Objective To explore the potential relationship between ubiquitination of transforming growth factor kinase 1(TAK1)/nuclear factor-κB(NF-κB)signaling pathway mediated by ring finger protein 99(RNF99)and septic acute respiratory distress syndrome(ARDS).Methods Plasmid and siRNA transfection were conducted to overexpress or knock down RNF99 in MLE12,and expressions of p65 phosphate and p65 protein were analyzed.The protein interaction between RNF99 and TRAF6 or TAK1 was analyzed by immunoprecipitation assay.Forty mice were randomly divided into WT plus PBS,WT plus LPS,RNF99 specific expression(TG)plus PBS,and TG plus LPS groups,with 10 mice in each group.Sepsis was induced by intraperitoneal injection of 30 mg/kg LPS.Results As compared with vector group,protein expression levels of TRAF6 and TAK1 in MLE12 cells decreased significantly in RNF99 group(P<0.05).Ubiquitinated TRAF6 protein increased in MLE12 cells with RNF99 knockdown.As compared with LPS plus vector group,phosphorylation level of p65 in MLE12 cells was signifi-cantly lower in LPS plus RNF99 group(P<0.05).As compared with si-NC group,protein expression levels of RNF99 and IκBα in si-RNF99 group decreased significantly(P<0.05).As compared with LPS plus si-NC group,phosphorylation level of p65 in LPS plus si-RNF99 group increased significantly(P<0.05).The staining percentage of CD68 macrophages in lung tissues was significantly lower in TG plus LPS group than in WT plus LPS group(P<0.05).Phosphorylation level of p65 in lung tissues was significantly lower in TG plus LPS group than in WT plus LPS group(P<0.05).Conclusion RNF99 regulates NF-κB signaling pathway by interacting with the key regulator of NF-κB signaling pathway(TRAF6/TAK1),and improves lung injury after intraperitoneal injection of LPS in mice.

Objective To observe the clinical efficacy of sivelestat sodium combined with ulinastatin in the treatment of sepsis-induced acute respiratory distress syndrome(ARDS).Methods One hundred and four patients with sepsis-induced ARDS had admitted to our hospital from January 2020 to May 2023 were selected and randomly divided into a control group(routine treatment plus sivelestat sodium)and combination group(routine treatment plus sivelestat sodium and ulinastatin)by a computer random number generator,52 in each group.Murray lung injury score(MLIS),sequential organ failure assessment(SOFA)score,extravascular lung water index(ELWI),arterial blood oxygen partial pressure/fraction of inspired oxygen(PaO2/FiO2),white blood cell count(WBC),neutrophil percentage(NEUT%),and levels of endothelial cell specific molecule-1(ESM-1),soluble urokinase-type plasminogen activator receptor(suPAR)and interleukin-6(IL-6)were compared between the two groups before and after treatment.The recovery speed,prognosis and adverse reactions were compared between the two groups.Results T-test showed there were no significant differences in MLIS score,SOFA score,ELWI,PaO2/FiO2,WBC,NEUT%,ESM-1,suPAR and IL-6 levels between the control group and the combination group before treatment(P>0.05).After treatment,MLIS score,SOFA score,ELWI,WBC,NEUT%,ESM-1,suPAR and IL-6 levels in the combination group were lower than those in the control group(P<0.05),and PaO2/FiO2 was higher than that in the control group(P<0.05).Time to mechanical ventilation and length of ICU stay in the combination group were shorter than those in the control group(P<0.05),and the 28-day mortality rate was lower than that in the control group(P<0.05).No serious adverse reactions occurred in both groups during the treatment period.Conclusion Sivelestat sodium combined with ulinastatin can reduce lung injury and inflammatory response,accelerate recovery speed,improve lung function and prognosis in patients with sepsis-induced ARDS,and the therapy has higher safety.

Objective To observe the effect of Xinmailong injection on microcirculation in patients with septic shock,so as to understand the application effect and value of Xinmailong injection in patients with septic shock.Methods A total of 82 patients with septic shock treated in the intensive care unit of our hospital were selected as the research objects and randomly divided into two groups.41 patients in the control group were treated with conventional septic shock cluster therapy,and 41 patients in the treatment group were treated with Xinmailong injection on the basis of cluster therapy in the control group.Then hemodynamic parameters(HR,CVP,CO,MAP,SVR),hemorheology(whole blood reducing viscosity,hematocrit,fibrinogen,platelet aggregation rate),microcirculation perfusion indexes(oxygenation index,blood lactate level,SCVO2,Pcv-aCO2)and 28-day survival rate were detected and compared between the two groups before and after treatment.Results Before treatment,there were no significant differences in hemodynamic parameters,blood rheology and microcirculation perfusion indexes between the two groups(P>0.05).After treatment,the above test results of the two groups were signifi-cantly improved,and the test results of the treatment group were significantly better than those of the control group,and the 28-day survival rate was also better than that of the control group.The differences were statistically significant(P<0.05).Conclusion Xinmailong injection can significantly improve the hemodynamic parameters,blood rheology and microcirculation perfusion indexes,and improve the 28-day survival rate of patients with sepsis.It has a positive clinical effect in the treatment of septic shock patients.

Objective To investigate whether the pituitary tumor transformation gene 1(PTTG 1)plays a role in colitis by regulating intestinal epithelial cells pyroptosis.Methods Ten PTTG 1 wild-type(WT)mice and Ten PTTG 1 knockout(KO)mice were randomly divided into 4 groups of 5 each,respectively PTTG1 WT control and experimental group,PTTG1 KO control and experimental group.The mice in the experimental group were given 3%dextran sodium sulfate(DSS)for 6 days to induce acute colitis,and the control group was given sterile double distilled water(ddH2O).The disease activity index of the respective group of mice was observed and recorded.Mouse colonic tissue were collected,and the expression levels of NLRP3,ASC,and GSDMD were determined by immuno-histochemistry and western blot.In HCoEpiC,PTTG1 expression was knocked down using shRNA,and the cells were subsequently treated with TNF-α to induce inflammation.Then,the expression of GSDMD was detected.Results The expression of PTTG1 was decreased in colonic mucosal tissue in mice with acute colitis(P<0.01).Compared with WT mice,the colitis was significantly aggravated in PTTG1 KO mice after 3%DSS treatment.The expression of pyroptosis-related proteins was significantly up-regulated in the colon mucosal tissues of PTTG1 KO experimental mice(P<0.05).After knocking down the expression of PTTG1 in HCoEpiC and TNF-α treatment,the expression levels of GSDMD were significantly up-regulated(P<0.05).Conclusion PTTG1 reduced pyroptosis in intestinal epithelial cells(IECs),while PTTG1 loss can enhance IEC pyroptosis,aggravating colonic inflammation.

Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue.Methods 84 Kunming mice were randomly divided into sham group,control group,VEGF group,HGF group and VEGF+HGF group,and the left lower limb ischemia model was established.The blood supply of ischemic tissue was observed by rheometer,and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA.Immunohistochemical staining was used to detect angiogenesis(CD31,SMA)in ischemic tissues.Safety was assessed by side effects during treatment in mice.Results After the successful modeling,the blood flow velocity of the left lower limb in each group decreased significantly.On the 7th day after operation,the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation(P<0.05),and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups(P<0.05).On the 28th day after operation,the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized,the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups,and both VEGF group and HGF group were significantly better than the control group(P<0.05).On the 7th,14th,and 28th days following surgery,HGF and VEGF protein levels in the Ad-HGF,Ad-VEGF,and Ad-VEGF-HGF groups were substantially greater than those in the control group(P<0.05).The expression level in the Ad-VEGF-HGF group peaked on the 14th day(all P<0.001)and subsequently declined to preoperative levels on the 28th day after operation.Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level,encour-aging angiogenesis after lower limb ischemia,increasing blood flow,and improving lower limb circulation.

Objective To investigate the expression of lncRNA SNHG8 in placenta accrete(PA)and its effect on trophoblast invasion and migration.Methods qRT-PCR was used to detect the expression of lncRNA SNHG8 in placenta tissue of 30 cases in PA group and 30 cases in control group,and the correlation between lncRNA SNHG8 expression and prenatal ultrasound score of 30 cases in PA group was analyzed.Transwell and scratch assay were used to detect the effect of lncRNA SNHG8 interference on the invasion and migration of human chorionic trophoblast cells(HTR8/SVneo cells),and western blot was used to detect the expression of MMP-2 and MMP-9.The downstream targets of lncRNA SNHG8 were predicted by StarBase software,and the expression of lncRNA SNHG8 was detected in placental tissues of the two groups.Dual luciferase reporter assay was used to detect the targeting relationship between lncRNA SNHG8 and miR-542-3p.Results Compared with that of the control group,the expression of lncRNA SNHG8 was up-regulated in the placenta tissue of the PA group(P<0.05),and it was positively correlated with prenatal ultrasound score.Interference with lncRNA SNHG8 inhibited the invasion and migration of trophoblast cells(P<0.05);the protein expression of MMP-9 and MMP-2 also decreased signifi-cantly(P<0.05).Biological prediction indicates that miR-542-3p had a binding site with lncRNA SNHG8,and miR-542-3p expression was down-regulated in PA placental tissue(P<0.05).Dual luciferase reporter assay confirmed that lncRNA SNHG8 could target miR-542-3p.Compared with si-SNHG8+inhibitor-NC,co-transfection of si-SNHG8 and miR-542-3p inhibitor enhanced the invasion and migration ability of trophoblast cells(P<0.05).Conclusion lncRNA SNHG8 is highly expressed in PA and is related to the severity of PA.LncRNA SNHG8 promotes the invasion and migration of trophoblast by regulating the level of miR-542-3p.The study suggests that lncRNA SNHG8 plays an important role in the invasion and migration of PA trophoblast cells,which is expected to be a clinical diagnostic biomarker and therapeutic target.

Objective To study the diagnostic value of miR-571 for liver fibrosis by detecting miR-571 expression in the peripheral blood of patients with liver fibrosis.Methods From December 2022 to September 2023,40 patients with liver fibrosis,40 patients with chronic hepatitis,and 40 healthy controls were chosen as research subjects.The expression level of miR-571 in peripheral blood was detected using a real-time quantitative polymerase chain reaction,and the relative expression of miR-571 in each group was evaluated.The Spearman correlation method was utilized to examine the relationship between miR-571 and clinical detection indices.To assess the capacity of miR-571 and the multivariate diagnostic model to identify liver fibrosis,binary logistic regression was used to create a multivariate diagnostic model,and ROC curves were generated.Results The expression of miR-571 was significantly higher in the liver fibrosis group than in the healthy control and hepatitis groups,and the difference was statistically significant(P<0.001).The expression level of miR-571 was positively connected with ALT,APRI score,and FIB-4 index(r = 0.23,0.30,0.22,P<0.05)and negatively correlated with PLT(r =-0.19,P<0.05)according to Spearman correlation analysis.Logistic regression research revealed that miR-571 and the FIB-4 index were independent risk factors for liver fibrosis.The AUC for miR-571 to diagnose fibrosis was 0.91(95%CI:0.85～0.96),while the AUC for miR-571 paired with the FIB-4 index was 0.94(95%CI:0.90～0.98).Conclusion MiR-571 expression was shown to be considerably higher in the peripheral blood of hepatic fibrosis patients,and the combined FIB-4 index offers some clinical diagnostic value for hepatic fibrosis.

Objective To investigate the clinical significance of ANCA in the disease activity of patients with SLE.Methods A total of 1 025 SLE patients were recruited and were divided into inactive and active groups according to SLEDAI score.Demographic characteristics,clinical symptoms,autoantibodies,routine laboratory tests and renal pathology were also recorded and compared between the two groups.Results All patients were divided into inactive group(n = 750)and active group(n = 250).The occurrence of renal,pulmonary,cutaneous,arthritis manifestations were significantly higher than those of the inactive group(all P<0.05).All patients were tested for ANCA,and the most common pattern being perinuclear or p-ANCA,the percentage of p-ANCA seroposi-tive increased greatly with the increased disease activity(P<0.05).The autoantibodies were further analyzed between the two groups,25 patients had reactivity to MPO,but no patient had reactivity to PR3.Also,there were significant differences in anti-dsDNA antibody,anti-nucleosome antibody between the two groups(P<0.05).In the active group,patients with p-ANCA seropositive exhibited higher serum beta-2-microglobulin(β2-MG),titers of anti-dsDNA antibody,SLEDAI scores,lower albumin,C3,and C4 levels(P<0.05).Meanwhile,p-ANCA was associated with IL-6,which increased significantly with the increase of SLEDAI score.In addition,patients with p-ANCA seropositive had more occurrence of lupus nephritis,but it had no association with the renal pathology.Conclusion The appearance of p-ANCA in SLE patients indicated more severe disease activity status.

Objective To develop a nomogram for predicting the risks of early anastomotic recurrence(EAR)after primary bowel resection in patients with Crohn's disease(CD).Methods The patients with CD under-going preoperative magnetic resonance enterography(MRE)and primary bowel resection were enrolled in this retrospective study and divided into an EAR group(18 patients)and EAR-free group(12 patients).The EAR group included the patients having an endoscopic Rutgeerts score of≥I2 month or the need for anastomotic resection within 12 months after surgery.All the 38 indexes including preoperative demographic characteristics,laboratory examina-tions,multi-parameter MRE features of the resected intestine and its adjacent mesentery,histological findings,and postoperative pharmacotherapy were analyzed.Least absolute shrinkage and selection operator(LASSO)regression and multivariate binary logistic regression analysis were performed to identify independent risk factors to be incorpo-rated into the nomogram for predicting the risks of early anastomotic recurrence and the prediction performance was evaluated.Results Mesenteric creeping fat index on MRE and comb sign were independent risks of EAR,with a concordance index of 0.882(95%CI:0.764～1).The calibration plot revealed a strong relationship between actual observation and predicted probability of EAR.Conclusions The preoperative MRE-based nomogram may be a potential tool for predicting EAR following surgery in patients with CD,which is beneficial to individual management in those patients.It provides reference for the formulation of early postoperative individualized drug adjuvant therapy in patients at high risk of EAR.

Objective To explore the correlation between intestinal dose and acute radiation enteritis(ARE)in patients with cervical cancer received concurrent chemoradiotherapy,and optimize the dose limit of intestinal tissue.Methods 158 cervical cancer patients received concurrent chemoradiotherapy from 2014 to 2019 were selected in this study.According to CTCAE 5.0,patients with ARE≥grade 2 were classified as ARE≥grade 2 group,otherwise classified as ARE0.7,P<0.05).Conclusions For patients with cervical cancer received concurrent chemoradiotherapy,the dose of bowelbag V5 and V40 should be considered to rationally optimize the dose of bowelbag in the radiotherapy plan,so as to reduce the incidence of ARE≥grade 2.