Presepsis clotting disease early warning system based on neutrophil extracellular trap net

Abstract

The invention discloses a sepsis blood clotting disease early warning system based on a neutrophil extracellular trapping net. Compared with a traditional detection method, the sepsis clotting disease early warning system has the advantages that the forms of the Nets in the peripheral blood glass slide are observed and counted through blood cell morphology, then the number of the Nets is calculated so as to evaluate the disease risk of an SIC patient, good clinical value is achieved, and compared with traditional peripheral blood MPO-NDA determination, the system has the advantages that the detection accuracy is high, and the detection cost is low. The method is simpler, more direct, quicker, more economical and higher in repeatability. When the score of the combined model exceeds a 0.35 threshold value, it is indicated that a patient has entered a pathological blood coagulation state turning point dominated by a neutrophil extracellular trap net (Nets), in this stage, the generation rate of microthrombus exceeds the clearing ability of an organism, and if anticoagulation treatment is delayed, the risk of organ function failure will be significantly increased.

Description

TECHNICAL FIELD

[0001] The present application relates to a sepsis coagulopathy early warning system based on neutrophil extracellular traps, belonging to the technical field of detection. BACKGROUND

[0002] Sepsis coagulopathy (SIC). According to statistics, about 40.0% to 60.0% of sepsis patients worldwide develop SIC, and in China, the rate is as high as 67.9%. If not intervened in time, it often progresses to disseminated intravascular coagulation (DIC), the mortality doubles, and the prognosis is extremely poor. Given the complex and diverse pathological process of SIC, exploring new biomarkers to improve the accuracy of its early diagnosis is still a top priority.

[0003] In infected organisms, activated neutrophils release DNA, histone, granular protein (such as myeloperoxidase, neutrophil elastase) and other components to form a network complex called neutrophil extracellular traps (Nets). Recent studies have shown that Nets play a key role in driving coagulation imbalance. It not only can directly activate platelets and coagulation factors, but also can interfere with the fibrinolytic system. For example, the histone released by Nets can directly induce platelet activation, aggregation and release of pro-coagulation microparticles by binding to Toll-like receptors (TLR2 / TLR4) and integrin receptors (GPIIb / IIIa) on the surface of platelets. At the same time, histone H3 can also bind to C-type lectin 2 (CLEC-2) on platelets, activate factor Xa, promote intravascular fibrin formation, and exacerbate platelet activation and retention in microvessels, leading to a decrease in circulating platelets, thereby significantly enhancing the coagulation response. On the other hand, the negatively charged DNA in Nets can connect and induce conformational changes in factor XII (FXII), effectively activating the endogenous coagulation cascade. In addition, Nets can also induce monocyte-derived extracellular vesicles to highly express tissue factor (TF) on their surface, thereby activating the coagulation system through the extrinsic coagulation pathway. The unique network structure of Nets itself also provides a stable scaffold for platelets, red blood cells and fibrin, accelerating the formation of microthrombi, and collectively leading to disseminated intravascular coagulation (DIC) and a pro-coagulation state in the body. As sepsis progresses, Nets continue to generate excess thrombin through the FXII / TF-dependent pathway, forcing activated protein C (APC) to continuously neutralize coagulation factors Va and VIIIa, ultimately leading to APC depletion, promoting the transition of sepsis-induced coagulopathy (SIC) from hypercoagulability to consumptive hypocoagulability, fully embodying the core driving role of Nets in the pathological development of SIC.

[0004] Currently, the scoring table included in the SIC diagnostic criteria mainly includes platelet count, PT or INR, D-dimer and SOFA score, etc. However, these indicators are easily disturbed by other factors, not only have low sensitivity for early hypercoagulability, but also change lag behind the actual coagulation activation process, and cannot timely reflect the early dynamics of microthrombus formation and coagulation imbalance. Moreover, SIC involves multiple links such as endothelial damage, platelet activation, coagulation factor consumption and fibrinolytic system disorder, and a single biomarker is difficult to fully reflect the dynamic pathological process and capture the initial stage of sepsis coagulation disorder, leading to delayed clinical intervention. Therefore, emerging biomarkers need to be combined to achieve early diagnosis and dynamic monitoring.

[0005] Current studies have confirmed that specific markers of neutrophil extracellular traps (Nets) such as MPO-DNA complexes, nucleosome H3.1 and cfDNA can be used to evaluate the degree of organ damage in sepsis patients through techniques such as enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, fluorescence spectroscopy, etc. More importantly, recent studies have found that there is a functional subpopulation of Nets in "smear cells" recognized by peripheral blood smear automatic analyzers - these traditionally considered artificial fragments of coagulation have typical Nets structure (DNA-protein complex characteristics) verified by confocal microscopy and flow cytometry. The inventors previously used an automated blood smear system to quantitatively identify the morphological characteristics of Nets-related smear cells and successfully evaluated the mortality risk of COVID-19 patients. However, there is currently no related report on the evaluation of SIC early risk (referring to the time window when the patient meets the Sepsis-3 standard but has not yet met the ISTH SIC diagnosis). This application aims to break through the time limitation of traditional diagnostic methods and achieve earlier warning of Nets-driven microthrombosis and coagulation imbalance, providing a key time window and decision basis for clinical intervention of SIC. SUMMARY

[0006] To solve the problems of the prior art, the present application provides a sepsis coagulopathy early warning system based on neutrophil extracellular traps, specifically a joint diagnostic model based on Nets (morphological quantification), antithrombin III (AT-III) and sequential organ failure score (SOFA). Compared with serological quantification, the Nets quantification method based on blood cell morphology not only has higher diagnostic efficiency, but also has the advantages of rapidity and economy, providing a powerful and efficient tool for early and accurate identification of SIC.

[0007] The present application is achieved by the following technical solutions:

[0008] The first object of the present application is to provide a sepsis coagulopathy early warning system based on neutrophil extracellular traps, which comprises a collection device and an evaluation device. The collection device is used to collect the proportion of neutrophil extracellular traps in white blood cells, the activity of natural anticoagulant substance antithrombin III and the SOFA score in the whole blood of a patient. The evaluation device calculates the joint index through the following prediction equation and evaluates the risk of sepsis coagulopathy in the patient according to the joint index:

[0009] Logit(SIC) model = 41.5 x NETs (%) - 0.068 x ATIII + 0.228 x SOFA - 0.683

[0010]

[0011] Wherein, Logit(SIC) is defined as a combined index for predicting the risk of sepsis-induced coagulopathy (SIC) by integrating NETs%, ATIII and SOFA score, NETs(%) represents the proportion of neutrophil extracellular traps in white blood cells, ATIII represents the activity of natural anticoagulant antithrombin III, SOFA represents SOFA score, and P(SIC) is the prediction probability of sepsis-induced coagulopathy (SIC); for example, when Logit(SIC) = 0.35, the prediction probability of suffering from sepsis-induced coagulopathy (SIC) is about 58.66%.

[0012] In an embodiment of the present application, the proportion of neutrophil extracellular traps in white blood cells is obtained by the following method:

[0013] Preparation and staining, collecting peripheral blood samples of patients, preparing blood smears from the peripheral blood samples, and performing Giemsa staining on the blood smears; and,

[0014] Picture acquisition, obtaining optical microscopic images of white blood cell morphology of the blood sample; and,

[0015] Information processing, counting the number of white blood cells N (白细胞数) and the number of neutrophil extracellular traps N (NETs) in the microscopic images according to the cell morphology; and,

[0016] Data processing, calculating the proportion of neutrophil extracellular traps in white blood cells NETs(%) according to the following formula:

[0017]

[0018] In an embodiment of the present application, images with reticular, segmented nucleus depolymerization and granular characteristics are counted as neutrophil extracellular traps.

[0019] In an embodiment of the present application, the peripheral blood sample is from a patient with sepsis.

[0020] In an embodiment of the present application, the activity of natural anticoagulant antithrombin III is obtained by detecting plasma in an anticoagulant tube with an ACL TOP Family instrument.

[0021] In an embodiment of the present application, the SOFA score is evaluated by a clinician according to the admission status of a patient, including the respiratory system, the cardiovascular system, the liver, the coagulation system, the kidney and the nervous system, and each system is scored according to the degree of functional failure.

[0022] In an embodiment of the present application, the total score of SOFA score is 24, and the higher the score, the more serious the organ failure.

[0023] In one embodiment of the present application, when the combined index is greater than or equal to 0.35, it indicates that the patient with sepsis has a higher risk of secondary coagulopathy; when the combined index is less than 0.35, the patient with sepsis has a lower risk of secondary coagulopathy.

[0024] A second object of the present application is to provide the use of the above-mentioned system in screening drugs for preventing or treating sepsis coagulopathy.

[0025] A third object of the present application is to provide an information data processing terminal for implementing the early warning system for sepsis coagulopathy based on neutrophil extracellular traps.

[0026] The present application has the following beneficial effects:

[0027] Compared with the traditional detection method, the early warning system for sepsis coagulopathy of the present application has good clinical value by observing and counting the morphology of Nets in the peripheral blood slide using blood cell morphology, and then calculating the number of Nets (number per 100 cells) to evaluate the risk of SIC patients. Compared with the traditional determination of peripheral blood MPO-NDA, it is simpler, more direct, faster, more economical and has better repeatability. When the combined model score exceeds the threshold of 0.35, it indicates that the patient has entered the turning point of pathological coagulation state dominated by neutrophil extracellular traps (Nets), at this stage, the generation rate of microthrombus exceeds the body's clearance capacity, and if the anticoagulant therapy is delayed, the risk of organ failure will be significantly increased. The application of the model can realize the transition of clinical intervention strategy from experience to precision. In emergency, Nets count > 7.5% can be used as a high-sensitivity (86%) preliminary screening indicator; subsequently, in the intensive care unit (ICU), through the combined model evaluation of antithrombin III activity without significant consumption and sequential organ failure assessment score without significant increase, the false positive rate is effectively reduced.

[0028] Nets-related markers not only have the potential to help stage sepsis coagulopathy, but also can develop stage-specific Nets intervention strategies, which is an important direction for future treatment of SIC. BRIEF DESCRIPTION OF DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description only some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of these drawings.

[0030] Figure 1 is the flowchart of the patients included;

[0031] Figure 2are morphological observation of neutrophilic NETs cells in SIC patients, A: degenerative lymphocytes; B: typical morphological changes of NETs in sequence: vacuolization, nuclear decondensation, granule release, chromatin extrusion and protrusion; images were captured by 100x magnification;

[0032] Figure 3 are A: NETs number in peripheral blood of non-SIC patients and SIC patients group; B: MPO-DNA content in peripheral blood serum of non-SIC patients and SIC patients group;

[0033] Figure 4 are comparison of each index level of three groups of sepsis patients; the results are shown as a box representing the 25th-75th percentile, including the median and individual values; A: NETs count, B: PT, C: TT, D: APTT, E: PLT, F: D-dimer, G: AT-III, H: organ failure assessment score (SOFA), I: WBC, J: CRP, K: PCT, L: simplified acute physiology score correlation; *p<0.05, **p<0.01, ***p<0.001; non-parametric multivariate analysis of variance, Mann-Whitney U test and Dunn post-hoc test were used for group and more than two group comparisons;

[0034] Figure 5 are binary Logistic regression analysis results of indicators with significant meaning, A: univariate regression results, B: multivariate regression results;

[0035] Figure 6 are divided into non-SIC group and SIC progression group according to whether coagulation dysfunction occurs within 72 hours after admission, A: combination model constructed by admission Nets, AT-III and SOFA score is related to the incidence of secondary coagulopathy in sepsis patients; the results are shown as a box representing the 25th-75th percentile, including the median and individual values; Mann-Whitney U test was used for group comparison; B: sepsis receiver operating characteristic (ROC) curve of the combination model of the admission group; AUC = area under the ROC curve; C: predictive value of MPO-DNA, NETs and combined model for sepsis patients with coagulation dysfunction. DETAILED DESCRIPTION

[0036] The application will be further described below in conjunction with specific examples. These examples are only used to illustrate the application and not used to limit the scope of the application. In addition, after reading the content taught by the application, those skilled in the art can make various modifications or changes to the application, and these equivalent forms also fall within the scope defined by the claims attached hereto.

[0037] Detection method:

[0038] (1) Peripheral blood cell analyzer:

[0039] Peripheral blood samples were collected within 2 hours after the patients were diagnosed with sepsis (met the Sepsis-3 criteria). NETs detection used an automated hematology slide preparation system (Sysmex SP-10) to prepare blood smears, which were dyed with Giemsa and analyzed by a fully automatic cell morphological analyzer (Sysmex DI-60) and CellaVision software (RL1019).

[0040] (2) Smear cell morphological analysis index:

[0041] 1. Lymphocyte smear cells are round or round-like cells without cytoplasm, with degenerative nuclei or swollen nuclei, and the structure is blurred and dyed uniformly blue-purple. Typical morphology is shown in Figure 2 Fig. A.

[0042] 2. The size of neutrophil smear cells is uneven, lacks a fixed shape, has a reticular structure, the nucleus is depolymerized with granular characteristics, and is dyed uniformly blue-purple. Typical morphology is shown in Figure 2 Fig. B.

[0043] (3) Neutrophil smear NETs count:

[0044] Neutrophil smear NETs count: 100 white blood cells (about 3 minutes) in peripheral blood smears were observed in a fully automatic cell morphological analyzer, and pictures with reticular, lobed nucleus depolymerization and granular characteristics were counted as NETs (%), strictly excluding lymphocyte smear cells (uniform blue-purple without granules) and mechanically damaged cells (tear-shaped edges), typical morphology is shown in Figure 2 Fig. A. NETs (%) = N(NETs) / N(white blood cell count) * 100%. Typical morphology is shown in Figure 2 Fig. B. The counting process follows the principle of blind method: two experienced blood morphologists independently reviewed the number of neutrophil smear cells in each 100 white blood cells (ICC) was 0.92 (95% CI: 0.87-0.95), and the third expert arbitrated the disagreement cases (>10% counting difference). Randomly selected 10% of the smears to evaluate the intra- / inter-group repeatability.

[0045] (4) Detection of natural anticoagulant substance antithrombin III activity:

[0046] The ACL TOP Family instrument used is based on the chromogenic substrate method, which detects the amount of nitroaniline PNA released continuously in the synthetic product by direct or indirect means, and evaluates the activity of the detected substance by the change in optical density.

[0047] (5) SOFA score:

[0048] Table 1

[0049]

[0050]

[0051] (6) MPO-DNA concentration detection:

[0052] According to the manufacturer's instructions, the level of myeloperoxidase-DNA complex (MPO-DNA) in human peripheral serum (within 2 hours of admission) was detected in the test sample. The microplate was coated with purified human myeloperoxidase-DNA complex (MPO-DNA) antibody to form a solid phase antibody, and then the myeloperoxidase-DNA complex (MPO-DNA) was added to the coated microplate, followed by the addition of HRP-labeled myeloperoxidase-DNA complex (MPO-DNA) antibody to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB was added for color development. TMB is converted to blue under the catalysis of HRP enzyme, and is converted to the final yellow under the action of acid. The color depth is positively correlated with the myeloperoxidase-DNA complex (MPO-DNA) in the sample. The absorbance (OD value) was measured at 450 nm wavelength by a microplate reader, and the concentration of human myeloperoxidase-DNA complex (MPO-DNA) in the sample was calculated by a standard curve. All samples were analyzed in duplicate.

[0053] (7) Basic information collection:

[0054] Basic clinical data of patients at admission were collected, such as admission temperature, heart rate, blood pressure, etc., and SOFA score was calculated according to the admission condition. Coagulation function (PT, INR, D-dimer, etc.), inflammatory markers (PCT, CRP), demographic characteristics, infection parameters, and vital signs were collected.

[0055] (8) Statistical analysis:

[0056] R 4.4.3, SPSS27 and Origin 2024 software were used for data processing. Continuous variables were represented by median (IQR), and categorical variables were described by frequency (percentage). Mann-Whitney U test (continuous variables) or χ 2 Fisher test (categorical variables) was used for comparison between groups. Binary Logistics regression analysis was used to analyze the risk factors for coagulopathy secondary to sepsis; according to whether the sepsis patients developed coagulopathy, the subjects were grouped, the receiver operating characteristic curve (ROC) was drawn, and the area under the ROC curve (AUC) was calculated. The significance level was set at α = 0.05, and all analyses were declared ethical approval (license number: KY24115) and informed consent.

[0057] The technical solution of the present invention will be described in detail below with reference to specific embodiments. In the following embodiments, unless otherwise specified, the reagents, materials and equipment used can be purchased commercially, prepared by conventional methods, or commonly used in the industry.

[0058] Example 1:

[0059] This prospective observational cohort study selected patients with sepsis admitted to Wuxi People's Hospital affiliated with Nanjing Medical University from May 2023 to May 2025 as study subjects. Inclusion criteria were: 1. Sepsis meeting the diagnostic criteria for sepsis 3.0 (SOFA score ≥2 points higher than baseline); 2. Age 18-80 years; 3. Not meeting the ISTH 2019 SIC diagnostic criteria at enrollment (ISTH SIC score <4 points); 4. Expected ICU stay ≥72 hours. Exclusion criteria included hematologic disorders, active connective tissue diseases, advanced malignancies, Child-Pugh B / C liver disease, end-stage renal disease, major surgery or severe trauma within 72 hours, active bleeding (24-hour transfusion >2 units of red blood cells), anticoagulation / antiplatelet therapy within 72 hours, or long-term immunosuppressant use. Based on the inclusion and exclusion criteria, a total of 147 patients were ultimately included as study subjects. The patient inclusion process is as follows: Figure 1 As shown, patients were divided into a non-SIC group (85 cases) and a SIC progression group (62 cases) based on their coagulation dysfunction within 72 hours of admission, according to the ISTH 2019 guidelines for SIC diagnosis. The 62 cases in the SIC progression group (sepsis with coagulation dysfunction) included 33 cases (22.4%) of sepsis-induced coagulopathy (SIC) and 29 cases (19.7%) of disseminated intravascular coagulation (DIC). Table 2 describes the main demographic and clinical characteristics of the three groups. Of the 85 patients (57.8%), 62 (42.2%) had sepsis with coagulation dysfunction, including 33 (22.4%) with SIC and 29 (19.7%) with DIC. Table 1 describes the main demographic and clinical characteristics of the three groups. Among them, the SOFA score and APACHE II score of patients in the sepsis coagulation disorder group were significantly higher than those in the sepsis without coagulation disorder group (P<0.05); from the admission data, the heart rate of patients in the sepsis with coagulation disorder group was significantly higher than that of patients in the sepsis without coagulation disorder group (P<0.05), while there were no differences in other vital signs upon admission; there were no significant differences in demographic characteristics and underlying diseases (all P>0.05): the median age was 64-72 years, the proportion of males was 36.4%-48.3%, and chronic diseases such as diabetes (41.4%-51.8%) and hypertension (40.6%-55.2%) were evenly distributed.

[0060] Table 2. Main clinical characteristics of patients

[0061]

[0062]

[0063] Morphological observation of NETs in blood smears of sepsis patients showed that in the blood smears of sepsis SIC progression group patients, we observed significant pathological changes in neutrophils. When the body is attacked by pathogens, neutrophils are stimulated by the external environment, begin to appear swollen and blurred edge characteristics, and also appear blurred nuclear chromatin structure, unevenly light and dark staining, and unclear nuclear outline, which means that NETs begin to form. When the neutrophil nuclear membrane ruptures and disintegrates, the reticular fibrillar material is released into the cytoplasm, causing the cell to lose its normal morphology. Further observation found that Figure 2 NETs released by neutrophils trapped red blood cells in the reticular structure (NETs), which showed chemotaxis to red blood cells, further confirming the role of NETs in capturing red blood cells.

[0064] The above non-SIC group (sepsis 85 people) and SIC progression group (sepsis combined with coagulopathy 62 people) were compared between groups at admission Nets marker, the results as shown in Figure 3 Nets marker levels were significantly correlated with the severity of sepsis coagulopathy and coagulation disorders, and Nets levels were significantly increased in a stepwise manner.

[0065] Example 2:

[0066] The levels of each index were compared among the three groups of sepsis patients, i.e. sepsis (non-SIC group), SIC and DIC, and the results are shown in Figure 4 As the disease progresses, sepsis patients progress to sepsis coagulopathy, and traditional coagulation indicators also deteriorate Figure 4 ), platelet (PLT) consumption leading to a significant decrease in PLT; prothrombin time (PT) significantly prolonged and its corresponding international normalized ratio (INR) significantly increased, thrombin time (TT) prolonged, indicating activation of the intrinsic and extrinsic coagulation pathways, and anticoagulant substance antithrombin III (AT-III) activity significantly decreased, suggesting anticoagulant substance consumption; the continuous increase of fibrinolysis marker D-dimer, but fibrinogen (FIB) in this study had no significant correlation with coagulation disorders in sepsis. The increase of NETs and systemic inflammatory markers C-reactive protein (CRP), procalcitonin (PCT) occurred basically synchronously. These results show that the increase of NETs level is closely related to the severity of coagulation dysfunction in sepsis patients and the enhancement of systemic inflammatory response, suggesting that NETs play a key role in the development of sepsis coagulopathy.

[0067] Subsequently, binary logistic regression analysis was performed on the statistically significant indicators, and the results are as follows: Figure 5 As shown, univariate analysis revealed that multiple indicators were significantly associated with septic coagulopathy. Elevated levels of extraneutrophilic nets (Nets) significantly increased disease risk, while prolonged prothrombin time (PT), prolonged activated partial thromboplastin time (APTT), prolonged thrombin time (TT), decreased antithrombin III (AT-III) activity, elevated C-reactive protein (CRP), and elevated sequential organ failure score (SOFA) were all independently associated with increased disease risk. Multivariate analysis further identified four independent predictors: the risk effect of Nets was stronger than in univariate analysis, indicating that its predictive value was independent of other confounding factors; D-dimer showed no significant effect in univariate analysis (*p*=0.460), but after adjusting for other variables, it showed independent predictive value; decreased AT-III activity consistently indicated high risk; CRP and SOFA score also maintained independent association. It is worth noting that the coagulation indicators (PT, APTT, TT) that were significant in the univariate analysis all lost statistical significance in the multivariate model (*p* all > 0.05), suggesting that their effects may be explained by core indicators such as Nets and AT-III.

[0068] To further screen for independent predictors of septic coagulopathy, we performed collinearity analysis on Nets, D-dimer, AT-III, CRP, and SOFA scores, finding no collinearity. Subsequently, we conducted a binary logistic regression analysis on these five indicators. Nets, D-dimer, AT-III, CRP, and SOFA scores constituted the core predictor combination for septic coagulopathy. Nets, AT-III, and SOFA scores were the core independent predictors of septic coagulopathy (Table 2). Among them, Nets showed the strongest predictive power after multivariate adjustment (OR increase of 59%), and its contribution to the pathological mechanism surpassed that of traditional coagulation indicators.

[0069] Table 3: Results of binary logistic regression analysis of Nets, D-dimer, AT-III, CRP, and SOFA scores. Univariate logistic regression analysis.

[0070]

[0071] Multivariate logistic regression

[0072]

[0073] Example 3:

[0074] This study evaluated the predictive efficacy of Nets (neutrophil extracellular trap-related markers) for septic coagulopathy (SIC) using a logistic regression model. The results are as follows: Figure 6 As shown, Nets exhibited some potential when used alone, but combining them with traditional indicators significantly improved their predictive value. Nets alone had an area under the curve (AUC) of 0.78 for predicting SIC, indicating high sensitivity (86%), meaning it was effective in identifying true SIC patients. However, its specificity was relatively low (56%), suggesting a possibility of false positives, with an overall prediction accuracy of 73%. Meanwhile, MPO-DNA, another marker of NETs in peripheral blood, showed a systematic decrease in key indicators such as accuracy, specificity, positive predictive value, and negative predictive value. Notably, the predictive performance of Nets was similar to that of the clinically used SOFA score (AUC 0.75, accuracy 73%).

[0075] The prediction equation was established by integrating NETs with traditional coagulation function indicators AT-III and SOFA scores: Logit(SIC) = 41.5 × NETs (%) - 0.068 × ATIII + 0.228 × SOFA - 0.683, forming a joint indicator model. Figure 6 When the combined indices (as shown in the diagram) are within a certain range, the predictive efficacy achieves a significant leap: the AUC increases dramatically to 0.91, and the accuracy reaches 83%. This combined model maintains high sensitivity (78%) while significantly improving specificity to 89% and achieving a positive predictive value as high as 90%, meaning that patients identified as positive by the model are highly likely to actually have septic coagulopathy. When the combined indices are ≥0.35, it indicates a higher risk of secondary coagulopathy in sepsis patients, and clinicians should be alert to changes in the patient's condition and implement aggressive anticoagulation therapy; when the combined indices are <0.35, the risk of secondary coagulopathy in sepsis patients is lower, but due to the rapid progression and changes in SIC, continued monitoring is still necessary.

[0076] These results clearly demonstrate that while Nets, when used alone, can serve as a primary screening indicator for septic coagulopathy, especially suitable for high-sensitivity screening scenarios, their predictive value is limited. However, incorporating them into a multi-indicator integrated model that includes AT-III and SOFA scores enables more accurate and reliable prediction and identification of septic coagulopathy, providing stronger decision support for early clinical intervention. Therefore, Nets are important biomarkers for predicting septic coagulopathy, playing a central role, particularly in multi-indicator combined strategies.

[0077] The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.

Claims

1. An early warning system for sepsis-induced coagulopathy based on neutrophil extracellular traps, characterized in that, The early warning system includes a data acquisition device and an assessment device. The data acquisition device collects data on the percentage of neutrophils with extracellular traps in white blood cells, the activity of the natural anticoagulant antithrombin III, and the SOFA score in the patient's whole blood. The assessment device calculates a combined indicator using the following predictive equation and assesses the patient's risk of septic coagulopathy based on this combined indicator: Logit(SIC)=41.5×NETs(%)-0.068×ATIII+0.228×SOFA-0.683 Among them, Logit(SIC) is a combined indicator for predicting the risk of sepsis-induced coagulopathy, NETs(%) represents the proportion of neutrophil extracellular traps in white blood cells, ATIII represents the activity of the natural anticoagulant antithrombin III, SOFA represents the SOFA score, and P(SIC) is the predictive probability of sepsis-induced coagulopathy.

2. The early warning system for sepsis-induced coagulopathy according to claim 1, characterized in that, The percentage of neutrophil extracellular trapping nets in white blood cells was obtained through the following method: Smear preparation and staining: Collect peripheral blood samples from the patient, prepare blood smears from the peripheral blood samples, and perform Giemsa staining on the blood smears; and, Image acquisition, obtaining optical microscopic images of white blood cell morphology from blood samples; and... Information processing, counting the number N of white blood cells in the microscopic image based on cell morphology. (白细胞数) The number N of extracellular traps for neutrophils (NETs) ;and, Data processing was performed to calculate the percentage of neutrophil extracellular traps (NETs) in white blood cells. The calculation formula is as follows:

3. The early warning system for sepsis and coagulopathy according to claim 2, characterized in that, Images exhibiting reticular, segmented nuclear disaggregation, and granular characteristics are classified as neutrophil extracellular traps.

4. The early warning system for sepsis and coagulopathy according to claim 2, characterized in that, The peripheral blood samples were from patients with sepsis.

5. The early warning system for sepsis-induced coagulopathy according to claim 1, characterized in that, The activity of the natural anticoagulant, antithrombin III, was determined by detecting plasma in anticoagulant tubes using the ACL TOP Family instrument.

6. The early warning system for sepsis-induced coagulopathy according to claim 1, characterized in that, The SOFA score is an assessment conducted by clinicians based on a patient's admission status, including the respiratory, cardiovascular, liver, coagulation, kidney, and nervous systems. Each system is scored according to the degree of its functional failure.

7. The early warning system for sepsis-induced coagulopathy according to claim 6, characterized in that, The SOFA score has a total score of 24 points. The higher the score, the more severe the organ failure.

8. The early warning system for sepsis-induced coagulopathy according to claim 1, characterized in that, When the combined index is ≥0.35, it indicates a higher risk of secondary coagulation disorders in sepsis patients; when the combined index is <0.35, the risk of secondary coagulation disorders in sepsis patients is lower.

9. The use of the system according to any one of claims 1 to 8 in screening drugs for the prevention or treatment of sepsis-related coagulopathy.

10. An information data processing terminal, the information data processing terminal being used to implement the early warning system for sepsis coagulopathy based on neutrophil extracellular traps as described in any one of claims 1 to 8.

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