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.

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 .

OBJECTIVE：To e stablish the method for simultaneous determination of 10 kinds of active components in Tibetan medicine Siwei jianghuang prescription ，and to optimize its decoction technology. METHODS ：HPLC method was adopted. Using soaking time ，the amount of added water ，decoction time and decoction times as factors ，comprehensive score of the contents of 10 kinds of components and solid extracts rate as response values ，one the basis of single factor test ，Box-Behnken response surface method was used to optimize its decoction technology. RESULTS ：The linear range of gallic acid ，corilagin，magnoflorine， ellagic acid ， hydrochloric jatrorrhizine ， hydrochloride palmatine ， hydrochloride berberine ， bisdemethoxycurcumin， demethoxycurcumin and curcumin were 0.280 6-1.683 6，0.289 6-1.737 6，0.320 8-1.924 8，0.116 0-0.696 0，0.018 9-0.113 5， 0.013 3-0.079 9，0.092 3-0.553 8，0.025 5-0.153 0，0.036 1-0.216 3，0.041 0-0.245 7 µg（all r were 0.999 9），respectively. The limits of quantitation were 0.28，14.48，3.21，11.60，1.89，4.44，0.46，0.26，0.36，0.41 ng，respectively. The limits of detection were 0.11，4.14，1.24，3.32，0.58，1.33，0.13，0.09，0.14，0.12 ng，respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％. The recoveries were 92.56％-103.69％（RSDs were 0.90％-3.81％，n＝6）. The optimal decoction technology included soaking 60 min，adding 8-fold（mL/g）water，decoction for twice ，lasting for 65 min each time. In 6 validation tests ，comprehensive scores were 3.323 2-3.422 4，and the absolute value of the relative error with the predicted value （3.437 4）was less than 2％.CONCLUSIONS：Established method is simple and repeatable ，and can be used for simultaneous determination of 10 kinds of active components in Siwei jianghuang prescription. Optimized decoction technology is stable and feasible.