Cytochrome c oxidase IV isoform 1 (COX4-1) regulates the proliferation, migration and invasion of trophoblast cells via modulating mitochondrial function.

INTRODUCTION: Spontaneous miscarriage is a common complication of early pregnancy. Previous studies have shown that mitochondrial function plays an important role in establishment of a successful pregnancy. Cytochrome c oxidase subunit 4 isoform 1 (COX4I1), a component of electron transport chain complex Ⅳ, is required for coupling the rate of ATP production to energetic requirements. However, there is very limited research on its role in trophoblast biology and how its dysfunction may contribute to spontaneous miscarriage. METHODS: Placental villi (7-10 weeks gestational age) collected from either induced termination of pregnancy or after spontaneous miscarriage were examined for expression of COX4I1. COX4I1 was knocked down by siRNA transfection of primary isolates of EVT cells. Real-time cell analysis (RTCA) and 5-Ethynyl-2'-deoxyuridine (EdU) were used to detect changes in proliferation ability after COX4I1 knockdown of EVT cells. Migration and invasion indices were determined by RTCA. Mitochondrial morphology was observed via MitoTracker staining. Oxidative phosphorylation, ATP production, and glycolysis in COX4I1-deficient cells and controls were assessed by a cellular energy metabolism analyzer (Seahorse). RESULTS: In placental villous tissue, COX4I1 expression was significantly decreased in the spontaneous miscarriage group. Knockdown of COX4I1 inhibited EVT cell proliferation, increased the migration and invasion ability and mitochondrial fusion of EVT cells. Mitochondrial respiration and glycolysis were impaired in COX4I1-deficient EVT cells. Knockdown of MMP1 could rescue the increased migration and invasion induced by COX4I1 silencing. DISCUSSION: Low expression of COX4I1 leads to mitochondrial dysfunction in EVT, resulting in altered trophoblast function, and ultimately to pregnancy loss.

Trophoblastic mitochondrial DNA induces endothelial dysfunction and NLRP3 inflammasome activation: Implications for preeclampsia.

AIMS: Preeclampsia (PE) is characterised by systemic vascular endothelium dysfunction. Circulating trophoblastic secretions contribute to endothelial dysfunction, resulting in PE; however, the underlying mechanisms remain unclear. Herein, we aimed to determine the potential correlation between the release of trophoblastic mitochondrial deoxyribonucleic acid (DNA) (mtDNA) and endothelium damage in PE. MATERIALS AND METHODS: Umbilical cord sera and tissues from patients with PE were investigated for inflammasome activation. Following this, trophoblastic mitochondria were isolated from HTR-8/SVneo trophoblasts under 21 % oxygen (O2) or hypoxic conditions (1 % O2 for 48 h) for subsequent treatments. Primary human umbilical veinendothelial cells (HUVECs) were isolated from the human umbilical cord and then exposed to a vehicle (phosphate-buffered saline [PBS]), mtDNA, hypo-mtDNA, or hypo-mtDNA with INF39 (nucleotide oligomerisation domain-like receptor family pyrin domain containing 3 [NLRP3]-specific inhibitor) for 12 h before flow cytometry and immunoblotting. The effects of trophoblastic mtDNA on the endothelium were further analysed in vivo using enzyme-linked immunosorbent assay (ELISA) and vascular reactivity assay. The effects of mtDNA on vascular phenotypes were also tested on NLRP3 knockout mice. RESULTS: Elevated interleukin (IL)-1ß in PE sera was accompanied by NLRP3 inflammasome activation in cord tissues. In vitro and in vivo experiments revealed that the release of trophoblastic mtDNA could damage the endothelium via NLRP3 activation, resulting in the overexpression of NLRP3, caspase-1 p20, IL-1ß p17, and gasdermin D (GSDMD); reduced endothelial nitric oxide synthase (eNOS) levels; and impaired vascular relaxation. Flow cytometric analysis confirmed that extensive cell death was induced by mtDNA, and simultaneously, a more pronounced pro-apoptotic effect was caused by hypoxia-treated trophoblastic mtDNA. The NLRP3 knockout or pharmacologic NLRP3 inhibition partially reversed tumour necrosis factor-α (TNF-α) and IL-1ß levels and endothelium-dependent vasodilation in mice. CONCLUSION: These findings demonstrate that trophoblastic mtDNA induced NLRP3/caspase-1/IL-1ß signalling activation, eNOS-related endothelial injury, and vasodilation dysfunction in PE.

[Characteristics of soil anti-scouribility in gully head wall of grass-covering on the gullied Loess Plateau, Northwest China]. / 黄土高塬沟壑区草地沟头立壁土壤抗冲性特征.

Vegetation restoration in the Chinese Loess Plateau has significantly changed soil erosion process of gully head wall. In order to investigate the characteristics and controlling factors of soil anti-scour properties of gully head covered by grasses, we carried out indoor undamaged soil trench scouring tests. By using barren gully head as the control, the physical and chemical properties and anti-scouring characteristics of soil in different soil layers (0-10, 10-20, 20-40, 40-60, 60-80, 80-100 cm) of the vertical wall of gully with grass cover were analyzed. The results showed that water-stable aggregate content and cohesion in barren and grass-covering gully head decreased with soil depth. Soil organic matter content and soil anti-scouribility coefficient in barren gully head decreased with soil depth, while the two indicators for gully head covered by grass increased firstly and then decreased with soil depth, with the maximum value (24.30 g·kg-1 and 58.86 L·g-1) in 10-20 cm soil layer. Meanwhile, the soil anti-scouring coefficient in each soil layer of grass-covering gully head was 1.7-9.3 times of that in soil layer of barren gully head. Soil organic matter content, water-stable aggregate content, cohesion and root length density all presented significantly positive correlation with soil anti-scouribility, among which soil organic matter content had the highest coefficient (r=0.98). Results of this study might provide basic data for the study of headcut erosion mechanism in Loess Plateau gully region, and scientific basis for effective control of soil and water loss in this region.

Interventions to reduce the cesarean delivery rate in a tertiary hospital in China.

INTRODUCTION: High cesarean delivery rate has been a global public health concern. This study assesses the effect of medical interventions and societal changes on cesarean delivery rates in a Chinese tertiary hospital. MATERIAL AND METHODS: A retrospective study including all live births ≥34-week gestation between 2008 and 2016 from Guangzhou Women and Children's Medical Center was divided into 5 stages: (1) no interventions; (2) patient-controlled epidural analgesia; (3) episiotomy restriction; (4) new labor management; (5) universal two-child policy. An interrupted time series design was used to measure the effect of interventions on overall cesarean rate, primary cesarean rate, maternal and neonatal outcomes. RESULTS: There were 126,609 deliveries including 49,092 cesarean deliveries and 77,517 vaginal deliveries in this period. Overall cesarean delivery rate declined after implementing patient-controlled epidural analgesia, episiotomy restriction and universal two-child policy. Primary cesarean rate decreased after implementing episiotomy restriction. Cesarean rate with previous cesarean dramatically increased, and maternal request cesarean rate decreased gradually. Low Apgar rate (score ≤7 at 5 min) increased after episiotomy restriction and maternal postpartum hemorrhage rate increased after new labor management. CONCLUSIONS: Patient-controlled epidural analgesia, episiotomy restriction and the universal two-child policy showed the most significant effects to reducing the cesarean rate.

Comparison between periapical radiography and cone beam computed tomography for the diagnosis of anterior maxillary trauma in children and adolescents.

BACKGROUND/AIMS: Under-estimating the damage caused by trauma to the dental structures may delay treatment. Timely and accurate diagnosis remains challenging in clinical practice. Radiography is an important modality for the diagnosis of traumatic injuries. The aim of this study was to compare the efficacy of periapical radiography and cone beam computed tomography for the diagnosis of trauma to the anterior maxillary dentoalveolar region in children and adolescents. MATERIAL AND METHODS: Images of patients who underwent both periapical radiography and cone beam computed tomography simultaneously because of trauma to the anterior maxillary region between January 2016 and January 2020 were analyzed retrospectively. Pairwise comparison between the receiver operating characteristic curves was performed to statistically compare the two methods for the diagnosis of crown fractures, root fractures, alveolar bone fractures and luxations, tooth resorption, and periapical radiolucencies. RESULTS: A total of 190 patients met the inclusion criteria. There were 120 (63.2%) males and 70 (36.8%) females, with a mean age of 11.1 years (range: 6-17 years). A crown fracture was observed in 144 teeth, while a root fracture was observed in 71 teeth. Alveolar fracture and luxation were observed in 44 incisors. During follow-up, tooth resorption and periapical radiolucencies were observed in 25 and 33 teeth, respectively. Pairwise receiver operating characteristic curve analysis revealed that cone beam computed tomography was significantly superior to periapical radiography for the diagnosis of root fractures, alveolar fractures and luxations, and tooth resorption (p < .05). However, no significant differences were found for the diagnosis of crown fractures and periapical radiolucencies (p > .05). CONCLUSIONS: Cone beam computed tomography in the low-dose mode was better for diagnosing root and bone fractures and resorption, but no different to periapical radiographs for crown fractures and periapical radiolucencies in pediatric patients.

Clinical effectiveness of orthodontic miniscrew implantation guided by a novel cone beam CT image-based computer aided design and computer aided manufacturing (CAD-CAM) template.

BACKGROUND: Although miniscrews are widely used in orthodontic treatment as temporary anchorage devices, their correct and safe placement has attracted little attention. This study aimed to introduce a novel cone beam CT (CBCT) image-based computer aided design and computer aided manufacturing (CAD-CAM) template for orthodontic miniscrew implantation and to evaluate the effectiveness of miniscrews implanted under the guidance of this template. METHODS: The CBCT scans of ten patients requiring miniscrews as anchorage were analyzed in NNT software to predetermine the insertion sites of miniscrew implants. The DICOM data of the scans, along with virtual miniscrews acquired in Solidworks software, were imported into Mimics software to construct three-dimensional (3D) images of teeth and bone and to determine the virtual position of miniscrews and 3D virtual templates were designed following consideration of the virtual implantation plans. A STL (Stereolithography) file of the virtual template was output, and the resin template was then fabricated with a stereolithographic appliance (SLA). 24 Miniscrews were then implanted guided by the template and clinical evaluation of their safety and stability, as well as their placement deviations, were made. A dental casts model and cephalometric analysis before and after orthodontic treatment were made to assess the dentomaxillofacial changes. RESULTS: All 24 miniscrews had no contact with adjacent roots. 18 miniscrews had a grade I safety score and six had a grade II. The miniscrews were stable at 1, 3, 6, and 9 months after implantation, although there was mild inflammation around two miniscrews. Implantation deviation of miniscrew in the crown was (1.03±0.65) and (1.26±0.72) mm in the apex, on average. Satisfactory dentomaxillofacial changes in 10 patients with these 24 miniscrews as anchorage were acquired. CONCLUSIONS: Miniscrews could be implanted in the targeted position safely and precisely when guided by the novel templates, and remained stable during orthodontic treatment. Patients treated with these miniscrews as anchorage in orthodontic treatment acquired satisfactory dentomaxillofacial changes.

Persistent hypoxia induced autophagy leading to invasiveness of trophoblasts in placenta accreta.

BACKGROUND: Excessive trophoblasts erosivity is the main pathological manifestation in placenta accreta. Similar to early pregnancy, trophoblasts of placenta accreta might have a similar anoxic state in abnormal continuous invasion, in which autophagy may also have some changes causing invasive ability in accreta. METHODS: Ten accreta placentas (placenta accreta group), as well as 10 non-accreta placentas (control group), were collected according to accreta criteria. The expression of hypoxia-induced autophage factors (HIF1α, Beclin 1, LC3B, and P62) and invasion-related markers (E-cadherin and MMP-9) were detected using immunohistochemical method. Comparison in scores grade was made between the two groups by Fisher's exact test and Spearman's test was used for correlation analysis. RESULTS: HIF1α was mainly expressed in cytomembrane of trophoblasts, in which moderate positive 50% (5/10) and strong positive 50% (5/10) in placenta accreta group compared to 50% (5/10) or weak positive 30% (3/10) in control group, with a significant statistically difference (p < .05). The negative, weak positive, moderate positive, and strong positive rates of Beclin-1 expression were 0, 10, 30, and 60% versus 60, 40, 0, and 0% in placenta accreta group and control group, respectively, statistically different (p < .05). The expression of LC3B was also statistically significant (0, 10, 20, 70% versus 50, 20, 30, 0%) between two groups, and P62 expression was also statistically different between two groups. The positive rates of E-cadherin expression were obviously negatively correlated with Beclin-1, LC3B, and P62 expression, while positive rates of MMP-9 expression were positively correlated with autophagy-associated markers. CONCLUSIONS: Hypoxia state might be involved in the occurrence of placental accreta, and persistent hypoxia state induced autophage disorders could cause down-regulated E-cadherin and down-regulated MMP-9, thus leading to more invasiveness of placenta trophoblasts.

Psychological impact in non-infectious disease specialists who had direct contact with patients with COVID-19.

BACKGROUND: The coronavirus disease 2019 (COVID-19) outbreak has become a pandemic. Obstetricians and midwives, among other medical staff, are tackling COVID-19 and are under immense psychological stress. AIMS: We aimed to survey the mental health of non-infectious disease specialist staff, specifically obstetricians and midwives, working in officially designated hospitals treating patients with COVID-19. METHOD: A nationwide online survey was conducted from 7 March to 17 March 2020 investigating the mental health of obstetricians and midwives (who were not themselves infected with COVID-19) working in hospitals treating patients with COVID-19. We used the 9-item Patient Health Questionnaire (PHQ-9), the 7-item Generalized Anxiety Disorder (GAD-7) scale and the 7-item Insomnia Severity Index (ISI) to assess their symptoms of depression, anxiety and insomnia. RESULTS: A total of 885 (41.6%), 609 (28.6%) and 729 (34.3%) obstetricians and midwives reported depression (PHQ-9 ≥ 5), anxiety (GAD-7 ≥ 5) and insomnia (ISI ≥ 8), respectively, during the COVID-19 pandemic. Regardless of whether or not they had direct contact with patients with COVID-19, obstetricians and midwives were more likely to report mild and moderate depression and anxiety during the COVID-19 pandemic when compared with before the pandemic. Those who had direct contact with patients with COVID-19 were more likely to report depression and insomnia than those who did not. Those who had sufficient protective equipment or training were less likely to report depression, anxiety and insomnia than those who did not. CONCLUSIONS: Our data suggest that non-infectious disease specialist staff have experienced varying, but increased levels of depression, anxiety and insomnia during this COVID-19 pandemic, which could be reduced by sufficient levels of protective equipment and occupational COVID-19 workplace training.

AQP8 and AQP9 expression in patients with polycystic ovary syndrome and its association with in vitro fertilization-embryo transfer outcomes.

The aim of the present study was to investigate aquaporin (AQP)8 and AQP9 expression in patients with polycystic ovary syndrome (PCOS) and its association with in vitro fertilization-embryo transfer (IVF-ET) outcomes. A total of 45 patients with PCOS undergoing IVF-ET (test group) and 50 patients with oviduct obstruction or ovarian cyst (control group) were assessed for the mRNA expression of AQP8 and AQP9 in ovarian tissues by reverse transcription-quantitative (RT-q)PCR. The levels of luteinizing hormone, anti-mullerian hormone and testosterone were determined, which were revealed to be significantly different between the two groups (P<0.05). The RT-qPCR results indicated that AQP8 expression in the control group was lower than that in the test group (t=37.75, P<0.01), whereas AQP9 expression in the control group was higher than that in the test group (t=19.59, P<0.01). The number of eggs obtained in the group with high AQP8 expression was significantly lower than that in the group with low AQP8 expression (t=2.64, P<0.01). The number of high-quality embryos in the high AQP8 expression group was not significantly different from that in the low AQP8 expression group (t=1.02, P>0.05). The pregnancy rate in patients with high AQP9 expression was higher than that in the low AQP9 expression group (P<0.05) and the abortion rate in the former was lower than that in the latter (P<0.05). In conclusion, AQP8 and AQP9 are differentially expressed in ovarian tissues of patients with PCOS vs. normal control subjects. The expression of AQP8 is closely associated with the occurrence and development of oocytes, whereas the expression of AQP9 is associated with the success rate of pregnancy in patients with PCOS.

Changes in the Expression of AQP4 and AQP9 in the Hippocampus Following Eclampsia-Like Seizure.

Eclampsia is a hypertensive disorder of pregnancy that is defined by the new onset of grand mal seizures on the basis of pre-eclampsia. Until now, the mechanisms underlying eclampsia were poorly understood. Brain edema is considered a leading cause of eclamptic seizures; aquaporins (AQP4 and AQP9), the glial water channel proteins mainly expressed in the nervous system, play an important role in brain edema. We studied AQP4 and AQP9 expression in the hippocampus of pre-eclamptic and eclamptic rats in order to explore the molecular mechanisms involved in brain edema. Using our previous animal models, we found several neuronal deaths in the hippocampal CA1 and CA3 regions after pre-eclampsia and that eclampsia induced more neuronal deaths in both areas by Nissl staining. In the current study, RT-PCR and Western blotting data showed significant upregulation of AQP4 and AQP9 mRNA and protein levels after eclamptic seizures in comparison to pre-eclampsia and at the same time AQP4 and AQP9 immunoreactivity also increased after eclampsia. These findings showed that eclamptic seizures induced cell death and that upregulation of AQP4 and AQP9 may play an important role in this pathophysiological process.

Osmotic water permeability diversification in primary trophoblast cultures from aquaporin 1-deficient pregnant mice.

AIM: Aquaporins (AQP) are water channel proteins, and some play an important role in maternal-fetal fluid exchange. The present study aimed to measure the osmotic water permeability in primary cultures of trophoblast cells from AQP1-deficient (AQP1(-/-) ) pregnant mice and to define the quantitative role of AQP1 in water transport across the trophoblast plasma membrane. MATERIAL AND METHODS: Trophoblast cells were obtained from placental tissue cell culture of AQP1(-/-) pregnant mice and were characterized by cytokeratin 7 immunostaining. The expression of the AQP1 gene in trophoblast cells of wild-type (AQP1(+/+) ) mice was confirmed by immunofluorescence. The osmotic water permeability of trophoblast plasma membranes was measured by a calcein fluorescence quenching method in response to osmotic gradients. RESULTS: A primary cell culture system for trophoblasts was successfully established. Immunofluorescence showed the expression of AQP1 in the trophoblast cell membrane of AQP1(+/+) mice. The osmotic water permeability of AQP1(-/-) trophoblast cells was significantly lower than that in AQP1(+/+) trophoblast cells, in response to both hypotonic and hypertonic challenges. CONCLUSION: The results suggest an important role of AQP1-mediated plasma membrane water permeability in maternal-fetal fluid balance and also provide a potential direction for the identification of therapeutic targets for the treatment of abnormalities in amniotic fluid volume.

Maternal-fetal fluid balance and aquaporins: from molecule to physiology.

Maternal-fetal fluid balance is critical during pregnancy, and amniotic fluid is essential for fetal growth and development. The placenta plays a key role in a successful pregnancy as the interface between the mother and her fetus. Aquaporins (AQPs) form specific water channels that allow the rapid transcellular movement of water in response to osmotic/hydrostatic pressure gradients. AQPs expression in the placenta and fetal membranes may play important roles in the maternal-fetal fluid balance.

Pregnant phenotype in aquaporin 8-deficient mice.

AIM: Aquaporin 8 (AQP8) is expressed within the female reproductive system but its physiological function reminds to be elucidated. This study investigates the role of AQP8 during pregnancy using AQP8-knockout (AQP8-KO) mice. METHODS: Homozygous AQP8-KO mice were mated, and the conception rate was recorded. AQP8-KO pregnant mice or their offspring were divided into 5 subgroups according to fetal gestational day (7, 13, 16, 18 GD) and newborn. Wild type C57 pregnant mice served as the control group. The number of pregnant mice, total embryos and atrophic embryos, as well as fetal weight, placental weight and placental area were recorded for each subgroup. The amount of amniotic fluid in each sac at 13, 16, and 18 GD was calculated. Statistical significance was determined by analysis of variance of factorial design and chi-square tests. RESULTS: Conception rates did not differ significantly between AQP8-KO and wild type mice. AQP8-KO pregnant mice had a significantly higher number of embryos compared to wild type controls. Fetal/neonatal weight was also significantly greater in the AQP8-KO group compared to age-matched wild type controls. The amount of amniotic fluid was greater in AQP8-KO pregnant mice than wild type controls, although the FM/AFA (fetal weight/amniotic fluid amount) did not differ. While AQP8-KO placental weight was significantly larger than wild type controls, there was no evidence of placental pathology in either group. CONCLUSION: The results suggest that AQP8 deficiency plays an important role in pregnancy outcome.