Sepsis has emerged as a major threat to human mortality. Increasing evidence explores the impact of mitochondrial metabolism on the prognosis of sepsis patients and its therapeutic potential. To enhance risk stratification and identify potential targets, we conducted a retrieval and analysis of differential expression of mitochondrial metabolism related genes (MMRG) between sepsis and normal samples from public databases. Immune infiltration analysis was preformed to gain comprehensive knowledge of features of the established risk model. Additionally, single-cell sequencing results suggested MTHFD2 may be a critical target in sepsis for regulating immune infiltration characteristics, potentially altering platelet metabolism pathways significantly, thereby influencing sepsis occurrence and progression. Utilizing molecular docking, we further screened Ginsenoside Rb1 (Grb1) as a key pharmacological target interacting with MTHFD2. Further animal experiments preliminarily indicated that Grb1 administration was associated with reduced MTHFD2 expression and improved organ function and survival in CLP-induced septic rats. These findings provide new insights and potential therapeutic targets for clinical treatment of sepsis in the future.

Sepsis is a systemic inflammatory syndrome that can cause coagulation abnormalities, leading to damage in multiple organs. Vascular endothelial cells (VECs) are crucial in the development of sepsis-induced coagulopathy (SIC). The role of Parthenolide (PTL) in regulating SIC by protecting VECs remains unclear. The study utilized septic rats and lipopolysaccharide (LPS)-stimulated VECs to simulate a SIC model and observe the therapeutic effects of PTL. Additionally, nanotechnology was employed to produce Nano-PTL (N-PTL), to observe whether it has advantages over PTL in treating SIC. PTL has been shown to mitigate lung injury in septic rats, significantly reduce tumor necrosis factor-α (TNF-α) levels, and increase survival rates. PTL treatment also enhances coagulation function, augments vascular endothelial cell (VEC) function, reduces mitochondrial fragmentation, and increases both mitochondrial oxygen consumption rate (OCR) and mitochondrial membrane potential (MMP), while inhibiting reactive oxygen species (ROS) production. By increasing BRD4/BCL-xL levels, PTL can prevent mitochondrial-mediated apoptosis in VECs, improve VEC function, and consequently ameliorate SIC. Additionally, nanotechnology-synthesized N-PTL further enhances the protective effects on VECs and coagulation function. This study clarifies the therapeutic effects and mechanisms of PTL on SIC, offering new strategies and directions for the treatment of sepsis.

Macrophages are present in all tissues and body compartments under homeostatic physiological conditions. Importantly, they play a key role in pathological inflammatory processes when disturbed. They can quickly produce large amounts of inflammatory cytokines in response to danger signals. Macrophages can recognize muramyl dipeptide (MDP) through nucleotide-binding oligomerization domain (NOD)-like receptors, subsequently activating the NF-κB signaling pathway and producing proinflammatory cytokines. Erbin can bind to NOD2 and inhibit MDP-induced NF-κB activation, thus participating in the regulation of inflammatory response. Stabilizing or enhancing Erbin expression is essential for suppressing inflammatory responses. In this study, we used a deubiquitination enzyme plasmid library to screen for a key deubiquitinase, VCPIP1, which interacts with Erbin and influences its stability through deubiquitination modification. We investigated whether VCPIP1 affects inflammation using MDP-stimulated RAW 264.7 and BMDMs cells. The results showed that VCPIP1 deficiency reduced Erbin expression and increased NF-κB phosphorylation. Additionally, VCPIP1 deficiency promoted the release of inflammatory factors (IL-1β, IL-6, and TNF-α) in RAW 264.7 cells and BMDMs. This study further expands the role of deubiquitinases (DUBs) in inflammation, providing new insights for the prevention and treatment of sepsis, tumors, immune diseases, and other inflammatory reactions.

Mounting attention has been focused on defects of the autophagy-lysosomal pathway in sepsis, however, the precise mechanisms governing the autophagy-lysosomal process in sepsis are poorly known. We have previously reported that Erbin deficiency aggravated the inflammatory response and organ injuries caused by sepsis. In the present study, we found that Erbin knockout impaired the autophagy process in both muramyl dipeptide (MDP)-induced bone marrow-derived macrophages (BMDMs) and sepsis mouse liver and lung, as detected by the accumulation of LC3-II and SQSTM1/p62, and autophagosomes. Pretreatment with autophagy inhibitor chloroquine (CQ) further aggravated inflammatory response and organ injuries in vivo and in vitro sepsis model. We also observed that the impaired lysosomal function mediated autophagic blockade, as detected by the decreased expression of ATP6V, cathepsin B (CTSB) and LAMP2 protein. Immunoprecipitation revealed that the C-terminal of Erbin (aa 391–964) interacts with the N-terminal of transcription factor EB (TFEB) (aa 1–247), and affects the stability of TFEB-14-3-3 and TFEB-PPP3CB complexes and the phosphorylation status of TFEB, thereby promote the nucleus translocation of TFEB and the TFEB target genes transcription. Thus, our study suggested that Erbin alleviated sepsis-induced inflammatory responses and organ injuries by rescuing dysfunction of the autophagy-lysosomal pathway through TFEB-14-3-3 and TFEB-PPP3CB pathway.

Background The tourniquet technique is often used in total knee arthroplasty (TKA). However, its effect on postoperative delirium (POD) in elderly patients undergoing TKA is unknown. Methods This prospective randomized controlled trial assessed the eligibility of 245 elderly patients. A total of 197 patients who met the inclusion criteria were randomly divided into a tourniquet group ( n = 98) and a non-tourniquet group ( n = 99). The primary outcome was the incidence of POD within 72 h after surgery. The secondary outcome was the quality of rehabilitation, including inflammatory reaction, postoperative pain, hypoproteinemia and anemia. Results Of 245 patients, 184 patients completed this clinical trial, with 92 cases in each group. There were 14 patients (15.22%) with POD in the tourniquet group and 5 patients (5.43%) in the non-tourniquet group (95% CI 1.076 to 9.067, P = 0.029). The changes in white blood cell count (WBC), the proportion of neutrophils (NEUT%), c-reactive protein (CRP), interleukin-6 (IL-6) and middle patellar circumference in the tourniquet group were higher than those in the non-tourniquet group ( P < 0.05). The visual analog scale (VAS) at rest and activity in the tourniquet group were higher than those in the non-tourniquet group ( F = 170.102, P < 0.001 F = 75.391, P < 0.001). There were 41 (44.57%) patients with hypoproteinemia in the tourniquet group and 26 (28.26%) in the non-tourniquet group (95% CI 1.106 to 3.765, P = 0.022). Conclusion The application of the tourniquet technique in elderly patients with TKA procedures increased the incidence of POD. This may be attributed to the increased inflammatory reaction, severe postoperative pain and hypoproteinemia caused by the tourniquet technique. Trial registration Clinical trial registration number: ChiCTR2100045711. Full date of the first registration: 23/04/2021.

目的:分析髋关节置换术后转入ICU治疗的危险因素和早期预后.方法:回顾性分析武汉大学中南医院2012年12月1日-2018年12月1日髋关节置换术患者,根据术后是否立即转入ICU分为未转入ICU组(n＝972)和转入ICU组(n=261).记录一般资料、术中情况和术后并发症等,分析转入ICU的危险因素和早期预后.结果:转入ICU患者术后心血管并发症、肺部感染、术后谵妄发生率偏高,住院时间延长;年龄、麻醉方式、ASA分级、术前合并COPD、肾功能不全、癌症、脑梗病史、红细胞输注量是术后转入ICU的独立危险因素(P＜0.05).结论:髋关节置换术后立即转入ICU并不改善早期预后,多种因素对术后转入ICU有影响,术前应针对性处理这些危险因素,减少围术期并发症.

Background: Previous studies showed that GTS-21, a selective alpha 7 nAchR agonist, can trigger anti-inflammatory effects and improve the survival of septic animals. However, whether GTS-21 affects autophagy responses remains unclear. Here, we tested the hypothesis that GTS-21 ameliorates sepsis-induced hepatic injury by modulating autophagy in mice. Method: C57BL/6 male mice were randomly separated and categorized into four groups: the sham group, and CLP group subjected to caecal ligation and puncture (CLP, a model of polymicrobial sepsis). The CLP + GTS-21 group was administered GTS-21 immediately after CLP challenge. alpha-Bungarotoxin (an alpha 7 nAchR antagonist) was injected before CLP was performed, and then, after CLP challenge, GTS-21 was administered to alpha-BGT + CLP + GTS-21 group. The hepatic tissue and blood samples were harvested 6 h after the operation. Results: CLP challenge increased TNF-alpha and IL-6 production, and hepatic enzyme alanine aminotransferase and aspartate transaminase levels. CLP also elevated the expression of hepatic LC3-II, sequestosome-1/p62, Atg7 and Atg5. The administration of GTS-21 inhibited pro-inflammatory cytokine production and hepatic enzymatic marker expression, promoted the expression of LC3-II, Atg7, Atg5, and decreased the expression of p62, which could be reversed by alpha-BGT treatment. Conclusion: Our findings suggested that alpha 7nAchR is involved in diminishing hepatic damage by inhibiting inflammatory responses and improving autophagy in mice with polymicrobial sepsis.

Renal ischemia-reperfusion (rI/R) is a risk factor for acute lung injury (ALI). Alveolar macrophages (AMs) activation mediated by rI/R-induced ALI is one of the pathogeneses associated with the development of ALI. In rI/R, α2-adrenergic receptor agonists have been indicated to be effective in decreasing urea nitrogen concentrations. In this study, we explored the underlying pathogenesis of the clinically obtainable α2-adrenergic receptor agonist dexmedetomidine (DEX) in protecting against rI/R -mediated AMs activation. We incubated AMs with the serum of sham and rI/R rats in the presence or absence of various concentrations of DEX. We used an enzyme-linked immunosorbent assay to detect the secretion levels of GSH, LDH, IL-18, IL-1β, and HMGB1 in the culture supernatant. We employed real-time polymerase chain reaction to assess the expression of NOX-4 mRNA, and western blotting to observe the protein levels of NOX-4, the NLRP3 inflammasome, AMPK, and eNOS. In addition, we used immunofluorescence to analyze ROS and MMP activity. Incubation of AMs with DEX suppressed rI/R-mediated cellular LDH production and ROS release. DEX also abolished the rI/R-mediated decrease in the activity of GSH and increased the levels of the rI/R-related NADPH oxidase protein NOX-4. Furthermore, DEX reduced the amelioration of the mitochondrial potential induced by rI/R. Our study showed that DEX inhibits rI/R-mediated levels of the NLRP3 inflammasome proteins ASC, NLRP3, HMGB1 and p20, and ameliorates rI/R-mediated AMPK signaling inactivation. Therefore, DEX reduces the levels of two mediators that are activated by the NLRP3 inflammasome: IL-18 and IL-1β. Finally, our study established that DEX mitigates the rI/R-mediated decrease in eNOS, demonstrating its protective functions against AMs activation. In conclusion, our study demonstrated that the protective action of DEX in AMs is induced through amelioration of HMGB1-NLRP3 inflammasome-AMPK signaling. Our results suggest that the anesthetic reagent DEX exerts beneficial effects to ameliorate rI/R-induced ALI.

目的 比较不同麻醉方式对老年患者髋关节置换术后转归的影响.方法 回顾性分析2012年12月至2018年10月行髋关节置换术患者566例,年龄≥75岁,根据麻醉方式分为两组:全身麻醉组(G组,n=233)和蛛网膜下腔阻滞组(S组,n=333).采用倾向评分将患者按1:1匹配,得到组间协变量均衡样本.比较两组术中情况、术后并发症、转入ICU例数和住院时间.结果 对患者一般资料进行匹配后,G组和S组各纳入患者225例.与G组比较,S组手术时间、麻醉时间、手术室停留时间明显缩短,心血管并发症、肺部感染、深静脉血栓形成/肺栓塞、术后谵妄、术后恶心呕吐、转入ICU率明显降低,住院时间明显缩短,术后头痛、尿潴留发生率明显升高(P<0.05).结论 对于老年髋关节置换术患者,蛛网膜下腔阻滞可以改善术后转归,加速术后康复进程.

Objective:To evaluate the role of ErbB2-interacting protein (Erbin) in muramyl dipeptide (MDP)-induced inflammatory responses in the macrophages of mice.Methods:Erbin gene knockout RAW264.7 cell line (Erbin -/ -RAW264.7) was constructed by CRISPR/CAS9 gene-editing technology.RAW264.7 cells were cultured in vitro.Each type of cells was divided into 2 groups ( n=16 each)by a random number table method: RAW264.7 group, RAW264.7 plus MDP group, erbin -/ -RAW264.7 group, and erbin -/ -RAW264.7 plus MDP group.In each MDP group, cells were incubated with 10 μg/ml MDP for 6 h, then immunofluorescence was used to determine the expression of nuclear factor kappa B (NF-κB) p65, and the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the culture medium were determined by enzyme-linked immunosorbent assay. Results:Compared with RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased( P<0.05), NF-κB p65 moved to the nucleus, and the red fluorescence area was increased in RAW264.7+ MDP group.Compared with RAW264.7+ MDP group and Erbin -/- RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased ( P<0.05), NF-κB p65 moved more markedly to the nucleus, and the red fluorescence area was increased in Erbin -/-RAW264.7+ MDP group. Conclusion:Erbin inhibits MDP-induced inflammatory responses in macrophages through inhibiting the activity of NF-κB p65 in mice.

BackgroundThe influence of anesthetic technique on perioperative outcomes for total hip arthroplasty remains poorly elucidated. We studied a sample of total hip arthroplasty recipients, hypothesizing that spinal anesthesia has a superior impact on perioperative outcomes.MethodsWe conducted a retrospective cohort study of patients undergoing total hip arthroplasty between December 1, 2012 and October 31, 2018 in Zhongnan Hospital of Wuhan University. The primary outcome was cardiorespiratory complications. Secondary outcomes were intraoperative hypotension, packed red blood cells (pRBCs) transfusion, prolonged hospital length of stay, intensive care unit (ICU) use, life-threatening event, and mortality. Multivariable regression analyses were used to identify the impact of anesthetic technique on perioperative outcomes.ResultsAmong the 1,233 patients, 561 had general anesthesia, and 672 had spinal anesthesia. Patients were averagely younger in general group than in spinal anesthesia group, (69.0 and 72.1 years, respectively; P < 0.001), with insignificant difference in comorbidity burden. When spinal anesthesia was used, the hospital length of stay, ICU times, and volume of pRBCs transfusion were significant decreased (P < 0.05). Life-threatening event and in-hospital mortality occurred frequently in general anesthesia, but with insignificant difference. After adjusting for covariates, spinal anesthesia was associated with 54.3% reduction in cardiorespiratory complications (adjusted odds ratio [OR]: 0.457, 95% confidence interval [CI]: 0.320–0.652; P < 0.001). Spinal anesthesia was favorably associated with decreased odds for intraoperative hypotension (OR: 0.653, 95% CI: 0.494–0.863; P = 0.003) and ICU use (OR: 0.371, 95% CI: 0.268–0.514; P < 0.001). The use of spinal anesthesia was not found to influence the risk of pRBCs transfusion (adjusted odds ratio [OR]: 0.823, 95% CI: 0.631–1.073; P = 0.149) and prolonged hospital length of stay (adjusted odds ratio [OR]: 0.886, 95% CI: 0.684–1.148; P = 0.360).ConclusionsCompared with general anesthesia, spinal anesthesia for total hip arthroplasty was associated with decreased rates of cardiorespiratory complications, intraoperative hypotension, and ICU use.

Objective To evaluate the effect of penehyclidine hydrochloride (PHC) on the expression of caveolin-1 (Cav-1) with lipopolysaccharide (LPS)-induced lung injury (LI) in rats.Methods Thirty SPF healthy male Sprague-Dawley rats,weighing 170-190 g,were divided into 3 groups (n =10each) using a random number table method:control group (group C),LPS-induced LI group (group LI)and PHCD group.LI was produced by injecting LPS 0.2 ml (5 mg/kg) via the trachea in anesthetized rats.PHCD 0.5 ml (2 mg/kg) was intraperitoneally injected at 1 h before establishing the model in group PHCD.Arterial blood samples were collected at 24 h after establishing the model for blood gas analysis and for determination of serum tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) concentrations by enzyme-linked immunosorbent assay.Rats were then sacrificed,and the lungs were removed.The main bronchus was lavaged,and the broncho-alveolar lavage fluid (BALF) was collected for calculation of the percentage of polymorphonuclear neutrophils (PMNs).Lung tissues were obtained for examination of pathological changes and for determination of myeloperoxidase (MPO) activity (by colorimetric assay),wet/dry weight ratio (W/D ratio),and expression of Cav-1 and nuclear factor kappa B (NF-sB) in nucleoprotein (by Western blot).Results Compared with group C,pH value and PaO2 were significantly decreased,the PaCO2,percentage of PMNs in BALF,W/D ratio and MPO activity were increased,the Car-1 expression was down-regulated,the expression of NF-κB in nucleoprotein was up-regulated,and the serum TNF-α and IL-1β concentrations were increased in group LI (P＜0.05).Compared with group LI,pH value and PaO2 were significantly increased,the PaCO2,percentage of PMNs in BALF,W/D ratio and MPO activity were decreased,the Cav-1 expression was up-regulated,the expression of NF-κB in nucleoprotein was down-regulated,and the serum TNF-α and IL-1β concentrations were decreased (P＜0.05),and the path ological changes of lung tissues were significantly attenuated in group PHCD (P＞0.05).Conclusion The mechanism by which PHC reduces LPS-induced LI may be related to up-regulating the expression of Cav-1 and mitigating inflammatory responses in lung tissues of rats.

Objective To evaluate the relationship between cholinergic anti-inflammatory pathway and autophagy during liver injury in septic mice.Methods SPF healthy male 32 C57BL/6 mice,aged 6-8 weeks,weighing 18-22 g,were divided into 4 groups (n=8 each) using a random number table method:sham operation group (group Sham),sepsis group (group Sep),α7nAChR agonist GTS-21 plus sepsis group (GTS-21+Sep group),and α7nACh antagonist α-BGT plus sepsis plus GTS-21 group (α-BGT+Sep+GTS-21 group).Sepsis was induced by cecal ligation and puncture.GTS-21 4 mg/kg was intraperitoneally injected immediately after operation in group GTS-21 +Sep.α-BGT 1 mg/kg was intraperitoneally injected at 15 min before operation,and GTS-21 4 mg/kg was intraperitoneally injected immediately after operation in group α-BGT+Sep+GTS-21.Blood samples were obtained at 6 h after surgery for determination of serum aspartate transaminase (AST) and alanine transaminase (ALT) concentrations by enzyme-linked immunosorbent assay.Liver tissues were obtained for determination of the expression of microtubule-associated protein-1 light chain 3-Ⅱ (LC3 Ⅱ) and sequestosome-1/p62 (by Western blot) and for examination of the number of autophagosomes in liver cells (under a transmission electron microscope).Results Compared with Sham group,the expression of LC3 Ⅱ and sequestosome-1/p62 was significantly up-regulated,the number of autophagosomes was increased,and the serum AST and ALT concentrations were increased in Sep group (P＜0.05).Compared with Sep group,the expression of LC3 Ⅱ was significantly up-regulated,the expression of sequestosome-1/p62 was down-regulated,the number of autophagosomes was increased,and the serum AST and ALT concentrations were decreased in GTS-21 +Sep group (P＜0.05).Compared with GTS-21 +Sep group,the expression of LC3 Ⅱ was significantly down-regulated,the expression of sequestosome-1/p62 was up-regulated,the number of autophagosomes was decreased,and the serum AST and ALT concentrations were increased in α-BGT+Sep+GTS-21 group (P＜0.05).Conclusion Activation of the cholinergic anti-inflammatory pathway can enhance the level of autophagy in liver cells and is involved in the endogenous protective mechanism of sepsis-induced liver injury in septic mice.

Objective To evaluate the effect of dexmedetomidine on acute lung injury (ALI) induced by double hit in rats. Methods Forty SPF male Sprague-Dawley rats, aged 8 weeks, weighing 215-225 g, were divided into 4 groups(n=10 each) using a random number table method: sham operation group (Sham group), double hit -induced ALI group (ALI group), double hit -induced ALI plus dexmedetomidine group (ALI+ Dex group) and double hit -induced ALI plus dexmedetomidine plus yohimbine group (ALI+ Dex+ Y group). ALI was induced by blunt chest trauma and hemorrhagic shock-resuscitation in pentobarbital sodium-anesthetized rats in ALI, ALI+ Dex and ALI+ Dex+ Y groups.In group ALI+ Dex, dexmedetomidine was intravenously infused for 10 min in a loading dose of 1 μg·kg-1·h-1 starting from the time point immediately after blunt chest trauma, followed by continuous infusion of 5 μg·kg-1·h-1 until the end of operation.Yohimbine 0.1 mg/kg was injected after giving the loading dose of dexmedetomidine and at 3 h after blunt chest trauma in ALI+ Dex+ Y group.Rats were sacrificed at 6 h after successful establishment of the model, blood samples were obtained for determination of concentrations of TNF-α and IL-6 in serum (by enzyme-linked immunosorbent assay), and lung tissues were obtained for determination of wet/dry weight (W/D) ratio, expression of Fas, FasL and caspase-3 in lung tissues (by SABC assay), cell apoptosis (by TUNEL), content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) and for examination of the pathological changes which were scored.The apoptosis index (AI) was calculated. Results Compared with Sham group, the lung injury score, AI, levels of Fas, FasL, caspase-3 and MDA, and concentrations of TNF-α and IL-6 in serum were significantly increased, and the SOD activity in lung tissues was decreased in ALI group (P<0.05). Compared with ALI group, the lung injury score, AI, levels of Fas, FasL, caspase-3 and MDA, and concentrations of TNF-α and IL-6 in serum were significantly decreased, and the SOD activity in lung tissues was increased in ALI+ Dex and ALI+ Dex+ Y groups (P<0.05). Compared with group ALI+ Dex, the lung injury score, AI, levels of Fas, FasL, caspase-3 and MDA, and concentrations of TNF-α and IL-6 in serum were significantly increased, and the SOD activity in lung tissues was decreased in group ALI+ Dex+ Y (P<0.05). Conclusion Dexmedetomidine reduces ALI induced by double hit in rats, and the mechanism may be partially related to activating alpha-2 adrenergic receptor and inhibiting inflammatory responses, lipid peroxidation and cell apoptosis. Key words: Dexmedetomidine; Thoracic injury; Shock, hemorrhagic; Acute lung injury

Ginsenoside Rg1 (G-Rg1) is an active ingredient of Panax ginseng, which has previously been reported to attenuate alcohol-induced hepatic damage; however, the underlying mechanisms remain largely unknown. The present study aimed to investigate the protective effects of G-Rg1 on alcohol-induced cell injury in vitro and on a rat model of alcoholic hepatitis in vivo. For the in vitro model, L-O2 cells were incubated with ethanol in the presence or absence of G-Rg1. For the in vivo model, rats were administered ethanol by intragastric injection and were treated with G-Rg1, or dexamethasone as a control. The results indicated that serum biochemical parameters, including alanine aminotransferase, aspartate aminotransferase and total bilirubin, as well as the expression of nuclear factor (NF)-kappa B pathway-associated inflammatory cytokines, including interleukin (IL)-6, tumor necrosis factor-alpha and IL-1 beta, were elevated in response to alcohol; however, they were significantly decreased by G-Rg1 treatment. Furthermore, NF-kappa B pathway activation was reduced by treatment with G-Rg1. G-Rg1 also decreased oxidative stress by inhibiting cytochrome P450 2E1 expression and reactive oxygen species production, and promoting glutathione peroxidase expression. Furthermore, G-Rg1 inhibited the expression levels of caspase-3 and -8, which may be associated with decreased hepatocyte apoptosis. These data suggested that G-Rg1 may protect hepatocytes against alcohol-induced injury, through preventing excessive inflammation and hepatocellular apoptosis.