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.

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.

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.

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: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.

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 explore the value of metagenomic next-generation sequencing (mNGS) in the pathogen diagnosis of liver abscess.Methods:A perspective study was performed in 35 hospitalized patients with liver abscess in Department of Emergency Medicine, Zhongshan Hospital, Fudan University from February 2020 to April 2021. Blood samples and abscess drainage fluid samples were detected by routine microbial culture and mNGS. Patients were divided into two groups according to whether they had septic shock or not. SPSS 25.0 was used for statistical analysis.Results:The overall positive rate of mNGS in blood samples and drainage fluid samples was significantly higher than that of routine microbial culture methods (blood: 67.6% vs. 15.2%, P<0.05; Drainage fluid: 100% vs. 55.2%, P<0.05). In 35 patients with liver abscess, 71.4% of the pathogens were Klebsiella pneumoniae. The sequence number of pathogenic pathogens detected by mNGS in abscess drainage fluid samples of patients in the shock group was significantly higher than that in the non-shock group ( P<0.05). Conclusions:The mNGS can quickly and accurately detect the pathogen of liver abscess, which can provide important etiological diagnostic for clinical treatment.

Objective:To investigate whether deletion of Gpr174 alleviates gut injuries during sepsis and identify the differential gut microbiota, metabolites and related metabolic pathways between C57BL/6 and Gpr174 -/- mice in physiological condition and sepsis. Methods:Twelve 8-week-old male C57BL/6 and Gpr174 -/- mice were randomly (random number) divided into the following four groups: Wildtype (WT) group, Gpr174 -/- (KO) group, Wildtype + CLP (WT+CLP) group and Gpr174 -/- + CLP (KO+CLP) group. Sepsis mice model was established by cecal ligation and puncture (CLP) as previously described. Feces were collected from C57BL/6 and Gpr174 -/- mice in normal condition and 3 days after CLP. The nucleic acid of stool was extracted and then amplified the V3 V4 region using TransStart FastPfu DNA Polymerase; 16S rDNA gene amplicons were sequenced on an Illumina MiSeq instrument. After demultiplexing and quality filtering, differential microbiota was identified. Metabonomics was determined by liquid chromatography (LC-MS). Endogenous metabolites were identified by the Feihn metabonomics database. Then, metabolic pathways were further analyzed via KEGG. Results:The principal component analysis (PCA) showed a cluster type distribution between the WT group and KO group . The difference between the WT+ CLP group and KO + CLP group was significantly reduced after CLP. The differential gut microbiota included Lactobacillus, Akkermansia, Bacteroides, Allobaculum, Bifidobacterium, Rikenella, Olsenella, Barnesiella, and the differential metabolites included L-Alanine, Hydroxypropionic acid, Oxoglutaric acid. The related signal pathways of differential metabolites were Glucose-Alanine Cycle, Alanine Metabolism and Propanoate Metabolism. Conclusions:Gpr174 deletion could alleviate gut injuries during sepsis and change the composition of gut microbiota and metabolites.

Objective:Dysregulated host immune responses contribute to the pathogenesis of sepsis. G protein-coupled receptor 174 (GPR174) was found to be involved in the immune responses and associated with the susceptibility to autoimmune diseases. This study aimed to investigate the association of GPR174 variants with sepsis susceptibility and the contribution of GPR174 in sepsis development.Methods:From May 2005 to December 2017, a total of 575 sepsis patients and 579 non-septic controls admitted to our Emergency ICU were enrolled in this case-control study. The non-synonymous SNP rs3827440 in GPR174 was genotyped using TaqMan Real-time PCR assays on ABI7900 platform. Then the correlation between rs3827440 and serum levels of interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-α) were investigated in septic patients. Gpr174-deficient mice were generated and subjected to cecal ligation and puncture (CLP). The concentrations of inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA).Results:Rs3827440 TT/T genotype in GPR174 was positively associated with sepsis risk after logistic regression analysis adjusted for sex [odds ratio ( OR) = 1.68, 95% confidence interval ( CI): 1.19-2.20, P = 0.0004]. IL-6 and TNF-α serum levels in female TT and male T allele carriers of septic patients were significantly higher than those in female CC and male C allele carriers ( P < 0.05). Preclinical validation of Gpr174 gene was performed in Gpr174 knockout (KO) mice using CLP models. Gpr174 KO mice had higher survival rate. Moreover, Gpr174 KO mice had significantly decreased serum concentrations of IL-1β, IL-6 and TNF-α compared with WT mice, while the levels of IL-10 was increased ( P < 0.01). Conclusions:GPR174 as a novel sepsis susceptibility gene in Chinese Han population is involved in the development and physiopathology of sepsis.

Objective:To compare the predictive value of the HEART, TIMI and GRACE scores for major adversecardiovascular events (MACEs) at 7 and 28 days in patients with actue non-ST-segment elevation myocardial infarction (NSTEMI).Methods:More than 12 000 patients with chest pain from the Emergency Department of Zhongshan Hospital Affiliated to Fudan University from October 2017 to October 2018 were studied, including 566 patients with cardiogenic chest pain, 105 patients with ST-segment elevation myocardial infarction (STEMI) excluded and 15 patients lost to follow-up. Finally, 109 patients with NSTEMI and 337 non-myocardial patients with cardiogenic chest pain were enrolled. NSTEMI patients were divided into subgroups according to whether MACEs occurred. LSD t-test, Mann-Whitney U test or χ2 test were used to analyze and compare the differences between the two subgroups about the baseline data, clinical data, HEART, TIMI and GRACE scores at the time of visit. Multivariate logistic regression analysis was used to explore the independent factors of MACEs at 7 and 28 days. And the predictive values of different scores for 7-day MACEs and 28-day MACEs were compared in NSTEMI patients through the receiver operating characteristic (ROC) curve. Results:Compared NSTEMI patients with non-myocardial patients with cardiogenic chest pain, we found a statistically significant differences in sex, past history of coronary heart disease,≥3 risk factors for atherosclerosis, electrocardiogram, high-sensitivity troponin T (hs-cTnT), creatinine value, past history of myocardial infarction, HEART score, TIMI score and GRACE score. In further subgroup analysis of NSTEMI patients who were divided according to whether MACEs occurred, we found previous history of stroke and increased hs-cTnT were statistically different in 7 days after the onset of the disease. The multivariate analysis showed that the previous history of stroke and increased hs-cTnT were independent factors for the occurrence of MACEs at 7 days after the onset of NSTEMI; The previous history of stroke and increased hs-cTnT, electrocardiogram ST segment depression and TIMI score were statistically different at 28 days after the onset of NSTEMI. The multivariate analysis showed that the previous history of stroke and TIMI score were independent factors for the occurrence of MACEs at 28 days after the onset of NSTEMI patients. ROC curve indicated that the predictive value of TIMI score (AUC=0.715, 95% CI: 0.482-0.948) was better than HEART (AUC=0.659, 95% CI: 0.414-0.904) and GRACE scores (AUC=0.587, 95% CI: 0.341-0.833)in predicting MACEs in NSTEMI patients. Conclusions:HEART score, TIMI score and GRACE score can be used to evaluate NSTEMI patients. There is an independent predictive value on TIMI score for the occurrence of 28-day MACEs in NSTEMI patients.