UNASSIGNED: This study aimed to assess variations in the absolute counts of various leukocyte subsets in the peripheral blood of women with pregnancies affected by preterm prelabour rupture of membranes (PPROM), in relation to the presence of intra-amniotic inflammation (IAI).
UNASSIGNED: The study included fifty-two women with singleton pregnancies experiencing PPROM. Absolute counts of different leukocyte subpopulations, such as granulocytes, monocytes, lymphocytes, T cells and their subsets, B cells and their subsets, and NK cells and their subsets, were measured in maternal peripheral blood samples using multicolour flow cytometry. IAI was identified by elevated concentrations of interleukin 6 (IL-6) in the amniotic fluid, which was collected through transabdominal amniocentesis.
UNASSIGNED: Women with IAI exhibited higher absolute counts of leukocytes (p = 0.003), granulocytes (p = 0.008), and monocytes (p = 0.009). However, the presence of IAI did not significantly affect the absolute counts of lymphocytes or their subpopulations.
UNASSIGNED: The study found that IAI is associated with changes in the absolute counts of leukocytes from the innate immunity compartment in the peripheral blood of women with pregnancies complicated by PPROM. Conversely, it does not significantly alter the counts of cells from the adaptive immune system. The changes observed may reflect the natural, temporal, and localised characteristics of IAI.
Preterm birth is the most serious complication in contemporary perinatal medicine. Preterm birth, which is defined as a labour before the completion of 37 weeks of pregnancy, is often accompanied by premature rupture of the amniotic membranes and drainage of amniotic fluid. Such a situation is often complicated by inflammation, which adversely affects the health of the foetus. A number of procedures and markers have been developed for the diagnosis of inflammation, but they are determined from hard-to-reach amniotic fluid. It is therefore appropriate to try to find reliable markers of inflammation in the much more accessible maternal peripheral blood. Such a marker can be increased numbers of leukocytes, which have been repeatedly investigated in this context. However, little attention is directed to other leukocyte populations and especially to various lymphocyte subpopulations. This study aimed to test changes in absolute counts of different types of leukocytes and lymphocyte subpopulations in women with premature rupture of membranes with respect to ongoing inflammation. The results of the study showed that inflammation is accompanied by increased numbers of leukocytes, granulocytes and monocytes, however, the results did not show significant changes in the number of lymphocytes and their subpopulations.

BACKGROUND: Despite extensive research, the identification of effective biomarkers for early prediction of preterm birth (PTB) continues to be a challenging endeavor. This study aims to identify amniotic fluid (AF) protein biomarkers useful for the early diagnosis of PTB.
METHODS: We initially identified the protein expression profiles in the AF of women with PTB (n = 22) and full-term birth (FTB, n = 22), from the First People\'s Hospital of Yunnan Province who underwent amniocentesis from November 2019 to February 2020, using mass spectrometry employing the data-independent acquisition (DIA) technique, and then analyzed differentially expressed proteins (DEPs). Subsequently, the least absolute shrinkage and selection operator (LASSO) and random forest analysis were employed to further screen the key proteins for PTB biomarker identification. The receiver operating characteristic (ROC) analysis, calibration plots, and decision curve analyses (DCA) were utilized to assess the discrimination and calibration of the key biomarkers.
RESULTS: A total of 25 DEPs were identified between the PTB and FTB groups, comprising 13 up-regulated and 12 down-regulated proteins. Three key protein biomarkers for early PTB diagnosis were identified: IL1RL1 (interleukin-1 receptor-like 1), APOE (apolipoprotein E), and NECTIN4 (nectin cell adhesion molecule 4). The results of the ROC analysis showed that the area under the curve (AUC) of the three proteins combined as a biomarker for early diagnosis of PTB was 0.913 (95% CI: 0.823-1.000), with a sensitivity of 0.864 and a specificity of 0.955, both superior to those of the individual biomarkers. Bootstrap internal validation revealed a concordance index (C-index) of 0.878, with a sensitivity of 0.812 and a specificity of 0.773, indicating the robust predictive performance of these biomarkers.
CONCLUSIONS: We identified three previously unexplored yet potentially useful protein biomarkers in AF for early PTB diagnosis: IL1RL1, APOE, and NECTIN4.

暂无摘要。

OBJECTIVE: To determine whether altered concentrations of various inflammation/immune-, acute phase-, extracellular matrix-, adhesion-, and serine protease-related proteins in the amniotic fluid (AF) are independently associated with microbial invasion of the amniotic cavity and/or intra-amniotic inflammation (MIAC/IAI), imminent spontaneous preterm delivery (SPTD; ≤7 days), and major neonatal morbidity/mortality (NMM) in women with early preterm prelabor rupture of membranes (PPROM).
METHODS: This was a retrospective cohort study involving 111 singleton pregnant women with PPROM (24-31 weeks) undergoing amniocentesis to diagnose MIAC/IAI. The following proteins were measured in stored AF samples by enzyme-linked immunosorbent assay (ELISA): APRIL, DKK-3, Gal-3BP, IGFBP-2, IL-8, VDBP, lumican, MMP-2, MMP-8, SPARC, TGFBI, TGF-β1, E-selectin, ICAM-5, P-selectin, haptoglobin, hepcidin, SAA1, kallistatin, and uPA.
RESULTS: Multivariate logistic regression analyses revealed that (i) elevated APRIL, IL-8, MMP-8, and TGFBI levels in the AF, reduced lumican and SPARC levels in the AF, and high percentages of samples above the lower limit of quantification for AF TGF-β1 and uPA were significantly associated with MIAC/IAI; (ii) elevated AF levels of IL-8 and MMP-8 were significantly associated with SPTD within 7 days; and (iii) elevated AF IL-6 levels were significantly associated with increased risk for major NMM, when adjusted for baseline covariates.
CONCLUSIONS: ECM (lumican, SPRAC, TGFBI, and TGF-β1)- and serine protease (uPA)-associated proteins in the AF are involved in the regulation of the host response to infection/inflammation in the amniotic cavity, whereas AF inflammation (IL-8, MMP-8, and IL-6)-associated mediators are implicated in the development of preterm parturition and major NMM in early PPROM.

UNASSIGNED: At the maternal-fetal interface in pregnancy, stress during pregnancy can lead to an increased vulnerability to later psychopathology of the fetus. Potential mediators of this association have scarcely been studied and may include early alterations of fetal brain-derived neurotrophic factor (BDNF). Amniotic fluid is of particular interest for effects on fetal endocrine alterations, as the assessment in amniotic fluid allows for measurements over a time integral. This study hypothesized that maternal psychometrics, socioeconomic status and glucocorticoids are related to BDNF levels in amniotic fluid at birth. The association of fetal BDNF with newborn anthropometrics was tested.
UNASSIGNED: Women near term who underwent elective cesarean section and their newborns were investigated (n = 37). Maternal psychometrics, socioeconomic status and glucocorticoids (the sum of cortisol and cortisone) in amniotic fluid at birth were analyzed for an association with fetal BDNF in amniotic fluid at birth. Newborn anthropometrics were assessed by length, weight, head circumference and gestational age at birth.
UNASSIGNED: In bivariate analysis, maternal psychometrics and socioeconomic status were not related to fetal BDNF in amniotic fluid at birth. The sum of cortisol and cortisone related to increased fetal BDNF in amniotic fluid at birth (r = 0.745, p < 0.001). BDNF in amniotic fluid was associated negatively with fetal birth weight per gestational age (r = -0.519, p < 0.001), length per gestational age (r = -0.374, p = 0.023), head circumference per gestational age (r = -0.508, p = 0.001), but not with gestational age at birth. In multiple regression analysis, the sum of cortisol and cortisone (p < 0.001) and birth weight per gestational age (p = 0.012) related to higher fetal BDNF levels in amniotic fluid at birth (R2 = 0.740, p < 0.001) when controlling for fetal sex and maternal age. Head circumference per gestational age predicted fetal BDNF with borderline significance (p = 0.058) when controlling for confounders.
UNASSIGNED: Glucocorticoids in amniotic fluid were positively associated with high fetal BDNF at birth, which may be an adaptive fetal response. Maternal psychological variables and socioeconomic status did not link to fetal BDNF. Birth weight and head circumference per gestational age were inversely associated with fetal BDNF at birth, which may represent a compensatory upregulation of BDNF in fetuses with low anthropometrics. Longitudinal studies are needed to assess the role of stress during pregnancy on later offspring development. The analysis of additional fetal growth factors and inflammation upon maternal stress in further biomaterials such as the placenta is warranted, to understand mechanistic alterations of how maternal stress links to fetal development and an increased vulnerability for psychopathology.

OBJECTIVE: To explore the clinical utility of nine inflammatory immune-, adhesion-, and extracellular matrix-related mediators in the plasma for predicting intraamniotic inflammation and/or microbial invasion of the amniotic cavity (IAI/MIAC) and composite neonatal morbidity and/or mortality (CNMM) in women with preterm premature rupture of membranes (PPROM) when used alone or in combination with conventional blood-, ultrasound-, and clinical-based factors.
METHODS: This retrospective cohort comprised 173 singleton pregnant women with PPROM (24 + 0 - 33 + 6 weeks), who underwent amniocentesis. Amniotic fluid was cultured for microorganisms and assayed for IL-6 levels. Plasma levels of AFP, CXCL14, E-selectin, Gal-3BP, kallistatin, progranulin, P-selectin, TGFBI, and VDBP were determined by ELISA. Ultrasonographic cervical length (CL) and neutrophil-to-lymphocyte ratio (NLR) were measured.
RESULTS: Multivariate logistic regression analyses revealed significant associations between (i) decreased plasma kallistatin levels and IAI/MIAC and (ii) decreased plasma progranulin levels and increased CNMM risk after adjusting for baseline variables (e.g., gestational age at sampling [or delivery] and parity). Using stepwise regression analysis, noninvasive prediction models for IAI/MIAC and CNMM risks were developed, which included plasma progranulin levels, NLR, CL, and gestational age at sampling, and provided a good prediction of the corresponding endpoints (area under the curve: 0.79 and 0.87, respectively).
CONCLUSIONS: Kallistatin and progranulin are potentially valuable plasma biomarkers for predicting IAI/MIAC and CNMM in women with PPROM. Particularly, the combination of these plasma biomarkers with conventional blood-, ultrasound-, and clinical-based factors can significantly support the diagnosis of IAI/MIAC and CNMM.

Background and Objectives: Epidemiological and microbiological-immunological studies have led to the conclusion that periodontal disease may be a risk factor for preterm birth. The aim of this study was to investigate and identify the relationship of some hematological cellular biomarkers characterizing the chronic oral focus of infection with pregnancy outcomes and their impact on those outcomes. Materials and Methods: Clinical and laboratory tests were conducted on 100 pregnant women, grouped by full-term or preterm births, with the assessment of the following markers: DMF, CPI and PIRI, PHP, microbiological examination of periodontal pockets and amniotic fluid, WBS count, WBCSI, LGI, and NMR. A statistical analysis was carried out with SPSS Statistics version 19.0. Results: Women with preterm labor had higher-grade caries (CSL > 0.3), while women with full-term childbirth had moderate caries (CSL < 0.3). A satisfactory level of oral hygiene efficiency was found in 50% (group 1) and 38.1% (group 2) of the expectant mothers. The periodontal status by the PIRI showed tissue lesions in 20.7% (group 1) and 92.9% (group 2) of the women. The WBCSI was 2.27 ± 0.82 and 2.15 ± 0.68, the NMR was 9.29 ± 5.119 and 11.62 ± 7.78, and the LGI was 3.54 ± 1.1 and 3.73 ± 0.81 in groups 1 and 2, respectively. Comparative analysis of bacterial contamination of the amniotic fluid revealed the predominance of Fusobacterium nucleatum (64.3%), Tannerella forsythia (57.1%), Prevotella intermedia (50%), Porphyromonas gingivalis (57.1%), Staphylococcus aureus (45.2%), and Candida albicans (50%) in women with premature birth. Conclusions: In women with preterm birth, the values of the indices characterizing a chronic oral focus of infection evoke more significant correlations with the timing of delivery, which indicates the significant role of an oral focus of infection. The presence of microbial invasion of amniotic fluid may indicate the role of periodontopathogenic bacteria in pregnant women diagnosed with a risk of preterm birth.

Antenatal administration of extracellular vesicles from amniotic fluid stem cells (AFSC-EVs) reverses features of pulmonary hypoplasia in models of congenital diaphragmatic hernia (CDH). However, it remains unknown which lung cellular compartments and biological pathways are affected by AFSC-EV therapy. Herein, we conducted single-nucleus RNA sequencing (snRNA-seq) on rat fetal CDH lungs treated with vehicle or AFSC-EVs. We identified that intra-amniotically injected AFSC-EVs reach the fetal lung in rats with CDH, where they promote lung branching morphogenesis and epithelial cell differentiation. Moreover, snRNA-seq revealed that rat fetal CDH lungs have a multilineage inflammatory signature with macrophage enrichment, which is reversed by AFSC-EV treatment. Macrophage enrichment in CDH fetal rat lungs was confirmed by immunofluorescence, flow cytometry, and inhibition studies with GW2580. Moreover, we validated macrophage enrichment in human fetal CDH lung autopsy samples. Together, this study advances knowledge on the pathogenesis of pulmonary hypoplasia and further evidence on the value of an EV-based therapy for CDH fetuses.

Objective: To explore the relationship between amniotic fluid and peripheral blood inflammatory factors and the pregnancy outcomes after emergency cervical cerclage, and to identify effective indicators for predicting adverse pregnancy outcomes after the procedure. Methods: A case-control study was conducted, including pregnant women who were hospitalized at Sun Yat-sen Memorial Hospital, from January 1, 2013, to July 31, 2019, and underwent emergency cervical cerclage due to cervical dilatation at gestational age between 16 and 28 weeks. A total of 85 pregnant women who underwent amniocentesis for the detection of amniotic fluid inflammatory factors during the perioperative period were included. Based on whether their baby was perinatal death, the participants were divided into the case group (28 cases with perinatal death) and the control group (57 cases with live births). Univariate logistic regression analysis was performed to identify risk factors associated with adverse pregnancy outcomes, followed by multivariate logistic regression analysis to establish a regression model and nomogram. Results: (1) The levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, IL-8, IL-10 in the amniotic fluid during the perioperative period and postoperative serum C-reactive protein (CRP) were significantly higher in the case group compared to the control group (all P<0.05). The case group underwent emergency cervical cerclage at an earlier gestational age compared to the control group, and their cervical dilation was greater than that of the control group (all P<0.05). However, there were no significant differences in the white blood cell counts, neutrophil percentage, and the level of preoperative CRP in the peripheral blood of pregnant women during the perioperative period (all P>0.05). (2) Univariate logistic regression analysis showed that the levels of amniotic fluid WBC, TNF-α, IL-1β, IL-2 receptor (IL-2R), IL-6, IL-8, IL-10, postoperative CRP in the peripheral blood, gestational age at cerclage and cervical dilation were associated with adverse pregnancy outcomes (all P<0.05). Multivariate regression analysis indicated that only the levels of amniotic fluid WBC and TNF-α were independent risk factors for perinatal death. (3) Based on clinical practice, a multivariate logistic regression model was constructed including the levels of amniotic fluid TNF-α, WBC, gestational age at cervical cerclage, and cervical dilation. A nomogram and calibration curve were plotted, which suggested its good predictive value for adverse pregnancy outcomes. Conclusions: During the perioperative period of emergency cervical cerclage, the levels of amniotic fluid WBC, TNF-α, IL-1β, IL-2R, IL-6, IL-8, IL-10 are associated with adverse pregnancy outcomes, with amniotic fluid WBC and TNF-α showing the closest relationship. However, there is no significant correlation between maternal peripheral hemogram during the perioperative period and adverse pregnancy outcomes. A model constructed by amniotic fluid TNF-α, WBC, cervical cerclage gestational age, and cervical dilation has a good predictive effect on adverse pregnancy outcomes.
目的： 探讨羊水和外周血中炎症因子水平与紧急子宫颈环扎术孕妇妊娠结局的关系，寻找预测术后不良妊娠结局的指标。 方法： 采用病例对照研究，收集2013年1月1日至2019年7月31日于中山大学孙逸仙纪念医院住院，妊娠16~28周因子宫颈外口扩张行紧急子宫颈环扎术的孕妇，选取其中围术期行羊膜腔穿刺术并检测羊水中炎症因子的孕妇共85例。根据是否抱婴回家，分为不良结局组（28例）与活产组（57例）。采用单因素logistic回归分析寻找不良妊娠结局的相关危险因素，进一步行多因素logistic回归分析建立预测不良妊娠结局的列线图。 结果： （1）与活产组比较，不良结局组孕妇行紧急子宫颈环扎术的孕周较早［分别为（23.7±1.8）、（22.9±1.9）周］，宫口扩张程度较大（中位数分别为2.0、3.0 cm），分娩孕周较早［分别为（32.8±4.0）、（25.2±2.0）周］、延长孕周时间较短（中位数分别为65.0、13.5 d），分别比较，差异均有统计学意义（P均<0.05）。（2）不良结局组紧急子宫颈环扎术围术期羊水中肿瘤坏死因子α（TNF-α）、白细胞介素（IL）1β、IL-6、IL-8、IL-10及术后外周血C反应蛋白（CRP）水平显著高于活产组（P均<0.05）；而环扎术前及术后孕妇外周血白细胞计数（WBC）、中性粒细胞百分比，以及术前CRP水平的差异均无统计学意义（P均>0.05）。（2）单因素logistic回归分析显示，羊水WBC、TNF-α、IL-1β、IL-2受体（IL-2R）、IL-6、IL-8、IL-10、术后外周血CRP、环扎术孕周及宫口扩张程度与不良结局相关（P均<0.05），多因素logistic回归分析显示，仅羊水WBC、TNF-α为不良结局的独立危险因素。（3）结合临床实践，综合羊水TNF-α、WBC、环扎术孕周及宫口扩张程度构建多因素logistic回归模型，绘制列线图及校准曲线，提示该多因素logistic回归模型对不良结局的预测价值良好，曲线下面积为0.811（95%CI：0.697~0.926），预测不良结局的敏感度为0.792，特异度为0.852，阳性预测值为0.679，阴性预测值为0.912。 结论： 紧急子宫颈环扎术围术期羊水WBC、TNF-α、IL-1β、IL-2R、IL-6、IL-8、IL-10与不良结局相关，其中羊水WBC及TNF-α关系最密切。而围术期母体外周血检查指标与不良结局无明显相关性。综合羊水TNF-α、WBC、环扎术孕周及宫口扩张程度构建的列线图对不良结局有良好的预测作用。.

Osteoarthritis (OA) is a degenerative disease that causes chronic pain and disability worldwide. This disease is mainly caused by IL-1β and TNF-α, which lead to cartilage degradation and inhibit the repair capacity of damaged cartilage. Recent studies have shown that amniotic fluid mesenchymal stem cells (AF-MSCs) secrete proteins that can effectively help in the treatment of cartilage damaged by OA. However, the underlying mechanism is still unclear. Therefore, the aim of this study was to investigate the effects and mechanisms behind the healing properties of the AF-MSC secretome (AFS-se) under OA conditions. This study involved growing chondrocyte progenitor cells (CPCs) and traumatized cartilage tissues in the presence of the cytokines IL-1β and TNF-α, which mimic OA conditions. AFS-se was then added to the culture medium to determine its effect on the CPCs and cartilage. Cell migration, endogenous cell outgrowth, the expression of chondrogenic and anabolic genes, and the mechanism of proteins in the NF-κB and MAPK signaling pathways were examined in this study. AFS-se inhibited the inflammatory effects of IL-1β and TNF-α by significantly reducing ERK phosphorylation in the MAPK signaling pathway and decreasing downstream proinflammatory COX2 products. The impaired CPCs recovered their ability to migrate, and endogenous CPCs in injured osteoarthritic cartilage were able to regrow in response to inflammatory stimuli. Additionally, the expression of anabolic genes such as Col I, Col II, and IGF1 was restored in defective CPCs. In conclusion, this study demonstrated that AFS-se has therapeutic effects on OA by inhibiting the inflammatory functions of IL-1β and TNF-α through protein phosphorylation in the MAPK pathway while also promoting the regenerative and self-repair functions of CPCs in traumatized cartilage.