Objective:To investigate expression of IL-36 in ulcerative colitis(UC),and to assess regulatory activity of IL-36 receptor antagonist(IL-36Ra)to monocytes function in UC patients.Methods:Seventy-two UC patients and twenty controls were enrolled between April 2021 and January 2023 in the First Affiliated Hospital of Xinxiang Medical University.Plasma and peripheral blood mononuclear cells(PBMCs)were isolated.Tissue infiltrating lymphocytes(TILs)were isolated from UC tissue and normal tis-sue.Monocytes from PBMCs and TILs were purified.Plasma IL-36α,IL-36β,IL-36γ and IL-36Ra levels were measured by ELISA.mRNA levels of IL-36 in tissue and IL-36 receptor subunits in monocytes were semi-quantified by real-time PCR.Purified monocytes were stimulated with recombinant IL-36Ra and co-culture with CaCO2 cells.Percentage of CaCO2 cell death was measured.Serum TNF-α,granzyme B and granzyme H levels were measured by ELISA.Mean fluorescence intensity(MFI)of Fas ligand(FasL)and TNF-related apoptosis-inducing ligand(TRAIL)in monocytes was measured by flow cytometry.Results:There were no significant differences of plasma IL-36α,IL-36β or IL-36γ levels between UC group and control group(P>0.05).Plasma IL-36Ra levels were lower in UC group compared with control group[(87.33±11.95)pg/ml vs(100.81±15.62)pg/ml,t=4.162,P<0.001].There were no significant differences of IL-36α,IL-36β or IL-36γ mRNA levels between UC tissue and normal tissue(P>0.05).IL-36Ra mRNA level was remarkably reduced in UC tissue compared with normal tissue(1.00±0.23 vs 1.36±0.21,t=5.500,P<0.001).There were no signifi-cant differences of IL-36 receptor subunits mRNA relative levels between UC group and control group,as well as between UC tissue and normal tissue(P>0.05).Peripheral monocytes-induced CaCO2 cell death was higher in UC group compared with control group[(14.45±3.33)%vs(10.92±2.87)%,t=2.965,P=0.005].Monocyes purified from UC tissue also mediated increased CaCO2 cell death compared with those from normal tissue[(17.41±4.19)%vs(14.31±3.02)%,t=2.327,P=0.027].TNF-α,granzyme B and granzyme H levels in supernatants were significantly elevated(P<0.05).However,there were no remarkable differences of FasL or TRAIL MFI in monocytes between UC group and control group,as well as between UC tissue and normal tissue(P>0.05).Recombi-nant IL-36Ra stimulation reduced CaCO2 cell death was decreased in both monocytes purified from peripheral blood and UC tissue in UC patients(P<0.05),TNF-α,granzyme B and granzyme H levels in supernatants were also down-regulated(P<0.05).There were no remarkable differences of FasL or TRAIL MFI in monocytes between cells with and without IL-36Ra stimulation(P>0.05).Conclu-sion:IL-36Ra may suppress monocytes cytotoxicity through down-regulation of TNF-α and granzyme secretions in UC patients.

Objective:To investigate expression of IL-36 in ulcerative colitis(UC),and to assess regulatory activity of IL-36 receptor antagonist(IL-36Ra)to monocytes function in UC patients.Methods:Seventy-two UC patients and twenty controls were enrolled between April 2021 and January 2023 in the First Affiliated Hospital of Xinxiang Medical University.Plasma and peripheral blood mononuclear cells(PBMCs)were isolated.Tissue infiltrating lymphocytes(TILs)were isolated from UC tissue and normal tis-sue.Monocytes from PBMCs and TILs were purified.Plasma IL-36α,IL-36β,IL-36γ and IL-36Ra levels were measured by ELISA.mRNA levels of IL-36 in tissue and IL-36 receptor subunits in monocytes were semi-quantified by real-time PCR.Purified monocytes were stimulated with recombinant IL-36Ra and co-culture with CaCO2 cells.Percentage of CaCO2 cell death was measured.Serum TNF-α,granzyme B and granzyme H levels were measured by ELISA.Mean fluorescence intensity(MFI)of Fas ligand(FasL)and TNF-related apoptosis-inducing ligand(TRAIL)in monocytes was measured by flow cytometry.Results:There were no significant differences of plasma IL-36α,IL-36β or IL-36γ levels between UC group and control group(P>0.05).Plasma IL-36Ra levels were lower in UC group compared with control group[(87.33±11.95)pg/ml vs(100.81±15.62)pg/ml,t=4.162,P<0.001].There were no significant differences of IL-36α,IL-36β or IL-36γ mRNA levels between UC tissue and normal tissue(P>0.05).IL-36Ra mRNA level was remarkably reduced in UC tissue compared with normal tissue(1.00±0.23 vs 1.36±0.21,t=5.500,P<0.001).There were no signifi-cant differences of IL-36 receptor subunits mRNA relative levels between UC group and control group,as well as between UC tissue and normal tissue(P>0.05).Peripheral monocytes-induced CaCO2 cell death was higher in UC group compared with control group[(14.45±3.33)%vs(10.92±2.87)%,t=2.965,P=0.005].Monocyes purified from UC tissue also mediated increased CaCO2 cell death compared with those from normal tissue[(17.41±4.19)%vs(14.31±3.02)%,t=2.327,P=0.027].TNF-α,granzyme B and granzyme H levels in supernatants were significantly elevated(P<0.05).However,there were no remarkable differences of FasL or TRAIL MFI in monocytes between UC group and control group,as well as between UC tissue and normal tissue(P>0.05).Recombi-nant IL-36Ra stimulation reduced CaCO2 cell death was decreased in both monocytes purified from peripheral blood and UC tissue in UC patients(P<0.05),TNF-α,granzyme B and granzyme H levels in supernatants were also down-regulated(P<0.05).There were no remarkable differences of FasL or TRAIL MFI in monocytes between cells with and without IL-36Ra stimulation(P>0.05).Conclu-sion:IL-36Ra may suppress monocytes cytotoxicity through down-regulation of TNF-α and granzyme secretions in UC patients.

Objective To investigate the effect of thalidomide combined with infliximab (IFX) in treatment of refractory inflammatory bowel disease (IBD) and its effects on insulin-like growth factor-1 (IGF-1) and transforming growth factor-β1 (TGF-β1). Methods A total of 120 patients with refractory IBD were randomly divided into experimental group and control group, with 60 cases in each group. The two groups were given conventional treatment (mesalazine), the control group was given IFX, and the experimental group was given IFX combined with thalidomide, continuous treatment for two months. The efficacy, intestinal flora disturbance rate, adverse reactions, Crohn's disease activity index (CDAI), Lewis score, serum IGF-1, TGF-β1 levels and nutritional status indexes[albumin (ALB), transferrin (Tf)]before and after treatment for 1 month and 2 months of the two groups were compared. Results The total effective rate of the experimental group was significantly higher than that of the control group (P < 0.05). After one month and two months of treatment, CDAI and Lewis scores of the experimental group were significantly lower than those of the control group (P < 0.05); the serum levels of IGF-1, TGF-β1 as well as ALB and Tf in the experimental group were significantly higher than those in the control group (P < 0.05). The improvement of intestinal flora disturbance rate in the experimental group was significantly better than that in the control group (P < 0.05). There was no significant difference in the incidence of oral and nasal mucosa dryness, throat discomfort, nausea and vomiting between two groups (P>0.05). Conclusion In the treatment of refractory IBD patients, thalidomide combined with IFX can regulate serum IGF-1 and TGF-β1 levels, effectively relieve clinical manifestations, inhibit inflammatory activities, and improve nutritional status and intestinal flora disorders of patients, and it has high safety.