was highly expressed in AS.Down-regulation of miR-142-5p using antagonists may reduce Th1/Th2 ratio and Th17/Treg ratio through up-regulation of SOCS1,there by improving the immune balance of AS mice and inhibiting the progression of AS.

Objective To explore the relationship between serum microRNA(miR)-122 and microRNA(miR)-558 levels and cardiac function and heart rate variability in patients with chronic heart failure(CHF).Methods A total of 100 patients with chronic heart failure who were hospitalized in Wuhan Dongxihu District Poeple's Hospital from January 2019 to January 2022 were regarded as the study group,and 100 healthy individuals who underwent physical examinations in the hospital during the same period served as the control group.The real-time fluorescence quantitative PCR method was applied to detect the expressiou levels of serum miR-122 and miR-558 in two groups.The heart rate variability and cardiac function indicators of the two groups were compared.Pearson method was applied to analyze the correlation between serum miR-122 and miR-558 expression levels,and heart rate variability and cardiac function indicators in patients with chronic heart failure.Logistic regression was applied to analyze the risk factors affecting chronic heart failure.Results Compared with the control group,the serum miR-122(0.86±0.19 vs 1.07±0.16),heart rate variability indicators,and left ventricular ejection fraction(LVEF)of patients with chronic heart failure were decreased(t=3.844～15.448),and the expression level of miR-558(1.28±0.27 vs 1.02±0.12)and left ventricular end diastolic volume index(LVEDVI),left ventricular end systolic volume index(LVESVI)were increased(t=6.794～9.358),and the differences were statistically significant(all P＜0.05).The expression level of serum miR-122(1.19±0.23,1.04±0.21)in patients with functional grades Ⅰ to Ⅱ was higher than that in patients with grades Ⅲ to Ⅳ(0.68±0.16,0.53±0.15),while the expression level of miR-558(0.93±0.24,1.21±0.26)was lower than that in patients with grade Ⅲ-Ⅳ(1.42±0.27,1.64±0.29),with significant differences(qmiR-122=3.751～14.568,qmiR-558=3.929～11.194.all P＜0.05).The serum miR-122 level in patients with chronic heart failure was positively correlated with heart rate variability indicators and LVEF(r=0.337～0.573),but negatively correlated with LVEDVI and LVESVI(r=-0.385,-0.323).Serum miR-558 was negatively correlated with heart rate variability indicators and LVEF(r=-0.646～-0.246),but positively correlated with LVEDVI and LVESVI(r=0.528,0.547).Serum miR-122[OR(95％CI)=0.528(0.328～0.850)]and miR-558[OR(95％CI)=2.845(1.364～5.933)]were influencing factors for chronic heart failure(P＜0.05).Conclusion The expression levels of serum miR-122 and miR-558 in patients with chronic heart failure were closely related to heart rate variability and cardiac function,indicating that serum miR-122 and miR-558 may be influencing factors of chronic heart failure.

Objective To explore the correlation between the expression of serum microRNA(miR)-103a and microRNA(miR)-497 and micro vascular disease(MVD)in patients with type 2 diabetes mellitus(T2DM).Methods Patients diagnosed with T2DM in Tangshan Union Medical College Hospital from May 2022 to April 2023 were selected as the study group(n=113),which were divided into type 2 diabetes microangiopathy(DMAP)group(n=51)and T2DM group(n=62)according to whether MVD was complicated.Healthy subjects in the same period were selected as control group(n=105).Clinical data were collected,and real-time fluorescence polymerase chain reaction(qRT-PCR)was applied to detect serum levels of miR-103a and miR-497.Logistic regression was applied to analyze the influencing factors of T2DM complicated with MVD.Receiver operating characteristic(ROC)curve was plotted to analyze the diagnostic value of serum miR-103a and miR-497 levels in T2DM complicated with MVD.Results The serum levels of miR-103a(0.62±0.13)and miR-497(0.79±0.14)in the study group were lower than those in the control group(0.96±0.16,1.03±0.18),with significant differences(t=17.273,11.031,all P＜0.05).The serum levels of miR-103a(0.53±0.08)and miR-497(0.69±0.10)in the DMAP group were lower than those in the T2DM group(0.69±0.10,0.87±0.13),with significant differences(t=9.247,8.108,all P＜0.05).The proportions of patients with a disease course of ≥ 8 years and concomitant hyperlipidemia,and the levels of FBG,TC,Scr and BUN in the DMAP group were higher than in the T2DM group,with significant differences(t/x2=8.294,15.342,-2.855,-5.659,-8.951,-3.880,all P＜0.05).The area under curve(AUC)of serum miR-103a,miR-497 and their combination in the diagnosis of T2DM with MVD were 0.899,0.897 and 0.970,respectively.The combined diagnosis effect was better than that of individual diagnosis(Z=2.268,2.267,all P＜0.05).The results of logistic regression analysis showed that FBG(OR=1.879,95％CI:1.262～2.797),TC(OR=2.141,95％CI:1.348～3.400),and Scr(OR=3.417,95％CI:1.569～7.440)were independent risk factors for T2DM with MVD(all P＜0.05),while miR-103a(OR=0.784,95％CI:0.648～0.948)and miR-497(OR=0.839,95％CI:0.750～0.938)were protective factors for T2DM with MVD(all P＜0.05).Conclusion The serum miR-103a and miR-497 were low in patients with T2DM complicated with MVD,which may be potential markers for the diagnosis of T2DM complicated with MVD.

Objective To investigate the relationship between the expression level of miRNA Let-7 and the deletion of DNA mismatch repair(dMMR)proteins in endometrial carcinoma(EC).Methods A total of 74 patients with EC who underwent radical surgery at Lianyungang Maternal and Child Health Hospital in Jiangsu Province from May 2016 to December 2022 were selected.Immunohistochemistry was used to detect the expression of dMMR proteins(including MLH1,PMS2,MSH2,and MSH6),while real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was performed to measure the relative expression levelof miRNA Let-7.According to the expression of dMMR proteins,EC patients were divided into the complete expression group(n=43)and the absent expression group(n=31).Logistic multivariate regression analysis was conducted to identify risk factors associated with dMMR protein deficiency,and receiver operating characteristic(ROC)curve was plotted to evaluate the predictive value of relevant factors.Results In 74 cases of EC,the expression level of miRNA Let-7 was higher in the myographic infiltration＜1/2 group than in the myographic infiltration ≥ 1/2 group,and the difference was significant(t=1.79,P=0.04).The deletion rate of dMMR protein expression was 41.89％,the deletion rate in patients aged＜55 years and miRNA Let-7 low expression group(＜0.715)were higher than those in ≥ 55 years and miRNA Let-7 high expression group(≥ 0.715),with significant differences(x2=3.92,4.50,all P＜0.05).Logistic regression analysis revealed that miRNA Let-7 expression level was an independent risk factor for dMMR expression deletion(P=0.012).Spearman correlation analysis demonstrated a negative correlation between miRNA Let-7 expression level and dMMR protein deletion(r=-0.247,P=0.034).ROC curve analysis indicated that the expression level of miRNA Let-7 had a certain predictive value for dMMR protein deletion in EC patients,with an AUC of 0.737,an optimal critical value of 0.77,a sensitivity and a specificity of 0.651 and 0.806,respectively.Conclusion The expression level of miRNA Let-7 was correlated with the deletion of dMMR proteins in patients diagnosed with EC.Moreover,it served as a potential risk factor for dMMR protein deficiency.It may offer valuable insights into predicting dMMR expression deletion.

Objective To investigate the regulatory effects of exosome microRNA-3614-5p(miR-3614-5p)derived from mesenchymal stem cells on the progression of preeclampsia(PE)in model rats and its related mechanisms.Methods Thirty-six SD rats(24 females and 12 males)were housed in cages at a female-to-male ratio of 2∶1 for natural conception.Twenty-four pregnant rats were randomly divided into sham group(sham group),PE model group(PE group)and exosome miR-3614-5p group(PE+exo group),with 8 rats in each group.The PE model was established by subcutaneous injection of 100 mg/kg NG-nitro-L-arginine methyl ester in PE group.PE model was constructed in PE+exo group.Meanwhile,160 μ g/ml exosome suspension(0.5 ml/individual/day)was intraperitoneally injected on the 14th day for 6 consecutive days,and the experiment lasted for 21 days.Sham group was given an equal amount of normal saline.Blood pressure and urinary protein concentration were measured on days 0,7,14 and 21 of pregnancy.The levels of miR-3614-5p,B lymphoblastoma-2(Bcl-2)and Bcl-associated X protein(Bax)mRNA were detected by RT-qPCR.The activity of Caspase-3,the levels of reactive oxygen species(ROS)and the content of malondialdehyde(MDA),glutathione(GSH)and ferrous ion(Fe2+)were detected by ELISA.Western blot was used to analyze the protein levels of the iron death-related protein glutathione peroxidase 4(GPx4)and solute carrier family 7 member 11(SLC7A11).Results Compared with the sham group,the expression of miR-3614-5p in the placental tissues(0.43±0.05 vs 1.01±0.07)and peripheral blood(0.51±0.07 vs 1.01±0.12)of rats in the PE group was down-regulated,with significant differences(t=19.070,10.180,all P＜0.01).Compared with supernatant liquid phase,miR-3614-5p in exosomes derived from MSCs was enriched.Compared with sham group,the diastolic blood pressure(175.43±6.02 mmHg vs 113.26±5.11 mmHg),systolic blood pressure(123.57±5.63 mmHg vs 82.63±5.26 mmHg)and urinary protein content(175.48±13.21 mg/ml vs 67.65±5.76 mg/ml)of rats in PE group were increased on the 21st day with statistical significante between groups(t=22.606,16.440,23.168,all P＜0.01).Compared with PE group,diastolic blood pressure(124.57±5.33 mmHg vs 175.43±6.02 mmHg),systolic blood pressure(89.76±3.88 mmHg vs 123.57±5.63 mmHg)and urinary protein content(97.69±7.23 mg/ml vs 175.48±13.21 mg/ml)in PE+exo group were decreased,and the differences between groups were significant(t=18.493,13.557,16.713,all P＜0.01).Compared with sham group,Caspase-3 activity(238.56％±13.22％vs 100.12％±5.93％),Bax level(3.18±0.71 vs 1.01±0.11),ROS level(387.65％±25.98％vs 100.51％±5.89％),MDA content(33.21±3.17 nmol/mg vs 14.83±2.69 nmol/mg)and Fe2+concentration(38.77±6.53 nmol/ml vs 17.51±3.15 nmol/ml)in placenta tissue of PE group were increased,while Bcl-2 level(0.47±0.08 vs 1.01±0.12),GSH content(4.12±1.22 nmol/mg vs 9.76±0.93 nmol/mg),GPX4 protein(0.48±0.06 vs 1.01±0.24)and SLC7A11 protein(0.51±0.11 vs 1.01±0.11)levels were decreased(t=6.459～32.863,all P＜0.01);Caspase-3 activity(117.35％±8.67％vs 238.56％±13.22％),Bax level(1.13±0.45 vs 3.18±0.71),ROS level(128.73％±14.37％vs 387.65％±25.98％),MDA content(18.13±3.89 nmol/mg vs 33.21±3.17 nmol/mg)and Fe2+concentration(19.05±3.45 nmol/ml vs 38.77±6.53 nmol/ml)in placental tissues of PE+exo group were decreased,while Bcl-2 level(1.04±0.11 vs 0.47±0.08),GSH content(7.86±1.07 nmol/mg vs 4.12±1.22 nmol/mg),GPX4 protein(0.98±0.14 vs 0.48±0.06)and SLC7A11 protein(1.11±0.09 vs 0.51±0.11)levels were increased compared with PE group,with significant differences between groups(t=6.093～29.633,all P＜0.01).Conclusion In the placental tissues and peripheral blood of PE rats,miR-3614-5p was down-regulated.Exosomes overexpressing miR-3614-5p derived from MSCs suppressed PE progression in rats by inhibiting ferroptosis.These results suggested that exosomes miR-3614-5p derived from MSCs may be a novel potential biomarker for PE treatment.

Objective To explore the intestinal flora distribution characteristics in patients with Hashimoto's thyroiditis(HT)and analyze the correlation between intestinal flora changes and HT and the predictive value on the occurrence of HT,so as to provide reference for clinical diagnosis and treatment of HT.Methods A total of 120 patients with HT admitted to Qinhuangdao Traditional Chinese Medicine Hospital from June 2022 to June 2023 were included in observation group,and 30 healthy physical examination subjects during the same period were enrolled as control group.Both groups performed intestinal flora detection and thyroid function detection.The α diversity and the relative abundance of intestinal flora at phylum level were compared between the two groups and among HT patients with different thyroid functions,and the correlation between α diversity of intestinal flora and relative abundance of intestinal flora at phylum level,serum levels of anti-thyroglobulin antibody(TgAb)and thyroid peroxidase antibody(TPOAb)and thyroid function was analyzed by spearman correlation analysis.Receiver operating characteristic(ROC)curve was used to analyze the predictive efficiency of α diversity and relative abundance of intestinal flora at phylum level on HT.Results Compared with control group,the intestinal flora α diversity indexes of shannon-wiener diversity index(shannon)and simpson diversity index(simpson)were lower in observation group[4.66±0.61 vs 5.21±0.46,0.89±0.06 vs 0.93±0.07],and the differences were statistically significant(r=4.617,3.156,P＜0.05).The relative abundance of Bacteroidetes in observation group was lower than that in control group(52.31％±2.02％vs 58.59％±2.11％),while the relative abundances of Firmicutes,Proteobacteria,Fusobacteria,Actinobacteria and Verrucomicrobia were higher than those in control group(37.15％±2.43％vs 33.46％±2.56％,3.36％±0.26％vs 2.79％±0.19％,1.02％±0.09％vs 0.65％±0.11％,1.01％±0.12％vs 0.57％±0.13％,0.82％±0.11％vs 0.41％±0.09％),and the differences were statistically significant(t=15.096,7.360,11.267,19.231,17.665,18.882,all P＜0.001).The levels of serum TgAb(169.12±10.23 IU/ml vs 18.59±1.78 IU/ml)and TPOAb(56.43±12.11 IU/ml vs 2.51±0.57 IU/ml)in observation group were higher compared with those in control group,and the differences were statistically significant(t=80.097,24.319,all P＜0.001).Among 120 HT patients in this study,there were 89 cases with normal thyroid function and 31 cases with abnormal thyroid function.The intestinal flora α diversity indexes of shannon(4.49±0.64 vs 4.83±0.58),and simpson(0.87±0.07 vs 0.91±0.05)in patients with abnormal thyroid function were lower than those in normal patients,and the differences were statistically significant(t=2.736,3.439,all P＜0.05).The relative abundance of Bacteroides was lower(49.10％±1.99％vs 55.52％±2.05％),while the relative abundances of Firmicutes,Proteobacteria,Fusobacteria,Actinobacteria and Verrucomicrobia were higher than those in normal people(38.87％±2.60％vs 35.43％±2.26％,3.60％±0.26％vs 3.12％±0.19％,1.15％±0.09％vs 0.89％±0.11％,1.16％±0.13％vs 0.86％±0.11％,0.97％±0.13％vs 0.67％±0.09％)],and the differences were statistically significant(t=15.128,7.016,9.849,14.576,12.464,14.148,all P＜0.001).Spearman correlation analysis showed that the intestinal flora α diversity indexes(shannon and simpson)and relative abundance of Bacteroides in patients with HT were negatively correlated with serum TgAb and TPOAb levels(r=-0.436,-0.427,-0.402;-0.419,-0.456,-0.416,all P＜0.001),but were positively correlated with thyroid function(r=0.401,0.409,0.487,all P＜0.001).The relative abundances of Firmicutes,Proteobacteria,Fusobacteria,Actinobacteria and Verrucomicrobia were positively correlated with serum TgAb and TPOAb levels(r=0.411,0.441,0.447,0.421,0.447;0.425,0.415,0.438,0.402,0.469,all P＜0.001),but were negatively correlated with thyroid function(r=-0.412,-0.417,-0.475,-0.463,-0.471,all P＜0.001).ROC curve analysis revealed that the best cut-off value,the area under curve(AUC),sensitivity,specificity,95％confidence interval(95％CI)and Youden index in shannon,simpson,combined detection of simpson and shannon,and the combined detection of relative abundance of intestinal bacteria at phylum level for predicting HT were 4.677,0.749,49.17％,96.67％,95％CI(0.672～0.816)and 0.458;0.940,0.644,80.00％,53.33％,95％CI(0.562～0.721)and 0.333;0.752,85.83％,50.00％,95％CI(0.664～0.810)and 0.462;0.743,96.67％,46.67％,95％CI(0.665～0.811)and 0.548,respectively Conclusion HT had intestinal flora imbalance which was related to thyroid cell destruction and thyroid dysfunction in patients with HT.Intestinal flora detection has predictive efficiency on HT.