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Objective To use disability - adjusted life years (DALYs) to assess unequal differences in the burden of disease due to diabetes across global socioeconomic groups. Methods Number of DALYs due to diabetes, age 一 standardized DALYs rates from the GBD database, 1990 -2019, and national Human Development Index(HDI) data from the Human Development Report 2021 - 2022 were collected. The relationship between age 一 standardized DALY rates and HDI was analyzed to understand socioeconomic differences in diabetes disease burden. Calculation of Annual Percentage Change(APC) and Average Annual Percentage Change(AAPC) to assess temporal trends in the burden of diabetes disease during recent decades. Results From 1990 一 2019, the number of DALYs due to diabetes globally increased from 28. 3 million to 69. 9 million, an increase of 146. 85%, with the fastest growth (206. 49%) in Lower middle 一 income countries. The age - standardized DALYs rates were negatively correlated with HDI(r = -0. 480, P <0. 001). Countries with medium HDI exhibit higher DALYs rates due to diabetes mellitus. From 1990 一 2019, the average annual increase in the rate of age 一 standardized DALYs was 0. 78% (AAPC =0. 78%, 95% CI = 0. 74% to 0. 81%), and the average annual increase in the age 一 standardized mortality rate was 0. 46% (AAPC = 0.46%, 95% CI = 0. 41% ~ 0. 51%). Conclusion The global burden of diabetes disease has increased substantially over the last few decades, with the fastest growth in Lower middle - income countries. The age 一 standardized burden was higher in developing countries. Although a slower increase in the burden of diabetes was observed, the persistence of the increase in the burden of disease suggests that more diabetes prevention programs and health - care services should be made available to developing countries. [ABSTRACT FROM AUTHOR]

[Objective] To study the effects of protein X (pX) on Toll-like receptors after fowl adenovirus serotype 4 (FAdV-4) infection of LMH cells, the present paper provided data reference for further explaining the pathogenic mechanism and immune response mechanism of FAdV-4 infection. [Method] FAdV-4 X gene coding sequence (CDS) was amplified by PCR, and linked to pEFl α-HA vector, and the recombinant expression plasmid pEFlα-HA-X was constructed. The recombinant expression plasmid pEFlα-HA-X and empty vector pEFlα-HA-were transfected into LMH cells respectively, after 24 hours, the transfected cells were stimulated with FAdV-4, and the treatments were marked as pEFlα-HA-X+ and pEFlα-HA+, without virus stimulation after transfection was set up as control and marked as pEFlα-HAX- and pEFlα-HA-. Western-blotting and indirect immunofluorescence were used to verified expression of recombinant protein X, real-time florescence quantitative PCR assays was used to detected the mRNA expression levels of Toll-like receptors (chTLRla, chTLRlb, chTLR2a, chTLR2b, chTLRJ, chTLR4, chTLR5, chTLR7, chTLR15, chTLR21) and effect factors (JFN-α, INF-β, IL-1β, IL-6, IL-8, IL-15). [Result] The open reading frame (ORF) of X gene was 540 hp, encoding 179 amino acids residues. Western-blotting showed that recombinant X protein reacted with HA tag mouse-derived monoclonal antibody with a single band at 27 kD and green fluorescence visualized by indirect immunofluorescence. Real-time florescence quantitative PCR showed that, compared with pEFlα-HA-group the mRNA transcription levels of chTLRla and chTLRl b in pEFl α-HA-X-group were extremely significantly up-regulated (P < 0. 01, the same as below), as 9. 76 and 12. 34 times as that of pEFl α-HA-group, mRNA transcription levels of chTLR2a, chTLR2b, chTLR5, chTLR7, chTLRl 5 and chTLR21 were significantly up-regulated (P < 0. 05, the same as below). After virus infection, the mRNA transcription levels of chTLRl a and chTLRl b in pEFlα-HA-X+ group were extremely significantly down-regulated by 60. 0% and 66. 7% compared with pEFlα-HA-X-group, and the mRNA transcription levels of chTLR2a, chTLR2b and chTLRJ were significantly increased by 2. 91, 2. 01 and 1. 47 times respectively, the transcription levels of other chTLRs were down-regulated but not significantly ( P > 0. 05). At the same time, the mRNA transcription levels of IFN-α, IFN-β, IL-1β in pEFlα-HA-X + group were significantly up-regulated compared with pEFlα-HA-X-group, mRNA transcription level of IL-8 was extremely significantly down-regulated. [Conclusion] Overexpression of pX in LMH cells and after infection with FAdV-4, pX can activate host immune response system, mRNA transcription level of Toll-like receptors (chTLR2a, chTLR2b, chTLRJ) and effector factors (JFN-α, INF-β, IL-1β) significantly up-regulate to inhibit viral replication, indicating that pX has the function of activating the natural immune system of host cells. [ABSTRACT FROM AUTHOR]

【Objective】The study aimed to explore the influence of fowl adenovirus serotype 4(FAdV-4) protein Ⅷ(pⅧ) on expression innate immune-associted genes of host, providing a reference for further study of the pathogenesis of FAdV-4.【Method】FAdV-4Ⅷ gene coding sequence(CDS) was amplified by PCR,linked to pEF1α-HA eukaryotic expression vector, and recombinant expression plasmid pEF1α-HA-Ⅷ was constructed. Recombinant expression plasmid pEF1α-HA-Ⅷ(experimental group) and empty plasmid pEF1α-HA(control group) were transfected into LMH cells, and the expression of protein Ⅷ was verified by Western blotting and indirect immunofluorescence, real-time florescence quantitative PCR was used to detected the expression levels of STING, MDA5, TBK1, MAVS, NF-κB, IRF7, IFN-α, INF-β, IL-6, IL-8, IL-15, IL-1β.【Result】The open reading frame(ORF) length of Ⅷ gene was 744 bp, encoding 247 amino acid residues, recombinant protein Ⅷ can react with HA-tag monoclonal antibody from mouse, a single band was obtained at 34 kD, and green fluorescence was observed by immunofluorescence detection, indicating that recombinant protein Ⅷ was well expressed in LMH cells. Compared with the control group, after the recombinant protein Ⅷ was expressed in LMH cells, the expression level of the mode receptor STING gene was significantly increased by 5.6 times(P＜0.05, the same as below), and the expression level of MDA5 gene was increased by 30%. The gene expression levels of adaptor protein TBK1 and MAVS genes were significantly increased by 1.5 and 2.4 times compared with the control group, respectively. The level of transcription factor IRF7 was significantly increased by 3.4 times, and the expressions of NF-κB genes were suppressed. Meanwhile, the expression levels of interferon(IFN-α and IFN-β) and interleukin(IL-6, IL-8, IL-15 and IL-1β) were up-regulated, and IFN-α and IFN-β were extremely significantly up-regulated 19.2 and 14.4 times(P＜0.01), respectively. IL-6, IL-8, IL-15 and IL-1β were significantly up-regulated 4.1, 2.1, 6.0 and 7.9 times, respectively.【Conclusion】Overexpression of FAdV-4 pⅧ leads to up-regulated expression of STING, MDA5, TBK1, MAVS, IRF7, IFN-α,IFN-β, IL-6, IL-8, IL-15 and IL-1β in LMH cells. The results indicate that protein Ⅷ can promote the natural immunity of host stimulated by FAdV-4 to a certain extent. [ABSTRACT FROM AUTHOR]

【Objective】The paper aimed to express the NS1 and VP2 proteins of chicken parvovirus（ChPV） in prokaryotes and prepare polyclonal antibodies against them, which would provide material basis for further study of the protein structure and function of chicken parvovirus（ChPV） and provide materials for the establishment of serological diagnostic methods. 【Method】In the study, the sequences of NS1 and VP2 gene of chicken parvovirus GX-Ch-PV-21 strain were synthesized. The prokaryotic recombinant expression vectors pET-32a-NS1 and pET-32a-VP2 were constructed, and the correct recombinant plasmids were transformed into transsetta competent cells for prokaryotic expression. The expression conditions were optimized, and the expressions of NS1 and VP2 proteins were identified by SDS-PAGE and Western-blotting. The expression products were immunized to SPF chickens to obtain polyclonal antibodies. The titer of chicken polyclonal antibodies was determined by enzyme linked immunosorbent assay（ELISA）,and the polyclonal antibodies were identified by Western-blotting and indirect immunofluorescence assay（IFA）. 【Result】SDS-PAGE analysis showed that the size of NS1 and VP2 protein bands was about 104 and 87 kD,respectively, which was consistent with the expected size. The optimal induction conditions were screened for the recombinant plasmids pET-32a-VP2 and pET-32a-NS1. The results showed that the optimal induction conditions for the expression of NS1 protein were as follows: The final concentration of isopropyl 1-thio-β-D-galactopyranoside（IPTG） was 1. 0 mmol/mL and the expression was induced overnight at 16 ℃. The optimal conditions for the induction of VP2 protein expression were as follows: The final IPTG concentration was 0. 8 mmol/mL and the expression was induced overnight at 20 ℃. The soluble analysis showed that both NS1 and VP2 proteins mainly existed in the form of inclusion bodies. The titers of polyclonal antibodies against NS1 and VP2 by indirect ELISA were 1∶10 240 000. Western-blotting analysis showed that His tag protein and its corresponding chicken polyclonal antibody could specifically bind to NS1 and VP2 proteins. The IFA analysis showed that both VP2 and NS1 polyclonal antibodies could specifically bind to ChPV. 【Conclusion】In the study, the NS1 and VP2 proteins of chicken parvovirus were successfully expressed, and the chicken anti-polyclonal antibody was successfully prepared, which provided material basis for further study of the structure and function of chicken parvovirus protein and materials for the establishment of serological diagnostic methods. [ABSTRACT FROM AUTHOR]