Objectives:The clinical efficacy of short-acting beta-blockers in the management of sepsis remains uncertain. In particular, the comparative effects of two commonly used agents-esmolol and landiolol-have not been clearly established. This network meta-analysis aims to systematically evaluate and compare the effects of esmolol, landiolol, and standard of care (SOC) on mortality in patients with sepsis.Data Sources:A systematic search of PubMed, Web of Science, Embase, MEDLINE, CENTRAL, ClinicalTrials.gov, preprints, and citation searching was conducted before April 15, 2025.Study Selection:Randomized controlled trials that enrolled adult patients (>= 18 yr) diagnosed with sepsis or septic shock and treated with beta-blockers and conducted in ICUs.Data Extraction:Data were extracted on study characteristics, enrolled patients' characteristics, administration strategies of drugs, and key clinical outcomes (including 28-d mortality, ICU length of stay, and other relevant endpoints).Data Synthesis:A total of 1165 records were identified through searches of five databases, registries, and relevant references up to April 15, 2025. Ten studies involving 1035 patients were included, after screening and eligibility assessment. Compared with esmolol, landiolol was associated with increased 28-day mortality (relative risk [RR], 1.57; 95% CI, 1.08-2.30; low certainty) and higher norepinephrine requirements (mean difference [MD], 0.17 mu g/kg/min; 95% CI, 0.02-0.32; low certainty). Esmolol significantly reduced 28-day mortality (RR, 0.69; 95% CI, 0.56-0.85; moderate certainty) and 24-hour heart rate (MD, -16.92 beats/min; 95% CI, -23.49 to -10.36; moderate certainty) compared with SOC. In contrast, landiolol increased norepinephrine use compared with SOC (MD, 0.09 mu g/kg/min; 95% CI, 0.01-0.18; moderate certainty).Conclusions:Among patients with sepsis treated with beta-blockers, esmolol probably improves clinical outcomes compared with SOC. However, the effect of landiolol remains uncertain due to the low certainty of evidence. Esmolol may confer a relative clinical advantage over landiolol, but further studies are needed to confirm this finding and elucidate the underlying mechanisms.

Sepsis is a fatal syndrome characterized by uncontrolled systemic inflammatory responses. Circular RNAs (circRNAs) are involved in the modulation of various pathophysiological processes, but their potential role in sepsis has largely been unexplored. In this study, we observed differential expression of circMLH3 between healthy volunteers and septic patients, and revealed the value of circMLH3 for sepsis diagnosis and prognostic prediction. Interestingly, we discovered a correlation between the expression level of circMLH3 and the degree of pyroptosis, a critical mechanism contributing to uncontrolled inflammation in sepsis patients. Knocking down circMLH3 alleviated macrophage pyroptosis whereas overexpressing circMLH3 aggravated pyroptosis. circMLH3 regulated macrophage pyroptosis by sponging miR-590-3p and subsequently modulating TAK1 expression. Furthermore, we found that the miR-590-3p/TAK1 axis inhibited the activation of pro-caspase-1 and the NLRP3 inflammasome. miR-590-3p overexpression had a protective effect by reducing macrophage pyroptosis, thereby alleviating sepsis-induced lung injury and systemic inflammatory responses. In conclusion, our study elucidated the circMLH3/miR-590-3p/TAK1 signaling pathway and identified its role in regulating mononuclear macrophage pyroptosis, thus providing potential novel targets and strategies for sepsis diagnosis and therapy. © 2024 The Authors

Early detection of acute kidney injury (AKI) may provide a crucial window of opportunity to prevent further injury, which helps improve clinical outcomes. This study aimed to develop a deep interpretable network for continuously predicting the 24-hour AKI risk in real-time and evaluate its performance internally and externally in critically ill patients. A total of 21,163 patients' electronic health records sourced from Beth Israel Deaconess Medical Center (BIDMC) were first included in building the model. Two external validation populations included 3025 patients from the Philips eICU Research Institute and 2625 patients from Zhongda Hospital Southeast University. A total of 152 intelligently engineered predictors were extracted on an hourly basis. The prediction model referred to as DeepAKI was designed with the basic framework of squeeze-and-excitation networks with dilated causal convolution embedded. The integrated gradients method was utilized to explain the prediction model. When performed on the internal validation set (3175 [15 %] patients from BIDMC) and the two external validation sets, DeepAKI obtained the area under the curve of 0.799 (95 % CI 0.791–0.806), 0.763 (95 % CI 0.755–0.771) and 0.676 (95 % CI 0.668–0.684) for continuousAKI prediction, respectively. For model interpretability, clinically relevant important variables contributing to the model prediction were informed, and individual explanations along the timeline were explored to show how AKI risk arose. The potential threats to generalisability in deep learning-based models when deployed across health systems in real-world settings were analyzed. © 2024

The development of acute respiratory distress syndrome (ARDS) in sepsis is associated with substantial morbidity and mortality. However, the molecular pathogenesis underlying sepsis-induced ARDS remains elusive. Neutrophil heterogeneity and dysfunction contribute to uncontrolled inflammation in patients with ARDS. A specific subset of neutrophils undergoing reverse transendothelial migration (rTEM), which is characterized by an activated phenotype, is implicated in the systemic dissemination of inflammation. Using single-cell RNA sequencing (scRNA-seq), it identified functionally activated neutrophils exhibiting the rTEM phenotype in the lung of a sepsis mouse model using cecal ligation and puncture. The prevalence of neutrophils with the rTEM phenotype is elevated in the blood of patients with sepsis-associated ARDS and is positively correlated with disease severity. Mechanically, scRNA-seq and proteomic analys revealed that inflamed endothelial cell (EC) released extracellular vesicles (EVs) enriched in karyopherin subunit beta-1 (KPNB1), promoting abluminal-to-luminal neutrophil rTEM. Additionally, EC-derived EVs are elevated and positively correlated with the proportion of rTEM neutrophils in clinical sepsis. Collectively, EC-derived EV is identified as a critical regulator of neutrophil rTEM, providing insights into the contribution of rTEM neutrophils to sepsis-associated lung injury. © 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.

Background: Sepsis is a life -threatening clinical syndrome caused by dysregulated host response to infection. The mechanism underlying sepsis -induced immune dysfunction remains poorly understood. Natural killer T (NKT) cells are cytotoxic lymphocytes that bridge the innate and adaptive immune systems, the role of NKT cells in sepsis is not entirely understood, and NKT cell cluster differences in sepsis remain unexplored. Methods: Mendelian randomization (MR) analyses were first conducted to investigate the causal relationship between side scatter area (SSC-A) on NKT cells and 28 -day mortality of septic patients. A prospective and observational study was conducted to validate the relationship between the percentage of NKT cells and 28 -day mortality of sepsis. Then, the single -cell RNA sequencing (scRNAseq) data of peripheral blood mononuclear cells (PBMCs) from healthy controls and septic patients were profiled. Results: MR analyses first revealed the protective roles of NKT cells in the 28 -day mortality of sepsis. Then, 115 septic patients were enrolled. NKT percentage was significantly higher in survivors (n = 84) compared to non -survivors (n = 31) (%, 5.00 +/- 3.46 vs 2.18 +/- 1.93, P < 0.0001). Patients with lower levels of NKT cells exhibited a significantly increased risk of 28 -day mortality. According to scRNA-seq analysis, NKT cell clusters exhibited multiple distinctive characteristics, including a distinguishing cluster defined as FOS + NKT cells, which showed a significant decrease in sepsis. Pseudo -time analysis showed that FOS + NKT cells were characterized by upregulated expression of crucial functional genes such as GZMA and CCL4 . CellChat revealed that interactions between FOS + NKT cells and adaptive immune cells including B cells and T cells were decreased in sepsis compared to healthy controls. Conclusion: Our findings indicate that NKT cells may protect against sepsis, and their percentage can predict 28 -day mortality. Additionally, we discovered a unique FOS + NKT subtype crucial in sepsis immune response, offering novel insights into its immunopathogenesis.

Current clinical approaches for septic shock increasingly incorporate bundle treatment, a multi-component approach that uses a collection of tests and agents to assist in the identification and treatment of infection. The present study analyzed completion rates of 3 h and 6 h bundle treatment among patients with septic shock in intensive care units (ICUs) of hospitals in Jiangsu Province from 2016 to 2020, using data from the Jiangsu Provincial Intensive Care Medical Quality Control Center. Current approaches and factors affecting treatment completion were assessed.The completion rates of 3 h and 6 h bundle treatment in ICUs of all medical units in Jiangsu Province and in ICUs of hospitals of different levels were recorded. Analyses show that the completion rate of 3 h and 6 h bundle treatment for patients with septic shock in ICUs in Jiangsu Province increased year by year from 2016 to 2020.The completion rate of 3 h bundle treatment increased from 69.82% (3 604/5 162) to 82.47% (8 915/10 775) (all P<0.001). The completion rate of 6 h bundle treatment increased from 62.69% (3 236/5 162) to 72.54% (7 816/10 775) (all P<0.001). In addition, year by year, the completion rate of 3 h bundle treatment in ICUs in tertiary hospitals increased, from 69.80% (3 596/5 152) to 82.23% (7 375/8 969), while the completion rate of 6 h bundle treatment increased from 62.69% (3 230/5 152) to 72.18% (6 474/8 969) (all P<0.001). Completion rates in secondary hospitals also increased year by year, from 80.00% (8/10) to 85.27% (1 540/1 806) for 3 h treatment and from 60.00% (6/10) to 74.31% (1 342/1 806) (all P<0.001) for 6 h treatment. Completion rates for 3 h treatment in first-tier cities (83.99% (2 099/2 499)) and second-tier cities (84.68% (3 952/4 667)) was higher than in third-tier cities (79.36% (2 864/3 609)). The completion rate of 6 h bundle treatment gradually decreased in first-line (77.19% (1 929/2 499)), second-line (74.37% (3 471/4 667)), and third-line (66.94% (2 416/3 609)) cities (all P<0.001). The data collectively show that from 2016 to 2020, the completion rate of bundle treatment in septic shock patients in ICUs in Jiangsu Province improved significantly.

脓毒症是全球医疗面临的巨大挑战，是重症医学亟待解决的重大难题。血管内皮损伤则是脓毒症发生发展过程中的关键环节，而细胞焦亡导致血管内皮损伤重要机制，遏制血管内皮焦亡是减缓脓毒症发生和进展的有效策略。①我们通过体内外敲除GSDMD阻断内皮细胞焦亡，发现阻断内皮细胞焦亡能够缓解脓毒症血管内皮细胞损伤，减轻全身炎症反应和器官损伤。②通过脓毒症患者全血转录组芯片及加权基因共表达网络分析发现lncRNA NBR2与焦亡关键分子GSDMD表达显著正相关；体外诱导内皮细胞焦亡后NBR2和GSDMD显著增加；沉默NBR2明显降低GSDMD表达同时抑制内皮细胞焦亡；提示抑制NBR2表达可能是减缓脓毒症内皮细胞焦亡的有效措施。③通过RNA-pulldown联合蛋白质谱筛选并验证介导NBR2靶向调控GSDMD表达的蛋白，筛选到HNRNPK蛋白介导NBR2和GSDMD互作。④采用染色质免疫共沉淀联合qPCR检测HNRNPK蛋白与GSDMD基因的相互作用，荧光素酶报告基因检测HNRNPK对GSDMD基因转录的调控作用，通过荧光原位杂交技术和免疫荧光检测NBR2与HNRNPK蛋白亚细胞分布和共定位情况，证实tLPS刺激NBR2和HNRNPK向细胞核内移位，结合到GSDMD基因上促进GSDMD基因转录，增强GSDMD表达，为脓毒症治疗提供新靶点。课题组最新研究发现，细胞焦亡时，会释放大量细胞外泌体，这些外泌体携带了执行细胞焦亡的GSDMD。⑤探究了脓毒症患者血浆外泌体中GSDMD与患者病情及预后的相关性，结果证实，血浆外泌体中GSDMD含量与脓毒症病情及预后密切相关，为评估脓毒症患者的病情及预后提供了重要指标。

BACKGROUND: Although the mean arterial pressure (MAP) target of 65mmHg was achieved, diastolic blood pressure (DBP) was still low in some septic shock patients. The effects of DBP on the prognosis and optimal target for patients with septic shock are unclear. We sought to investigate the relationship between DBP and 28-day mortalityin septic shock patients.; METHODS: In this retrospective observational study, we obtained data from the Chinese Database in Intensive Care (CDIC). We included patients with an admission diagnosis of septic shock and shock was controlled. DBP was measured every 1h, and the mean DBP during the first 24h (mDBP24h) was recorded. The primary outcome was 28-day mortality. Multivariable logistic regression determined the relationship between mDBP24h and 28-day mortality.; RESULTS: In total, 1251 patients were finally included. The 28-day mortality of included septic shock patients was 28.3%. The mDBP24h, not mSBP24h, was higher among 28-day survivors compared with non-survivors. 28-day mortality was inversely associated with mDBP24h (unadjusted OR 0.814 per 10mmHg higher mDBP24h, P=0.003), with a stepwise increase in 28-day mortality at lower mDBP24h. The 28-day mortality of patients with mDBP24h<59mmHg had an absolute risk reduction of 9.4% (P=0.001). And mDBP24h<59mmHg was the remaining high risk factor inversely associated with 28-day mortality after multivariable adjustment (adjusted OR 1.915, 95% CI 1.037-3.536, P=0.038), while mMAP24h and mSBP24h were not.; CONCLUSION: In patients with septic shock after initial resuscitation, we observed an inverse association between mDBP24h and 28-day mortality. The poor outcomes in patients with mDBP24h<59mmHg provide indirect evidence supporting a further DBP goal of 59mmHg for patients with septic shock after MAP of 65mmHg was achieved. © 2023. BioMed Central Ltd., part of Springer Nature.

第一章GSDMD介导的内皮细胞焦亡在脓毒症血管内皮损伤的作用及机制研究目的:探讨GSDMD介导的内皮细胞焦亡在脓毒症血管内皮损伤的作用及机制。方法:将LPS转染（tLPS）至脐静脉内皮细胞EA.hy926内诱导内皮细胞焦亡,或分别将对照及GSDMD干扰质粒转染至内皮细胞中,再诱导细胞焦亡,进而采用光镜观察焦亡细胞的形态改变,流式细胞术检测焦亡细胞的比例,乳酸脱氢酶（LDH）细胞毒性检测试剂盒检测LDH释放,ELISA检测IL-1β水平,RTqPCR检测炎性Caspase和GSDMD mRNA表达,Western Blot或免疫荧光检测炎性Caspase和GSDMD蛋白表达和剪切。采用尾静脉注射含sh Gsdmd的腺相关病毒干扰小鼠体内Gsdmd表达,通过盲肠结扎穿孔术（CLP）构建脓毒症小鼠模型,采用透射电镜检测肺血管内皮细胞改变,称重检测小鼠肺湿重体重比,BCA法检测肺泡灌洗液中蛋白含量,Evans蓝实验检测肺血管通透性,ELISA检测血浆中IL-1β、IL-18和TNF-α分泌,HE染色检测多器官损伤情况。结果:tLPS刺激脐静脉内皮细胞,导致内皮细胞焦亡,LDH释放显著增加,IL-1β分泌明显增多,炎性Caspase和GSDMD mRNA和蛋白表达均明显增多,炎性Caspase和GSDMD剪切明显增多。转染GSDMD干扰质粒后,GSDMD mRNA和蛋白表达均显著降低,内皮细胞焦亡明显缓解,LDH释放显著减少,IL-1β分泌明显减少。脓毒症小鼠肺血管内皮细胞焦亡增多,肺湿重体重比升高,肺泡灌洗液中蛋白增多,肺组织中Evans蓝增多,血浆中炎症因子显著增多,肺脏、肾脏、肝脏和心脏器官损伤严重,而尾静脉注射含sh Gsdmd的腺相关病毒显著缓解肺血管内皮细胞焦亡,改善肺组织水肿,减少蛋白渗出,改善肺血管通透性,降低血浆中IL-1β和IL-18水平,缓解多个器官损伤。结论:GSDMD介导内皮细胞焦亡导致脓毒症血管内皮损伤、多器官功能障碍。第二章LncRNA NBR2通过GSDMD调控脓毒症血管内皮细胞焦亡的作用及机制研究目的:探讨lncRNA在脓毒症血管内皮细胞焦亡中的作用及机制。方法:采用基因芯片检测脓毒症患者血液中lncRNA和mRNA表达谱,加权基因共表达网络分析筛选与细胞焦亡相关的lncRNAs,tLPS刺激脐静脉内皮细胞EA.hy926诱导内皮细胞焦亡,RT-qPCR检测候选lncRNAs表达。通过小干扰RNA干扰候选lncRNAs表达筛选调控细胞焦亡的lncRNA。分别将lncRNA NBR2过表达质粒、GSDMD干扰质粒转染至内皮细胞中,并给予tLPS刺激,通过RT-qPCR检测候选lncRNAs、NBR2和GSDMD表达,Western Blot或免疫荧光检测炎性Caspase和GSDMD蛋白表达,观察内皮细胞焦亡的形态改变,LDH细胞毒性检测试剂盒检测LDH释放,ELISA检测IL-1β分泌。采用核质分离试剂盒分离细胞质和细胞核,RT-qPCR分别检测NBR2表达,采用荧光原位杂交（FISH）检测NBR2亚细胞定位。结果:加权基因共表达网络分析提示12个lncRNAs与GSDMD等细胞焦亡相关分子表达密切相关,其中4个lncRNAs在tLPS刺激的内皮细胞中的表达显著升高。通过si RNA分别干扰这4个lncRNAs表达,发现干扰lncRNA NBR2时,tLPS所致的内皮细胞焦亡形态改变明显减少,LDH释放显著减少,IL-1β分泌明显减少,GSDMD mRNA和蛋白表达均显著降低。NBR2包含1371个核苷酸,无蛋白编码能力,符合lncRNA的基本特征。FISH和核质分离检测结果表明NBR2在细胞核中表达高于细胞质中,tLPS刺激增强NBR2表达,促进NBR2向核内移位。过表达NBR2能够促进GSDMD mRNA和蛋白表达,促进内皮细胞焦亡、LDH释放、IL-1β分泌,而干扰GSDMD能够阻断NBR2促进GSDMD表达和细胞焦亡的作用。结论:lncRNA NBR2通过靶向调控GSDMD表达,介导脓毒症血管内皮细胞焦亡。第三章LncRNA NBR2靶向调控GSDMD基因表达的机制研究目的:探讨NBR2靶向调控GSDMD表达的分子机制。方法:首先通过RNA纯化染色质分离实验（ChIRP）捕获NBR2结合的蛋白质,联合液质联用串联质谱（LC-MS/MS）技术和Western Blot检测和验证NBR2结合的关键蛋白HNRNPK,通过RNA结合蛋白免疫沉淀（RIP）联合RT-qPCR验证NBR2和HNRNPK的相互作用。然后,采用含sh HNRNPK载体的慢病毒感染内皮细胞,同时给予tLPS刺激内皮细胞,RT-qPCR检测候选NBR2、HNRNPK和GSDMD表达,Western Blot或免疫荧光检测HNRNPK和GSDMD蛋白表达,观察内皮细胞焦亡形态,LDH细胞毒性检测试剂盒检测LDH释放,ELISA检测IL-1β分泌。再采用染色质免疫共沉淀（Ch IP）联合qPCR检测HNRNPK蛋白与GSDMD基因的相互作用,荧光素酶报告基因检测HNRNPK对GSDMD基因转录的调控作用。最后,使用干扰HNRNPK的慢病毒感染内皮细胞并予以tLPS刺激,通过FISH和免疫荧光检测NBR2与HNRNPK蛋白亚细胞分布和共定位情况,并利用核质分离,分别采用RT-qPCR检测细胞质和细胞核中NBR2的表达,Western Blot检测细胞质和细胞核中HNRNPK蛋白的表达。结果:ChIRP-MS检测到42个蛋白质可能与NBR2结合,同时筛选可能与GSDMD基因结合的蛋白,发现HNRNPK蛋白可能同时结合NBR2和GSDMD基因,ChIRP-Western Blot结合RIP-RT-qPCR结果双向证实HNRNPK与NBR2的相互作用。tLPS导致内皮细胞焦亡、LDH释放增多、IL-1β分泌增多,GSDMD表达增多;而干扰HNRNPK表达明显缓解内皮细胞焦亡、减少LDH释放、降低IL-1β水平,GSDMD表达减少,阻断NBR2促进GSDMD表达和细胞焦亡的作用。Ch IP-qPCR结果表明HNRNPK与GSDMD基因相互作用,荧光素酶报告基因结果表明HNRNPK结合到GSDMD基因上促进GSDMD基因转录,增强GSDMD表达。共定位实验表明,tLPS刺激NBR2和HNRNPK向细胞核内移位,干扰HNRNPK表达,NBR2核细胞移位减少。核质分离检测结果也证实tLPS刺激NBR2和HNRNPK向细胞核内移位,干扰HNRNPK表达,NBR2核细胞移位减少。结论:lncRNA NBR2招募并结合HNRNPK蛋白,转移至细胞核内,靶向调控GSDMD表达,介导胞内LPS引起的内皮细胞焦亡。