Objective To explore the dynamic changes of gut microbiota metabolites in septic patients following admission, as well as the correlations between these metabolites, the gut microbiota, and the prognosis of septic patients. Methods A total of 119 fecal samples were collected from 23 septic patients, 16 non-septic patients admitted to the Emergency Intensive Care Unit (ICU), and 20 healthy controls at Zhongshan Hospital, Fudan University, from January to August 2019. The 16S rRNA gene sequencing technology was applied to analyze the microbiome, while ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used for metabolomics research. The R software was used to analyze the gut microbiota and metabolite data. Based on 180-day survival status after admission, the sepsis group was divided into the survival group (n＝15) and the death group (n＝8) to analyze differential metabolites between the two groups. Spearman correlation coefficients were used to assess correlations between gut microbiota and metabolites. Results In the first week of ICU stay, gut microbiota metabolites such as nicotinic acid, methylsuccinic acid, and glutaric acid were significantly lower in septic patients than in healthy controls (P＜0.05), whereas tryptophan, histidine, valine, and pyroglutamic acid were higher in septic patients (P＜0.05). The methylsuccinic acid and phenylacetic acid levels in the first week were lower in the death group than those in the survival group (P＜0.05), and the levels of methylsuccinic acid, phenylacetic acid, and glutaric acid were lower in the third week (P＜0.05). Further analysis indicated that methylsuccinic acid was closely associated with sepsis prognosis. These differential metabolites involved in metabolic pathways such as phenylalanine metabolism and β-alanine metabolism. Most differential amino acids were positively correlated with opportunistic pathogens but negatively correlated with normal gut microbiota. Conversely, metabolites such as nicotinic acid, phenylacetic acid, methylsuccinic acid, and glutaric acid were negatively correlated with opportunistic pathogens and positively correlated with normal gut microbiota. Conclusions Significant dynamic changes occur in gut microbiota metabolites in septic patients, with methylsuccinic acid, phenylacetic acid, and glutaric acid potentially playing important roles in determining patient prognosis.

Objective:To assess the real-world triage performance of a dual-marker strategy (DMS) combining copeptin and high-sensitivity cardiac troponin T (hs-cTnT) in patients presenting with chest pain and suspected non-ST-segment elevation myocardial infarction (NSTEMI).Methods:It was conducted a prospective study of 277 consecutive chest pain patients admitted to the Emergency Department of Zhongshan Hospital, Fudan University, between July and August 2023. Admission levels of copeptin and hs-cTnT were measured. The safety, efficacy, and triage efficiency of the DMS (defined as copeptin <10 pmol/L and hs-cTnT <0.014 ng/mL) for excluding NSTEMI were evaluated based on final diagnoses and clinical outcomes.Results:Among 277 patients, 141 (50.9%) had cardiogenic diseases (51 NSTEMI, 37 unstable angina pectoris [UAP], 11 myocardial bridges, and 42 non-coronary artery disease), 29 (10.5%) had non-cardiac conditions, and 107 (38.6%) had low-risk chest pain of unknown etiology. A total of 103 patients (37.2%) were DMS-negative (copeptin and hs-cTnT both below cutoff), including 0 NSTEMI cases, 2 UAP cases, 1 myocardial bridge, 6 non-coronary artery diseases, 4 non-cardiac conditions, and 90 low-risk cases. The DMS demonstrated a negative predictive value (NPV) of 100% for excluding NSTEMI, with no major adverse cardiac events (MACE) observed in DMS-negative patients during 30-day follow-up. Real-world data revealed that only 42.2% of suspected NSTEMI patients received a second troponin test (timing: 1 hour—5.9%, 2 hours—23.9%, ≥3 hours—70.1%). The DMS enabled safe and efficient triage of 37.2% of chest pain patients at 0-hour, outperforming other strategies in applicability and feasibility ( P < 0.05). Conclusions:In real-world clinical practice, the DMS (copeptin combined with hs-cTnT) optimally complements guideline-recommended hs-cTnT algorithms. It provides a simple, rapid, and safe approach to managing acute chest pain, demonstrating superior applicability for improving emergency triage efficiency.

Objective·To explore the diagnostic,therapeutic,and prognostic value of metagenomics next-generation sequencing(mNGS)in patients with pneumonia-induced sepsis.Methods·This study consisted of a multicenter,prospective,non-randomized controlled trial and a diagnostic test.Patients with pneumonia-induced sepsis who were hospitalized in four hospitals across China were enrolled between March 2020 and October 2021.All patients met the Sepsis-3 criteria issued by the Society of Critical Care Medicine and the European Society of Intensive Care Medicine,as well as the clinical diagnostic standard of pneumonia.Enrolled patients were assigned based on their preference to either the conventional test-only group[receiving only conventional test(CMT)]or the combined mNGS test group(receiving CMT and mNGS concurrently).The primary outcome was the 7-day all-cause mortality rate,and secondary outcomes included the changes in SOFA and APACHE Ⅱ scores from baseline to day 7,28-day all-cause mortality rate,the composite endpoint of mechanical ventilation or death within 28 d,28 d ventilation-free days,28 d hospital-free days,and the average daily hospitalization cost.Propensity score matching was used to balance covariates between the two groups.Kaplan-Meier curves were plotted and Cox proportional hazards models were built to compare the risk of death between the two groups.Pathogen detection results from infection site samples in the combined mNGS test group were used for the diagnostic test.The clinically-adjudicated causative pathogens was used as the reference standard.The results of traditional pathogen detection and mNGS detection were compared respectively with the reference standard.The positive percent agreement,negative percent agreement,positive predictive value,and negative predictive value between the two methods and the reference standard were calculated.McNemar's χ2 test was used to evaluate the causative pathogen detection capabilities of the two methods.Results·A total of 533 patients were enrolled,of whom 311 opted for additional mNGS testing,while 222 received only conventional pathogenetic testing.In the non-randomized controlled trial,after propensity score matching to balance covariates,the 7-day all-cause mortality was lower in the combined mNGS test group compared to the conventional test-only group[4.8%vs 8.6%,HR 0.37(95%CI 0.15?0.91),P=0.031].Additionally,the 28-day ventilation-free days were increased in the combined mNGS test group(19.9 d vs 18.4 d,P=0.041).No significant difference was observed between the two groups in terms of 28-day all-cause mortality or the average daily hospitalization costs.In the diagnostic test,compared to the reference standard,the positive percent agreement of mNGS with the clinical composite judgment for causative pathogens was higher than that of CMT[91.9%(95%CI 87.7%?95.0%)vs 56.1%(95%CI 49.7%?62.4%),P<0.001].Conversely,the negative percent agreement of mNGS was lower than that of CMT[29.2%(95%CI 18.6%?41.8%)vs 69.2%95%CI 56.6%?80.1%),P<0.001].The negative predictive value of nNGS was higher than that of CMT[48.7%(95%CI 32.4%?65.2%)vs 29.4%(95%CI 22.3%?37.3%),P=0.001].Conclusion·In patients with pneumonia-induced sepsis,mNGS of infection site samples demonstrated a higher detection rate of causative pathogen compared to CMT.Furthermore,the combination of mNGS with CMT may help reduce the 7-day all-cause mortality,suggesting that mNGS has clinical value and potential for application in the management of sepsis caused by pulmonary infections.

Objective·To explore the diagnostic,therapeutic,and prognostic value of metagenomics next-generation sequencing(mNGS)in patients with pneumonia-induced sepsis.Methods·This study consisted of a multicenter,prospective,non-randomized controlled trial and a diagnostic test.Patients with pneumonia-induced sepsis who were hospitalized in four hospitals across China were enrolled between March 2020 and October 2021.All patients met the Sepsis-3 criteria issued by the Society of Critical Care Medicine and the European Society of Intensive Care Medicine,as well as the clinical diagnostic standard of pneumonia.Enrolled patients were assigned based on their preference to either the conventional test-only group[receiving only conventional test(CMT)]or the combined mNGS test group(receiving CMT and mNGS concurrently).The primary outcome was the 7-day all-cause mortality rate,and secondary outcomes included the changes in SOFA and APACHE Ⅱ scores from baseline to day 7,28-day all-cause mortality rate,the composite endpoint of mechanical ventilation or death within 28 d,28 d ventilation-free days,28 d hospital-free days,and the average daily hospitalization cost.Propensity score matching was used to balance covariates between the two groups.Kaplan-Meier curves were plotted and Cox proportional hazards models were built to compare the risk of death between the two groups.Pathogen detection results from infection site samples in the combined mNGS test group were used for the diagnostic test.The clinically-adjudicated causative pathogens was used as the reference standard.The results of traditional pathogen detection and mNGS detection were compared respectively with the reference standard.The positive percent agreement,negative percent agreement,positive predictive value,and negative predictive value between the two methods and the reference standard were calculated.McNemar's χ2 test was used to evaluate the causative pathogen detection capabilities of the two methods.Results·A total of 533 patients were enrolled,of whom 311 opted for additional mNGS testing,while 222 received only conventional pathogenetic testing.In the non-randomized controlled trial,after propensity score matching to balance covariates,the 7-day all-cause mortality was lower in the combined mNGS test group compared to the conventional test-only group[4.8%vs 8.6%,HR 0.37(95%CI 0.15?0.91),P=0.031].Additionally,the 28-day ventilation-free days were increased in the combined mNGS test group(19.9 d vs 18.4 d,P=0.041).No significant difference was observed between the two groups in terms of 28-day all-cause mortality or the average daily hospitalization costs.In the diagnostic test,compared to the reference standard,the positive percent agreement of mNGS with the clinical composite judgment for causative pathogens was higher than that of CMT[91.9%(95%CI 87.7%?95.0%)vs 56.1%(95%CI 49.7%?62.4%),P<0.001].Conversely,the negative percent agreement of mNGS was lower than that of CMT[29.2%(95%CI 18.6%?41.8%)vs 69.2%95%CI 56.6%?80.1%),P<0.001].The negative predictive value of nNGS was higher than that of CMT[48.7%(95%CI 32.4%?65.2%)vs 29.4%(95%CI 22.3%?37.3%),P=0.001].Conclusion·In patients with pneumonia-induced sepsis,mNGS of infection site samples demonstrated a higher detection rate of causative pathogen compared to CMT.Furthermore,the combination of mNGS with CMT may help reduce the 7-day all-cause mortality,suggesting that mNGS has clinical value and potential for application in the management of sepsis caused by pulmonary infections.

Objective To explore the changes in the expression of CX3CR1 in natural killer (NK) cells from peripheral blood mononuclear cells (PBMC) in sepsis patients and its association with gut microbiota. Methods A total of 24 sepsis patients were selected from January 2020 to January 2021 at Zhongshan Hospital, Fudan University, and 10 healthy volunteers were recruited in January 2021 as healthy controls. Fecal samples and peripheral blood were collected from sepsis patients on the first and fourth days of hospitalization. Sequencing of the V3-4 region of the 16S rDNA gene of gut microbiota was performed using the Illumina MiSeq platform. The peripheral blood samples were isolated by positively selected magnetic beads, and the CD3^－CD56^＋NK cells were identified by flow cytometry. The mRNA expression of CX3CR1 was detected by qPCR, and the changes in CX3CR1 expression and its correlation with gut microbiota were analyzed. Results Compared with healthy control group, the Shannon diversity index of the gut microbiota and the proportion of Firmicutes in sepsis patients decreased; compared with admission day, the Shannon diversity of the gut microbiota in sepsis patients on the fourth day of hospitalization significantly decreased, the proportion of Proteobacteria on phylum level and the relative abundance of Enterococcus and Klebsiella on genus level significantly increased. The CX3CR1 expression of PBMC-NK cells in sepsis patients on the fourth day was significantly lower than that on the admission day (P＜0.001). Compared with surviving patients, CX3CR1 expression in non-surviving patients significantly decreased on the fourth day (P＜0.05). Spearman correlation analysis showed that CX3CR1 expression of PBMC-NK cells was positively correlated with the quantity of gut microbiota and the Shannon diversity index (P＜0.01). Conclusions The expression of CX3CR1 in PBMC-NK cells in sepsis patients decreases with disease progression, and is related to prognosis. Furthermore, its expression is found to be closely related to the gut microbiota.

Establishing a comprehensive mechanism for the initiation and review of investigator-initiated trial(IIT) plays an important role in ensuring the scientific validity of clinical research and improving research quality.Since 2021, Zhongshan Hospital affiliated to Fudan University had actively explored improvements in the project management of IIT. The hospital had established a standardized grading review management process, developed an integrated clinical research management system, established a three-level clinical research training system, built a methodological support platform, and formulated research plan templates, gradually formed a standardized grading project approval review management mode. As of February 2024, the hospital had completed 400 quick reviews and more than 400 expert letter reviews based on the integrated clinical research management system. The efficiency and quality of IIT project approval had been improved. At the same time, over 40 academic salons and forums had been held, cultivating a group of young clinical research talents, providing data management training for more than 30 clinical departments, and promoting the improvement of the quality of research protocol. In the future, hospitals should further optimize their information systems, expand the influence of their training systems, enhance the capabilities of their methodological support platforms, and improve the efficiency of the application of clinical research protocol templates, so as to escort the establishment and implementation of high-quality clinical research projects and provide references for other hospitals′ IIT project management.

BACKGROUND:Disseminated intravascular coagulation(DIC)is associated with increased mortality in sepsis patients.In this study,we aimed to assess the clinical ability of sepsis-induced coagulopathy(SIC)and sepsis-associated coagulopathy(SAC)criteria in identifying overt-DIC and pre-DIC status in sepsis patients. METHODS:Data from 419 sepsis patients were retrospectively collected from July 2018 to December 2022.The performances of the SIC and SAC were assessed to identify overt-DIC on days 1,3,7,or 14.The SIC status or SIC score on day 1,the SAC status or SAC score on day 1,and the sum of the SIC or SAC scores on days 1 and 3 were compared in terms of their ability to identify pre-DIC.The SIC or SAC status on day 1 was evaluated as a pre-DIC indicator for anticoagulant initiation. RESULTS:On day 1,the incidences of coagulopathy according to overt-DIC,SIC and SAC criteria were 11.7％,22.0％and 31.5％,respectively.The specificity of SIC for identifying overt-DIC was significantly higher than that of the SAC criteria from day 1 to day 14(P＜0.05).On day 1,the SIC score with a cut-off value＞3 had a significantly higher sensitivity(72.00％)and area under the curve(AUC)(0.69)in identifying pre-DIC than did the SIC or SAC status(sensitivity:SIC status 44.00％,SAC status 52.00％;AUC:SIC status 0.62,SAC status 0.61).The sum of the SIC scores on days 1 and 3 had a higher AUC value for identifying the pre-DIC state than that of SAC(0.79 vs.0.69,P＜0.001).Favorable effects of anticoagulant therapy were observed in SIC(adjusted hazard ratio[HR]=0.216,95％confidence interval[95％CI]:0.060-0.783,P=0.018)and SAC(adjusted HR=0.146,95％CI:0.041-0.513,P=0.003). CONCLUSION:The SIC and SAC seem to be valuable for predicting overt-DIC.The sum of SIC scores on days 1 and 3 has the potential to help identify pre-DIC.

Objective:To investigate the clinical characteristics of patients with acute aortic dissection (AAD) through a retrospective and observational study, and to construct an early warning model of AAD that could be used in the emergency room.Methods:The data of 11 583 patients in the Emergency Chest Pain Center from January to December 2019 were retrospectively collected from the Chest Pain Database of Zhongshan Hospital Affiliated to Fudan University. Inclusion criteria: patients with chest pain who attended the Emergency Chest Pain Center between January and December 2019. Exclusion criteria were 1) younger than 18 years, 2) no chest/back pain, 3) patients with incomplete clinical information, and 4) patients with a previous definite diagnosis of aortic dissection who had or had not undergone surgery. The clinical data of 9668 patients with acute chest/back pain were finally collected, excluding 53 patients with previous definite diagnosis of AAD and/or without surgical aortic dissection. A total of 9 615 patients were enrolled as the modeling cohort for early diagnosis of AAD. The patients were divided into the AAD group and non-AAD group according to whether AAD was diagnosed. Risk factors were screened by univariate and multivariate logistic regression, the best fitting model was selected for inclusion in the study, and the early warning model was constructed and visualized based on the nomogram function in R software. The model performance was evaluated by accuracy, specificity, sensitivity, positive likelihood ratio and negative likelihood ratio. The model was validated by a validation cohort of 4808 patients who met the inclusion/exclusion criteria from January 2020 to June 2020 in the Emergency Chest Pain Center of the hospital. The effect of early diagnosis and early warning model was evaluated by calibration curve.Results:After multivariate analysis, the risk factors for AAD were male sex ( OR=0.241, P<0.001), cutting/tear-like pain ( OR=38.309, P<0.001), hypertension ( OR=1.943, P=0.007), high-risk medical history ( OR=12.773, P<0.001), high-risk signs ( OR=7.383, P=0.007), and the first D-dimer value ( OR=1.165, P<0.001), Protective factors include diabetes( OR=0.329, P=0.027) and coronary heart disease ( OR=0.121, P<0.001). The area under the ROC curve (AUC) of the early diagnosis and warning model constructed by combining the risk factors was 0.939(95 CI:0.909-0.969). Preliminary validation results showed that the AUC of the early diagnosis and warning model was 0.910(95 CI:0.870-0.949). Conclusions:Sex, cutting/tear-like pain, hypertension, high-risk medical history, high-risk signs, and first D-dimer value are independent risk factors for early diagnosis of AAD. The model constructed by these risk factors has a good effect on the early diagnosis and warning of AAD, which is helpful for the early clinical identification of AAD patients.

Objective:Evaluation of combined inflammatory and coagulation markers for early identification of DIC in septic patients.Methods:This study was a single-center, retrospective, observational study involving 356 patients with sepsis. Sepsis was defined by the diagnostic criteria of Sepsis version 3.0. Definition of DIC was from the International Society on Thrombosis and Hemostasis (ISTH) DIC Score. Inflammatory biomarkers, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β,2R,6,8,10, etc. and biomarkers of coagulation, like platelet (PLT), international normalized ratio (INR), D-dimer, fibrinogen (Fib), etc. were included in this study.Results:Among 356 patients with sepsis, 301 patients did not develop DIC (non-DIC) during hospitalization, 32 patients had DIC on the day of admission (overt-DIC), and 23 patients developed DIC within 1 week of admission (pre-DIC). Compared to non-DIC patients, pre-DIC patients had lower platelet counts and fibrinogen ( P < 0.05), higher levels of INR and D-dimer ( P < 0.05), higher levels of cytokines (TNF-α、IL-1β、IL-2R、IL-8、IL-10) and procalcitonin ( P < 0.05), higher APACHEⅡ and SOFA scores ( P < 0.05). Using receiver operating characteristics (ROC) analysis, we found that some biomarkers of coagulation and inflammation could discriminate pre-DIC from non-DIC patients. The area under the curve (AUC) of INR in the ROC analysis was 0.773 (95% CI: 0.696-0.851), the AUC of IL-2R was 0.700 (95% CI: 0.599-0.798) which is highest among inflammation markers, the highest AUC was obtained from the combination of platelets, INR, Fib, D-dimer and IL-2R (AUC = 0.843; 95% CI: 0.758-0.928). Kaplan-Meier survival curve suggested that high level of IL-2R (> 1064.5 U/mL) was a valuable predictor of 28-day mortality in septic patients. Conclusion:Inflammatory marker, IL-2R, is related to the occurrence of DIC in septic patients and has predictive value for pre-DIC. Combination of coagulation (platelets, INR, Fib, D-dimer) and inflammatory markers (IL-2R) can help to identify pre-DIC state in septic patients.

Objective:To analyze the clinical features of patients with pyogenic liver abscess (PLA) and the application of mNGS in PLA, thus to provide reference for clinical diagnosis and treatment.Methods:The demographic and clinical data of 549 patients with liver abscess admitted to Zhongshan Hospital Affiliated to Fudan University from December 2015 to June 2020 were analyzed retrospectively. According to the detection of Klebsiella pneumoniae in 246 patients with positive etiological test results, the patients were divided into two groups: KPLA group and nKPLA group, and clinical characteristics of the two groups were compared. At the same time, the application value of mNGS in PLA was analyzed.Results:Among the 549 patients, the main clinical symptom of PLA was fever ( n= 503, 91.6%) and other clinical symptoms included chills and abdominal pain. Most patients had a single abscess ( n= 464, 84.5%) located in the right lobe ( n = 368, 67.0%), with a size between 5 and 10 cm ( n= 341, 62.1%). A total of 246 patients had positive etiological test results, including 202 KPLA patients which was the main pathogen of liver abscess. The prevalence of diabetes and fatty liver was higher in KPLA patients ( P < 0.05), but there were more culture of liver positive factors in nKPLA patients ( P < 0.001). Among the 109 patients with traditional microbiological results, 92 patients were suspected to KPLA (Klebsiella pneumoniae), of which 14 patients (15.2%) were multidrug resistant (MDR) infection; 17 patients were suspected to nKPLA, of which 10 patients (58.8%) were MDR infection; the incidence of MDR infection in patients with nKPLA was significantly higher than that in patients with KPLA ( P < 0.05). The positive rate of mNGS in plasma was 85.2%, the positive rate of traditional microbial culture in plasma was 14.8%, the positive rate of mNGS in pus was 96.2% and traditional microbial culture in pus was 65.4%. The positive rate of traditional culture was significantly lower than that of mNGS ( P < 0.05). Conclusions:PLA is usually manifested as fever, single and at the right lobe of the liver. Klebsiella pneumoniae is the most common pathogenic bacteria of PLA, which is more common in patients with diabetes and fatty liver, while non-Klebsiella pneumoniae is relatively more common in patients with culture of liver positive factors. The positive detection rate of mNGS is high, which has a unique advantage in pathogen detection.