1.The role of adeno-associated virus-mediated knockdown of OTUD6B in mouse liver glucose metabolism
Yunan SHI ; Kai WANG ; Tingchao TAN ; Xiang WANG ; Dongming LIU ; Xiuyun WANG
Acta Universitatis Medicinalis Anhui 2026;61(9):1505-1512
ObjectiveTo investigate the knockdown efficiency of the liver-specific knockdown recombinant adeno-associated virus (AAV) thyroid-binding globulin (TBG) (AAV-TBG-shOTUD6B) targeting the ovarian tumor domain 6B (OTUD6B) of the ubiquitin-proteasome system in the liver of C57BL/6J mice and its regulatory effect on liver glucose metabolism. MethodsAn effective siOTUD6B sequence was designed and verified. The DNA sequences of non-specific interference RNA negative control (shNC) and shOTUD6B were inserted into the AAV vector backbone containing the promoter TBG to construct and purify AAV-TBG-shNC and AAV-TBG-shOTUD6B viruses. AAV-TBG-shNC and AAV-TBG-shOTUD6B were injected into mice via the tail vein to target the liver. After 8 weeks of high-fat diet (HFD) feeding, the levels of OTUDB protein and mRNA in the mice liver, as well as blood glucose levels, glucose tolerance, insulin sensitivity, and protein expression levels of phosphorylated protein kinase B (p-AKT) and phosphorylated glycogen synthase kinase-3β (p-GSK-3β) were detected. ResultsCompared with the AAV-TBG-shNC group, the OTUD6B protein level in the liver of the AAV-TBG-shOTUD6B group decreased, the mRNA level of OTUD6B in the liver decreased (P<0.01), the fasting blood glucose levels of the mice after 4 hours of fasting and 16 hours of fasting both decreased (P<0.05, P<0.01), the protein level of p-GSK-3β in the liver decreased (P<0.05), glucose tolerance was enhanced (P<0.05), insulin sensitivity was enhanced (P<0.01), and the protein level of p-AKT in the liver increased (P<0.05). ConclusionThe construction of a recombinant adeno-associated virus for liver OTUD6B knockdown is successfully completed. OTUD6B plays an important role in regulating the homeostasis of liver glucose metabolism. Liver-specific knockdown of OTUD6B can improve the blood glucose levels of mice induced by HFD and enhance insulin sensitivity.
2.Analysis of differential expression of lipid metabolism-related proteins in adipose tissue exosomes from obese mice
Xinzhe Zhuo ; Kai Wang ; Yunan Shi ; Xiang Wang ; Jiao Yu ; Jiali Li ; Yidan Liu ; Xiuyun Wang
Acta Universitatis Medicinalis Anhui 2025;60(11):2069-2075
Objective:
To analyze differences in the expression levels of lipid metabolism-related proteins in adi- pose tissue exosomes between obese mice and wild-type mice using proteomic techniques .
Methods:
Wild-type (WT) and obese (ob/ob) model mice of the same age were selected , with 8 mice per group . Adipose tissue from both groups was minced and cultured for 48 hours . Conditioned media was collected , and exosomes were isolated u- sing differential centrifugation . Liquid chromatography-tandem mass spectrometry ( LC-MS/MS) was utilized for proteomic analysis to screen differentially expressed proteins ( DEPs) . Gene ontology ( GO) enrichment analysis and kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed on the DEPs . Heatmaps were generated to visualize DEPs expression patterns , and Western blot was employed to validate DEPs expression levels .
Results:
Exosomes were successfully extracted from the culture supernatant of adipose tissues from mice in the WT group and the ob/ob group . Mass spectrometry analysis identified a total of 25 629 peptides and 3 376 proteins . Compared with the WT group , there were 699 proteins with high expression and 632 proteins with low expression in the exosomes derived from adipose tissues of ob/ob mice . Both GO and KEGG analyses showed that DEPs were mainly enriched in metabolic pathways . Heatmap analysis visualized the expression patterns of metabolism-related DEPs , and the expression levels of lipid metabolism-related proteins such as acyl-CoA syn- thetase long-chain family member 1 (ACSL1) , apolipoprotein E (ApoE) , and albumin (Alb) changed significant- ly in the obese state (P < 0. 05) . Western blot verification results showed that the expression of ApoE and Alb pro- teins in the adipose tissue-derived exosomes of ob/ob mice decreased ( P < 0. 01) .
Conclusion
In the obese state , the expression levels of lipid metabolism-related proteins in mouse adipose tissue exosomes are significantly altered . These differentially expressed proteins may thus serve as potential molecular targets for treating obesity and its associated metabolic complications .
3.Analysis of pathogenic bacteria and its drug resistance in chronic suppurative otitis media
Yuzhi WANG ; Yumei WU ; Zongzhen ZHANG ; Fumin ZHA ; Linhong TIAN ; Xiuyun XIANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(06):-
OBJECTIVE To study the pathogenic bacteria and its resistance to antibiotics in chronic suppurative otitis media. METHODS Secretion from 132 cases with chronic suppurative otitis media were cultured. Drug sensitivity tests were done to separate pathogenic bacteria. RESULTS 152 strains pathogenic bacteria were isolated from middle ear secretion. The main pathogenic bacteria were staphylococcus aureus (49 strains), pseudomonas aeruginosa (25 strains) and fungus (5 strains). Staphylococcus aureus was sensitive to fluorquinolones, and the sensitivity rate was 79.6 %. Pseudomonas aeruginosa was sensitive to fluoroquinolones and floxacin, and the sensitivity rates were 84 % and 64 %. CONCLUSION Staphylococcus aureus and pseudomonas aeruginosa are main pathogens of chronic suppurative otitis media in our region. Both of them are sensitive to fluorquinolones. Meanwhile, pseudomonas aeruginosa is also sensitive to ceftazidime of the third cephalosporins. So doctors should choose the sensitive antibiotics for treatment of the chronic suppurative otitis media, that may avoid the bacteria with drug resistance come into being. The fungus can not be neglected as the pathogen in chronic suppurative otitis media.


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