Objective To observe the effect of supplemental Revolvin D1 (RvD1) on Toll-like receptor 4 (TLR4) in skeletal muscle of type 2 diabetic mice.Methods 35 male C57BL/6 mice were randomly divided into control group (NC group) and high glucose and high fat diet group.After 8 weeks,mice in high glucose and high fat diet group were given intraperitoneal injection of streptozotocin (STZ) 100 mg/ kg.Then they were randomly divided into two groups:Type 2 diabetes group (T2DM group) and type 2 diabetes + RvD1 intervention group (T2DM + RvD1 group).Mice in T2DM group mice were injected with phosphate buffer saline 0.2 ml and T2DM + RvD1 group mice were injected with Revolvin D1 100 ng/day respectively.The levels of fasting blood glucose,serum insulin and inflammatory factors were detected.The mRNA expression level of TLR4 was detected by real-time quantitative polymerase chain reaction (RT-qPCR) method,and the expression of TLR4 protein was detected by Western blot.Results The levels of insulin resistance index,interleukin(IL)-6 and tumor necrosis factor-α (TNF-α) in T2DM group and T2DM + RvD1 group increased (P ＜ 0.05).Compared with the T2DM group,the levels of insulin resistance index,IL-6 and TNF-α in T2DM + RvD1 group decreased (P ＜0.05).The expression of TLR4 protein in T2DM group and T2DM + RvD1 group was higher than that in NC group (P ＜ 0.05).The expression of TLR4 protein in T2DM + RvD1 group,was lower than that in T2DM group (P ＜0.05).The mRNA level of TLR4 in mice was consistent with the above results by RT-qPCR.Conclusions Moderate supplementation of RvD1 can not only decrease the level of inflammatory factors in type 2 diabetic mice,but also reduce the expression of TLR4 and insulin resistance in skeletal muscle of type 2 diabetic mice.

Objective:To investigate the clinical effects of perforator flaps in the repair of limb defects.Methods:Anterolateral thigh perforator flap (ALTP), deep inferior epigastric perforator flap (DIEP), posterior in- terosseous artery perforator flap (PIAP), thoracodorsal artery perforator flap (TDAP), medial sural artery perforator flap (MSAP) and radial collateral artery perforator flap (RCAP) were used to repair 56 cases of limb wounds from Novem- ber, 2016 to October, 2018. The sizes of soft tissue defect were ranged from 1.5 cm×1.5 cm to 10.0 cm×24.0 cm, while the sizes of flap were ranged from 2.0 cm×2.0 cm to 11.0 cm×25.0 cm. The recipient sites were all sutured di- rectly. The flaps survived and wound healing were observed postoperatively. Appearance, texture, recovery of the limb function, shape and function of donor site were followed-up regularly.Results:One case of venous crisis occurred on the 3rd day after inferior epigastric artery perforator flap surgery, and survived after exploratory surgery with the necrosis in the distal part of the flap, which healed after dressing. Other 55 flaps all survived. Both recipient site and donor site healed in primary union. The color of the flaps was normal. The skins were soft, and the functions of the limbs recovered well. Only a linear scar remained in the donor area without dysfunction.Conclusion:The perforator flap surgery is reliable for limbs repair and can be promoted and applied.

Objective To investigate the mechanism of Yes-associated protein 1(YAP1)ameliorating aristolochic acid 1(AAI)-induced liver injury in mice based on untargeted metabolomics techniques.Methods There were 83-week-old male hepatocyte-specific Yap1 gene knockout mice(genotyped as Yap1Flox/Flox,Albumin-Cre,aka.Yap1LKO)were randomly selected as the Yap1LKO+AAI group,and 8 Yap1Flox control mice as the Yap1Flox+AAI group.Both groups were injected intraperitoneally with AAI at a dose of 2.5 mg·kg-1·d-1 for 14 consecutive days.Genotypes were identified by tail PCR;serum alanine transaminase(ALT)and aspartate transaminase(AST)activities were determined by microplate assay;histopathological changes of liver tissue were observed by HE staining;and the protein expression of YAP1 in liver tissue was determined by immunohistochemistry.The untargeted metabolomics approach was used to analyze the liver tissue differential metabolites,and the samples were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbit trap high-resolution mass spectrometry,and the differential metabolites were screened by principal component analysis(PCA),Partial least square-discriminant analysis(PLS-DA),and orthogonal partial least squares-discriminant analysis(OPLS-DA);using HMDB database and METLIN database to identify metabolites,and the pathway enrichment of differential metabolites was analyzed by KEGG database.Results(1)After 14 days of AAI induction,the increase of body mass in Yap1LKO mice was lower than that in Yap1Flox mice,but there was no statistical significance(P＞0.05).On day 14,compared with the Yap1Flox+AAI group,the serum ALT and AST enzyme activities in the Yap1LKO+AAI group of mice were significantly increased(P＜0.05),and the histopathological damage of the liver was significantly aggravated.The livers of the Yap1Flox mice had a positive protein expression of YAP1,whereas the Yap1LKO mice did not have a positive protein expression of YAP1.(2)A total of 139 differential metabolites with significant changes(VIP＞1 and P＜0.05)were screened by metabonomic analysis;compared with Yap1LKO+ AAI group,62 liver metabolites in Yap1Flox+AAI group were up-regulated,including choline,taurine,hypotaurine,α-linolenic acid,eleostearic acid,chenodeoxycholic acid and so on.Seventy-seven metabolites were down-regulated including glycerophosphocholine,L-phosphatidylcholine,L-glutamine,L-serine,L-glutathione,5-methionine,phenylalanine,glucose 6-phosphate,lactic acid,uric acid glycosides,etc..KEGG-enriched pathways were mainly choline metabolism,glycerophospholipid metabolism,insulin resistance,glutathione metabolism,etc..Conclusion Hepatocyte-specific Yap1 gene knockout exacerbated AAI-induced liver injury in mice,and YAP1 was involved in the regulation of choline metabolism and glycerophospholipid metabolism through the up-regulation of unsaturated fatty acids,such as choline and taurine,which ameliorated AAI-induced liver injury in mice.

The effect of anti-programmed cell death 1 (anti-PD-1) immunotherapy is limited in patients with hepatocellular carcinoma (HCC). Yes-associated protein 1 (YAP1) expression increased in liver tumor cells in early HCC, and Akkermansia muciniphila abundance decreased in the colon. The response to anti-PD-1 treatment is associated with A. muciniphila abundance in many tumors. However, the interaction between A. muciniphila abundance and YAP1 expression remains unclear in HCC. Here, anti-PD-1 treatment decreased A. muciniphila abundance in the colon, but increased YAP1 expression in the tumor cells by mice with liver tumors in situ. Mechanistically, hepatocyte-specific Yap1 knockout (Yap1^LKO) maintained bile acid homeostasis in the liver, resulting in an increased abundance of A. muciniphila in the colon. Yap1 knockout enhanced anti-PD-1 efficacy. Therefore, YAP1 inhibition is a potential target for increasing A. muciniphila abundance to promote anti-PD-1 efficacy in liver tumors. Dihydroartemisinin (DHA), acting as YAP1 inhibitor, increased A. muciniphila abundance to sensitize anti-PD-1 therapy. A. muciniphila by gavage increased the number and activation of CD8⁺ T cells in liver tumor niches during DHA treatment or combination with anti-PD-1. Our findings suggested that the combination anti-PD-1 with DHA is an effective strategy for liver tumor treatment.