Objective To explore the association between 15 current used pesticides (CUPs) in maternal placenta and the risk of fetal neural tube defects (NTDs). Methods A case-control study was conducted among mothers of 308 controls and 150 NTD cases. Concentrations of 15 CUPs in placental tissues were measured by liquid chromatography-mass spectrometry. Multivariate logistic regression and weighted quantile sum (WQS) regression were used to explore the association between placental CUPs and the risk of NTDs. Questionnaire survey was used to collect information on maternal pesticide active exposure history and dietary habits during pregnancy. Results The overall detection rate of 15 CUPs was 89.7% in this study. Multivariate logistic regression showed that high level of flumorph (OR=2.58, 95%CI[1.10-5.90]) was significantly associated with a higher risk of NTDs, and high levels of pyraclostrobin (OR=0.28, 95%CI[0.08-0.86]) and imidacloprid (OR=0.38, 95%CI[0.15-0.90]) were significantly associated with lower risk of NTDs, but the associations were no longer significant after Bonferroni correction. WQS regression did not show any significant combined exposure effect of CUP mixtures on the risk of fetal NTDs (OR=0.69, 95%CI[0.14-3.45]). Conclusion Exposure to CUPs during pregnancy was quite common in this study, but no statistical association was found between placental CUPs and the risk of fetal NTDs.

Objective To study the change of serum metabolite profiles in pregnant mice with embryos occurring neural tube closure failure, and the possible role of histone modification in the development of neural tube defects(NTDs) induced by benzo[a]pyrene(BaP). Methods Pregnant ICR mice were classified into BaP exposure group and control group, which were treated by intraperitoneal injection of BaP or corn oil at gestational day(GD) 7-9. The serum of pregnant mice and embryos were collected at GD 9.5. Based on the phenotype of the embryos, the BaP exposure group was further classified into BaP_NTDs group and BaP_nonNTDs group. Ultra-performance liquid chromatography-mass spectrometry was used to detect serum metabolites, and the differences of metabolites between the two groups were compared. We further annotated metabolic pathways of the different serum metabolites in Kyoto Encyclopedia of Genes and Genomes(KEGG) database. RNA-seq and CUT&Tag were used to assay the genome-wide gene expression and the histone modifications of H3K27me3 and H3K27ac of embryonic neural tissues to explore the possible genes and histone modifications responsible for the NTDs based on the identified metabolic pathways. Results Compared to the control group, 44 differential metabolites were found in BaP_NTDs group. Compared to the BaP_nonNTDs group, 11 differential metabolites were found in BaP_NTDs group. The LysoPC(0:0/18:2(9Z,12Z)) was uniquely up-regulated in BaP_NTDs group. Pentose phosphate pathway was enriched both for the BaP_NTDs/control differential metabolites and BaP_nonNTDs/control differential metabolites. Further, the RNA-seq and CUT&Tag data of embryonic neural tissues revealed that the expression of Aldob in the BaP_NTDs group, one of the key genes involved in the pentose phosphate pathway, was 31.06 fold-change of that in the control group, and 9.68 fold-change of that in the BaP_nonNTDs group. They both had a higher peak of H3K27ac modification in the promoter of the gene. Conclusion LysoPC(0:0/18:2(9Z,12Z)) might be the key differential metabolite in the serum of pregnant mice with NTD embryos induced by BaP, and the histone modifications of Aldob in pentose phosphate pathway might be one of the underlying mechanisms.

Objective To explore the effects of the interventions on improving caregivers' family rearing practices of left-behind children(LBC)under 3 years old. Methods The Integrated Early Childhood Development(IECD) project was implemented in 80 villages of four counties in Shanxi and Guizhou provinces from July 2014 to December 2016, which provided integrated interventions of feeding and responsive care for children under 3 years old. A quasi-experimental design was applied in our study to evaluate the effect of intervention on the family rearing attitude and practice of caregivers of left-behind children under three years old.. The baseline and post-intervention survey data of the Integrated Early Childhood Development(IECD) project were used to carry out the study in July-Sep of 2013 and 2016, respectively. The baseline survey included 695 cases in the intervention group, 478 cases in the control group, 740 cases in the post-intervention survey and 514 cases in the control group. Adifference-in-difference model was used to estimate the effects analyze the relationship between intervention dose and nursing practice, and control for confounding factors such as sociodemographic characteristics. Results After two years' intervention, the proportion of undertaking responsive caregiving among caregivers in the intervention group increased from 74.1%(515/695) at baseline to 79.2%(585/739) after intervention with an increase of 5.1%,. The proportion of responsive care in the control group decreased by 29.3% over the same period. After controlling for confounding factors with difference-difference model, the intervention significantly improved the response care received by left-behind children(OR:13.11; 95%CI:7.06-25.35). The proportion of undertaking violent disciplinary practices decreased by 10.9%(baseline:52.7%(366/695); post-intervention:41.8%(309/739) in the intervention group, which was lower than that in the control group(aOR:0.57; 95%CI:0.40-0.82). But no significant improvement was found in the ownership rate of books, toys, experiencing inadequate supervision, and the caregivers' attitude towards physical discipline of left-behind children. Compared to the frequency of intervention less than once a month, the intervention at a frequency of at least once a month but less than once a week was associated with improving responsive care and violent care practices(P<0.05). Conclusion The integrated interventions effectively improved the caregivers' responsive care and violent disciplinary behaviors. Intervention should be at a frequency of no less than once a month, to promote good rearing care practices by caregivers.

Objective To explore factors that influence the follow-up compliance of high-risk infants and improve the follow-up and management strategy of the high-risk infants. Methods A total of 675 high-risk infants who were under 2 years old and admitted to the outpatient department of the Children`s Health Department of a hospital in Hohhot from 2019 to 2021 were included. Results In total, 49.6%(335/675) of the high-risk infants had high follow-up compliance. Multivariable logistic regression analysis showed that several factors including local residence, multiple high-risk factors, higher mother's education, and multiple caregivers were associated with high follow-up compliance. Conclusion Popularizing parenting knowledge, enhancing parents' health care awareness, strengthening the training of health care providers especially those of primary health care providers, improving child health care capacity, and providing accessible and convenient health care services might improve the follow-up compliance as well as health of high-risk infant.

Objective To describe the characteristics of pre-hospital emergencies, disease parity, and cause of death parity among children and adolescents aged 3-17 years in Yantai in the past 6 years. Methods All case data of children and adolescents aged 3-17 years from 2016-2021 in the pre-hospital emergency electronic medical record system of Yantai 120 Emergency Command Center were selected for analysis, and the student population data of each year provided by Yantai Education Bureau were used as the denominator to calculate the prevalence and mortality of children's pre-hospital emergencies, and to describe the distribution characteristics of different time(year), different regions(urban and rural), and different populations(gender and school periods). Results The annual prevalence of pre-hospital emergencies was 501.8-600.2/100,000 for boys and 361.7-431.6/100,000 for girls aged 3-17 years in 2016-2021, increasing yearly between 2016-2019, decreasing in 2020 and rebounding in 2021. Among the school periods, senior students had the highest annual prevalence(919.9-1058.6/100,000), while junior students had the lowest annual prevalence(275.5-357.6/100,000). Longkou and the Development Zone had the highest annual prevalence(>500/100,000), while Haiyang and Qixia had lower annual prevalence(304.2-467.3/100,000). The mortality rate for boys showed a decline followed by stabilization, with a fluctuation range of 7.5-10.0/100,000, and for girls showed a rise followed by a decline and stabilization, with a fluctuation range of 3.6-5.6/100,000. Accidental injuries accounted for 56.1% of pre-hospital emergencies, with drowning, traffic injuries and fall injuries as the top three causes of death; roads and homes were the main incident sites. The gender differences, school periods and urban and rural area differences for different injuries and different diseases were statistically significant. Conclusion The overall prevalence of pre-hospital emergencies among children and adolescents aged 3-17 years in Yantai City in 2016-2021 for both genders showed a fluctuating upward trend, and the mortality rate was stable, with increased prevention and control efforts for the types of such as accidental injuries, enhanced school-related health and safety education, and joint efforts of family, school, and community to prevent and reduce the occurrence of emergencies especially injuries.

Objective Using murine and cell models to explore the effects of Li₂CO₃ on the cell cycle during mouse embryonic neural development. Methods According to the Li₂CO₃-induced NTDs murine mode which we have established, 350mg/kg Li₂CO₃ was injected intraperitoneally into the C57BL/6J pregnant mice on embryonic day 7.5. The development of the offspring were observed. Li₂CO₃ in different concentrations(0.5,1,1.5, 2, 2.5, 3, 5 and 10mM) were used to treat the NE4Cs and NIH3T3. Cell survival, proliferation and cell cycle were detected by methylthiazolyldiphenyl tetrazolium bromide, 5-ethynyl-2-deoxyuridine labeling and flow cytometry. The quantitative real-time polymerase chain reaction and western blotting used to detect the expressions of cell cycle key genes. Results 350mg/kg Li₂CO₃ affected embryonic development. The incidence of NTDs was 32.7%(18/55). Treatment with the low concentration of Li₂CO₃ in NE4Cs and NIH3T3 cells increased cell survival, reduced the cells in G1 phase, most cells were in S and G2/M phases and promoted cell proliferation. Both in mice and cells models, the expressions of cell cycle key genes cyclin A, cyclin D1, Cdk2 and Cdk4 increased after Li₂CO₃ treatment(P<0.01). Conclusion Li₂CO₃ disturbed the cell cycle to cause abnormal cell proliferation leading to NTDs by up-regulating the expression of cell cycle key genes cyclin A, cyclin D1, Cdk 2 and Cdk 4.

Objective To conduct genetic analysis and preimplantation genetic testing for monogenic disorders(PGT-M) in a family with Angelman syndrome. Methods We included a family with Angelman syndrome who visited the Reproductive Department of Zhongshan Hospital Affiliated to Fudan University in April 2023. Methylation-specific multiplex ligation-dependent probe amplification(MS-MLPA) method was used to verify the SNRPN u5 deletion and methylation abnormalities in the 15q11.2-q13 region of the family. The ASA array was used to detect the single nucleotide polymorphism(SNP) sites of the family and construct a haplotype. In vitro fertilization was performed using intracytoplasmic sperm injection technology, followed by embryo biopsy on the third day of cleavage stage. Single cell whole genome amplification, second-generation sequencing, and SNP linked haplotype analysis were used to detect whether the embryo carried SNRPN u5 genetic variations in the 15q11.2-q13 imprinted region and chromosomal copy number variations(CNVs) of 4 Mb or more. Results The results showed that the proband and the younger brother had SNRPN u5 copy number deletion and methylation abnormalities in the 15q11.2-q13 imprinting region. The copy number deletion was inherited from the mother, and the mother had a 15q11.2-q13 SNRPN u5 copy number deletion. However, the mother's methylation status was normal. After successfully constructing a haplotype in the family, one cleavage embryo was obtained through clinical ovulation induction. The PGT-M results showed that the embryo carried pathogenic SNRPN u5 mutations inherited form the mother and had multiple chromosome copy number abnormalities. The embryo transfer is thus not recommended. Conclusion PGT-M technology can assist in reproductive intervention and prevent the birth of Angelman syndrome patients in their families, and is an important way to assist families with monogenic genetic diseases in eugenics.

Objective To explore the effect of circadian rhythm disruption on the expression of rhythm gene CLOCK in endometrial cancer and its possible promoting mechanism during the disease progression. Methods According to the patient's age, gravidity and parity, occupational history, educational background, environmental factors, and other background matching, 6 cases of endometrioid adenocarcinoma with a history of night shift work were selected as the abnormal rhythm group, and 6 cases without a history of night shift work were selected as the normal rhythm group. qPCR and Western blot were used to detect the expression of CLOCK mRNA and protein in the two groups. Immunohistochemistry was used to detect the expression of CLOCK and epithelial-mesenchymal transformation related phenotypic proteins(E-cadherin and vimentin) in 32 cases of endometrioid adenocarcinoma, than analyzed the relationship between the expression and clinicopathological features, and the correlation between CLOCK expression and epithelial-mesenchymal transformation related phenotypic protein. Results The results of qPCR and Western blot showed that the relative expression levels of CLOCK mRNA and protein in abnormal rhythm group were significantly lower than those in normal rhythm group(P<0.05). Immunohistochemistry indicated that the negative rate of E-cadherin and CLOCK, the positive rate of vimentin were higher in the tumors with night shift work history, FIGO stage II-III and deep muscle infiltration, and the difference was statistically significant (P<0.05).The correlation between CLOCK expression and E-cadherin, vimentin in endometrioid adenocarcinoma tissue specimens was statistically significant, (P<0.05). Conclusion The disruption of circadian rhythm caused by night shift down-regulates the expression of CLOCK gene in endometrioid adenocarcinoma tissues. The involvement of CLOCK gene in epithelial-mesenchymal transformation process of tumor tissues is a possible mechanism of endometrioid adenocarcinoma progression and may be an important factor affecting prognosis.