Objective To analyze the transcriptome sequencing results of Nelson Bay virus(NBV)-infected murine RAW264.7 mac-rophages,and to screen for differentially expressed genes(DEGs)to provide a theoretical basis for exploring the mechanism of innate immune response in reovirus infection.Methods RAW264.7 cells were infected with the NBV-Miyazaki virus strain at a multiplicity of infection(MOI)of 30.We used transcriptome sequencing technologies,with q＜0.05 and|log2FC|≥ 1,for screening the DEGs in the infection and control groups.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases were used for enrichment analysis of DEGs.Results A total of 442 genes were differentially expressed in the infection group,of which 381 genes were significantly upregulated and 61 genes were significantly downregulated.In the GO analysis,the enrichment of DEGs was primarily related to the innate immune response,defense response to viruses,cytokine production,and cell response to cytokine stimulation.In the KEGG analysis,the enrichment of DEGs were primarily related to the Toll-like receptor,retinoid acid inducible gene Ⅰ-like receptor,PI3K/Akt,and other signaling pathways.Conclusion RAW264.7 macrophages infected with the NBV-Miyazaki virus can activate pattern recognition receptors;promote the release of cytokines,chemokines,and other immune-related factors;and enhance antibody-dependent cell-mediated cytotoxicity to exert an immune effect.This study provides a theoretical basis for exploring the mechanisms of innate immu-nity during NBV-Miyazaki virus infection.

Objective To analyze the transcriptome sequencing results of Nelson Bay virus(NBV)-infected murine RAW264.7 mac-rophages,and to screen for differentially expressed genes(DEGs)to provide a theoretical basis for exploring the mechanism of innate immune response in reovirus infection.Methods RAW264.7 cells were infected with the NBV-Miyazaki virus strain at a multiplicity of infection(MOI)of 30.We used transcriptome sequencing technologies,with q＜0.05 and|log2FC|≥ 1,for screening the DEGs in the infection and control groups.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases were used for enrichment analysis of DEGs.Results A total of 442 genes were differentially expressed in the infection group,of which 381 genes were significantly upregulated and 61 genes were significantly downregulated.In the GO analysis,the enrichment of DEGs was primarily related to the innate immune response,defense response to viruses,cytokine production,and cell response to cytokine stimulation.In the KEGG analysis,the enrichment of DEGs were primarily related to the Toll-like receptor,retinoid acid inducible gene Ⅰ-like receptor,PI3K/Akt,and other signaling pathways.Conclusion RAW264.7 macrophages infected with the NBV-Miyazaki virus can activate pattern recognition receptors;promote the release of cytokines,chemokines,and other immune-related factors;and enhance antibody-dependent cell-mediated cytotoxicity to exert an immune effect.This study provides a theoretical basis for exploring the mechanisms of innate immu-nity during NBV-Miyazaki virus infection.

Objective:To discuss the host proteins that interact with the Nelson Bay orthoreovirus(NBV)σNS protein in the fibroblasts L929 of the mice,and to clarify the effect of host proteins on the viral replication.Methods:The bait plasmid pGBKT7-S3 expressing σNS protein was constructed,and sequencing technology was used to verify the accurate expression of the bait plasmid in the Y2H yeast cells.The pGBKT7-S3 and pGADT7 plasmids were separately and jointly transformed into the Y2HGold yeast cells,plated on solid medium,and the colony growth was observed to confirm that the σNS protein was non-toxic to the yeast cells and could not self-activate the reporter gene.The bait plasmid pGBKT7-S3 was hybridized with the cDNA library in fibroblasts L929 of the mice,and the plasmids encoding the interacting proteins were extracted from the positive clones.The positive sequencing results were screened for the proteins interacting with NBV σNS through the Uniprot database.Gene Ontology(GO)functional enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis,and STRING bioinformatics analysis were performed on the interacting proteins.Results:A total of 61 positive clones were successfully screened.The DNA sequencing analysis and BLAST alignment removed 23 positive clones that did not match the database or were similar in sequence.The positive sequencing results identified 38 proteins interacting with NBV σNS through the Uniprot database.Thirty-one proteins were involved in cellular biological processes;thirty-six proteins were cellular anatomical components;thirty-one proteins had binding functions.Five proteins were part of the mitochondrial respiratory chain;seven proteins were ribosomal proteins and components of the ribosomal subunits;two proteins were involved in iron metabolism homeostasis.The GO functional enrichment analysis results showed that the interacting proteins were enriched in biological processes(BP)such as cellular processes,metabolic processes,biological regulation,localization,and response to stimuli;the cellular components were mainly cellular anatomical components and protein-containing complexes;the molecular functions were concentrated in binding,catalytic activity,structural molecule activity,and transporter activity.The KEGG signaling pathway enrichment analysis results showed that the proteins were highly enriched in translation,folding,sorting,and degradation pathways of genetic information processing and were mainly associated with the digestive system in the organism;they were linked to various viral infections and cancers.The STRING analysis results showed that the interacting proteins included ribosomal proteins,protein modification proteins,metabolic proteins,and immune proteins.Conclusion:The host proteins that interact with NBV σNS protein are successfully screened,and these host proteins play important roles in viral replication.

Objective:To investigate the maternal pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) during pregnancy at advanced maternal age for their second child at advanced maternal age, and to explore the relationship with neonatal outcomes.Methods:This study involved 1 965 women of advanced maternal age who delivered the second child in the Northwest Women's and Children's Hospital from July 1 to December 31, 2017. Clinical data of these women and their newborns were collected through the electronic medical record information system. According to pre-pregnancy BMI, all subjects were divided into four groups: underweight group (<18.5 kg/m 2, n=139), normal weight group (18.5-23.9 kg/m 2, n=1 342), overweight group (24.0-27.9 kg/m 2, n=404) and obese group (≥28.0 kg/m 2, n=80). According to the GWG standard recommended by the American Institute of Medicine (IOM) in 2009, they were also divided into three groups: inadequate GWG group ( n=478), normal GWG group ( n=884) and excessive GWG group ( n=603). Mann-Whitey U test, Chi-square test or Fisher's exact test were used as statistical methods. Effects of pre-pregnancy BMI and GWG on gestational age and birth weight of the newborns were analyzed by binary and multi-class logistic regression models. Results:The median pre-pregnancy BMI of the 1 965 women was 22.1 (20.3-23.9) kg/m 2 and patients with abnormal pre-pregnancy BMI accounted for 31.7% (623/1 965). Their median GWG was 13.0 (10.0-16.0) kg and 55.0% (1 081/1 965) of them were abnormal. Compared with normal pre-pregnant weight women, overweight and obesity subjects were associated with increased risks of preterm birth ( OR=2.100, 95% CI: 1.398-3.156), low birth weight infants (LBWI) ( OR=3.187, 95% CI: 1.892-5.367) and macrosomia ( OR=1.758, 95% CI: 1.182-2.614); pre-pregnancy underweight reduced the incidence of large for gestational age (LGA) infants ( OR=0.476, 95% CI: 0.236-0.960). Compared with the normal GWG group, the inadequate GWG group had increased risks of preterm birth ( OR=2.316, 95% CI: 1.530-3.505) and LBWI ( OR=1.850, 95% CI: 1.103-3.104), while the excessive GWG group showed increased risks of macrosomia ( OR=1.828, 95% CI: 1.225-2.726) and LGA infants ( OR=1.955, 95% CI: 1.448-2.640), but a reduced risk of LBWI ( OR=0.359, 95% CI: 0.193-0.667) and small for gestational age infants ( OR=0.452, 95% CI: 0.240-0.852). Conclusions:Both abnormal pre-pregnancy BMI (underweight, overweight and obese) and GWG (inadequate and excessive) have adverse effects on neonatal outcomes in women of advanced age in pregnancy for their second baby. Weight management should be addressed during the whole pregnancy, including both adjusting the pre-pregnancy BMI to normal range and maintaining reasonable GWG, so as to reduce potential adverse outcomes in newborns.

Objective To establish the normal parameters of psychometric measures such as the number connection tests A(NCT-A)and digit symbol tests(DST)in assessment of minimal hepatic en- cephalopathy(MHE).Methods One hundred and sixty healthy volunteers(aged 25 to 64 years;educa- tional level＞9 years)were divided into＜35 ys,35～44 ys,45～54 ys and 55～64 ys groups.All of the healthy volunteers were assessed with NCT-A and DST to establish the normal value of age-related parameters,which can be used for diagnosis of MHE in patients with liver cirrhosis.Two standard devi- ation of the normal mean was used as a diagnostic criterion for MHE.One hundred and six cirrhotic patients were assessed with these parameters.Results The parameters of NCT-A were(25.1?4.6) sec in＜35 ys group,(32.1?6.8) sec in 35～44 ys group,(38.6?7.1)sec in 45～54 ys group or (49.3?6.3)sec in 55～64 ys group.The scores of DST were 49.9?4.7 in＜35 ys group,44.6?4.8 in 35～44 ys group,38.5?5.0 in 45～54 ys group or 35.4?4.7 in 55～64 ys group.Thirty one out of 106 cirrhotic patients were diagnosed as MHE based on these parameters.Conclusion The NCT- A and DST are psychometric assessments for diagnosis of MHE.Age-based normal paramerters of NCT- A and DST are needed to be established.