Objective To investigate the mechanism of fisetin inhibiting the activation of microglia NOD-like receptor family protein 3(NLRP3)inflammasome in microglia and alleviating cognitive impairment after sepsis.Methods C57BL/6 mice were used to establish the sepsis model by cecal ligation and puncture.Mice were divided into four groups:the sham group,the sepsis group,the sepsis+caspase-1 knockout group(sepsis+Cas-1-/-group)and the sepsis+fisetin group.Evans blue was used to detect the permeability of blood-brain barrier(BBB).Morris water maze was used to evaluate the cognitive function of mice.Western blot assay and immunofluorescence double staining were used to detect the expression of NLRP3 inflammasome-related proteins including caspase-1,N-terminal fragment of the GSDMD(GSDMD-N),interleukin(IL)-1β,IL-18 and mitophagy-related proteins(Pink1,Parkin and LC3-Ⅱ)in brain tissue and microglia.Results Compared with the sham group,expression levels of caspase-1,GSDMD-N,IL-1β and IL-18 were significantly increased in the sepsis group(P＜0.05).Compared with the sepsis group,expression levels of caspase-1,GSDMD-N,IL-1β and IL-18 were significantly decreased in the sepsis+Cas-1-/-group(P＜0.05).The expression levels of Pink1,Parkin and LC3-Ⅱ were significantly higher in the sepsis+fisetin group than those of the sepsis group(P＜0.05),and expression levels of caspase-1,GSDMD-N,IL-1β and IL-18 were significantly lower(P＜0.05).After fisetin intervention,the permeability of BBB was decreased and the cognitive impairment(decreased escape latency and increased frequencies of crossing the platform)was alleviated in the sepsis+fisetin group compared with those of the sepsis group(P＜0.05).Conclusion Fisetin may alleviate central inflammation and cognitive impairment after sepsis by inhibiting the activation of microglial NLRP3 inflammasome through activating mitophagy.

Strongyloidiasis stercoralis can cause disease when larvae invade the human body through the skin or mucosa and can also infect a host when the host ingests its eggs.[1] Strongyloidiasis lacks characteristic manifestations, and its clinical symptoms are related to the immune response of the host and the degree of infection. Immunodeficient patients with underlying disease or who are receiving long-term corticosteroid treatment are more prone to developing severe disease.[2] The present study reports a case of Strongyloides stercoralis-induced sepsis and acute respiratory distress syndrome (ARDS) in a patient with Guillain-Barré syndrome.

Objective To analyse whether Resveratrol can inhibit the inflammatory response in septic cardiomyopathy by modulating the mammalian target of rapamycin-transcription factor EB(mTOR-TFEB)signaling pathway.Methods A total of 32 clean-grade male Sprague-Dawley(SD)rats were selected and divided into sham operation group(Sham group),sepsis model group[cecal ligation and puncture(CLP)group],Resveratrol group(Res group)and mTOR inhibitor group(Torin1 group),with 8 rats in each group.Sepsis was induced by CLP.The sham group only underwent laparotomy and did not perform CLP.In the Torin1 group,20 mg·kg-1·d-1 of mTOR inhibitor Torin l injection was injected intraperitoneally 10 days before molding,once a day for 10 days.The Sham group and the CLP group were given the same amount of normal saline through the same route before molding.The Res group was injected intraperitoneally with 60 mg/kg Resveratrol injection before molding.At 6 hours after model establishment,the heart function was evaluated by echocardiography;Serum cardiac troponin I(cTnI)levels were detected by enzyme linked immunosorbent assay(ELISA).The mRNA expressions of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The protein expressions of phosphorylation-mTOR(p-mTOR),TFEB(nuclear),TFEB(total)and light chain 3-Ⅱ(LC3-Ⅱ)of myocardial tissue were detected by Western blotting.The interaction between Resveratrol and mTOR was simulated using a network pharmacology approach.Results Compared with the sham group,the left ventricular ejection fraction(LVEF)and left ventricular short-axis shortening rate(FES)were significantly reduced in the CLP group[LVEF:0.37±0.02 vs.0.69±0.01,FES(%):17.8±3.1 vs.33.9±2.8,both P<0.01],serum cTnI levels were elevated(ng/L:1 935.96±47.78 vs.87.95±7.98,P<0.01),the mRNA expression levels of IL-6 and TNF-α were significantly up-regulated[IL-6 mRNA(2-ΔΔCt):26.10±2.08 vs.1.61±0.03,TNF-α mRNA(2-ΔΔCt):12.80±1.51 vs.1.26±0.22,both P<0.01),serum levels of inflammatory cytokines IL-6 and TNF-α mRNA were increased[IL-6 mRNA(2-ΔΔCt):26.10±2.08 vs.1.61±0.03,TNF-α mRNA(2-ΔΔCt):12.80±1.51 vs.1.26±0.22,both P<0.01),the expression level of p-mTOR protein in myocardial tissue was significantly increased(p-mTOR/β-actin:0.60±0.07 vs.0.30±0.05),while the protein levels of TFEB(total),TFEB(nuclear)and LC3-Ⅱwere significantly decreased[TFEB(total)/β-actin:0.52±0.08 vs.0.80±0.09,TFEB(nuclear)/H3:0.36±0.06 vs.0.09±0.07,LC3-Ⅱ/β-actin:0.25±0.08 vs.0.48±0.08,all P<0.01];Compared with the CLP group,the Res group and the Torin1 group had significantly higher LVEF and FES[LVEF:0.66±0.02,0.67±0.03 vs.0.37±0.02;FES(%):32.5±3.5,33.7±3.3 vs.17.8±3.1,both P<0.01],serum cTnI level was decreased(ng/L:1 216.88±36.66,1 225.78±35.64 vs.1 935.96±47.78,both P<0.01),mRNA levels of serum inflammatory cytokines IL-6 and TNF-α were decreased[IL-6 mRNA(2-ΔΔCt):3.91±0.46,4.14±0.39 vs.26.1±2.08,TNF-α mRNA(2-ΔΔCt):4.67±1.16,5.16±1.25 vs.12.8±1.51,both P<0.01],the p-mTOR protein expression in myocardial tissue was significantly decreased(p-mTOR/β-actin:0.22±0.05,0.24±0.06 vs.0.60±0.07,all P<0.01),the protein levels of TFEB(total),TFEB(nuclear),LC3-Ⅱwere significantly increased[TFEB(total)/β-actin:0.86±0.06,0.84±0.07 vs.0.52±0.08;TFEB(nuclear)/H3:0.96±0.08,0.86±0.07 vs.0.36±0.06;LC3-Ⅱ/β-actin:0.57±0.07,0.55±0.06 vs.0.25±0.08,all P<0.01].Network pharmacology was used to verify that resveratrol binds well to mTOR.Conclusion Resveratrol enhanced cardiac autophagy via the mTOR-TFEB pathway,thereby attenuating myocardial inflammation and subsequent cardiac injury in septic rats.

Objective To analyse whether Resveratrol can inhibit the inflammatory response in septic cardiomyopathy by modulating the mammalian target of rapamycin-transcription factor EB(mTOR-TFEB)signaling pathway.Methods A total of 32 clean-grade male Sprague-Dawley(SD)rats were selected and divided into sham operation group(Sham group),sepsis model group[cecal ligation and puncture(CLP)group],Resveratrol group(Res group)and mTOR inhibitor group(Torin1 group),with 8 rats in each group.Sepsis was induced by CLP.The sham group only underwent laparotomy and did not perform CLP.In the Torin1 group,20 mg·kg-1·d-1 of mTOR inhibitor Torin l injection was injected intraperitoneally 10 days before molding,once a day for 10 days.The Sham group and the CLP group were given the same amount of normal saline through the same route before molding.The Res group was injected intraperitoneally with 60 mg/kg Resveratrol injection before molding.At 6 hours after model establishment,the heart function was evaluated by echocardiography;Serum cardiac troponin I(cTnI)levels were detected by enzyme linked immunosorbent assay(ELISA).The mRNA expressions of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The protein expressions of phosphorylation-mTOR(p-mTOR),TFEB(nuclear),TFEB(total)and light chain 3-Ⅱ(LC3-Ⅱ)of myocardial tissue were detected by Western blotting.The interaction between Resveratrol and mTOR was simulated using a network pharmacology approach.Results Compared with the sham group,the left ventricular ejection fraction(LVEF)and left ventricular short-axis shortening rate(FES)were significantly reduced in the CLP group[LVEF:0.37±0.02 vs.0.69±0.01,FES(%):17.8±3.1 vs.33.9±2.8,both P<0.01],serum cTnI levels were elevated(ng/L:1 935.96±47.78 vs.87.95±7.98,P<0.01),the mRNA expression levels of IL-6 and TNF-α were significantly up-regulated[IL-6 mRNA(2-ΔΔCt):26.10±2.08 vs.1.61±0.03,TNF-α mRNA(2-ΔΔCt):12.80±1.51 vs.1.26±0.22,both P<0.01),serum levels of inflammatory cytokines IL-6 and TNF-α mRNA were increased[IL-6 mRNA(2-ΔΔCt):26.10±2.08 vs.1.61±0.03,TNF-α mRNA(2-ΔΔCt):12.80±1.51 vs.1.26±0.22,both P<0.01),the expression level of p-mTOR protein in myocardial tissue was significantly increased(p-mTOR/β-actin:0.60±0.07 vs.0.30±0.05),while the protein levels of TFEB(total),TFEB(nuclear)and LC3-Ⅱwere significantly decreased[TFEB(total)/β-actin:0.52±0.08 vs.0.80±0.09,TFEB(nuclear)/H3:0.36±0.06 vs.0.09±0.07,LC3-Ⅱ/β-actin:0.25±0.08 vs.0.48±0.08,all P<0.01];Compared with the CLP group,the Res group and the Torin1 group had significantly higher LVEF and FES[LVEF:0.66±0.02,0.67±0.03 vs.0.37±0.02;FES(%):32.5±3.5,33.7±3.3 vs.17.8±3.1,both P<0.01],serum cTnI level was decreased(ng/L:1 216.88±36.66,1 225.78±35.64 vs.1 935.96±47.78,both P<0.01),mRNA levels of serum inflammatory cytokines IL-6 and TNF-α were decreased[IL-6 mRNA(2-ΔΔCt):3.91±0.46,4.14±0.39 vs.26.1±2.08,TNF-α mRNA(2-ΔΔCt):4.67±1.16,5.16±1.25 vs.12.8±1.51,both P<0.01],the p-mTOR protein expression in myocardial tissue was significantly decreased(p-mTOR/β-actin:0.22±0.05,0.24±0.06 vs.0.60±0.07,all P<0.01),the protein levels of TFEB(total),TFEB(nuclear),LC3-Ⅱwere significantly increased[TFEB(total)/β-actin:0.86±0.06,0.84±0.07 vs.0.52±0.08;TFEB(nuclear)/H3:0.96±0.08,0.86±0.07 vs.0.36±0.06;LC3-Ⅱ/β-actin:0.57±0.07,0.55±0.06 vs.0.25±0.08,all P<0.01].Network pharmacology was used to verify that resveratrol binds well to mTOR.Conclusion Resveratrol enhanced cardiac autophagy via the mTOR-TFEB pathway,thereby attenuating myocardial inflammation and subsequent cardiac injury in septic rats.

Objective:To evaluate the predictive effect of sepsis-induced coagulopathy (SIC) score level on the prognosis of septic patients under sepsis 3.0 criteria.Methods:A retrospective single-center observational study was conducted on the septic patients admitted to the department of critical care medicine and the department of emergency in Guangdong Provincial People's Hospital from August 2016 to July 2021. The baseline data, laboratory indexes and SIC scores of the patients were collected on the first and fourth (4th) day after hospitalization. Whether the patients were survival within 30 days after enrollment was recorded. Univariate and multivariate Logistic regression were used to analyze the independent risk factors for 30-day mortality in septic patients. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of SIC score on the 30-day prognosis of septic patients.Results:A total of 173 patients met the inclusion criteria including 111 (64%) survivors and 62 (36%) non-survivors. There were significant differences in lymphocyte count (LYM), sequential organ failure assessment (SOFA), oxygenation index (PaO 2/FiO 2) and cardiovascular SOFA score between the survival group and the non-survival group. And there were no significant differences in other indexes. On the first day of admission, there were statistically significant differences in PaO 2/FiO 2, cardiovascular SOFA score, LYM, SIC score between the non-survival group and the survival group. There were significant differences in international normalized ratio (INR), prothrombin activity (PTA), prothrombin time (PT), PaO 2/FiO 2, cardiovascular SOFA score, LYM, C-reactive protein (CRP) and procalcitonin (PCT) between the two groups on the 4th day after admission. The mortality of septic patients increased with the increase of SIC score. Binary Logistic regression analysis showed that SIC score and LYM on the 4th day after admission were independent risk factors for 30-day mortality in septic patients (both P < 0.05). The ROC curve showed that SIC score had a certain predictive value for the 30-day prognosis of septic patients [area under the ROC curve (AUC) = 0.712, 95% confidence interval (95% CI) was 0.629-0.794, P < 0.001]. The predictive value of SIC score combined with LYM was better than that of the two alone (AUC = 0.748, 95% CI was 0.688-0.828, P < 0.001). Conclusions:The SIC score has a certain predictive value for the 30-day prognosis of septic patients. The predictive value of SIC score combined with LYM is better than that of the two alone, which is expected to be a potential indicator for early assessment of the condition and prognosis of septic patients.

OBJECTIVE: To explore whether β1 receptor blocker could decrease the myocardial inflammation through the Toll-like receptor 4/nuclear factor-ΚB (TLR4/NF-ΚB) signaling pathway in the sepsis adult rats. METHODS: Sixty male Wistar rats (250-300 g) aged 3 months old were allocated to four groups by random number table (n = 15): sham operation group (S group), sepsis model group (CLP group), β1 receptor blocker esmolol intervention group (ES group), and inhibitor of the TLR4 E5564 intervention group (E5564 group). The rat sepsis model was established by cecal ligation and puncture (CLP); S group of rats underwent only an incision. Rats in S group, CLP group and E5564 group were subcutaneous injected with 0.9% sodium chloride (NaCl) 2.0 mL/kg. Besides, the rats in ES group were injected with esmolol (15 mg×kg^-1×h^-1) by micro pump through the caudal vein. The rats in E5564 group were injected with E5564 (0.3 mg×kg^-1×h^-1) by micro pump through the caudal vein 1 hour before the CLP surgery. Samples were collected 6 hours after the modelling in each group. The average arterial pressure (MAP) and cardiac output index (CI) were monitored by PU electrical conduction ECG monitor. The levels of serum cardiac troponin I (cTnI), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). The expressions of TLR4, NF-ΚB p65, IL-1β, TNF-α in myocardial tissue was detected by Western Blot. RESULTS: There was no significant difference in MAP in each group. Compared with the S group, the CI in the CLP group was significantly decreased, the levels of serum cTnI, IL-1β, TNF-α were significantly increased, the protein expressions of myocardial tissue TLR4, NF-ΚB p65, IL-1β and TNF-α were significantly increased. Compared with the CLP group, the CI in the ES group and E5564 group were significantly increased (mL×s^-1×m^-2: 58.6±4.3, 58.9±4.4 vs. 41.2±3.9, both P < 0.01), the levels of serum cTnI, IL-1β and TNF-α were significantly decreased [cTnI (μg/L): 1 113.81±26.64, 1 115.74±25.90 vs. 1 975.96±42.74; IL-1β (ng/L): 39.6±4.3, 38.9±4.4 vs. 61.2±3.9; TNF-α (ng/L): 43.1±2.8, 48.7±2.6 vs. 81.3±4.4, all P < 0.01], the protein expressions of myocardial tissue NF-ΚB p65, IL-1β, TNF-α were significantly decreased (NF-ΚB p65/β-actin: 0.31±0.03, 0.43±0.04 vs. 0.85±0.08; IL-1β/β-actin: 0.28±0.05, 0.32±0.03 vs. 0.71±0.06; TNF-α/β-actin: 0.18±0.04, 0.28±0.03 vs. 0.78±0.07, all P < 0.01), but there was no significant difference in protein expression of TLR4 (TLR4/β-actin: 0.89±0.07, 0.87±0.09 vs. 0.95±0.09, both P > 0.05). There was no significant difference in CI, the levels of serum cTnI, IL-1β, TNF-α, and the protein expressions of myocardial tissue TLR4, NF-ΚB p65, IL-1β, TNF-α between ES group and E5564 group (all P > 0.05). CONCLUSIONS β1 receptor blocker esmolol may inhibit myocardial inflammatory response in sepsis adult rats through TLR4/NF-ΚB signaling pathway, thereby alleviating sepsis-induced myocardial injury.
Animals ; Inflammation/prevention & control* ; Interleukin-1beta ; Male ; Myocardium/pathology* ; NF-kappa B/metabolism* ; Propanolamines/pharmacology* ; Rats ; Rats, Wistar ; Sepsis/drug therapy* ; Signal Transduction/drug effects* ; Toll-Like Receptor 4/metabolism* ; Tumor Necrosis Factor-alpha

ObjectiveTo investigate whether esmolol could improve clinical outcome and tissue oxygen metabolism by controlling heart rate (HR) in patients with septic shock.Methods A single-center double-blinded randomized controlled trial was conducted. The patients suffering from septic shock received 6-hour early goal directed therapy (EGDT) with pulmonary artery wedge pressure≥ 12 mmHg (1 mmHg = 0.133 kPa) or central venous pressure (CVP)≥ 12 mmHg requiring norepinephrine to maintain mean arterial pressure (MAP)≥ 65 mmHg and HR≥95 bpm admitted to intensive care unit (ICU) of Guangdong General Hospital from September 2013 to September 2014 were enrolled. They were randomly divided into esmolol group and control group by computer-based random number generator. All patients received conventional basic treatment, while those in the esmolol group received in addition persistent esmolol infusion by micro pump with dosage of 0.05 mg·kg-1·min-1 with the dosage adjusted to maintain HR lower than 100 bpm within 24 hours. The patients in control group did not receive drug intervention for HR. The primary end-points consisted of length of stay in ICU and 28-day mortality. The secondary end-points included hemodynamic parameters [HR, MAP, CVP, cardiac index (CI), stroke volume index (SVI), systemic vascular resistance index (SVRI)] and tissue oxygen metabolism parameters [central venous oxygen saturation (ScvO2), lactate level (Lac)]before and 24, 48, 72 hours after the treatment.Results A total of 48 patients with septic shock were enrolled with 24 patients in esmolol group and 24 in control group.① The primary end-points: compared with control group, the length of stay in the ICU in the esmolol group was significantly shortened (days: 13.75±8.68 vs. 21.70±6.06,t = 3.680, P = 0.001), and 28-day mortality was significantly lowered [25.0% (6/24) vs. 62.5% (15/24),χ2 = 6.857,P = 0.009].② The secondary end-points: there were no significant difference in the hemodynamic and tissue metabolism parameters before treatment between two groups. No significant difference was found between before and after treatment of all above parameters in control group. HR and Lac in the esmolol group were obviously declined, SVI, SVRI, ScvO2 were gradually increased, but no significant difference in MAP, CVP, and CI was found. Compared with the control group, HR in the esomolol group was significantly lowered (bpm: 84.4±3.5 vs. 111.2±7.2,P< 0.01), SVRI and ScvO2 were significantly increased from 24 hours [SVRI (kPa·s·L-1·m-2): 137.9±1.6 vs. 126.9±1.3, ScvO2: 0.652±0.017 vs. 0.620±0.017, bothP< 0.01]; SVI was significantly increased (mL/m2: 39.9±2.2 vs. 36.8±1.7,P< 0.01) and Lac level significantly declined from 48 hours (mmol/L: 2.8±0.3 vs. 3.4±0.3,P< 0.01).Conclusion The results demonstrate that HR controlled by a titrated esmolol infusion given to septic shock patients was associated with an improvement in tissue metabolism, reduction in the length of ICU stay and lowering of 28-day mortality.