Objective:To analyze the infiltration of immune cells in different molecular subtypes of endometrial cancer(EC)at the protein level and explore the correlation between immune infiltration and clinicopathological features.Methods:Clinical data from 75 mismatch repair-deficient(MMR-D)EC cases and 74 mismatch re-pair-proficient(MMR-P)EC cases with comparable differentiation levels,diagnosed at Sichuan Provincial People's Hospital between January 2013 and January 2021,were selected.Molecular classification of EC was con-ducted according to the ProMisE standard.Immunohistochemistry(IHC)staining and fluorescent multiplexed im-munohistochemistry(F-mIHC)techniques were used to quantify the distribution of CD8+T cells,regulatory T cells(Tregs),and macrophages.The clinicopathological features and immune infiltration characteristics of different subtypes were investigated.Results:Among 149 samples,75 were MMR-D,8 were POLE-mutated(POLEmut),13 were p53-mutated(p53abn),and 53 were p53 wild-type(p53wt).The infiltration characteristics of CD8+T cells,Tregs,and macrophages were no statistically significant differences correlated with age and pathological stage(P>0.05).Among the molecular subtypes of EC,MMR-D tumors demonstrated a significantly higher densi-ty of CD8+T cells and Tregs compared to MMR-P tumors(P<0.05).Further comparison across the four EC mo-lecular subtypes revealed that the MMR-D subtype,which had a favorable prognosis,displayed a significantly high density of CD8+T cells compared to the p53abn subtype and the p53wt subtype(P<0.05).Additionally,the Tregs density of the MMR-D tumors was significantly higher than that of the p53wt tumors,in both the tumor area(P<0.05)and the stroma area(P<0.05).The POLEmut type tumors,which had the best prognosis,showed a high infiltration of CD8+T cells and Tregs cells compared to the p53abn and p53wt types,although there was no signifi-cant statistical difference(P>0.05).The p53wt subtype,which had a moderate prognosis,exhibited low density of CD8+T cells(P<0.05)and Tregs(tumor area,P<0.01,stroma area,P<0.05)compared to MMR-D tumors.The p53abn subtype,which was associated with the worst prognosis,exhibited higher infiltration of M2-like macro-phages than the MMR-D subtype(P<0.05)and the p53wt subtype(P<0.05)in tumor region,and lower CD8+T cell density than the MMR-D subtype(P<0.05).No significant statistical difference was observed in the density of M1-like macrophages across the four molecular subtypes in either the tumor or stroma areae(P>0.05).Conclu-sions:There was no statistically significant relation found between infiltration characteristics of immune cells and clinicopathological features.The types and distribution of infiltrated immune cells were different across these EC molecular subtypes.This indicated that ECs with different molecular subtype exhibited different features of immune infiltration which is closely associated with patient prognosis.

Objective:To investigate the factors for adverse pregnancy outcomes and the value of throm-boelastography(TEG)in patients with prethrombotic state(PTS)during pregnancy.Methods:A total of 183 preg-nant women with PTS treated at the Department of Obstetrics,Xiangya Third Hospital,Central South University,from January 2020 to September 2023 were retrospectively analyzed.These women were divided into two groups based on pregnancy outcomes:the favorable pregnancy outcome group(97 cases)and the adverse pregnancy outcome group(86 cases).The risk factors for adverse pregnancy outcomes were evaluated by univariate and multivariate analyses,and the diagnostic value of TEG was assessed using the receiver operating characteristic(ROC)curve and Kaplan-Meier method.Results:The results of univariate analysis indicated that,compared to-the favorable pregnancy outcome group,the positive rates of antinuclear antibody(ANA),anticardiolipin antibody(ACA),and anti-beta2 glycoprotein antibody 1(β2-GP1)were higher in the adverse pregnancy outcome group(P<0.05),and lower utilization rates of hydroxychloroquine and low-molecular heparin(P<0.05),and shorter clotting reaction time in the thromboelastography(TEG-R)(P<0.05).Multivariate Logistic regression analyses revealed that TEG-R,ANA,β2-GP1 and gestational age at treatment initiation were all independent risk factors for adverse pregnancy outcome in pregnant women with PTS during pregnancy(P<0.05).ROC curve analysis showed the area under the curve for TEG-R was 0.632(95%Cl 0.551-0.741).At a TEG-R value of 4.65 min,the specificity was 0.948 and the sensitivity was 0.302.Kaplan-Meier survival curve analysis revealed that the gesta-tional weeks of pregnant women with an TEG-R>4.65 min were significantly longer than those with an TEG-R≤4.65 min.These results were statistically significant(P<0.01).Conclusions:ANA,ACA,β2-GP1,and TEG are valuable for predicting and evaluating adverse pregnancy outcomes in PTS patients.The TEG-R value≤4.65 min may serve as a critical threshold to guid anticoagulant therapy during pregnancy.

Objective:To explore the relationship between serum soluble myeloid cell trigger receptor-1(sTREM-1)messenger RNA(mRNA),high mobility group protein B1(HMGB1)mRNA levels and intestinal flora and the occurrence of preterm premature rupture of membranes(PPROM)in gestational diabetes mellitus(GDM)patients.Methods:98 GDM pregnant women with GDM combined with PPROM who underwent regular prenatal check ups and delivered at the First Hospital Affiliated to Hebei North University from June 2022 to May 2024 were selected as the GDM combined with PPROM group,102 GDM pregnant women without PPROM were selected as the GDM without PPROM group,and 108 normal pregnant women were selected as the control group.Intestinal flora,serum sTREM-1 mRNA,HMGB1 mRNA levels of the three groups of pregnant women were collected and compared;the correlation between serum sTREM-1 mRNA and HMGB1 mRNA levels and intestinal flora in GDM pregnant women,the factors affecting the occurrence of PPROM in pregnant women with GDM,and the predictive value of serum sTREM-1 mRNA and HMGB1 mRNA in the occurrence of PPROM in GDM pregnant women were analyzed.Results:Compared with control group,colony number of Enterococcus,Enterobacter,Bacteroides and Fusobacterium were significantly increased in GDM without PPROM group and GDM combined with PPROM group(P＜0.05),the colony number of Bifidobacterium and Lactobacillus was significantly de-creased(P＜0.05).The gestational age at delivery of GDM combined with PPROM group was significantly lower than that of control group and GDM without PPROM group(P＜0.05).Compared with GDM without PPROM group,the colony number of Bifidobacterium and Lactobacillus were significantly decreased(P＜0.05),colony number of Enterococcus,Enterobacter,Bacteroides and Fusobacterium were significantly increased in GDM com-bined with PPROM group(P＜0.05).The serum levels of sTREM-1 mRNA and HMGB1 mRNA in control group,GDM without PPROM group,GDM combined with PPROM group were increased sequentially(P＜0.05).Serum sTREM-1 mRNA was positively correlated with HMGB1 mRNA in GDM pregnant women(r＞0,P＜0.05);serum sTREM-1 mRNA and HMGB1 mRNA were positively correlated with the colony numbers of Enterococcus,Enter-obacter,Bacteroides and Fusobacterium(r＞0,P＜0.05),and negatively correlated with the colony numbers of Bifidobacterium and Lactobacillus(r＜0,P＜0.05).Enterococcus,Enterobacter,Bacteroides,Fusobacterium,sTREM-1 mRNA and HMGB1 mRNA were independent risk factors for PPROM in GDM pregnant women(OR＞1,P＜0.05),while Bifidobacterium and Lactobacillus were independent protective factors for PPROM in GDM pregnant women(OR＜1,P＜0.05).The area under curve(AUC)of predicting PPROM in GDM pregnant women by combining serum sTREM-1 mRNA and HMGB1 mRNA(0.922)was higher than the AUC predicted by serum sTREM-1 mRNA and HMGB1 mRNA alone(0.829,0.841)(P＜0.05).Conclusions:Serum sTREM-1 mRNA and HMGB1 mRNA were highly expressed in GDM pregnant women,and the increase trend of serum both was more obvious in GDM pregnant women with PPROM,both of which were related to intestinal flora imbalance and the occurrence of PPROM.The combination of sTREM-1 mRNA and HMGB1 mRNA has high efficacy in predicting the occurrence of PPROM.

Objective:To explore the feasibility of measuring the angle of progress(AOP)and the fetal head-perineum distance(HPD)by ultrasound to predict the duration of the second stage of labor.Methods:A to-tal of 135 full-term singleton primiparas who gave birth through vagina in Shandong Provincial Hospital Affiliated to Shandong First Medical University from September 2022 to June 2024 were selected.They all received medication for labor analgesia during labor,and the fetal position was determined as occipital anterior position by ultrasound when the cervix was fully dilated.AOP and HPD were measured during the contraction interval.All parturients were divided into group A(duration of the second stage of labor≤60 min),group B(60 min＜duration of the sec-ond stage of labor≤120 min)and group C(duration of the second stage of labor＞120 min).The differences of AOP and HPD among these groups were compared,and their predictive efficacy for vaginal delivery within 60min and 120min was evaluated.Results:①The AOP in group A was higher than that in group B.The HPD in group A was lower than that in group B.The differences were statistically significant(P＜0.01).The AOP in group B was higher than that in group C.The HPD in group B was lower than that in group C.The differences were statistically significant(P＜0.05).②The area under the ROC curve for AOP and HPD to predict a vaginal delivery within 60 min was 0.872 and 0.914,and the optimal cut-off value was 126.25°and 4.52 cm,respectively(P＜0.01).③The area under the ROC curve for AOP and HPD to predict a vaginal delivery within 120 min was 0.901 and 0.801,and the optimal cut-off value was 116.40°and 4.69 cm,respectively(P＜0.01).Conclusions:It is feasible to pre-dict the duration of the second stage of labor duration by measuring AOP and HPD using intrapartum ultrasound.

Objective:To investigate the relationship between IUA occurrence and circRNA_0003031 by con-structing a mouse model of intrauterine adhesion(IUA).Methods:A mouse model of IUA was established by me-chanical curettage and lipopolysaccharide infection.The mice were divided into control group(NC group),IUA group,IUA+si-circRNA_0003031 group,and IUA+si-circRNA_0003031+miR-93-5p inhibitor group.The number of endometrial glands and the degree of fibrosis were determined using HE and Masson staining,respectively.Western blot test was used to detect the expression of ephthelial mesenchymal transition(EMT)marker proteins(vimentin,E-cadherin,N-cadherin),fibrosis-related molecules(collagen Ⅰ,collagen Ⅲ),α-smooth muscle actin(α-SMA),Transforming growth factor β1(TGF-β1),and receptors(TGF-β1R),Smad2,P-Smad2].Results:Compared with the IUA group,the number of endometrial glands increased,the degree of endometrial fibrosis de-creased in the IUA+si-circRNA_0003031 group(P＜0.05),and the protein expression of vimentin,N-cadherin,α-SMA,collagen Ⅲ,collagen Ⅰ,P-Smad2,Smad2,TGF-β1R,and TGF-β1 was down-regulated and E-cadherin was up-regulated(P＜0.05).Compared with the IUA+si-circRNA_0003031 group,in the IUA+si-circRNA_0003031+miR-93-5p inhibitor group,the number of endometrial glands was reduced and the degree of endometrial fibrosis was increased(P＜0.05),the protein expression of vimentin,N-cadherin,α-SMA,collagen Ⅲ,collagen Ⅰ,P-Smad2,Smad2,TGF-β1R,and TGF-β1 was up-regulated and E-cadherin was down-regulated,and the differences were statistically significant except for P-Smad 2(P＜0.05).Conclusions:CircRNA_0003031 may promote the oc-currence and development of IUA by inhibiting the expression of miR-93-5p and relieving its inhibitory effect on the TGFβ1/Smad2 signaling pathway.

Objective:To compare the long-term efficacy and incidence of complications between robotic-assis-ted sacrocolpopexy(RASC)and traditional laparoscopic sacrocolpopexy(LSC)in the treatment of pelvic organ prolapse(POP),and to explore the clinical efficacy of RASC.Methods:Clinical data of 258 POP patients who un-derwent RASC or LSC in the First Affiliated Hospital of Zhengzhou University from June 2014 to September 2023 were collected and retrospectively analyzed.They were divided into RASC group(109 cases)and LSC group(149 cases)according to the surgical procedure.The postoperative and intraoperative conditions between the groups were statistically analyzed and compared,the long-term complications were followed up.The scores of the pelvic floor dysfunction questionnaire(PFDI-20)and pelvic floor dysfunction impact questionnaire(PFIQ-7)were com-pared between groups and within groups to reflect the long-term postoperative efficacy of patients,so as to evalu-ate the surgical effect.Results:①There were no significant differences between the two groups after surgery in the incidence of long-term complications such as recurrence of prolapse,exposure to mesh,polyps of stump of vagina,difficulty defecation,new urinary incontinence,osteomyelitis,lumbago,lower abdominal pain(P＞0.05);②There were statistically significant differences in operation time,intraoperative blood loss,intestinal recovery time and postoperative hospital stay between the two groups(P＜0.05);③In the two groups,PFDI-20 and PFIQ-7 scores at 6 months,1 year,3 years,5 years,and more than 5 years after surgery were significantly different from those before surgery(P＜0.05).However,there was no statistically significant difference in the PFDI-20 and PFIQ-7 scores between the two groups at each postoperative time point(P＞0.05);④There were statistically sig-nificant differences in POP-Q indicator points Aa,Bb,C,Ap,and Bp between each group at 6 months and 1 year after surgery compared to preoperative levels(P＜0.05).However,there was no statistically significant difference in POP-Q indicator point values between the two groups at each time point(P＞0.05);⑤The operations in both groups were successfully completed(100％)without conversion to laparotomy,and the postoperative recovery was satisfactory.Conclusions:Compared with LSC,RASC has the advantages of less intraoperative bleeding,shorter operation time,faster recovery of intestinal function,and shorter hospital stay.Meanwhile,RASC and LSC can achieve similar results in long-term efficacy and postoperative long-term complications.RASC can be a major choice for the treatment of pelvic organ prolapse patients.

Objective:To investigate the expression of miR-769-5p in serum exosomes of epithelial ovarian cancer(EOC)and its mechanism of influence on EOC cell proliferation,migration,and invasion.Methods:The expression level of miR-769-5p in serum exosomes from the malignant group,benign ovarian disease group,and healthy control group was detected using reverse transcription quantitative polymerase chain reaction(RT-qPCR).Lipofectamine 3000 transfection was used to construct overexpression and knockdown cell lines,which were divided into four groups:miR-769-5p mimics group,mimics-NC group,miR-769-5p inhibitor group,and inhibi-tor-NC group.The effects of miR-769-5p on cell proliferation,migration,and invasion were assessed using CCK-8,wound healing,and Transwell assays.Transcriptome sequencing was conducted on EOC cells overex-pressing miR-769-5p,and potential target genes were predicted using public databases.The interaction between miR-769-5p and its target gene was validated by dual-luciferase reporter assay.RT-qPCR and Western blotting were used to further confirm the regulatory effect of miR-769-5p on the target gene.Results:The expression level of miR-769-5p in serum exosomes from the malignant group was significantly lower than that in the healthy control and benign ovarian disease groups(P＜0.05).Compared with the mimics-NC group,the miR-769-5p mimics group exhitied lower OD450 values,migration rates,and numbers of invading cells in both SKOV3 and OVCAR3 EOC cell lines(P＜0.05).Compared with the inhibitor-NC group,cells in the miR-769-5p inhibitor group exhibited increased OD450 values,migration rates,and invasion counts(P＜0.05).The dual-luciferase reporter assay showed that co-transfection of miR-769-5p mimics with pEZX-FR02-TFAM-WT significantly reduced the luciferase activity compared to the mimics-NC group(P＜0.01),whereas co-transfection of miR-769-5p mimics with pEZX-FR02-TFAM-MUT showed no significant difference in luciferase activity compared to the control(P＞0.05).These findings indicate that miR-769-5p can directly bind to the 3'-UTR region of TFAM.Furthermore,compared with the mimics-NC group,EOC cells transfected with miR-769-5p mimics showed significantly decreased TFAM mRNA and protein expression levels(P＜0.05).Conclusion:miR-769-5p is significantly downregulated in EOC serum exosomes and may serve as a promising biomarker for early tumor diagnosis.Moreover,miR-769-5p may inhibit the proliferation,migration,and invasion of ovarian cancer cells by targeting TFAM.

Objective:To investigate the clinical application value of expanded non-invasive prenatal testing(NIPT-plus)in screening for fetal chromosome copy number variations(CNV).Methods:From January 2021 to December 2023,141 pregnant women who voluntarily underwent amniocentesis at the Prenatal Diagnostic Centre of Urumqi Maternal and Child Health Hospital due to NIPT-plus suggesting a high risk of fetal CNV were selected.Amniotic fluid samples were collected for fetal chromosome karyotyping and chromosome microarray analysis(CMA).Pregnant women who underwent the above tests signed an informed consent form,and all cases were followed up forpregnancy outcome.Results:Among 141 NIPT-plus screen positive pregnant women,41 true posi-tive cases were detected by chromosomal karyotype analysis and CMA.The positive predictive value(PPV)for NIPT-plus screening for CNV was 29.08％(41/141).There was no statistically significant difference(P＞0.05)in the PPV of CNV detected by NIPT plus among different ages,indications and variant types.However,the PPV of CNV size＜10 Mb was significantly higher than that of CNV size≥ 10 Mb,and the difference was statistically signif-icant(39.62％vs.22.73％,P＜0.05).Among the 41 true positive cases,in addition to CNV,the CMA also detec-ted 7 cases of regions of Homozygosity(ROH),accounting for 17.07％(7/41)of the cases,two of which involved imprinted genes located on chromosomes 6 and 7.All continued pregnancy after genetic counselling and no signif-icant abnormalities were seen at neonatal follow-up after birth.Conclusions:NIPT-plus screening for fetal CNV has some clinical value,especially for CNV with fragment size＜10Mb,but accuracy needs to be further improved;CMA as a molecular diagnostic technique can detect ROH in cases where NIPT-plus suggests CNV abnormali-ties,and the combined use of the two techniques also opens new avenues for screening and diagnosis of prenatal imprinted diseases.

Objective:To investigate the clinical significance of prenatal screening and genetic analysis in the diagnosis of fetal aberrant right subclavian artery(ARSA).Methods:The ultrasonographic features of ARSA fetu-ses detected by prenatal ultrasound at the University of Hong Kong-Shenzhen Hospital from October 2017 to March 2022 were retrospectively analyzed.The fetuses were divided into isolated ARSA group and complicated ARSA group.Their genetic analysis results and pregnancy outcome were analyzed.Results:Among 30,260 preg-nant women,185 fetuses were diagnosed with ARSA by prenatal ultrasound screening,with an incidence of 0.6％;5 fetuses(2.6％)were diagnosed by ultrasound in the first trimester,and the remaining were diagnosed by fetal grade Ⅲ structural ultrasound examination at 20～24 weeks' gestation.Among them,158 fetuses(85.4％)had isolated ARSA,and 27(14.6％)had complicated ARSA.Among fetuses with ARSA and other structural abnormal-ities,cardiovascular system accounted for the highest proportion(44.4％),followed by nervous system(22.2％)and urinary system(22.2％).Through genetic analysis,8.3％(4/48)fetuses with isolated ARSA and 40.0％(4/10)fetuses with complicated ARSA were found to have chromosomal numerical or structural abnormalities,with statis-tically significant difference between the two groups(P=0.024).Genetic analysis was completed in 48 isolated ARSA,and the positive rate of pathogenic copy number variants(CNV)was 4.2％(2/48),which was not signifi-cantly different from the pathogenic CNV incidence rate of 0.4％(1/239)in elderly pregnant cases during the same period(P=0.074).The Down syndrome positive likelihood ratio(LR+)for isolated ARSA was 2.5 and the Down syndrome LR+for complicated ARSA was 49.6.Conclusions:Complicated ARSA is often associated with cardiovascular abnormalities and is more likely to develop Down syndrome than isolated ARSA.Although the inci-dence of pCNV in isolated ARSA is slightly higher than the natural incidence,the correlation between pCNV and i-solated ARSA has not been clearly determined by the current sample size.

Objective:To construct a risk prediction model for Vaginal Intraepithelial Neoplasia Grade 2 or Worse(VaIN 2+)lesions,and to establish a nomogram for individual diagnosis of VaIN 2+and risk stratification,so as to provide guidance for the treatment of vaginal lesions.Methods:A total of 248 women diagnosed with VaIN through colposcopic biopsy at the Center for Gynecologic and Cervical Diseases,First Affiliated Hospital of Nanjing Medical University,from January 2021 to January 2024 were included in this study.Based on the gold standard established by histological and pathological findings,these patients were categorized into a lower VaIN 2 group and a VaIN 2+group.Univariate comparative analysis was performed on the two groups.Multivariate Logis-tic regression analysis was used to determine the risk factors of VaIN 2+and to construct a diagnostic model.The nomogram model was established by using R Studio software.The discrimination,calibration and clinical practical value of the model were evaluated by the area under the receiver operating characteristic(ROC)curve and cali-bration curve.Results:Univariate analysis identified that HPV type,cervical lesion grade,acetowhite change,vagi-nal lesion duration,vaginal lesion location,and cervical lesion duration as influencing factors for diagnosing VaIN 2+(P<0.1).Multivariate binary Logistic regression analysis indicated that HPV16/18 positivity,cervical lesion grade≥CIN 2,thick acetowhite change,vaginal lesion duration≥5 years,and vaginal lesion location at the upper 1/3 of the vagina were independent risk factors for diagnosing VaIN 2+(OR>1,P<0.05),while cervical lesion duration<3 years was a protective factor(OR<1,P<0.05),with acetowhite change having the greatest impact(OR4.54).A regression model was established based on the multivariate binary Logistic regression analysis,with an AUC of 0.813.A nomogram model was constructed and internally validated,yielding a consistency index(C-index)of 0.81.Patients were stratified into risk groups using the X-tile software,with higher total scores indi-cating a greater risk of developing VaIN 2+.Conclusions:The nomogram model constructed in this study can in-dividually predict the risk of VaIN 2+lesions in patients,with high accuracy and clinical practicability.