Enriched environment for offspring improves learning and memory impairments induced by sevoflurane exposure during the second trimester of pregnancy.

Studies in animals indicate that sevoflurane exposure in the second trimester of pregnancy has harmful effects on the learning and memory of offspring. Whether an enriched environment can reverse the damage of sevoflurane exposure in the second trimester of pregnancy on the learning and memory of rat offspring remains unclear. In this study, rats at 14 days of pregnancy were exposed to 3.5% sevoflurane for 2 hours and their offspring were treated with an enriched environment for 20 successive days. We found that the enriched environment for offspring increased nestin and Ki67 levels in hippocampal tissue, increased hippocampal neurogenesis, inhibited glycogen synthase kinase 3ß activity, and increased the expression of cell proliferation-related ß-catenin and apoptosis-related Bcl-2, indicating that an enriched environment reduces sevoflurane-induced damage by increasing the proliferation of stem cells in the hippocampus. These findings suggest that an enriched environment can reverse the effects of sevoflurane inhaled by rats during the second trimester of pregnancy on learning and memory of offspring. This study was approved by the Animal Ethics Committee of Shengjing Hospital of China Medical University (approval No. 2018PS07K) on January 2, 2018.

Effects of mild intrauterine hypoperfusion in the second trimester on memory and learning function in rat offspring.

Mild intrauterine hypoperfusion (MIUH) is a serious pathological event that affects the growth and development of fetuses and offspring. MIUH can lead to growth restriction, low birth weight, neurodevelopmental disorders, and other adverse clinical outcomes. To study the effects of MIUH on learning and memory function in offspring, a model of MIUH was established by placing a coil (length 2.5 mm, diameter 0.24 mm) on the uterine artery and ovarian uterine artery of Sprague-Dawley rats in the second trimester of pregnancy (day 17). Next, 120 mg/kg lithium chloride (the MIUH + Li group) or normal saline (the MIUH group) was injected intraperitoneally into these rats. In addition, 120 mg/kg lithium chloride (the Li group) or normal saline (the SHAM group) was injected intraperitoneally into pregnant rats without coil placement. The Morris water maze was used to detect changes in learning and memory ability in the offspring at 4 weeks after birth. In the MIUH group, the escape latency and journey length before reaching the platform were both increased, and the number of times that the platform was crossed and the activity time in the target quadrant within 90 seconds were both decreased compared with the SHAM group. Immunofluorescence double staining and western blot assays demonstrated that hippocampal nestin and Ki67 (both cell-proliferation-related proteins) expression was significantly downregulated in the MIUH group compared with the SHAM group. Furthermore, western blot assays were conducted to investigate changes in related signaling pathway proteins in the brains of offspring rats, and revealed that glycogen synthase kinase 3ß (GSK3ß) expression was upregulated and ß-catenin expression was downregulated in the MIUH group compared with the SHAM group. In addition, compared with the MIUH group, the expression levels of p-GSK3ß and ß-catenin were upregulated in the MIUH + Li group. These results suggest that MIUH may affect learning and memory function in rat offspring by regulating the GSK3ß signaling pathway. The experimental procedures were approved by Animal Ethics Committee of Shengjing Hospital of China Medical University (approval No. 2018PS07K) in June 2018.

Rare De Novo IGF2 Variant on the Paternal Allele in a Patient With Silver-Russell Syndrome.

Silver-Russell syndrome (SRS) is a rare, well-recognized disorder characterized by growth restriction, including intrauterine and postnatal growth. Most SRS cases are caused by hypomethylation of the paternal imprinting center 1 (IC1) in chromosome 11p15.5 and maternal uniparental disomy in chromosome 7 (UPD7). Here, we report on a Chinese family with a 4 year old male proband presenting with low birth weight, growth retardation, short stature, a narrow chin, delayed bone age, and speech delays, as a result of a rare molecular etiology. Whole-exome sequencing was conducted, and a novel de novo IGF2 splicing variant, NM_000612.4: c.157+5G > A, was identified on the paternal allele. In vitro functional analysis by RT-PCR and Sanger sequencing revealed that the variant leads to an aberrant RNA transcript lacking exon 2. Our results further confirm the IGF2 variant mediates SRS and expand the pathogenic variant and phenotypic spectrum of IGF2-mediated SRS. The results indicate that, beyond DNA methylation and UPD7 and CDKN1C variant tests, IGF2 gene screening should also be considered for SRS molecular diagnoses.

A small supernumerary marker chromosome resulting in mosaic partial tetrasomy 4q26-q31.21 in a foetus with multiple congenital malformations.

A parental diagnosis was performed for an unborn foetus of a healthy couple, who was due for ultrasound detection of multiple malformations and abnormal amniotic fluid karyotypes. For an accurate diagnosis, routine G-banding analysis and next generation sequencing (NGS) were carried out. Finally, conventional cytogenetic analysis suggested that the foetus had a karyotype of47,XX,+mar[52]/46,XN, meanwhile NGS also revealed a partial tetrasomy of 27.84Mb from 4q26-q31.21 (117,385,735-145,225,759), and G-banding analysis excluded the couple to have carried the 4q26-q31.21 duplication. We have identified a de novo mosaic small supernumerary marker chromosomes (sSMC) derived from 4q26-q31.21 in a foetus with hemivertebra, polydactyly, abnormal ears, and heart and ventricular septal defect.

High-concentration sevoflurane exposure in mid-gestation induces apoptosis of neural stem cells in rat offspring.

Sevoflurane is the most commonly used volatile anesthetic during pregnancy. The viability of neural stem cells directly affects the development of the brain. However, it is unknown whether the use of sevoflurane during the second trimester affects the survival of fetal neural stem cells. Therefore, in this study, we investigated whether exposure to sevoflurane in mid-gestation induces apoptosis of neural stem cells and behavioral abnormalities. On gestational day 14, pregnant rats were anesthetized with 2% or 3.5% sevoflurane for 2 hours. The offspring were weaned at 28 days and subjected to the Morris water maze test. The brains were harvested to examine neural stem cell apoptosis by immunofluorescence and to measure Nestin and SOX-2 levels by western blot assay at 6, 24 and 48 hours after anesthesia as well as on postnatal day (P) 0, 14 and 28. Vascular endothelial growth factor (VEGF) and phosphoinositide 3-kinase (PI3K)/AKT pathway protein levels in fetal brain at 6 hours after anesthesia were assessed by western blot assay. Exposure to high-concentration (3.5%) sevoflurane during mid-gestation increased escape latency and path length to the platform, and it reduced the average duration spent in the target quadrant and platform crossing times. At 6, 24 and 48 hours after anesthesia and at P0, P14 and P28, the percentage of Nestin/terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells was increased, but Nestin and SOX-2 protein levels were decreased in the hippocampus of the offspring. At 6 hours after anesthesia, VEGF, PI3K and phospho-AKT (p-AKT) levels were decreased in the fetal brain. These changes were not observed in animals given low-concentration (2%) sevoflurane exposure. Together, our findings indicate that exposure to a high concentration of sevoflurane (3.5%) in mid-gestation decreases VEGF, PI3K and p-AKT protein levels and induces neural stem cell apoptosis, thereby causing learning and memory dysfunction in the offspring.

[Fetal ventriculomegaly: diagnosis using magnetic resonance imaging and its prognosis].

OBJECTIVE: To evaluate the diagnostic value of magnetic resonance imaging (MRI) on fetal ventriculomegaly identified through prenatal ultrasonography and the outcomes of these newborns were followed up. METHODS: From March 2006 to July 2008, MRI was performed on 135 pregnant women whose fetuses diagnosed as fetal ventriculomegaly at an average of 32 gestational weeks in Shengjing Hospital Affiliated to China Medical University. Mild ventriculomegaly was defined when the width of unilateral or bilateral fetal cerebral ventricle triangle was 10-15 mm, moderate ventriculomegaly 16-20 mm and severe ventriculomegaly >20 mm. We introduced the Denver developmental screening test (DDST) to follow-up the mild ventriculomegaly and normal babies, confirmed by MRI, at 6-12 months after birth and a case-control study was conducted. The intelligence and growth of these infants were analyzed. RESULTS: (1) Diagnostic rate of fetal ventriculomegaly through MRI: Among the 135 gravidas, 60 (44.4%) showed isolated ventriculomegaly, 5(3.7%) complicated with ventricular hemorrhage; 12 (8.9%) complicated with agenesis of corpus callosum (ACC) and 2 (1.5%) complicated with cerebellar hypoplasia, while 56 (41.5%) were normal. Seventy-nine cases had fetal ventriculomegaly on MRI and 15.2% (n = 12) of them complicated with ACC. (2) Degree of fetal ventriculomegaly on MRI: Among the 60 isolated ventriculomegaly cases, 55 (91.7%) were mild and 5 (8.3%) moderate ones. Among the 5 cases complicated with ventricular hemorrhage, one was mild ventriculomegaly, and 4 moderate or severe cases. Among the 12 cases with ACC, 8 (66.7%) were moderate ventriculomegaly and 4 (33.3%) severe cases. The 2 cases with cerebellar hypoplasia were both moderate ventriculomegaly fetuses. (3) Follow-up at 6-12 months after birth: thirty out (case group) of the 55 isolated ventriculomegaly cases, 38 out of the 56 normal babies and 42 babies with normal MRI results were followed up, and the later 80 cases were taken as control. Four infants (13.3%) in the case group and 10 (12.5%) in the control group showed abnormal or suspected results in DDST (P > 0.05), the rest babies were all normal. (4) Clinical outcomes of the 79 ventriculomegaly fetuses diagnosed by MRI: thirty mild ventriculomegaly babies and 5 moderate ones were born at term and showed normal at follow ups. However, 7 gravidas were not compliant, 6 pregnancies were terminated, and 12 were lost. Three of the 12 cases with ACC continued the pregnancy, and postnatal MRI of the babies showed the same with the prenatal MRI, 8 pregnancies were induced and one was lost. All of the 5 fetuses with ventricular hemorrhage were induced and the prenatal diagnosis was confirmed by autopsy. One of the 2 fetuses with cerebellar hypoplasia was term delivered and diagnosed as cerebral palsy at the age of 6 months, and the other one was induced. CONCLUSIONS: MRI is an indispensable complementary diagnostic method for fetal ventriculomegaly diagnosed through ultrasound. The development of intelligence and growth of babies born with mild isolated ventriculomegaly is the same as normal ones.

[MRI diagnosis and analysis of 104 cases of fetal ventriculomegaly by ultrasonography].

OBJECTIVE: To evaluate the diagnostic value of MRI in the cases suspected of ventriculomegaly by prenatal ultrasonography. METHODS: 104 patients of suspected fetal ventriculomegaly (VM) diagnosed by ultrasonography were included from the Shengjing Hospital, China Medical University from March 2006 to October 2007. All cases were divided into 4 groups based on the standard of Gaglioti: 10 - 12 mm (66 cases), 13 - 15 mm (22 cases), 16 - 20 mm (14 cases), and 21 - 25 mm (2 cases); they included 75 cases of single intracerebroventricular expansion and 29 cases of double intracerebroventricular expansion. All of them were subjected to MRI scan within 48 h of ultrasonographic examination to determine the prenatal diagnosis by MRI pregnancy outcomes. RESULTS: Among the 26 072 cases who received prenatal ultrasonography, 104 cases (0.39%) were VM. (1) MRI detected 3 cases (5%) in 10 - 12 mm group: one case of cerebellar hypoplasia, vascular malformation, chest and abdominal anomalies each; 5 cases (23%) in 13 - 15 mm group: one case of agenesis of corpus callosum (ACC), cerebral hemorrhage, cerebral hemorrhage with cerebral meningocele, cerebral meningocele, intracranial mass meningocele each; 6 cases(43%)in 16 - 20 mm group: 4 cases of ACC, one case of intraventricular hemorrhage and ACC combined with ventricular hemorrhage each; 2 cases in 21 - 25 mm group: one case of ACC and intraventricular hemorrhage each. (2) MRI detected 4 cases (5%) among 75 unilateral VM cases and 12 cases (41%) among 29 bilateral VM cases. The differences were significant (P < 0.01). MRI diagnosis rate was 15.38% (16 cases). Follow-up of the outcomes of the pregnancy showed induction of labor in 15 cases (14%) all of which were the same as MRI results on autopsy, full-term delivery of 88 cases, of which all the neonates were healthy. CONCLUSIONS: When the expansion width is above 16 mm or bilateral VM is suspected by ultrasonography, we suggest MRI examination to determine fetal central nervous system disease.