Objective:To construct a lentiviral vector overexpression of micrRNA-29b and investigate the biological characteristics in mouse neuronal cell lines GT1-7.Methods:We chemically synthesized two oligonucleotide single-stranded,complete the comple-mentary by bridging extension into DNA double-stranded to form miR-29b precursor structure.The restriction enzyme digested vector plasmid FUGW was ligated to the precursor structure ofmiR-29b by homologous recombination to construct the corresponding lentiviral vector of microRNA-29b overexpression,and the stable cells were obtained in the mouse neuronal cell line GT1-7 by bleomycin drag screening.RT-PCR was used to detect the expression level of related genes at mRNA transcription level,Results:The recombinant lentiviral expression plasmid f-F-miR-29b was successfully constructed,and the expression level was about 30 times higher than that of the control group.The expressions of DCX,Vdac1 and pten were inhibited,have no changes in sex developmental related genes LH-β,kiss-l,Inshulin,IGF-I,GPR54,GnRH and leptin-R.Conclusion:Using the method of lentivirus screening,the microRNA-29b overexpressing stably transformed cells was successfully obtained in mouse neuron GT1-7 cells,which laid a foundation for the study of biological characteristics ofmicroRNA-29b.

Objective Deletion detection and annotation of 18 lines from the population of specific chromosome 1 substitution strains ( PCSSs) derived from Chinese wild mice based on whole genome re-sequencing data. Methods Whole genome re?sequencing of the 18 lines were performed on the Illumina Hiseq platform. SpeedSeq software was used to detect the deletion after read alignment. Further annotation was obtained using SnpEff software. Results 13803 dele?tions were identified among the 18 lines, the length of deletion was ranged from 51bp to 70 kb, among them nearly 50%were less than 500 bp. Through functional annotation,we found most of the variants were located in intronic (50. 361%) and intergenic (28. 745%) regions. However, we also identified 31 protein coding genes harboring loss?of?function dele?tions. Among them, 3 genes were associated with human diseases, 7 genes were participated in 11 KEGG pathways. Conclusion The chromosome 1 of PCSSs harbors abundant deletion mutations which can be used as genetic markers in genetic studies.

Purpose Polymorphisms of candidate gene Agouti was analyzed in order to reveal the molecular mech -anisms of coat color difference in chromosome engineering mice .Methods Firstly, differences of mouse coat color was detected by color measurement spectrophotometer .Then, candidate gene Agouti was found by whole genome scanning based on DNA chip.Finally, cDNA and amino acid sequence polymorphisms were analyzed , as well as the influence of protein properties and function after mutation was predicted by bioinformatics software .Results There are five SNPs in the Agou-ti cDNA sequences , resulting in three missense mutations in the amino acid sequence of Agouti signaling protein .Bioinfor-matics analysis revealed that one βsheet deletion in the secondary structure of the mutant protein , as well as tertiary struc-ture changed , leading to decrease of binding ability .Conclusion A novel missense mutation is found in candidate Agouti gene.It plays critical role in receptor binding activity , and may reflect on mice coat color changing from light gray to dark gray eventually .

Objective To establish a high throughput general multiple competitive polymerase chain reaction ( cPCR) detecting method of copy number variations ( CNVs) for the population of chromosome 1 substitution strains from wild mice.Method The selected 14 loci, including 11 CNVs on chromosome 1 and internal control loci on other three chromosmes (Chr 7, Chr 19 and Chr X), were detected based on the universal fluorescent primer multiple competitive pol-ymerase chain reaction.All specific cloned plasmids were constructed as competitors.Results Altogether 11 CNVs were designed in one panel, and the copy of Chr X accurately reflects the gender.Conclusions A rapid and high-throughput fluorescent multiplex cPCR assay is established which can be used for detection of copy number variations on chromosome 1 in mice.

Objective To analyze the growth phenotype and blood biochemical parameters of chromosome 1 substi-tution mouse strain(CSS1), and investigate their potential of QTL mapping .Methods Body weight, body length, tail length, organ weight of the CCS1 mice were measured at different days to create a growth curve while blood biochemical in -dexes were measured at about the 80th day.Results The CCS1 mice were different from C57BL/6 mice in several inde-xes.Compared with the C57BL/6 mice during different developmental stages , six strains including B6-Chr1KM mice were significantly different in body weight .There were five strains including B6-Chr1CM mice significantly different with C57BL/6 mice in body length, and all of the CSS1 mice were significantly different from C57BL/6 mice in tail length.Part of CCS1 mice were significantly different from C57BL/6 mice in the weight of liver, spleen, kidney and brain.The ALT of female B6-Chr1CM mice was significantly higher than that in the C 57BL/6 mice.The ALP of female B6-Chr1HZ mice was signifi-cantly higher than that in the male C57BL/6 and B6-Chr1KM mice, and was significantly lower than that in the C57BL/6 mice.The TB of male B6-Chr1CM, B6-Chr1SMX and B6-Chr1HZ mice was significantly higher than that of the C 57BL/6 mice.The TG of male B6-Chr1SMX mice and male B6-Chr1TW mice was significantly higher than that in the C 57BL/6 mice. Conclusions The phenotype of Chr1 CSS mice is quite different from commonly used inbred strain C 57BL/6 mice.CCS1 mice show great potential in QTL mapping for their characteristic growth phenotype and blood biochemical indexes .

Objective The DNA methylation microhaplotype(DMH)refers to the combination of multiple methylation sites within a very short range,and these haplotypes show wide diversity.We carried out screening and validation of age-related DMHs in mouse blood.Methods We initially constructed a theoretical dataset of DMHs based on the mouse reference genome.We then screened age-related DMHs by Spearman's rank correlation analysis,using high-throughput sequencing information for DNA methylation in mouse blood from a network database.Finally,cross-validation was performed using a validation dataset.Results A total of 6787 142 DMH sites were identified within 50 bp in the mouse genome,including 98.64%of single-digit CpG sites.A total of 5835 age-associated DMHs were screened in 58 mouse blood samples(|rho|＞0.5,P＜0.01),accounting for 0.086%of DMHs.Finally,we validated the top 100 age-associated DMHs with high correlation in 95 independent samples,Resultsing in 44 loci.Conclusions The age-associated DMHs screened in this study may be useful in future studies of apparent age prediction using mouse blood and in aging studies.

Objective To analyze the epidemic characteristics of varicella and the genetic characteristics of varicella zoster virus (VZV) in Yangzhou in 2021, and to provide a theoretical basis for the scientific prevention and control of varicella in Yangzhou. Methods Descriptive epidemiological analysis was carried out on the varicella outbreaks reported in Yangzhou in 2021. Throat swabs or herpes fluid samples from varicella cases in 2021 were collected, and the viral nucleic acid was detected by real-time fluorescent quantitative PCR. The genotype and evolutionary relationship of the virus strain were determined according to the 6 SNPs in the ORF22 gene fragment sequence. Results In 2021, there were 20 varicella outbreaks in Yangzhou, involving 147 cases, all of which occurred in kindergartens and primary and secondary schools, and the peak incidence was in the age group of 4-7 years old. The high incidence time of the outbreaks was from May to July, and from November to January of the next year. The varicella vaccination rate of the cases was low, and all were 1-dose vaccination. The gene sequencing results of 8 samples were J/clade 2, and 3 of them had A-C synonymous mutation at position 37997 in ORF22 sequence. Conclusion In 2021, varicella outbreaks in Yangzhou occurred mainly in kindergartens and schools. Preschool children are susceptible, all of which are caused by J/clade 2 varicella-zoster virus. It is suggested to strengthen the monitoring and management of the varicella epidemic situation in schools in the city, and at the same time incorporate the varicella vaccine into the routine immunization program of the city and strengthen 2 doses of varicella vaccination.