Aortic dissection (AD) is a life-threatening cardiovascular emergency characterized by high acute mortality. While immune dysregulation is known to drive AD pathogenesis, the specific involvement of T-cell exhaustion-related genes (TEXRGs) remains largely elusive. Differentially expressed TEXRGs (DETEXRGs) between AD and control samples were identified using transcriptomic datasets. Functional enrichment and Mendelian randomization (MR) analyses were performed to investigate potential causal associations with AD risk. Key diagnostic biomarkers were selected through integrated machine learning algorithms. Immune infiltration landscapes were characterized via gene set enrichment analysis (GSEA), while single-cell RNA sequencing (scRNA-seq) was employed to elucidate cell-type-specific expression profiles. Furthermore, the transcription factor (TF)-regulatory network and drug-gene interaction map were constructed. Finally, the expression levels of candidate genes were validated using quantitative real-time PCR (qRT-PCR). A total of 270 DETEXRGs were identified, which were predominantly enriched in cytokine-mediated signaling pathways and viral infection-related processes. MR analysis identified six genes significant causal associations with AD susceptibility. Among these, CASP4 and FPR1 were prioritized as core biomarkers through integrated machine learning algorithms. Immune infiltration analysis revealed a significantly altered immune landscape in AD tissues, characterized by the enrichment of eight immune cell subtypes that positively correlated with the expression of the identified biomarkers. Furthermore, scRNA-seq analysis localized FPR1 expression primarily to macrophages and monocytes. Finally, qRT-PCR validation confirmed significantly elevated expression levels of FPR1, PLSCR1, and other candidate genes in AD samples. This study underscores the critical involvement of T-cell exhaustion-related mechanisms in the pathogenesis of AD and robust diagnostic biomarkers. These findings offer novel insights for early risk stratification and provide a theoretical foundation for the development of targeted immunotherapeutic strategies in aortic dissection.

Objective:To externally validate previously published prognostic models developed exclusively from pre-extracorporeal cardiopulmonary resuscitation (ECPR) variables in a contemporary ECPR cohort. Methods:We conducted a bicenter retrospective external validation of four published pre-ECPR prognostic models (Lee, RESCUE-IHCA, CHIU-S1, and CHIU-S2) in adult patients treated with ECPR between January 2015 and December 2024. Model performance was evaluated for in-hospital survival and favorable neurological outcome (FNO; Cerebral Performance Category 1-2) in the overall cohort, in-hospital cardiac arrest (IHCA), and cardiac-origin cardiac arrest (Cardio_CA) subgroups. Discrimination (the area under the receiver operating characteristic curve, AUROC), calibration, overall model fit (Brier score), and decision curve analysis (DCA) were assessed. For point-based CHIU scores, validation focused on observed outcome rates across predefined risk strata. Results:Among 214 patients, 79.0% (169/214) had IHCA; survival to discharge was 45.8% and FNO occurred in 24.8%. Discrimination for survival was modest across models (overall ECPR AUROC 0.608-0.709; IHCA 0.586-0.672; Cardio_CA 0.591-0.689) but was higher for FNO (overall ECPR 0.709-0.764; IHCA 0.696-0.744; Cardio_CA 0.698-0.718). The Lee model showed poor calibration with slopes far below 1, whereas RESCUE-IHCA model underestimated survival but demonstrated better calibration (slopes close to 1), higher overall accuracy (lower Brier scores) and broader clinical utility (wider net-benefit ranges in DCA). CHIU models provided limited risk separation between adjacent strata. Conclusions:In this external validation, pre-ECPR models showed modest performance, with better discrimination for neurological outcome than for survival. RESCUE-IHCA showed the most favorable overall performance. Future studies should develop and validate more robust, transportable tools.

Timely pathogen identification is crucial for guiding antimicrobial therapy in severe pneumonia requiring veno-venous extracorporeal membrane oxygenation (vv-ECMO). Compared with slow and often insensitive conventional culture, metagenomic next-generation sequencing (mNGS) enables more rapid and comprehensive pathogen detection and may improve clinical management. This study aimed to evaluate the clinical impact of mNGS compared with conventional culture in non-immunocompromised patients undergoing vv-ECMO for severe pneumonia. The retrospective study explored non-immunocompromised patients with severe pneumonia who received vv-ECMO support between January 2017 and June 2023. A total of 151 patients were categorized into the mNGS group and the non-mNGS group, based on whether they underwent mNGS testing. Furthermore, they were stratified into the Death group and Survive group according to their survival status at day 30. Demographics, laboratory test results, pathogens, antibiotic treatment, and clinical outcomes data were recorded and analyzed. The positivity rate identified through the mNGS method (73.3

Myocarditis is inflammatory injury of the myocardium and causes approximately 30,000 deaths globally each year. Fulminant myocarditis is an extremely severe form of myocarditis. Currently, the clinical evaluation of myocarditis and fulminant myocarditis is primarily based on symptoms, ECG findings, and biochemical markers. Cardiac magnetic resonance and endomyocardial biopsy can provide further support for the diagnosis, but both have limitations in routine practice. Recent studies have shown that cell-free DNA (cfDNA) has distinct methylation patterns depending on the organ of origin, suggesting new possibilities for tracking specific types of organ damage. The core mechanism of fulminant myocarditis is a cytokine storm, leading to multiorgan damage, differing from clinically suspected myocarditis. We performed Genome-wide cfDNA methylation detection on plasma from 20 healthy donors and 22 patients (fulminant myocarditis: clinically suspected myocarditis = 9:13, COVID-19 positive: COVID-19 negative = 14:8) and found that cfDNA can be used to specifically identify early multiorgan damage caused by fulminant myocarditis, and its AUC is superior to traditional biochemical indicators such as troponin, high-sensitivity troponin, and lactate dehydrogenase. This is critically important for the timely clinical recognition and treatment of this condition. Furthermore, our study findings suggest that SARS-CoV-2 infection may exacerbate the severity of myocarditis and multiorgan damage. In summary, cfDNA shows great potential as a noninvasive, early, and sensitive biomarker for reflecting disease severity and systemic injury in fulminant myocarditis, which may help guide earlier risk stratification and intervention.

Background This study aimed to evaluate the clinical applicability of metagenomic next-generation sequencing (mNGS) in non-immunocompromised patients undergoing veno-venous extracorporeal membrane oxygenation (vv-ECMO) for severe pneumonia. Methods The retrospective study explored non-immunocompromised patients with severe pneumonia who received vv-ECMO support between January 2017 and June 2023. A total of 151 patients were categorized into the mNGS group and the non-mNGS group, based on whether they underwent mNGS testing. Furthermore, they were stratified into the Death group and Survive group according to their survival status at day 30. Demographics, laboratory test results, pathogens, antibiotic treatment, and clinical outcomes data were recorded and analyzed. Results The positivity rate identified through the mNGS method (73.3%) was notably higher than that obtained through conventional culture (43.1%, P < 0.001). mNGS exhibited superior capabilities in identifying co-infections compared to conventional culture (70.9% vs. 30.1%, P < 0.001). Furthermore, the 30-day mortality rate within the mNGS group demonstrated a significant decrease compared to the no-mNGS group (P = 0.045). Additionally, Kaplan-Meier survival analysis yielded comparable outcomes in both groups (P = 0.035). Factors protecting against adverse outcomes encompassed prolonged hospital stay time, extended ECMO duration, lower Acute Physiology and Chronic Health Evaluation II (APACHE II) scores, successful ECMO weaning, and the application of mNGS. Antibiotic adjustments were implemented in 29 patients (58%) in the mNGS group and 39 patients (38.6%) in the no-mNGS group, with a markedly higher adjustment ratio in the former, demonstrating statistical significance (P = 0.024). Following adjustment of the treatment plan, the mNGS group demonstrated reduced APACHE II scores compared to the no-mNGS group after 5 days of treatment (P = 0.006). Conclusions The mNGS technique, with its superior pathogen detection rate, emerges as a promising tool for microbiological diagnosis and antibiotic management, ultimately contributing to enhanced patient outcomes.

Background Stanford type A aortic dissection (TAAD), as one of the most lethal cardiovascular disease, may be mediated by cellular ferroptosis, which exacerbates structural modification and dysfunction of endothelial and smooth muscle cells. Hypoxia inducible factor-1 (HIF-1), as a transcription factor has shown participate in the course of ferroptosis. However, the mechanism involved has not been uncovered clearly. Methods We collected clinical basic information and image data of TAAD patients received surgical operation for aortic replacement and non- TAAD patients who suffered from other surgeries including cardiac transplantation or aortic valve replacement. Then aortic tissues being replaced from each group was analyzed through RNA-sequencing to reveal underlying molecular mechanism participated in TAAD. Then, according to genes and signaling pathway selected, qRT-PCR and western blot were further implemented to verify the expression quantity of proteins. Results Baseline characteristics including age, gender, and other variables were non-differential between two groups. Computed tomography angiography (CTA) showed that linear low-density structure referring to tearing intima was floated in the full-course of aortic lumen which was separated into the true lumen and the false lumen. Disordered cells arrangement and fractured fibrous tissue were displayed in the H&E staining and Masson staining. RNA-seq found that more than 5000 differential genes were expressed in TAAD patients if |log 2 FC|≥1 and P value ≤ 0.5. Gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEEG) analysis revealed that genes related cellular ferroptosis and HIF-1 signaling pathway were expressed differentially and significantly during the development of TAAD. Moreover, the proteins of SLC7A11 and GPX4 which related ferroptosis were up-regulated, as well as the HIF-1 protein and its downstream protein of transferrin receptor 1(Trf1), which were in line with the results of RT-PCR. The amount of Fe 2+ was greater in aorta from TAAD patients than that from non-TAAD patients. Conclusion Cellular feroptosis plays a vital role during the development of TAAD, and the HIF-1/Trf/Fe 2+ pathway was involved in the regulation of cellular ferroptosis.

Background and purposeOur previous research discovered that cinnamamide derivatives are a new type of potential cardioprotective agents myocardial ischemia-reperfusion (MIR) injury, among which Compound 10 exhibits wonderful beneficial action in vitro. However, the exact mechanism of Compound 10 still needs to be elucidated.Experimental approachThe protective effect of Compound 10 was determined by detecting the cell viability and LDH leakage rate in H9c2 cells subjected to H2O2. Alterations of electrocardiogram, echocardiography, cardiac infarct area, histopathology and serum myocardial zymogram were tested in MIR rats. Additionally, the potential mechanism of Compound 10 was explored through PCR. Network pharmacology and Western blotting was conducted to monitor levels of proteins related to autophagic flux and mTOR, autophagy regulatory substrate, induced by Compound 10 both in vitro and in vivo, as well as expressions of Sirtuins family members.Key resultsCompound 10 significantly ameliorated myocardial injury, as demonstrated by increased cell viability, decreased LDH leakage in vitro, and declined serum myocardial zymogram, ST elevation, cardiac infarct area and improved cardiac function and microstructure of heart tissue in vivo. Importantly, Compound 10 markedly enhanced the obstruction of autophagic flux and inhibited excessive autophagy initiation against MIR by decreased ATG5, Rab7 and increased P-mTOR and LAMP2. Furthermore, Sirt1 knockdown hindered Compound 10’s regulation on mTOR, leading to interrupted cardiac autophagic flux.Conclusions and implicationsCompound 10 exerted cardioprotective effects on MIR by reducing excessive autophagy and improving autophgic flux blockage. Our work would take a novel insight in seeking effective prevention and treatment strategies against MIR injury.

ObjectivesMetagenomic next-generation sequencing (mNGS) is emerging as a novel diagnostic technology for various infectious diseases; however, limited studies have investigated its application in etiological diagnosis of early onset pneumonia (EOP) among patients undergoing extracorporeal cardiopulmonary resuscitation (ECPR) following in-hospital cardiac arrest (IHCA), The clinical significance of early mNGS in predicting short-term prognosis of IHCA patients after ECPR remains unclear.MethodsThis retrospective study included 76 patients with IHCA who underwent ECPR at the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2022. Baseline characteristics and etiological data of all patients during their hospitalization were collected and statistically analyzed. The primary outcome of this study was the diagnosis of EOP, while the secondary outcomes included successful extracorporeal membrane oxygenation (ECMO) weaning and survival at discharge. Additionally, the characteristics of bronchoalveolar lavage fluid (BALF) flora in these patients were compared by analyzing both mNGS results and culture results.ResultsMultivariate logistic regression were employed to analyze the predictors of ECMO weaning failure, mortality at discharge, and the incidence of EOP. Ultimately, patients with lower SOFA scores on admission [OR (95%CI): 1.447 (1.107-1.890), p=0.007] and those who underwent early mNGS testing within 48 hours after ECPR [OR (95%CI): 0.273 (0.086-0.865), p=0.027] demonstrated a higher probability of successful weaning from ECMO. Patients with higher SOFA scores on admission [OR (95%CI): 2.404 (1.422-4.064), p=0.001], and elevated lactate levels [OR (95%CI): 1.176 (1.017-1.361), p=0.029] exhibited an increased likelihood of mortality at discharge. Furthermore, early mNGS detection [OR (95%CI): 0.186 (0.035-0.979), p=0.047], and lower CRP levels (48h-7d after ECMO) [OR (95%CI):1.011 (1.003-1.019), p=0.006] were associated with a reduced incidence of EOP. In addition, the pathogens detected by mNGS within 48 hours after ECPR were mainly oral colonizing bacteria and viruses, and viruses were in the majority, while all BALF cultures were negative. In contrast, between 48 hours and 7 days after ECPR, BALF cultures were positive in all EOP patients.ConclusionsEarly mNGS testing to identify microbial flora facilitates timely adjustment of antibiotic regimens, thereby reducing the incidence of EOP and improving short-term prognosis in patients undergoing ECPR following IHCA.

ObjectiveTo evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) in sepsis and bloodstream infection (BSI).MethodsA retrospective analysis of patients diagnosed with sepsis and BSI at the First Affiliated Hospital of Zhengzhou University from January 2020 to February 2022 was conducted. All the patients underwent blood culture and were divided into mNGS group and non-mNGS group according to whether mNGS was performed or not. The mNGS group was further divided into early group (< 1 day), intermediate group (1–3 days), and late group (> 3 days) according to the time of mNGS inspection.ResultsIn 194 patients with sepsis and BSI, the positive rate of mNGS for identifying pathogens was significantly higher than that of blood culture (77.7% vs. 47.9%), and the detection period was shorter (1.41 ± 1.01 days vs. 4.82 ± 0.73 days); the difference was statistically significant (p < 0.05). The 28-day mortality rate of the mNGS group (n = 112) was significantly lower than that of the non-mNGS group (n = 82) (47.32% vs. 62.20%, p = 0.043). The total hospitalization time for the mNGS group was longer than that for the non-mNGS group (18 (9, 33) days vs. 13 (6, 23) days, p = 0.005). There was no significant difference in the ICU hospitalization time, mechanical ventilation time, vasoactive drug use time, and 90-day mortality between the two groups (p > 0.05). Sub-group analysis of patients in the mNGS group showed that the total hospitalization time and the ICU hospitalization time in the late group were longer than those in the early group (30 (18, 43) days vs. 10 (6, 26) days, 17 (6, 31) days vs. 6 (2, 10) days), and the ICU hospitalization time in the intermediate group was longer than that in the early group (6 (3, 15) days vs. 6 (2, 10) days); the differences were statistically significant (p < 0.05). The 28-day mortality rate of the early group was higher than that of the late group (70.21% vs. 30.00%), and the difference was statistically significant (p = 0.001).ConclusionsmNGS has the advantages of a short detection period and a high positive rate in the diagnosis of pathogens causing BSI and, eventually, sepsis. Routine blood culture combined with mNGS can significantly reduce the mortality of septic patients with BSI. Early detection using mNGS can shorten the total hospitalization time and the ICU hospitalization time of patients with sepsis and BSI.

目的:探讨宏基因二代测序( metagenomic next-generation sequencing, mNGS)技术在肾综合征出血热（Hemorrhagic fever with renal syndrome，HFRS）重症患者中的诊断价值。方法:回顾郑州大学第一附属医院2020年1月至2022年6月最终确诊为HFRS且完成mNGS的重症患者，收集完善相应的临床资料并进行随访，以mNGS是否检出汉坦病毒为标准将患者分为mNGS阳性组和阴性组，比较这两组从出现症状到送检的时间中位数，同时评价mNGS在不典型症状的HFRS重症患者中的检出率。结果:共纳入15例HFRS重症且完成mNGS的患者，其中男性14例，女性1例，年龄中位数为33岁（14~62岁）。所有HFRS患者中有9例mNGS结果为阳性，6例为阴性。mNGS阳性组从出现症状到送检的中位时间为8 d（2~12 d），阴性组为8 d（4~9 d），两组差异无统计学意义（ Z=-0.659， P=0.510）。在15例HFRS重症患者中不典型症状患者为6例，其mNGS结果均为阳性。所有患者均完成电话随访，随访中位时间为1年。其中3例死亡，11例肾功能恢复正常。 结论:mNGS技术可辅助诊断肾综合征出血热，对于诊断不明重症患者早期检测可减少漏诊。对临床考虑肾综合征出血热，mNGS未检测到汉坦病毒患者，仍需进一步检查，以明确诊断。

Objectives:This study aims to explore the pathogen-detected effect of mNGS technology and its clinical application in non-immunocompromised patients with severe pneumonia supported by vv-ECMO.Methods:A retrospective analysis was conducted on a cohort of 50 non-immunocompromised patients who received vv-ECMO support for severe pneumonia between January 2016 and December 2022. These patients were divided into two groups based on their discharge outcomes: the deterioration group (Group D), which included 31 cases, and the improvement group (Group I), consisting of 19 cases. Baseline characteristics and clinical data were collected and analyzed.Results:Among the 50 patients enrolled, Group D exhibited a higher prevalence of male patients (80.6% vs. 52.6%, p < 0.05), more smokers (54.8% vs. 21.1%, p < 0.05), and were older than those in Group I (55.16 ± 16.34 years vs. 42.32 ± 19.65 years, p < 0.05). Out of the 64 samples subjected to mNGS detection, 55 (85.9%) yielded positive results, with a positivity rate of 83.7% (36/43) in Group D and 90.5% (19/21) in Group I. By contrast, the positive rate through traditional culture stood at 64.9% (74/114). Among the 54 samples that underwent both culture and mNGS testing, 23 (42.6%) displayed consistent pathogen identification, 13 (24.1%) exhibited partial consistency, and 18 (33.3%) showed complete inconsistency. Among the last cases with complete inconsistency, 14 (77.8%) were culture-negative, while two (11.1%) were mNGS-negative, and the remaining two (11.1%) presented mismatches. Remarkably, mNGS surpassed traditional culture in pathogen identification (65 strains vs. 23 strains). Within these 65 strains, 56 were found in Group D, 26 in Group I, and 17 were overlapping strains. Interestingly, a diverse array of G+ bacteria, fungi, viruses, and special pathogens were exclusive to Group D. Furthermore, Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae were more prevalent in Group D compared to Group I. Importantly, mNGS prompted antibiotic treatment adjustments in 26 patients (52.0%).Conclusions:Compared with the conventional culture, mNGS demonstrated a higher positive rate, and emerges as a promising method for identifying mixed pathogens in non-immunodeficient patients with severe pneumonia supported by vv-ECMO. However, it is crucial to combine the interpretation of mNGS data with clinical information and traditional culture results for a comprehensive assessment.

ObjectivesInfection is one of the important causes of death in intensive care unit (ICU) patients. At present, there are few articles focused on the detailed analysis of pathogenic microorganisms detected in different therapy periods of critically ill patients supported by extracorporeal membrane oxygenation (ECMO). MethodsFrom October 2020 to October 2022, ECMO-assisted patients who underwent multiple times of both metagenomic next-generation sequencing (mNGS) test and conventional culture were enrolled continuously in the First Affiliated Hospital of Zhengzhou University. The baseline data, laboratory test results, and pathogenic microorganisms detected by mNGS and traditional culture in different time periods were recorded and analyzed. ResultsIn the present study, 62 patients were included finally. According to whether the patients survived at discharge, they were divided into the survivor group (n = 24) and the non-survivor group (n = 38). Then, according to the different types of ECMO support, they were divided into the veno-venous ECMO (VV ECMO) group (n = 43) and the veno-arterial ECMO (VA ECMO) group (n = 19). The summit period of specimens of traditional culture and mNGS detection of ECMO patients was 7 days after admission, and the largest number of specimens of surviving patients appeared after ECMO withdrawal. The total number of traditional culture specimens was 1,249, the positive rate was 30.4% (380/1,249), and the positive rate of mNGS was 79.6% (82/103). A total of 28 kinds of pathogenic microorganisms were cultured from conventional culture, and 58 kinds of pathogenic microorganisms were detected by mNGS, including Mycobacterium, Rickettsia, and Chlamydia psittaci. In conventional culture, the most frequent Gram-negative bacteria, Gram-positive bacteria, and fungi were Klebsiella pneumoniae, Corynebacterium striatum, and Candida glabrata, and those with the highest frequency of occurrence in mNGS detection were Acinetobacter baumannii, Enterococcus faecium, and Aspergillus flavus. ConclusionsThroughout the whole treatment process, different kinds of suspicious biological specimens of high-infection-risk ICU patients supported by ECMO should undergo both mNGS detection and traditional culture early and repeatedly.

Platelet transfusion is a life-saving therapy to prevent bleeding; however, the availability of platelets for transfusion is limited by the markedly short shelf life owing to the development of platelet storage lesions (PSLs). The mechanism of PSLs remains obscure. Dissection of the intracellular biological changes in stored platelets may help to reduce PSLs and improve platelet transfusion efficiency. In the present study, we explore the changes of stored platelets at room temperature under constant agitation. We found that platelets during storage showed an increased reactive oxygen species (ROS) generation accompanied with receptor shedding, apoptosis, and diminished platelet aggregation. ROS scavenger reduced platelet shedding but also impaired platelet aggregation. Autophagy is a conserved catabolic process that sequesters protein aggregates and damaged organelles into lysosomes for degradation and platelets' own intact autophagic system. We revealed that there exist a stable autophagic flux in platelets at the early stage of storage, and the autophagic flux in platelets perished after long-term storage. Treatment stored platelets with rapamycin, which stimulates autophagy in eukaryotic cells, markedly ameliorated PSLs, and improved platelet aggregation in response to extracellular stimuli.

包覆层界面脱粘极易使药柱局部应力集中,影响药柱正常燃烧,使其运行存在严重安全隐患.针对药柱结构在加工或服役过程中粘接界面产生分层、脱胶等缺陷所带来的结构性能差、耐久性能低等安全问题,基于数字敲击检测理论对粘结结构内预置的不同种类缺陷进行研究,得知脱粘弹性系数k2直接影响脱粘缺陷程度,而其与敲击碰撞产生的应力持续时间呈负相关性,因此利用ANSYS软件中显示动力学模块对敲击检测模型进行仿真分析,由数据结果提取出的应力持续时间界定影响包覆层粘接质量的不同因素对其产生的影响关系与程度.实验表明敲击检测能力会因结构中局部缺陷大小、缺陷深度、缺陷位置、敲击角度的不同产生相关性影响.

Objectives Extracorporeal membrane oxygenation (ECMO) patients with or without transport both have high hospital mortality rate and there are few data on adult VA-ECMO transport patients. Hence, this study was designed to analyze factors that affect the outcomes of patients with ECMO transport. Methods This study retrospectively enrolled 126 ECMO patients transferred from regional hospital to the First Affiliated Hospital of Zhengzhou University by our ECMO team during June 2012 to Sept 2020. Data were calculated and analyzed. Results The median distance of transportation was 141 (76–228) km, the median transport time consuming was 3 (1.3–4) h, the percentage of complications during transport was 40.5% (except for bleeding on cannula site, and no one death during transport), and the survival rate in hospital was 38.9%. Compared with survivors, the non-survivors were older and showed higher SOFA score, longer time with ECMO assisted, longer time in ICU and in hospital. However, after divided into VA-ECMO and VV-ECMO groups, the older age showed no significant difference between survivors and non-survivors groups of VA-ECMO patients. Moreover, the Cox regression survival analysis showed that higher SOFA score and lactate level indicated higher ICU mortality of VA-ECMO patients while higher SOFA score, higher lactate level, older age and lower MAP after transportation (<70mmHg) indicated higher ICU mortality of VV-ECMO patients. However, there was no significant difference of comorbidities and complications in survivors and non-survivors groups of ECMO patients. Conclusions The transportation for ECMO patients can be feasible performed although life-threatening complications might occur. The SOFA score and the lactate level could be used to evaluate the risk of ICU mortality of transportation ECMO patients. Besides, lower MAP after transportation (<70mmHg) had potential predictive value for short-term outcome of VV-ECMO patients.

Abstract Background The anti-coagulation protocol of patients with hemorrhage risk primary disease who need extracorporeal membrane oxygenation (ECMO) supported is controversial. This study evaluated the feasibility of a new anti-coagulation strategy, that is heparin-free after 3000 IU heparin loaded in veno-venous ECMO (VV ECMO) supported acute respiratory failure patients with hemorrhage risk. Methods A retrospective study was performed in a series of hemorrhage risk patients supported with VV ECMO at the First Affiliated Hospital of Zhengzhou University, between June 2012 to Sept 2020. A total of 70 patients received a low heparin bolus of 3000 units for cannulation but without subsequent, ongoing heparin administration. Patients were divided into survival (n = 25) and non-survival group (n = 45). Data of coagulation, hemolysis and membrane lung function were calculated and analyzed. The complications of patients were recorded. Finally, the binary Logistic regression was conducted. Results The longest heparin-free time was 216 h, and the mean heparin-free time was 102 h. Compared with survivors, the non-survivors were showed higher baseline SOFA score and lower platelet counts in 0.5 h, 24 h, 48 h and 96 h after ECMO applied. However, there was no significant differences between survivors and non-survivors in ACT, APTT, INR, D-dimer, fibrinogen, LDH, blood flow rate, Δp and Ppost-MLO2 (all p < 0.05) of all different time point. Moreover, only the baseline SOFA score was significantly associated with mortality (p < 0.001, OR(95%CI): 2.754 (1.486–5.103)) while the baseline levels of ACT, APTT, INR, platelet, D-dimer, fibrinogen and LDH have no association with mortality. The percentage of thrombosis complications was 54.3% (38/70) including 3 oxygenator changed but there was no significant difference of complications in survival and non-survival groups (p > 0.05). Conclusions The anticoagulation protocol that no heparin after a 3000 units heparin bolus in VV ECMO supported acute respiratory failure patients with hemorrhage risk is feasible.

Background: The anti-coagulation protocol of patients with hemorrhage risk primary disease who need extracorporeal membrane oxygenation (ECMO) supported is controversial. This study evaluated the safety of heparin-free after 3000IU heparin loaded in veno-venous ECMO (VV ECMO ) supported acute respiratory failure patients with hemorrhage risk. Methods: A retrospective study was performed in a series of hemorrhage risk patients supported with VV ECMO at the First Affiliated Hospital of Zhengzhou University, between June 2012 to Sept 2020. A total of 70 patients received a low heparin bolus of 3000 units for cannulation but without subsequent, ongoing heparin administration. Patients were divided into survival (n=25) and non-survival group (n=45). Data of coagulation, hemolysis and membrane lung function were calculated and analyzed. The complications of patients were recorded. Finally, the binary Logistic regression was conducted. Results: The longest heparin-free time was 216 hours, and the mean heparin-free time was 102 hours. The percentage of thrombosis complications was 54.3% (38/70) including 3 oxygenator changed but there was no significant difference of complications in survival and non-survival groups (p>0.05). Compared with survivors, the non-survivors were showed higher baseline SOFA score and lower platelet counts in 0.5 hour, 24 hours, 48 hours and 96 hours after ECMO applied. However, there was no significant differences between survivors and non-survivors in ACT, APTT, INR, D-dimer, fibrinogen, LDH, blood flow rate, Δp and P post-ML O 2 (all p<0.05) of all different time point. Moreover, only the baseline SOFA score was significantly associated with mortality (p<0.001, OR(95%CI): 2.754 (1.486-5.103)) while the baseline levels of ACT, APTT, INR, platelet, D-dimer, fibrinogen and LDH have no association with mortality. Conclusions: The anticoagulation protocol that no heparin after a 3000 units heparin bolus in VV ECMO supported acute respiratory failure patients with hemorrhage risk is safe.

Objectives There are few studies of metagenomic next-generation sequencing (mNGS) in immunocompromised patients assisted by veno-venous extracorporeal membrane oxygenation (vv-ECMO). The present study is aimed to investigate the pathogen-detected effect and clinical therapy value of mNGS technologies in immunocompromised patients assisted by vv-ECMO. Methods Our study retrospectively enrolled 46 immunocompromised patients supported by vv-ECMO from Jan 2017 to June 2021 at the First Affiliated Hospital of Zhengzhou University, respectively. Patients were divided into the deterioration group (Group D) (n = 31) and improvement group (Group I) (n = 15) according to their outcomes. Baseline characteristics and etiological data of patients during hospitalization of 2 groups were compared. The pathogens detected by mNGS and antibiotic regimens guided by mNGS in immunocompromised patients assisted by vv-ECMO were analyzed. Results Compared with Group I, the deterioration patients showed a higher percentage of chronic obstructive pulmonary disease (COPD) (32.3% vs. 6.7%, p < 0.01) and were significantly older (47.77 ± 16.72 years vs. 32 ± 15.05 years, p < 0.01). Within 48 h of being ECMO assisted, the consistency of the samples detected by traditional culture and mNGS at the same time was good (traditional culture vs. mNGS detection, the positive rate of bronchoalveolar lavage fluid (BALF) culture: 26.1% vs. 30.4%; the positive rate of blood sample culture: 12.2% vs. 12.2%, p > 0.05). However, mNGS detected far more pathogen species and strains than conventional culture (30 strains vs. 78 strains, p < 0.01); the most popular pathogen was Klebsiella pneumoniae. Parts of patients had their antibiotic treatment adjustments, and the improvement patients showed less usage of broad-spectrum antibiotics. Conclusions mNGS may play a relatively important role in detecting mixed pathogens and personalized antibiotic treatment in immunocompromised patients assisted by vv-ECMO.

Objective Reports on negative results of metagenomic next-generation sequencing (mNGS) are scarce. We aimed to explore the diagnostic value of negative results in bronchoalveolar lavage fluid (BALF) mNGS and how to deal with the negative results in patients with severe respiratory disease. Methods A retrospective analysis was performed on patients suspected severe community-acquired pneumonia who were admitted to the respiratory intensive care unit of the First Affiliated Hospital of Zhengzhou University from January 2020 to December 2021. According to the final diagnosis as the reference standard, the negative results of mNGS were divided into a true negative group and a false negative group. For enrolled patients, we recorded their demographic data, imaging results, laboratory results, therapeutic processes, and prognoses. Results A total of 21 patients were enrolled in this study, including 16 true negative patients and 5 false negative patients. In the true negative group, interstitial lung diseases were the most and neoplastic diseases were following. In addition to mNGS, 9 patients underwent pathological examination, 7 patients were finally diagnosed by medical history, autoantibodies, and point-of-care (POC) ultrasound. 14 patients eventually discontinued antibiotics, 2 patients underwent antibiotic de-escalation, the average interval time of treatment adjustment was 3.56 ± 2.00 days. In the false negative group, the leading missed pathogen was fungi, followed by tuberculosis bacilli. In contrast to 2 patients underwent pathological examination, 3 patients were confirmed by routine microbiological tests. Conclusions Negative results of BALF mNGS can help to rule out infection, but missed diagnoses may also exist. It should be re-evaluated with other clinical informations. Pathological examination or repeated mNGS may be viable options when the diagnosis cannot be confirmed.

Objectives The metagenomic next-generation sequencing (mNGS) test is useful for rapid and accurate detection and identification of pathogenic microorganisms. The aim of the present study was to investigate the factors associated with in-hospital mortality in pneumocystis pneumonia (PCP) patients with mNGS-assisted diagnosis. Methods Our study enrolled 154 patients with mNGS-positive PCP from August 2018 to February 2022 at the First Affiliated Hospital of Zhengzhou University respectively. Patients were divided into the survivor group (n=98) and the death group (n=56) according to whether in-hospital death occurred. Baseline characteristics, patients’ pre-hospital symptoms and patients’ CT imaging performance during hospitalization were carefully compared between the two groups. Risk factors for the occurrence of in-hospital death were sought by selecting indicators that were significantly different between the two groups for modelling and performing multiple logistic regression analysis. Results Compared with the in-hospital death patients, the survivors were younger and had higher levels of albumin (ALB) (age: 50.29 ± 14.63 years vs 59.39 ± 12.27 years, p<0.001; ALB: 32.24 ± 5.62 g/L vs 29.34 ± 5.42g/L, p=0.002; respectively), while the levels of lactate dehydrogenase (LDH) and C-reactive protein CRP were lower (LDH: 574.67 ± 421.24 U/L vs 960.80 ± 714.94 U/L, p=0.001; CRP: 54.97 ± 55.92 mg/L vs80.45 ± 73.26 mg/L, p= 0.018; respectively). Multiple logistic regression analysis revealed that age, the baseline LDH and CRP levels were all positively associated with high in-hospital mortality [age: OR(95%CI): 1.115 (1.062-1.172), p<0.001; LDH: OR(95%CI): 1.002 (1.001-1.003), p<0.001; CRP: OR(95%CI): 1.008 (1.000-1.017), p=0.045; respectively] while the platelet counts was negatively associated with it [OR(95%CI): 0.986 (0.979-0.992), p<0.001]. Conclusions Old age, high baseline levels of LDH and CRP and low platelet counts were risk factors of the in-hospital mortality in mNGS positive PCP patients.