Objective To investigate the mechanism by which AGE receptor(RAGE)antagonist FPS-ZM1 inhibits AGE accumulation and promotes wound repair in rats with diabetic foot ulcer(DFU).Methods A total of 30 SD rats were divided into normal control(Con)group,sham operation group(Sham),DFU group,FPS-ZM1 treatment group(DFU+FPS-ZM1),and phosphate buffer saline treatment group(DFU+PBS),with 6 rats in each group.Masson staining was used to evaluate the wound surface structure.ELISA was used to detect the expression of AGE,TNF-α,matrix metalloproteinase 9(MMP-9),and VEGF proteins in serum and tissues,and Western blot method was used to test the expression of RAGE,VEGF receptor(VEGFR),CD31,and nuclear factor κB(NF-κB)proteins.Results Compared with the Con and Sham groups,the collagen fibers had less indigo staining,disordered arrangement and sparse distribution in DFU group.Compared with the DFU and DFU+PBS groups,the DFU+FPS-ZM1 group showed obvious blue staining of collagen fibers in the wound granulation tissue,and the number of deposition layers was relatively neat and orderly.Compared with Sham group,the expressions of AGE,TNF-α and MMP-9 proteins,NF-κB and RAGE proteins in serum and tissues were increased(P<0.05),and the expressions of VEGF protein and CD31 and VEGFR proteins in serum and tissues were decreased in DFU group(P<0.05).Compared with DFU group,the expressions of AGE,TNF-α and MMP-9 proteins,NF-κB and RAGE proteins in serum and tissues decreased(P<0.05),and the expression of VEGF protein and CD31 and VEGFR proteins in serum and tissues increased in DFU+FPS-ZM1 group(P<0.05).Compared with DFU+FPS-ZM1 group,the expressions of AGE,TNF-α and MMP-9 proteins,NF-κB and RAGE proteins in serum and tissues increased(P<0.05),and the expressions of VEGF protein,CD31 and VEGFR proteins in serum tissues decreased in DFU+PBS group(P<0.05).Conclusions The RAGE antagonist FPS-ZM1 down-regulates MMP-9 and up-regulates VEGF by inhibiting the NF-κB inflammatory signaling pathway,and promotes DFU wound healing.

Objective To investigate the mechanism by which AGE receptor(RAGE)antagonist FPS-ZM1 inhibits AGE accumulation and promotes wound repair in rats with diabetic foot ulcer(DFU).Methods A total of 30 SD rats were divided into normal control(Con)group,sham operation group(Sham),DFU group,FPS-ZM1 treatment group(DFU+FPS-ZM1),and phosphate buffer saline treatment group(DFU+PBS),with 6 rats in each group.Masson staining was used to evaluate the wound surface structure.ELISA was used to detect the expression of AGE,TNF-α,matrix metalloproteinase 9(MMP-9),and VEGF proteins in serum and tissues,and Western blot method was used to test the expression of RAGE,VEGF receptor(VEGFR),CD31,and nuclear factor κB(NF-κB)proteins.Results Compared with the Con and Sham groups,the collagen fibers had less indigo staining,disordered arrangement and sparse distribution in DFU group.Compared with the DFU and DFU+PBS groups,the DFU+FPS-ZM1 group showed obvious blue staining of collagen fibers in the wound granulation tissue,and the number of deposition layers was relatively neat and orderly.Compared with Sham group,the expressions of AGE,TNF-α and MMP-9 proteins,NF-κB and RAGE proteins in serum and tissues were increased(P<0.05),and the expressions of VEGF protein and CD31 and VEGFR proteins in serum and tissues were decreased in DFU group(P<0.05).Compared with DFU group,the expressions of AGE,TNF-α and MMP-9 proteins,NF-κB and RAGE proteins in serum and tissues decreased(P<0.05),and the expression of VEGF protein and CD31 and VEGFR proteins in serum and tissues increased in DFU+FPS-ZM1 group(P<0.05).Compared with DFU+FPS-ZM1 group,the expressions of AGE,TNF-α and MMP-9 proteins,NF-κB and RAGE proteins in serum and tissues increased(P<0.05),and the expressions of VEGF protein,CD31 and VEGFR proteins in serum tissues decreased in DFU+PBS group(P<0.05).Conclusions The RAGE antagonist FPS-ZM1 down-regulates MMP-9 and up-regulates VEGF by inhibiting the NF-κB inflammatory signaling pathway,and promotes DFU wound healing.

ObjectiveTo investigate the effect of Xinfeng capsules on immunoinflammatory indicators in patients with rheumatoid arthritis （RA） due to spleen deficiency and dampness exuberance. MethodA total of 102 patients were randomly divided into control group and observation group according to the random number table method， with 51 cases in each group. All patients were treated with methotrexate tablets， while those in the observation group received additional Xinfeng capsules. The course of treatment in both groups was 12 weeks. The 28-joint disease activity score （DAS28）， visual analogue scale （VAS） scores， morning stiffness time， erythrocyte sedimentation rate （ESR）， high-sensitivity C-reactive protein （hs-CRP）， rheumatoid factor （RF）， anti-cyclic citrullinated peptide （anti-CCP） antibody， vascular endothelial growth factor （VEGF）， and serum amyloid A （SAA） of the two groups before and after treatment were compared. The efficacy and incidence of adverse events were compared between the two groups. The Apriori association rule model and random walk model were constructed to evaluate the effect of Xinfeng capsules in improving hs-CRP， ESR， RF， SAA， VEGF， and anti-CCP. ResultThere were no dropouts in this study. There was no statistical difference in the indicators between the two groups before treatment. After 12 weeks of treatment， the total effective rate in the observation group was 90.19% （46/51）， which was higher than 74.51% （38/51） in the control group （χ²＝4.320，P<0.05）. DAS28， VAS score， and morning stiffness time in the observation group were improved compared with those in the control group （P<0.05）. Apriori association rule model results showed that the application of Xinfeng capsules in the observation group had a strong correlation with the reduction of RF， ESR， hs-CRP， SAA， and VEGF. The results of the random walk model showed that the improvement coefficients of hs-CRP， ESR， RF， SAA， and VEGF in the observation group were all better than those of the control group， and the improvement coefficient of anti-CCP in the control group was better than that of the observation group. The improvement degree of hs-CRP， ESR， RF， SAA， and VEGF in the observation group was superior to that of the control group （P<0.05）. The incidence of adverse reactions in the observation group was lower than in the control group （χ²＝4.057，P<0.05）. ConclusionOn the basis of the treatment with methotrexate tablets， Xinfeng capsules can effectively improve the immunoinflammatory level in RA due to spleen deficiency and dampness exuberance and reduce the incidence of adverse reactions.

Objective Using network pharmacology to explore the mechanism of action of tripterygium wilfordii-white peony in the treatment of rheumatoid arthritis(RA)and validating it using an adjuvant arthritis rat model.Methods Obtain the effective ingredients of tripterygium wilfordii and white peony through TCMSP database and predict the action targets;Obtain the disease targets of RA using DisGeNET and GeneCards online database;Draw the network of"tripterygium wilfordii-white peony-active ingredient-RA"and screen the core effective components using the software Cytoscape 3.9.1;Construct the protein-protein interaction(PPI)network using String database and screen the core targets using the CytoNCA plug-in,using the Metascape database for gene enrichment analysis of common targets,and using Auto Dock Tools and Pymol software for molecular docking of core active ingredients and targets.Enzyme linked immunosorbent assay(ELISA)detection of serum tumor necrosis factor-α(TNF-α),interleukin(IL)-4,IL-6,and IL-17 levels in adjuvant arthritis rats,Western blot was used to detect the expression of phosphorylated phosphatidylinositol 3-hydroxy kinase(p-PI3K)and phosphorylated AKT protein(p-AKT),validate the results of network pharmacology analysis.Results There were a total of 61 effective ingredients in the treatment of RA with tripterygium wilfordii-white peony,with 116 targets and 191 signaling pathways involved.Animal experiments have shown that compared with the model group rats,the serum cytokine TNF-α,IL-6 and IL-17 of rats in tripterygium wilfordii-white peony group and tripterygium wilfordii extract group were significantly decreased(P<0.05),IL-4 significantly increased(P<0.05);The expression of p-PI3K and p-AKT in synovial tissue was significantly reduced(P<0.05);Compared with rats in tripterygium wilfordii extract group,the TNF-α,IL-6,IL-17,p-PI3K and p-AKT of rats in tripterygium wilfordii-white peony group were significantly reduced(P<0.05),while IL-4 was significantly increased(P<0.05).Conclusion Tripterygium wilfordii-white peony can inhibit the overexpression of phosphatidylinositol 3-hydroxy kinase(PI3K)/protein kinase B(AKT)signaling pathway,regulate cytokine release,exert anti-inflammatory effects,and ultimately treat RA through multiple active ingredients and targets.

Objective:To explore the long-term preservation value and repair effect of normothermic machine perfusion (NMP) on clinically discarded kidneys.Methods:A case of clinical discarded donor kidney was collected, and NMP was carried out in vitro for 9 hours with recovered blood. The dynamic changes of renal appearance, blood gas and biochemistry analysis of perfusate and renal pathology were recorded. Results:In the second to fifth hour of NMP, the appearance of renal was pink and ex vivo normothermic perfusion assessment score (EVNP) was grade Ⅰ. While, the sixth hour and beyond of NMP, the appearance of kidney turned to dark red and EVNP was grade Ⅲ. The renal perfusion blood flow maintained above 150 ml/min in the first 6 hours and decreased significantly after that, and at the end, was only 50 ml/min. During the whole process of perfusion, urine output was maintained at about 100 ml/h. PO 2 remained above 100 mmHg in the first 5 hours of perfusion and from the 6th hour, was lower than 80 mmHg and continued to decline, and was close to 0 at the end of perfusion. The results showed that although the K + concentration changes in blood and urine in the first 5 hours of NMP had a good consistency, the lactic acid level had been rising. In addition, there was no significant change in the histopathology at the fourth hour of perfusion compared with that before zero-point puncture, and the fibrinous thrombus in glomeruli was improved compared with that before perfusion. However, at the sixth hour after perfusion and before the end of perfusion, the pathological changes of renal tissue were significantly worse. There were a large of thrombosis in glomerular blood vessels, renal tubular atrophy and acute tubular necrosis. Conclusions:NMP can realize the evaluation of extended criteria donors before transplantation, and it proves the feasibility and repair potential of NMP in kidney to a certain extent. At the same time, NMP also provides a new way to expand the source of donor kidney and to pre-treat organ in vitro.