Existence of SARS-Cov-2 in the Peritoneal Fluid / Existência de SARS-Cov-2 no líquido peritoneal

Abstract Objective To determine the existence of SARS-CoV-2 in the peritoneal fluid to assess the risk of exposure through surgical smoke and aerosolization threatening healthcare workers during abdominal surgery. Background SARS-CoV-2 is a respiratory virus and possible ways of viral transmission are respiratory droplets, close contact, and fecal-oral route. Surgeries pose risk for healthcare workers due to the close contact with patients. Aerosolized particles may be inhaled via the leaked CO2 during laparoscopic procedures and surgical smoke produced by electrocautery. Methods All the data of 8 patients, who were tested positive for COVID-19, were collected between August 31, 2020 and April 30, 2021. Recorded clinicopathologic data included age, symptoms, radiological and laboratory findings, antiviral treatment before surgery, type of surgery and existence of the virus in the peritoneal fluid. Nasopharyngeal swab RT-PCR was used for the diagnosis. COVID-19 existence in the peritoneal fluid was determined by RT-PCR test as well. Results All 8 COVID-19 positive patients were pregnant, and surgeries were cesarean sections. 1 of the 8 patients was febrile during surgery. Also only 1 patient had pulmonary radiological findings specifically indicating COVID-19 infection. Laboratory findings were as follows: 4 of 8 had lymphopenia and all had elevated D-dimer levels. Peritoneal and amniotic fluid samples of all patients were negative for SARS-CoV-2. Conclusion SARS-CoV-2 exposure due to aerosolization or surgical fumes does not seem to be likely, provided the necessary precautions are taken.

Resumo Objetivo Determinar a existência de SARS-CoV-2 no fluido peritoneal para avaliar o risco de exposição através da fumaça cirúrgica e aerossolização que ameaçam os profissionais de saúde durante a cirurgia abdominal. Contexto O SARS-CoV-2 é um vírus respiratório e as possíveis formas de transmissão viral são gotículas respiratórias, contato próximo e rota fecal-oral. As cirurgias representam risco para os profissionais de saúde devido ao contato próximo com os pacientes. As partículas aerossolizadas podem ser inaladas através do CO2 vazado durante os procedimentos laparoscópicos e a fumaça cirúrgica produzida pela eletrocauterização. Métodos Todos os dados de 8 pacientes, que foram testados positivos para COVID-19, foram coletados entre 31 de agosto de 2020 e 30 de abril de 2021. Dados clinicopatológicos registrados incluíam idade, sintomas, achados radiológicos e laboratoriais, tratamento antiviral antes da cirurgia, tipo de cirurgia e existência do vírus no fluido peritoneal. O diagnóstico foi feito através do swab nasofaríngeo RT-PCR. A existência de COVID-19 no fluido peritoneal foi determinada pelo teste de RT-PCR também. Resultados Todas as 8 pacientes positivas para COVID-19 estavam grávidas, e as cirurgias eram cesarianas. 1 das 8 pacientes estava com febre durante a cirurgia. Também apenas 1 paciente tinha achados radiológicos pulmonares especificamente indicando infecção por COVID-19. Os achados laboratoriais foram os seguintes: 4 de 8 tinham linfopenia e todas apresentavam níveis elevados de D-dímero. Amostras de fluido peritoneal e líquido amniótico de todas as pacientes foram negativas para SARS-CoV-2. Conclusão A exposição ao SARS-CoV-2 devido à aerossolização ou fumaças cirúrgicas não parece ser provável, desde que sejam tomadas as precauções necessárias.

Whole exome sequencing analysis and prenatal diagnosis in children with neurodevelopmental disorders / 中华预防医学杂志

To explore the application value of whole exome sequencing (WES) in the diagnosis of prenatal and postnatal neurodevelopmental disorders (NDDs). A total of 70 patients diagnosed with NDDs who underwent WES at the Medical Genetics Center of the Maternal and Child Health Hospital of Hubei Province between June 2020 and July 2021 were retrospectively analyzed. Genomic DNA was extracted from peripheral blood samples and amniotic fluid. WES-based copy number variant (CNV) analysis was integrated into the routine WES data analysis pipeline. The results showed that a molecular diagnosis rate could be made in 21/70 (30%) cases. Of 21 positive cases, 14 (23%) cases were detected by single-nucleotide variant/small insertion/deletion (SNV/Indel) analysis, of which 12 variants were novel, 6 (9.8%) cases were detected by WES-based CNV analysis, and 1 (1.6%) case was detected by a combination of both. The diagnostic yield of WES combined with CNV analysis was higher than that of SNV/Indel analysis alone (30%, 21/70 vs. 20%, 14/70). Of the 28 prenatally diagnosed cases, 6 cases were found to have inherited parental variation for NDDs, 10 cases were found not to have the same pathogenic variation as the proband, and the remaining 12 cases were found to have no pathogenic or likely pathogenic variation that could explain the NDDs phenotype. Clinical follow-up showed that 5 families opted for abortion and the remaining had no current abnormalities. In conclusion, WES may be an effective method to clarify the genetic etiology and prenatal diagnosis of NDDs, which is helpful in assessing the prognosis to aid clinical management and reproductive guidance.

The evolution of approach in the resuscitation of neonates born with meconium-stained amniotic fluid: a tale of two countries, China and U.S.A., in the past 60 years / 中国当代儿科杂志

Neonates born through meconium-stained amniotic fluid (MSAF) may develop complications including meconium aspiration syndrome, persistent pulmonary hypertension of newborn and death. The approach to the resuscitation of these neonates has significantly evolved for the past few decades. Initially, under direct visualization technique, neonates with MSAF were commonly suctioned below the vocal cords soon after delivery. Since 2015, Neonatal Resuscitation Program (NRP®) of the American Academy of Pediatrics has recommended against "routine" endotracheal suctioning of non-vigorous neonates with MSAF but favored immediate resuscitation with positive pressure ventilation via face-mask bagging. However, the China neonatal resuscitation 2021 guidelines continue to recommend routine endotracheal suctioning of non-vigorous neonates born with MSAF at birth. This review article discusses the differences and the rationales in the approach in the resuscitation of neonates with MSAF between Chinese and American NRP® guidelines over the past 60 years.

Prenatal diagnosis for a fetus with 5p deletion syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a fetus with club foot detected upon mid-pregnancy ultrasonography.@*METHODS@#Amniotic fluid of the fetus and peripheral blood samples of its parents were collected and subjected to G-banding karyotype analysis and copy number variation sequencing (CNV-seq). The result was verified by fluorescence in situ hybridization (FISH).@*RESULTS@#The fetus and its parents all had a normal karyotype. CNV-seq analysis revealed that the fetus has harbored a 23.12 Mb on chromosome 5 and a 21.46 Mb duplication on chromosome 7. FISH assay has verified that its mother has carried a cryptic t(5;7)(p14.3;q33) translocation.@*CONCLUSION@#CNV-seq combined with FISH can effectively detect cryptic chromosome aberrations, and can help to reduce severe birth defects and provide a basis for prenatal genetic counseling.

Amniotic Sludge and Prematurity: Systematic Review and Meta-analysis / Sludge amniótico e prematuridade: revisão sistemática e metanálise

Abstract Objective To perform a systematic review and meta-analysis of studies on maternal, fetal, and neonatal outcomes of women with singleton pregnancies, after spontaneous conception, and with the diagnosis of amniotic sludge before 37 weeks of gestational age. Data Sources We conducted a search on the PubMed, Cochrane, Bireme, and Theses databases until June 2022. Selection of Studies Using the keywords intra-amniotic sludge or fluid sludge or echogenic particles, we found 263 articles, 132 of which were duplicates, and 70 were discarded because they did not meet the inclusion criteria. Data Collection The articles retrieved were analyzed by 2 reviewers; 61 were selected for full-text analysis, 18 were included for a qualitative analysis, and 14, for a quantitative analysis. Data Synthesis Among the maternal outcomes analyzed, there was an increased risk of preterm labor (95% confidence interval [95%CI]: 1.45-2.03), premature rupture of ovular membranes (95%CI: 1.99-3.79), and clinical (95%CI: 1.41-6.19) and histological chorioamnionitis (95%CI: 1.75-3.12). Regarding the fetal outcomes, there was a significant increase in the risk of morbidity (95%CI: 1.80-3.17), mortality (95%CI: 1.14-18.57), admission to the Neonatal Intensive Care Unit (NICU; 95%CI: 1.17-1.95), and neonatal sepsis (95%CI: 2.29-7.55). Conclusion The results of the present study indicate that the presence of amniotic sludge is a risk marker for preterm delivery. Despite the heterogeneity of the studies analyzed, even in patients with other risk factors for prematurity, such as short cervix and previous preterm delivery, the presence of amniotic sludge increases the risk of premature labor. Moreover, antibiotic therapy seems to be a treatment for amniotic sludge, and it may prolong pregnancy.

Resumo Objetivo Realizar revisão sistemática e metanálise de estudos que avaliaram os desfechos maternos, fetais e neonatais em gestantes de gravidez única, após concepção espontânea, e com o diagnóstico de sludge amniótico antes de 37 semanas de idade gestacional. Fontes dos dados Realizou-se uma pesquisa nas bases de dados PubMed, Cochrane, Bireme e Teses até junho de 2022. Seleção dos estudos Usando as palavras-chave intra-amniotic sludge ou fluid sludge ou echogenic particles, foram encontrados 263 artigos, 132 dos quais eram duplicatas, e 70 foram descartados por não corresponderem aos critérios de inclusão. Coleta de dados Os artigos encontrados foram analisados por 2 revisores; 61 foram selecionados para análise de texto completo, 18 foram incluídos em uma análise qualitativa e 14, em uma análise quantitativa. Síntese dos dados Entre os desfechos maternos analisados, houve aumento do risco de trabalho de parto prematuro (intervalo de confiança de 95% [IC95%]: 1.45-2.03), rotura prematura de membranas ovulares (IC95%: 1.99-3.79), e corioamnionite clínica (IC95%: 1.41-6.19) e histológica (IC95%: 1.75-3.12). Em relação aos desfechos fetais, houve aumento significativo do risco de morbidade (IC95%: 1.80-3.17), mortalidade (IC95%: 1.14-18.57), admissão em Unidade de Tratamento Intensivo (UTI) neonatal (IC95%: 1.17-1.95) e sepse neonatal (IC95%: 2.29-7.55). Conclusão Os resultados do presente estudo indicam que a presença de sludge amniótico é um marcador de risco para parto prematuro. Apesar da heterogeneidade dos estudos analisados, até mesmo em pacientes com outros fatores de risco para prematuridade, como colo curto e trabalho de parto prematuro anterior, a presença de sludge amniótico aumenta o risco de trabalho de parto prematuro na gestação. Além do mais, a antibioticoterapia parece ser um tratamento para o sludge amniótico, e pode ser capaz de prolongar a gravidez.

Corioamnionitis Histológica Asociada a Muerte Fetal / Histological Chorioamnionitis Associated with Fetal Death

INTRODUCCIÓN. Anualmente ocurren más de 2 millones de muertes fetales a nivel mundial, siendo fundamental el estudio anatomopatológico placentario para disminuir el número de muertes inexplicadas. OBJETIVO. Revisar la literatura existente acerca de corioamnionitis histológica, los criterios para establecer su diagnóstico, su presencia y posible asociación en estudios de causas de muerte fetal. METODOLOGÍA. Se realizaron búsquedas en bases de datos electrónicas para recopilar estudios de causas de muerte fetal que incluyeron corioamnionitis histológica. RESULTADOS. Se encontraron 13 estudios que evaluaron mortalidad fetal y que entre sus causas incluyeron corioamnionitis histológica. DESARROLLO. El estudio microscópico placentario en muertes fetales es esencial al investigar una muerte fetal. Las anomalías placentarias son la causa más común de muerte fetal, la corioamnionitis aguda es la lesión inflamatoria más frecuente. Se detallaron los criterios más relevantes para definir corioamnionitis aguda histológica pero aún no se establece un consenso. Estudios de causas de muerte fetal en años recientes han reportado corioamnionitis histológica entre 6,3% y 41,3% de casos. Las alteraciones inflamatorias del líquido amniótico son una causa importante de muerte fetal, siendo la corioamnionitis la más frecuente en este grupo. CONCLUSIÓN. En estudios para determinar las causas de muerte fetal se evidenció corioamnionitis aguda histológica en hasta el 41,3% de casos, por lo que podría estar asociada a dicho evento. Sin embargo, es necesario establecer un sistema de estadiaje de corioamnionitis histológica mediante un panel de expertos a nivel mundial.

INTRODUCTION. Annually more than 2 million fetal deaths occur worldwide, being fundamental the placental anatomopathological study to reduce the number of unexplained deaths. OBJECTIVE. To review the existing literature on histological chorioamnionitis, the criteria to establish its diagnosis, its presence and possible association in studies of causes of fetal death. METHODOLOGY. Electronic databases were searched to collect studies of causes of fetal death that included histologic chorioamnionitis. RESULTS. Thirteen studies were found that evaluated fetal mortality and that included histologic chorioamnionitis among their causes. DEVELOPMENT: Placental microscopic study in fetal deaths is essential when investigating a fetal death. Placental abnormalities are the most common cause of fetal death, acute chorioamnionitis being the most frequent inflammatory lesion. The most relevant criteria for defining histologic acute chorioamnionitis have been detailed but consensus has not yet been established. Studies of causes of fetal death in recent years have reported histologic chorioamnionitis in between 6,3% and 41,3% of cases. Inflammatory changes in the amniotic fluid are an important cause of fetal death, with chorioamnionitis being the most frequent in this group. CONCLUSIONS. In studies to determine the causes of fetal death, histological acute chorioamnionitis was evidenced in up to 41,3% of cases, so it could be associated with this event. However, it is necessary to establish a histological chorioamnionitis staging system by means of a worldwide panel of experts.

Parámetros del líquido amniótico para diagnóstico de respuesta inflamatoria en parto pretérmino / Amniotic fluid parameters for the diagnosis of inflammatory response in preterm delivery

OBJETIVO: Evaluar el rendimiento del Gram, la glucosa y los leucocitos en líquido amniótico para el diagnóstico de respuesta inflamatoria fetal y materna en pacientes con parto pretérmino. MÉTODO: Estudio de rendimiento de pruebas diagnósticas. Se incluyeron 63 pacientes a quienes se les realizó amniocentesis por sospecha de infección intraamniótica. Se estudió la placenta y se comparó con el Gram, la glucosa y el recuento de leucocitos en líquido amniótico para ver su relación con la respuesta inflamatoria. Se evaluaron la sensibilidad, la especificidad, las razones de verosimilitud (LR, likelihood ratio), los valores predictivos y el valor de kappa. RESULTADOS: Las pruebas con mejor rendimiento fueron en conjunto la glucosa 50/mm3 en líquido amniótico, con una especificidad del 94,3% (intervalo de confianza del 95% [IC95%]: 84,6-98,1), LR + 8,83 (IC95%: 2,5-31,2) y kappa de 0,48 (IC95%: 0,15-0,82). También se consideró la propuesta de un nuevo punto de corte para el recuento de leucocitos en líquido amniótico en la respuesta inflamatoria fetal. CONCLUSIONES: La combinación del recuento de leucocitos en líquido amniótico y los valores de glucosa mejora el rendimiento para el diagnóstico de respuesta inflamatoria fetal en comparación con la histopatología de la placenta, lo que proporciona información útil para el enfoque de los recién nacidos.

OBJECTIVE: To evaluate the performance of Gram, glucose and leukocytes in amniotic fluid for the diagnosis of fetal and maternal inflammatory response in patients with preterm delivery. METHOD: A diagnostic performance test study was carried out. Sixty-three patients with preterm labor were included who underwent amniocentesis due to suspected intra-amniotic infection. Histopathology of the placenta was studied and compared with the Gram result, glucose and leukocyte count in amniotic fluid, and their relationship with the maternal and fetal inflammatory response. Sensitivity, specificity, likelihood ratios, predictive values, and kappa were evaluated. RESULTS: The tests with the best performance were overall glucose 50/mm3 in amniotic fluid for the diagnosis of the fetal inflammatory response, with a specificity of 94.3% (95% confidence interval [95% CI]: 84.6-98.1%), likelihood positive ratio 8.83 (95% CI: 2.5-31.2) and kappa of 0.48 (95% CI: 0.15-0.82). A new cut-off point for leukocyte count in amniotic fluid to diagnose fetal inflammatory response was proposed. CONCLUSIONS: The combination of amniotic fluid leukocyte count and amniotic fluid glucose values improves performance for the diagnosis of inflammatory response compared with placental histopathology, providing useful information for newborns approach.

PCR em tempo real no diagnóstico da toxoplasmose congênita / Real-time PCR in the diagnosis of congenital toxoplasmosis

O diagnóstico da toxoplasmose congênita apresenta limitações sendo, portanto, necessárias novas opções de exames. A análise do líquido aminiótico pela PCR em tempo real já se mostrou eficaz para confirmação da infecção fetal. No entanto, o seu desempenho em outras amostras biológicas ainda não está claro. O objetivo deste estudo é avaliar a PCR em tempo real no sangue da mãe e do recém-nascido assim como no líquido amniótico e placenta, no diagnóstico da toxoplasmose congênita. Esse é um estudo descritivo de gestantes com toxoplasmose acompanhadas no Rio de Janeiro, Brasil. Foi realizada PCR em tempo real em amostras de sangue materno, líquido amniótico, placenta e sangue dos recém-nascidos e o exame histopatológico das placentas. Também foram coletados dados clínicos e laboratoriais dos recém-nascidos. Foram acompanhadas 116 gestantes e analisadas 298 amostras. Uma (0,9%) gestante apresentou PCR positiva no sangue, três (3,5%) no líquido amniótico, uma (2,3%) na placenta e nenhum recém-nascido apresentou PCR positiva no sangue. O estudo histopatológico foi sugestivo de infecção por toxoplasmose em 24 (49%) placentas. Seis (5,2%) recém-nascidos foram diagnosticados com toxoplasmose congênita e apenas os casos com PCR positiva no líquido amniótico tinham associação do resultado da PCR com o diagnóstico de infecção congênita. Tanto as amostras de sangue materno quanto as de sangue dos recém-nascidos e placenta, não demonstraram ser promissoras no diagnóstico da toxoplasmose congênita. Novos estudos são necessários para avaliar o real papel do diagnóstico molecular em outros materiais biológicos que não o líquido amniótico.

The diagnosis of congenital toxoplasmosis has limitations so new options are needed. Real-time PCR analysis of amniotic fluid has proven effective for confirming fetal infection. However, its performance in other biological samples still needs to be determined. This study aims to evaluate the real-time PCR role in the blood of the mother and newborn as well as in the amniotic fluid and placenta, in congenital toxoplasmosis diagnosis. It is a descriptive study of pregnant women with toxoplasmosis followed in Rio de Janeiro, Brazil. Real-time PCR was performed on maternal blood, amniotic fluid, placenta, and newborn blood samples. In addition, a histopathological examination of the placentas was performed and data from the babies were collected. One hundred and sixteen pregnant women were followed and 298 samples were analyzed. One (0.9%) pregnant woman had positive PCR in the blood, three (3.5%) in the amniotic fluid, one (2.3%) in the placenta, and any newborn had positive PCR in the blood. The histopathological study suggested toxoplasmosis infection in 24 (49%) placentas. Six (5.2%) newborns were diagnosed with congenital toxoplasmosis and only the cases with positive PCR in amniotic fluid associated with the diagnosis of congenital infection. Neither maternal nor newborn blood and placenta samples have not shown promise in diagnosing congenital toxoplasmosis. Further studies are needed to evaluate the fundamental role of molecular diagnostics in others biological materials than amniotic fluid.

Clinical features of severe meconium aspiration syndrome (MAS) and early predicting factors for severe MAS in neonates with meconium-stained amniotic fluid / 中国当代儿科杂志

OBJECTIVES@#To study the clinical features of severe meconium aspiration syndrome (MAS) and early predicting factors for the development of severe MAS in neonates with meconium-stained amniotic fluid (MSAF).@*METHODS@#A total of 295 neonates who were hospitalized due to Ⅲ° MSAF from January 2018 to December 2019 were enrolled as subjects. The neonates were classified to a non-MAS group (n=199), a mild/moderate MAS group (n=77), and a severe MAS group (n=19). A retrospective analysis was performed for general clinical data, blood gas parameters, infection indicators, and perinatal clinical data of the mother. The respiratory support regimens after birth were compared among the three groups. The receiver operating characteristic (ROC) curve and multivariate logistic regression analysis were used to investigate predicting factors for the development of severe MAS in neonates with MSAF.@*RESULTS@#Among the 295 neonates with MSAF, 32.5% (96/295) experienced MAS, among whom 20% (19/96) had severe MAS. Compared with the mild/moderate MAS group and the non-MAS group, the severe MAS group had a significantly lower 5-minute Apgar score (P<0.05) and a significantly higher blood lactate level in the umbilical artery (P<0.05). Compared with the non-MAS group, the severe MAS group had a significantly higher level of interleukin-6 (IL-6) in peripheral blood at 1 hour after birth (P<0.017). In the severe MAS group, 79% (15/19) of the neonates were born inactive, among whom 13 underwent meconium suctioning, and 100% of the neonates started to receive mechanical ventilation within 24 hours. Peripheral blood IL-6 >39.02 pg/mL and white blood cell count (WBC) >30.345×109/L at 1 hour after birth were early predicting indicators for severe MAS in neonates with MSAF (P<0.05).@*CONCLUSIONS@#Meconium suctioning cannot completely prevent the onset of severe MAS in neonates with MSAF. The neonates with severe MAS may develop severe respiratory distress and require mechanical ventilation early after birth. Close monitoring of blood lactate in the umbilical artery and peripheral blood IL-6 and WBC at 1 hour after birth may help with early prediction of the development and severity of MAS.

A retrospective cohort study of tracheal intubation for meconium suction in nonvigorous neonates / 中国当代儿科杂志

OBJECTIVES@#To study the feasibility of tracheal intubation for meconium suction immediately after birth of nonvigorous neonates born through meconium-stained amniotic fluid (MSAF).@*METHODS@#A retrospective cohort study was performed on nonvigorous neonates born through MSAF who were admitted to the Department of Neonatology, Zhecheng People's Hospital. The neonates without meconium suction who were admitted from July 1, 2017 to June 30, 2018 were enrolled as the control group. The neonates who underwent meconium suction from July 1, 2018 to June 30, 2019 were enrolled as the suction group. The two groups were compared in terms of the mortality rate and the incidence rates of neonatal meconium aspiration syndrome (MAS), persistent pulmonary hypertension of the newborn, pneumothorax, and pulmonary hemorrhage.@*RESULTS@#There were 80 neonates in the control group and 71 in the suction group. There were no significant differences between the two groups in the incidence rates of MAS (11% vs 7%), persistent pulmonary hypertension of the newborn (5% vs 4%), pneumothorax (3% vs 1%), and death (0% vs 1%). Compared with the control group, the suction group had a significantly lower proportion of neonates requiring oxygen inhalation (16% vs 33%, P<0.05), noninvasive respiratory support (25% vs 41%, P<0.05) or mechanical ventilation (10% vs 23%, P<0.05) and significantly shorter duration of noninvasive ventilation [(58±24) hours vs (83±41) hours, P<0.05] and length of hospital stay [6(4, 8) days vs 7(5, 10) days, P<0.05].@*CONCLUSIONS@#Although tracheal intubation for meconium suction immediately after birth may shorten the duration of respiratory support for mild respiratory problems, it cannot reduce the incidence rate of MAS, mortality rate, or the incidence rate of serious complications in nonvigorous infants born through MSAF.

Enlarged multicystic dysplastic kidneys with oligohydramnios during infancy caused by NPHP3 gene mutation / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical features and genomic abnorm ality of a fetus enlarged multicystic dysplastic kidneys with oligohydramnios caused by NPHP3 gene mutation.@*METHODS@#The fetuse was found to have multicystic dysplastic kidneys with oligohydramnios upon ultrasonography during the second trimester. Following induced abortion, fetal tissue was collected for the extraction of DNA, chromosomal microarray analysis (CMA) and whole exome sequencing (WES). Sanger sequencing was used to verify the suspected variants in the family.@*RESULTS@#Antenatal ultrasound examination at 19 weeks showed "polycystic" kidneys with Oligohydramnios. Delivery was by induced labour because of the critically low amniotic fluid volume. Testing of CMA was normal. WES showed a compound heterozygous mutation of c.1817G>A, p.W606X; c.432dupA, p.E145Rfs*18 mutations are novel mutations in this study.@*CONCLUSION@#The research may further expand the NPHP3 gene mutation spectrum. Enlarged multicystic dysplastic kidneys with oligohydramnios caused by NPHP3 gene mutation at least include one or two splice site mutation, frameshift mutation or nonsense mutation foetal poor prognosis.

Application of genomic copy number variation detection technology in prenatal diagnosis of 7617 pregnant women with serological screening abnormalities during the second trimester of pregnancy / 中华医学遗传学杂志

OBJECTIVE@#To analyze the genomic variation characteristics of fetal with abnormal serological screening, and to further explore the value of copy number variation (CNV) detection technology in prenatal diagnosis of fetal with abnormal serological screening.@*METHODS@#7617 singleton pregnant women who underwent amniocentesis for prenatal diagnosis solely due to abnormal Down's serological screening were selected. According to the results of serological screening, the patients were divided into high risk group, borderline risk group and single abnormal multiple of median (MOM) group. CMA and CNV-Seq were used to detect the copy number variation of amniotic fluid cell genomic DNA and combined with amniotic fluid cell karyotype analysis for prenatal diagnosis. Outpatient revisit combined with telephone inquiry was used for postnatal follow-up.@*RESULTS@#Among 7617 amniotic fluid samples, aneuploidy was detected in 138cases (1.81%) by CMA and CNV-Seq, 9 cases of aneuploid chimerism were detected by amniotic fluid cell karyotype analysis, and 203 cases of fetus carrying pathogenic and likely pathogenic CNV (P/LP CNV) were detected, the variant of uncertain significance (VUS) was detected in 437 cases (5.7%), the overall abnormal detection rate was 10.33%. The detection rate of aneuploidy by CMA and CNV-Seq in three group were 123 cases (2.9%), 13 cases (1.3%) and 2 cases (0.4%), respectively,and showing no significant difference (χ 2=7.469, P=0.024). The detection rate of pathogenic and likely pathogenic CNV in three group were 163cases (2.6%); 24 cases (2.6%) and 16 cases (3.3%), respectively, and showing no significant difference (χ 2=0.764, P=0.682). The CMA reported 2.9% (108/3729)P/LP CNV, and CNV-seq reported 2.4% (95/3888)P/LP CNV, both tests showed similar detective capabilities (χ 2=1.504, P=0.22).The most popular P/LP CNV in this cohort were Xp22.31 microdeletion, 16p13.11 microduplication /microdeletion, 22q11.21 microduplication /microdeletion. In fetuses with P/LP CNV CNV, 59 fetuses were terminated pregnancy, and 32 of 112 fetuses born had abnormal clinical manifestations. Non-medically necessary termination of pregnancy occurred in 11 fetuses carrying VUS CNV, 322 fetuses carrying VUS CNV were born, 4 of them presented abnormal clinical manifestations.@*CONCLUSION@#Compared with the traditional chromosome karyotype, CMA and CNV-Seq can improve the detection rate of pathogenic and likely pathogenic CNV. CMA and CNV-seq can be used for first tier diagnosis of pregnant women in the general population with abnormal Down's serological screening.

Differential expression profile of miRNAs in amniotic fluid exosomes from fetuses with Down syndrome / 南方医科大学学报

OBJECTIVE@#To investigate the role of miRNAs in amniotic fluid exosomes in growth and development of fetuses with Down syndrome (DS).@*METHODS@#Amniotic fluid were collected from 20 fetuses with DS and 20 normal fetuses (control) to extract amniotic exosome miRNA. MicroRNA sequencing technique was used to identify the differentially expressed miRNAs between the two groups, for which gene ontology (GO) and pathway analysis was performed. Three differentially expressed miRNAs with the strongest correlation with DS phenotype were selected for qPCR verification. Dual luciferase reporter assay was used to verify the activity of let-7d-5p for targeted regulation of BACH1.@*RESULTS@#We identified 15 differentially expressed miRNAs in DS as compared with the control group, among which 7 miRNAs were up-regulated and 8 were down-regulated. Target gene prediction results showed that the differentially expressed miRNAs targeted 17 DS-related genes. GO analysis revealed that the main functions of the target genes involved protein binding, protein transport, ATP binding, transferase activity and synapses. Pathway analysis revealed that the functional pathways were closely related with the development of the nervous system. qPCR results showed that the expression levels of miR-140-3p and let-7d-5p were significantly lower in DS group than in the control group (P < 0.05), as was consistent with miRNA sequencing results; the expression level of miR-4512 was significantly higher in DS group than in control group (P < 0.05), which was contrary to miRNA sequencing results. The results of double luciferase reporter gene assay confirmed that let-7d-5p was capable of targeted regulation of BACH1 expression.@*CONCLUSION@#Let-7d-5p in amniotic fluid exosomes may promote oxidative stress events in the brain of fetuses with DS by regulating BACH1 expression.

Variedades de cultivo de líquido amniótico para el diagnóstico in vitro de poliploidías / Amniotic fluid culture varietys for poliploydies in vitro diagnosis

RESUMEN Fundamento: El cultivo celular permite el análisis directo de las células vivas mediante un microscopio. El estudio de las células contenidas en el líquido amniótico, mediante técnicas de cultivo, detecta anomalías en número y morfología de los cromosomas, que pueden relacionarse con enfermedades genéticas. Objetivo: Caracterizar las variedades de cultivo de líquido amniótico para el diagnóstico in vitro de poliploidías. Metodología: Se realizó un estudio descriptivo transversal, en el Centro Provincial de Genética Médica de Camagüey, en el periodo de noviembre de 2016 a abril de 2018.La población de estudio estuvo constituida por 1571 muestras útiles de líquido amniótico obtenidas por amniocentesis, en gestantes en el segundo trimestre, evaluadas en consulta multidisciplinaria con criterios clínicos de estudios cromosómicos según lo establecido en el diagnóstico prenatal citogenético, previo consentimiento informado. Se utilizaron 20 mL de líquido amniótico para la siembra de células fetales, y se aplicaron tres variantes de cultivo abierto (directo, centrifugado y expandido). Se determinó el complemento cromosómico en cada variedad. Resultados: Predominó el complemento cromosómico normal. Las tetraploidías prevalecieron en el cultivo expandido. El índice mitótico fue similar en las tres variedades de cultivo y el cultivo directo tuvo el más bajo índice de poliploidías. Conclusiones: El cariotipo normal fue predominante. Las tetrapolidías fueron las alteraciones más frecuentes y prevalecieron en el cultivo expandido. En el cultivo directo se presentó el más bajo índice de errores inducidos in vitro.

ABSTRACT Background: Cell culture allows direct analysis of live cells under a microscope. The cell study contained in amniotic fluid, by culture techniques, detects abnormalities in chromosome number and morphology, which can be related to genetic diseases. Objective: To describe amniotic fluid culture strains for the in vitro diagnosis of polyploidy. Methodology: A cross-sectional descriptive study was conducted at the Camagüey Provincial Center of Medical Genetics, from November 2016 to April 2018.The study population consisted of 1571 useful amniotic fluid samples obtained by amniocentesis, in pregnant women in the second trimester, evaluated by multidisciplinary discussion with clinical criteria for chromosomal studies as established in the cytogenetic prenatal diagnosis, prior informed consent. 20 mL of amniotic fluid were used for fetal cell seeding, and three open culture strains (direct, centrifuged and expanded) were applied. Chromosomal complement was determined in each variety. Results: Normal chromosome complement was predominant. Tetraploidy prevailed in the expanded culture. The mitotic index was similar in the three culture strains and the direct culture had the lowest polyploidy index. Conclusions: Normal karyotype was predominant. Tetraploidy were the most frequent modifications and prevailed in the expanded culture. Direct culture had the lowest rate of the in vitro induced errors.

Muerte fetal por infección bacteriana ascendente: método diagnóstico, una revisión narrativa. ¿Por qué el método que incluye estudio placentario, evaluación de datos clínicos y de laboratorio es eficiente en identificar la infección bacteriana ascendente como causa de muerte fetal? / Fetal death from ascending bacterial infection: diagnostic method, a narrative review. Why is the method that includes placental study, evaluation of clinical and laboratory data efficient in identifying ascending bacterial infection as the cause of fetal death?

Resumen Esta revisión narrativa incluye estudios publicados sobre métodos de clasificación de mortinatos y su eficiencia para identificar la infección bacteriana ascendente (IBA) como causa de muerte fetal (MF), mediante búsqueda en PubMed, Cochrane, Embase, ScienceDirect, Wiley Online Library, Scielo. Muchos niños mueren antes de nacer en todo el mundo y no se ha logrado reducir la MF porque los métodos empleados no han sido los adecuados y porque no se diagnostica la IBA, la causa más frecuente de MF en un hospital público de Chile. Los sistemas que utilizan los datos clínicos, de laboratorio y estudio placentarios, INCODE, CORM, son los más eficientes para identificar la IBA como origen de la MF. Se ha demostrado que los marcadores específicos de infección/inflamación placentaria, corioamnionitis histológica/funisitis aguda son de mayor eficiencia para diagnosticar la IBA que la autopsia fetal, que el cultivo de líquido amniótico es más eficiente que el cultivo de la placenta para detectar invasión microbiana de la cavidad amniótica y que la muestra de sangre de cordón es eficiente para el diagnóstico etiológico de la infección. El conocimiento de la IBA como causa inicial de MF, ayuda a elaborar guías y normas de prevención de la MF por esta condición.

Abstract This narrative review includes published studies of stillbirth classification methods and their efficiency in identifying ascending bacterial infection (ABI), as a cause of fetal death (FD), by searching PubMed, Cochrane, Embase, ScienceDirect, Wiley Online Library, Scielo. Many children die before birth around the world and it has not been possible to reduce FD because the methods used have not been adequate and because ABI, the most frequent cause of FD in a public hospital in Chile, is not diagnosed. Systems using clinical, laboratory and placental study data, INCODE, CORM, are more efficient in identifying ABI as the origin of FD. Specific markers of infection/placental inflammation, histologic chorioamnionitis/acute funitis have been shown to be more efficient in diagnosing ABI than fetal autopsy, that amniotic fluid culture is more efficient than placental culture for detect microbial invasion of the amniotic cavity and that the cord blood sample is efficient for the etiological diagnosis of the infection. The knowledge of the ABI as the initial cause of FD helps to develop guidelines and norms for preventing FD due to this condition.

Assessment of fetal lung maturity using analysis of amniotic fluid from ewes that gave birth prematurely and at term / Avaliação da maturidade pulmonar fetal utilizando a análise de líquido amniótico de ovelhas que deram à luz prematuramente e a termo

The aim of this study was to evaluate the lung maturity of premature and full-term lambs by analyzing amniotic fluid using the following methods: Clements test, Nile blue cytology test, hematoxylin-Shorr stain, lamellar body count, and radiographic tests. The use of these methods is intended to identify high-risk newborns and provide immediate clinical intervention after birth. Altogether, 56 animals (24 ewes and 32 lambs) were included in the study and divided into 3 groups. Group I consisted of 8 ewes that were at approximately 145 days of gestation; this group delivered 10 lambs naturally. Group II consisted of 8 ewes that were at 138 days' gestation; this group delivered 11 lambs by cesarean section. Group III consisted of 8 ewes at 138 days' gestation; this group was administered intramuscular dexamethasone (16mg/animal) 36 hours prior to a cesarean section. Group III delivered11 lambs. Cytological tests were performed using a microscope with a maximum magnification of 1000x, while the Clements test was visually observed by one of the researchers. Amnioticfluid lamellar body counts were measured using transmission electron microscopy. Among the staining methods, hematoxylin-Shorr was reliable, and Group III had a greater number of orangeophilic cells when compared to Group II, probably due to corticoid administration. The Clements test showed pulmonary maturity in approximately 20% of Group I lambs and Group II showed 9.1% of bubbles; however, Group III had the highest pulmonary maturity percentage (36.4%). The lamellar bodies were measured, and all groups had sizes between 0.019 and 0.590μm. Radiographic evaluation revealed that the majority of lambs presented some level of pulmonary radiodensity, indicating an acinar pattern at birth. These results are in line with the expectations of each group. We found that the normal group showed greater pulmonary maturity, whereas Group II presented pulmonary immaturity, which is expected because this group comprised lambs born prematurely and Group III showed pulmonary maturity almost comparable to the normal delivery group (Group I). This is due to the fact that although these animals are premature, the use of dexamethasone helped in pulmonary maturation. Therefore, these pulmonary maturity tests are considered effective when more than one technique is used and can be used routinely in the care of a pregnant ewe in labor, where a simple collection of amniotic fluid can predict a high-risk pregnancy and alert the veterinarian if the newborn needs intensive supportive treatment.(AU)

O objetivo deste estudo foi avaliar a maturidade pulmonar de cordeiros prematuros e a termo por meio da análise do líquido amniótico utilizando os seguintes métodos: teste de Clements, teste de citologia do azul do Nilo, coloração de hematoxilina-Shorr, contagem de corpos lamelares e testes radiográficos. Um desses métodos tem por objetivo identificar recém-nascidos de alto risco e fornecer intervenção clínica imediata após o nascimento. Ao todo, 56 animais (24 ovelhas e 32 cordeiros) foram incluídos no estudo e divididos em 3 grupos. O grupo I foi composto por 8 ovelhas com aproximadamente 145 dias de gestação; este grupo deu à luz 10 cordeiros naturalmente. O Grupo II foi composto por 8 ovelhas com 138 dias de gestação; este grupo deu à luz 11 cordeiros por cesariana. Grupo III consistiu de 8 ovelhas com 138 dias de gestação; este grupo recebeu dexametasona intramuscular (16 mg / animal) 36 horas antes de uma cesariana. O Grupo III entregou 11 cordeiros. Os testes citológicos foram realizados em microscópio com aumento máximo de 1000x, enquanto o teste de Clements foi observado visualmente por um dos pesquisadores. A contagem de corpos lamelares de líquido amniótico foi medida usando microscopia eletrônica de transmissão. Dentre os métodos de coloração, o hematoxilina-Shorr foi confiável, sendo que o Grupo III apresentou maior número de células orangeofílicas quando comparado ao grupo II, provavelmente devido à administração de corticóide. O teste de Clements mostrou maturidade pulmonar em aproximadamente 20% dos cordeiros do Grupo I e o Grupo II apresentou 9,1% de bolhas; entretanto, o Grupo III apresentou o maior percentual de maturidade pulmonar (36,4%). Os corpos lamelares foram medidos e todos os grupos apresentaram tamanhos entre 0,019 e 0,590μm. A avaliação radiográfica revelou que a maioria dos cordeiros apresentava algum grau de radiodensidade pulmonar, indicando padrão acinar ao nascimento. Esses resultados estão alinhados com as expectativas de cada grupo. Verificamos que o grupo normal apresentou maior maturidade pulmonar, enquanto o Grupo II apresentou imaturidade pulmonar, o que é esperado por se tratar de cordeiros nascidos prematuramente e o Grupo III apresentou maturidade pulmonar quase comparável ao grupo de parto normal (Grupo I). Isso se deve ao fato de que, embora esses animais sejam prematuros, o uso da dexametasona auxiliou na maturação pulmonar. Portanto, esses testes de maturidade pulmonar são considerados eficazes quando mais de uma técnica são utilizadas e podem ser usadas rotineiramente no cuidado de uma ovelha gestante em trabalho de parto, onde uma simples coleta de líquido amniótico pode prever uma gravidez de alto risco e alertar o veterinário se o recém-nascido precisa de tratamento de suporte intensivo.(AU)

Avaliação da maturidade pulmonar de cabritos nascidos a termo e prematuros / [Evaluation of lung maturity of term and premature kid]

Avaliou-se a maturidade pulmonar de cabritos no líquido amniótico de suas mães pela coloração de Shor, pelo azul de Nilo e pela contagem de corpos lamelares, bem como a vitalidade e os níveis de glicose e lactato séricos em cabritos nascidos a termo e prematuros. Para tanto, foram utilizados 32 cabritos, divididos em três grupos, a saber: grupo I: cabritos nascidos de cesarianas com 149 dias de gestação; grupo II: cabritos nascidos de cesarianas com 143 dias de gestação; e grupo III: cabritos nascidos de cesarianas com 143 dias de gestação, oriundos de mães que receberam, por via intramuscular, 20mg/cabra de dexametasona, 36 horas antes da cirurgia eletiva. A coloração de Shorr e a contagem de corpos lamelares demonstraram ser métodos diagnósticos promissores para a avaliação da maturidade pulmonar em neonatos caprinos. Contudo, a administração de dexametasona nas cabras no período antenatal não influenciou a maturidade fetal. Constatou-se, entretanto, que a avaliação física do paciente, logo após o nascimento, também se mostra fundamental no que tange à percepção da vitalidade e da viabilidade de cabritos recém-nascidos.(AU)

Pulmonary maturity of goats in their amniotic fluid was evaluated by Shor, Nile blue staining, and lamellar body count, as well as vitality and serum glucose and lactate levels in term and premature goats. Twenty-four kids were divided into three groups: group I: comprised of eight animals born by cesarean section with 149 days of gestation; group II: comprised of eight animals born by cesarean section with 143 days of gestation; and group III: comprised of eight animals born by cesarean section with 143 days of gestation, in which the does received intramuscular dexamethasone (20mg/goat) 36 hours prior to elective cesarean section. Shorr staining and lamellar body count have shown to be promising diagnostic methods for the assessment of lung maturity in goat neonates. However, the administration of dexamethasone to goats during antenatal period did not influence fetal maturity. It was verified that the physical evaluation of the patient, shortly after birth, is fundamental for the perception of vitality and viability of newborn goats.(AU)

Trisomía 9, trisomía 13 y trisomía 18: resultados del análisis citogenético prenatal, Hospital Clínico Universidad de Chile, años 2000-2017 / Trisomy 9, trisomy 13 and trisomy 18: results of the prenatal cytogenical analysis, Hospital Clínico Universidad de Chile, years 2000-2017

INTRODUCCIÓN: En Chile, la norma técnica de la Ley N° 21.030 de 2017 considera tres aneuploidías como letales; las trisomías 9, 13 y 18, cuyo diagnóstico se confirma con un cariograma. No existe a la fecha registro nacional de frecuencia prenatal de estas patologías. OBJETIVO: Determinar la frecuencia de trisomías 9, 13 y 18 en los estudios citogenéticos prenatales en muestras de células obtenidas con amniocentesis y cordocentesis, procesados en el Laboratorio de Citogenética del Hospital Clínico Universidad de Chile. MATERIALES Y MÉTODOS: Estudio descriptivo y retrospectivo de los resultados de cariograma de líquido amniótico (LA) y sangre fetal (SF), procesados desde enero de 2000 a diciembre de 2017. RESULTADOS: Se incluyeron 2.305 muestras (402 de SF y 1.903 de LA), de ellas 442 (19%) fueron trisomías letales (TL), dentro de ellas fueron TL libres 416 (95%), TL estructurales 15 (2,7%) y mosaicos 11 (2,3%). La trisomía 18 fue en ambos tipos de muestra la más frecuente (73,5%), seguida de trisomía 13 (24,2%) y trisomía 9 (2,3%). Se desglosan resultados conforme al tipo de TL, muestra, motivo de derivación, edad materna y edad gestacional. CONCLUSIONES: El cariograma confirma el diagnóstico de aneuploidías y aporta datos relevantes para el consejo genético. La cromosomopatía letal más frecuente fue la trisomía 18. Se observó que uno de cada cinco cariogramas referidos por anomalías congénitas y/o marcadores de aneuploidía revelaban una TL.

INTRODUCTION: In Chile, the technical standard of Law No. 21,030 of 2017 considers three aneuploidies as lethal; trisomies 9, 13 and 18, whose diagnosis is confirmed with a Karyotype. To date there is not a national registry of prenatal frequency of these pathologies. OBJECTIVE: To determine the frequency of trisomies 9, 13 and 18 in prenatal cytogenetic studies in samples of cells obtained with amniocentesis and cordocentesis, processed in the Cytogenetics Laboratory of the Universidad de Chile Clinical Hospital. MATERIALS AND METHODS: Descriptive and retrospective study of the results of karyotypes of amniotic fluid (LA) and fetal blood (SF) processed from January 2000 to December 2017. Results: 2,305 samples (402 of SF and 1,903 of LA) were included, of which 438 (19%) were lethal trisomies (TL), corresponding to free TL 416 (95%), structural TL 12 (2,7%) and mosaics 10 (2.3%). Trisomy 18 was the most frequent in both types of sample (73,5 %), followed by trisomy 13 (24,2%) and trisomy 9 (2.3%). RESULTS are shown according to the type of TL, sample, reason for referral, maternal age and gestational age. CONCLUSIONS: The karyotype confirms the diagnosis of aneuploidies and provides relevant data for genetic counseling. The most frequent lethal chromosomopathy was trisomy 18. It was observed that one in five karyotypes referred for congenital anomalies and / or aneuploidy markers revealed a TL.

Maternal and fetal parameters in pregnant woman undergoing tocolysis with nifedipine / Parâmetros maternos e fetais em gestante submetida a tocólise com nifedipina

Abstract Objectives: to evaluate the effects of nifedipine with tocolysis under maternal and fetal parameters. Methods: a cohort study with 40 pregnant women admitted at a high-risk pregnancy ward to inhibit premature labor between September/2010 to May/2012. Nifedipine was used as a 20mg sublingual attack dose and maintained 20mg every six and eight hours orally. The variables of the analysis were fetal heart rate (FHR), maternal heart rate (MHR), systolic blood pressure (SBP) and diastolic blood pressure (DBP), and amniotic fluid index (AFI). All the variables were evaluated prior to administrating nifedipine and approximately after 6 hours and every 24 hours, until hospital discharge. Results: there were no modification of the FHR (p=0.48) and the SBP (p=0.29). The MHR increased after 24 hours, but with no statistical difference (p=0.08), returning to similar levels as at admission within 48 hours. The DBP decreased at 6 (p=0.04) to 72 hours, being stable afterwards. The AFI decreased significantly at 24, 48 and 72 hours. Conclusions: the use of high doses of nifedipine with tocolysis causes a decrease of the maternal's diastolic blood pressure and consequently decreases the amniotic fluid index, but probably without any clinical repercussions.

Resumo Objetivos: avaliar os efeitos da nifedipina utilizada na tocólise sobre os parâmetros maternos e fetais. Métodos: estudo de coorte incluindo 40 gestantes admitidas na enfermaria de alto risco para inibição do trabalho de parto prematuro entre setembro/2010 a maio/2012. Utilizou-se a nifedipina sublingual na dose de ataque de 20mg e uma manutenção de 20mg por via oral a cada seis e oito horas. As variáveis avaliadas foram os batimentos cardio-fetais (BCF), frequência cardíaca materna (FCM), pressão arterial sistólica (PAS) e diastólica (PAD) e índice de líquido amniótico (ILA). Todas as variáveis foram avaliadas antes da administração da nifedipina e aproximadamente após 6h e cada 24h até alta hospitalar. Resultados: não houve modificação dos BCF (p=0,48) e da PAS (p=0,29). A FCM aumentou após 24h, mas sem significância estatística (p=0,08) retornando a níveis similares ao da admissão com 48h. A PAD diminuiua partir de 6h (p = 0,04)até 72h, mantendo-se constante. O ILA diminuiu significativamente em 24h, 48h e 72h. Conclusão: a utilização de altas doses de nifedipina para tocóliseocasio na diminuição dos níveis pressóricos diastólicos maternos e consequentemente diminuição do ILA, mas provavelmente sem repercussões clínicas.

Detección de un mosaico de trisomía 21 en líquido amniótico / Detection of a mosaicism of trisomy 21 in amniotic liquid

Resumen Se analizó un resultado con alteración cromosómica tomado de una base de datos conformada por un total de 4755 muestras de líquido amniótico extraídos mediante amniocentesis con indicación de su médico tratante, riesgo sérico y edad materna avanzada. En este reporte se presenta la detección de un mosaico de trisomía 21 en líquido amniótico, mediante la técnica de Banda G donde se analizaron 20 metafases. Los resultados obtenidos documentan una composición cromosómica 47, XY+21 y 46, XY con una relación 9:11 respecto a las metafases analizadas, confirmándose así el diagnóstico del Síndrome de Down secundario a mosaico.

Abstract A result with chromosomal alteration was analyzed from a database consisting of a total of 4755 samples of amniotic fluid extracted by amniocentesis with indication of the attending physician, serum risk and advanced maternal age. This report presents the detection of a mosaicism of trisomy 21 in amniotic fluid, using G- Banding where 20 metaphases were analyzed. The results obtained document a chromosomal composition 47, XY + 21 and 46, XY with a 9:11 ratio with respect to the metaphases analyzed, confirming the diagnosis of Down syndrome secondary to mosaicism.