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Author(s): Tie-Ning Zhang 1,2,3, Xin-Mei Huang 4, Xin-Yi Zhao 1,2,3, Wei Wang 3, Ri Wen 3, Shan-Yan Gao 1,2,* Introduction Currently, the global prevalence of diabetes is increasing among women [...], Background Pre-gestational diabetes mellitus (PGDM) has been known to be a risk factor for congenital heart defects (CHDs) for decades. However, the associations between maternal PGDM and gestational diabetes mellitus (GDM) and the risk of specific types of CHDs and congenital anomalies (CAs) in other systems remain under debate. We aimed to investigate type-specific CAs in offspring of women with diabetes and to examine the extent to which types of maternal diabetes are associated with increased risk of CAs in offspring. Methods and findings We searched PubMed and Embase from database inception to 15 October 2021 for population-based studies reporting on type-specific CAs in offspring born to women with PGDM (combined type 1 and 2) or GDM, with no limitation on language. Reviewers extracted data for relevant outcomes and performed random effects meta-analyses, subgroup analyses, and multivariable meta-regression. Risk of bias appraisal was performed using the Cochrane Risk of Bias Tool. This study was registered in PROSPERO (CRD42021229217). Primary outcomes were overall CAs and CHDs. Secondary outcomes were type-specific CAs. Overall, 59 population-based studies published from 1990 to 2021 with 80,437,056 participants met the inclusion criteria. Of the participants, 2,407,862 (3.0%) women had PGDM and 2,353,205 (2.9%) women had GDM. The meta-analyses showed increased risks of overall CAs/CHDs in offspring born to women with PGDM (for overall CAs, relative risk [RR] = 1.99, 95% CI 1.82 to 2.17, P < 0.001; for CHDs, RR = 3.46, 95% CI 2.77 to 4.32, P < 0.001) or GDM (for overall CAs, RR = 1.18, 95% CI 1.13 to 1.23, P < 0.001; for CHDs, RR = 1.50, 95% CI 1.38 to 1.64, P < 0.001). The results of the meta-regression analyses showed significant differences in RRs of CAs/CHDs in PGDM versus GDM (all P < 0.001). Of the 23 CA categories, excluding CHD-related categories, in offspring, maternal PGDM was associated with a significantly increased risk of CAs in 21 categories; the corresponding RRs ranged from 1.57 (for hypospadias, 95% CI 1.22 to 2.02) to 18.18 (for holoprosencephaly, 95% CI 4.03 to 82.06). Maternal GDM was associated with a small but significant increase in the risk of CAs in 9 categories; the corresponding RRs ranged from 1.14 (for limb reduction, 95% CI 1.06 to 1.23) to 5.70 (for heterotaxia, 95% CI 1.09 to 29.92). The main limitation of our analysis is that some high significant heterogeneity still persisted in both subgroup and sensitivity analyses. Conclusions In this study, we observed an increased rate of CAs in offspring of women with diabetes and noted the differences for PGDM versus GDM. The RRs of overall CAs and CHDs in offspring of women with PGDM were higher than those in offspring of women with GDM. Screening for diabetes in pregnant women may enable better glycemic control, and may enable identification of offspring at risk for CAs.

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[Objective] The prokaryotic expression and immunogenicity of goose tembusu virus (GTMUV % E protein domain III were identified in order to lay the foundation for further researches on the biological function and subunit vaccine development of E protein domain III in GTMUV. [Method] According to homologous gene of E protein domain III in GTMUV JS804 strain from GenBank, a pair of primers with Flag tag sequence and enzyme digestion sites at both ends was designed to amplify E protein domain III gene(EIII) by using PCR technique. The PCR product(EIII) was inserted into prokaryotic expression vector pET32a to construct recombinant expression plasmid which then was transformed into E.coli BL21 (DE3). Through induction with IPTG, the fusion protein was expressed in order to identify its immunoreactivity by using Western blotting. [Result] The obtained EIII gene fragment with Flag tag sequence was about 430 bp in length by PCR amplification and then inserted into pET32a to construct expression plasmid pET32a-EIII successfully. The positive recombinant strain was induced with IPTG for 5 h, the expressed fusion protein existed as inclusion body in! E. coli and the molecular weight was about 33.0 kD. The identification results of Western blotting showed that fusion protein could react specifically with Flag McAb and mouse PcAb against tembusu virus to detect out target bands. [Conclusion]The E protein domain III of GTMUV can be successfully expressed in E.coli BL21 (DE3), and the obtained fusion protein has good immunogenicity. Therefore, it can be used for development of GTMUV diagnostic antigen and subunit vaccine. [ABSTRACT FROM AUTHOR]

[Objective] The prokaryotic expression and immunogenicity of goose tembusu virus (GTMUV) E protein domain I were identified in order to lay the foundation for further researches on biological characteristics and immunological function of E protein domain I in GTMUV. [Method] The encoding gene(EI% of GTMUV E protein domain I was artificially synthesised and was inserted into vector pGEX-4t-1 with GST tag. Then the recombinant plasmid was transformed into Escherichia coli. Through induced expression, the fusion protein was obtained and its immunogenicity was identified by using Western blotting. [Result] The obtained gene fragment E1 was 411 bp in length. Through inserting into vector pGEX-4t-1, the recombinant expression plasmid pGEX-4t-1-EI was constructed successfully. When positive recombinant strain was induced with 1 mmol/L IPTG, the expression quantity of fusion protein reached the peak after 5 hours. The fusion protein existed as inclusion body in E. coli, and the molecular weight was about 41.0 kD. The identification result of Western blotting showed that fusion protein could react specifically with GST McAb and E protein positive serum to detect out target bands. [Conclusion] The E protein domain I of GTMUV can be successfully expressed in E. coli, and the obtained fusion protein has good immunogenicity. Therefore it can be used for research & development of GTMUV serology detection kit. [ABSTRACT FROM AUTHOR]