Method for predicting process of chronic hepatitis B related diseases based on ratio of PLG to PT

Abstract

The invention discloses a chronic hepatitis B related disease process prediction method based on the ratio of PLG to PT. The method comprises the following steps: S1, sample collection: collecting sodium citrate anticoagulant venous blood of a to-be-predicted object; s2, index detection: detecting the plasminogen level in the plasma by adopting a chromophoric substrate method, and detecting the prothrombin time in the plasma by adopting a coagulation method; s3, specific value calculation: calculating the specific value of PLG to PT, wherein the specific value is used as a core prediction index of the chronic hepatitis B related end-stage liver disease process; and S4, process judgment: judging a disease process stage of the to-be-predicted object according to the PLG / PT ratio in combination with a preset threshold value. The method has the beneficial effects that the ratio of the PLG to the PT is used as a core prediction index. The specific value integrates dual information of liver synthesis function and blood coagulation fibrinolysis system imbalance, the scientificity of indexes is ensured, the design logic of the specific value is different from that of any existing verification method, and the crossing from single-dimensional detection to multi-dimensional integration evaluation is realized.

Description

[0001] This invention relates to the field of chronic hepatitis B disease progression prediction technology, specifically to a method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio. Background Technology

[0002] Chronic hepatitis B-related end-stage liver disease is a severe stage of HBV infection progression, encompassing chronic hepatitis B-related cirrhosis and liver failure. It is characterized by insidious onset, rapid progression, poor prognosis, and high mortality.

[0003] Currently, clinical prediction of the progression of chronic hepatitis B-related end-stage liver disease mainly relies on two types of technical means: one is the detection of traditional liver function indicators such as ALT, AST, ALB and HBV serological markers. These methods can only reflect some liver function or viral activity status, and cannot accurately characterize the degree of disease progression. They also have a significant lag and are difficult to identify the trend of disease deterioration in the early stage. The second is clinical assessment models such as the MELD scoring system. These models are mostly constructed based on different etiological populations and have limited applicability in chronic hepatitis B-related end-stage liver disease, especially in predicting cirrhosis and liver failure.

[0004] The core pathological feature of chronic hepatitis B-related end-stage liver disease is severely impaired liver function accompanied by an imbalance in the coagulation and fibrinolytic systems. As the main site of synthesis of coagulation factors and fibrinolytic proteins, abnormal liver function directly leads to disorders of the coagulation and fibrinolytic systems—a pathological mechanism that has been widely confirmed. While existing studies have found a correlation between coagulation-related indicators and liver disease progression, most use single-indicator detection, which only reflects local changes in the coagulation or fibrinolytic systems and cannot comprehensively reflect the intrinsic link between impaired liver function and the imbalance in the coagulation and fibrinolytic systems. Therefore, the predictive specificity and accuracy are both unsatisfactory.

[0005] Existing technologies include patented solutions for predicting the progression of chronic hepatitis B-related liver diseases. For example, Chinese patent CN105586426A discloses a detection method based on the rs2069852 locus marker of the IL-6 gene, which analyzes the AG genotype at this gene locus to predict the risk of chronic hepatitis B progressing to cirrhosis. This method relies on gene polymorphism detection, which is complex and time-consuming, and has very limited ability to assess the natural progression of the disease. The IL-6 gene polymorphism it relies on is only related to the chronic inflammation-mediated liver fibrosis process, but cannot reflect the acute pathological changes in the liver failure stage. Mechanistically, this marker lacks a direct correlation with the pathological characteristics of liver failure.

[0006] Furthermore, the existing technology has not yet recognized the technical approach of using the ratio of plasminogen glutathione (PLG) to prothrombin time (PT) as a core predictive indicator.

[0007] PLG, a precursor of plasminogen lysate (PLG), is produced by the liver. Its levels are directly related to liver synthetic function. Liver disease patients primarily exhibit prolonged progesterone (PT), reflecting both liver synthetic function and the liver's role in regulating the coagulation system. The PLG-PT ratio integrates information on both liver synthetic function and imbalances in the coagulation and fibrinolytic systems. However, current assessments of chronic liver disease progression lack highly predictive laboratory indicators, making it difficult to identify different stages of chronic hepatitis B-related end-stage liver disease and failing to meet the needs of precise clinical diagnosis and treatment. Therefore, developing a novel core indicator based on the PLG-PT ratio for predicting the progression of chronic hepatitis B-related diseases, entirely different from existing validation methods, has become an urgent clinical need. Summary of the Invention

[0008] The purpose of this invention is to provide a method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio in order to solve the above-mentioned problems and overcome the shortcomings of the prior art, as detailed below.

[0009] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio, comprising the following steps: S1. Sample collection: Collect sodium citrate anticoagulated venous blood from the subjects to be predicted, and separate the plasma after centrifugation at 3000 r / min for 15 min; S2. Indicator detection: Plasminogen levels in plasma were detected using the chromogenic substrate method, and prothrombin time in plasma was detected using the coagulation method. All tests were performed on the STGAO fully automated coagulation analyzer. S3. Ratio Calculation: Calculate the ratio of PLG to PT, which serves as a core predictive indicator for the progression of chronic hepatitis B-related end-stage liver disease. S4. Progress Judgment: Based on the PLG / PT ratio and a preset threshold, determine the disease progression stage of the subject to be predicted. The progression stages include chronic hepatitis B, chronic hepatitis B-related cirrhosis, and chronic hepatitis B-related liver failure.

[0010] In some embodiments of the present invention, the preset threshold in step S4 is determined in the following manner: Clinical samples were collected from healthy individuals, patients with chronic hepatitis B, patients with chronic hepatitis B-related cirrhosis, and patients with chronic hepatitis B-related liver failure. PLG and PT were measured and the PLG / PT ratio was calculated. ROC curve analysis was used to determine the critical thresholds corresponding to each stage of the process.

[0011] In some embodiments of the present invention, the criteria for judging the PLG / PT ratio corresponding to each process stage are as follows: When the PLG / PT ratio is between the first threshold and the second threshold, the object to be predicted is determined to be in the chronic hepatitis B stage. When the PLG / PT ratio is between the second and third thresholds, the subject to be predicted is determined to be in the stage of chronic hepatitis B-related cirrhosis. When the PLG / PT ratio is between the third and fourth thresholds, the subject to be predicted is determined to be in the stage of chronic hepatitis B-related liver failure.

[0012] In some embodiments of the present invention, in step S1, the amount of sodium citrate anticoagulated venous blood collected is 2-3 ml, and the plasma is separated and stored at 2-8°C for no more than 4 hours.

[0013] In some embodiments of the present invention, in step S3, the PLG is expressed as an activity level in U / ml, the PT is expressed as time in seconds, and the PLG / PT ratio is expressed in "U·s". -1 The results are presented in units of measurement.

[0014] In some embodiments of the present invention, in step S2, the reaction temperature for detecting PLG using the chromogenic substrate method is controlled at 37±0.5℃, and the reagent activation time for detecting PT using the coagulation method is 1-2 minutes.

[0015] The beneficial effects are: 1. This invention uses the ratio of PLG to PT as the core predictive indicator, abandoning the approach of existing technologies that rely on a single liver function indicator, a single coagulation indicator, or complex scoring models. This ratio integrates dual information on the imbalance of liver synthetic function and the coagulation and fibrinolytic system. The significant negative correlation between PLG and PT is p<0.001, R0. 2 =0.648 ensures the scientific nature of the indicator. Its design logic is different from any existing verification method, realizing a leap from single-dimensional detection to multi-dimensional integrated evaluation.

[0016] 2. The PLG / PT ratio demonstrates significantly better predictive efficacy for all stages of chronic hepatitis B-related end-stage liver disease than existing single indicators such as PLG alone (AUC=0.810) and ALB alone (limited efficacy). In particular, it shows the best predictive efficacy for chronic hepatitis B-related liver failure (AUC=0.9458), addressing the shortcomings of existing technologies in predicting cirrhosis and liver failure. The predictive AUC for chronic hepatitis B is 0.821, and the predictive AUC for chronic hepatitis B-related cirrhosis is 0.6139, both demonstrating stable predictive capabilities.

[0017] 3. The detection methods used, namely the chromogenic substrate method and the coagulation method, are both clinically mature technologies that can be completed using a standard fully automated coagulation analyzer without requiring special experimental conditions or equipment. The plasma preservation conditions and detection parameters are clearly defined, ensuring the reliability of the results. The PLG / PT ratio calculation logic is clear, avoiding the tedious calculations of complex scoring models, making it suitable for batch sample testing and easy to promote and apply in medical institutions at all levels.

[0018] 4. The PLG / PT ratio showed significant differences between healthy individuals and disease groups, as well as between different stages of the disease, clearly distinguishing between stages of chronic hepatitis B, cirrhosis, and liver failure. In particular, the PLG / PT ratio showed a significant decreasing trend across all groups of chronic hepatitis B-related end-stage liver disease. This regular change ensures accurate characterization of disease progression and solves the problem of difficulty in identifying early disease progression with existing technologies.

[0019] 5. The PLG / ALB ratio showed a significant difference only between the chronic hepatitis B group and the cirrhosis and liver failure group (p<0.05), while there were no statistically significant differences among the other groups (p>0.05). In contrast, the PLG / PT ratio showed significant differences in all stages of HBV-ESLD, providing more comprehensive coverage.

[0020] 6. The predictive power of PLG / PT in chronic hepatitis B-related cirrhosis (AUC=0.6139) and liver failure (AUC=0.9653) was significantly higher than that of the MELD scoring system (AUC=0.5738 and 0.8581, respectively), clarifying the superior division of labor between the two in different disease stages. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 This is a schematic diagram illustrating the correlation analysis of PLG, ALB, PT, and PLT counts in patients with chronic hepatitis B-related end-stage liver disease according to the present invention. Figure 2 This is a schematic diagram showing the expression of PLG / PT and PLG / ALB in chronic hepatitis B-related end-stage liver disease according to the present invention; Figure 3 This is a schematic diagram of ROC curve analysis of the diagnostic efficacy of PLG, PLG / PT, and PLG / ALB for chronic hepatitis B-related end-stage liver disease in this invention. Figure 4 This is a schematic diagram illustrating the changes in plasma coagulation-related indicators in patients with chronic hepatitis B-related end-stage liver disease according to the present invention; Figure 5 This is a schematic diagram illustrating the changes in plasma liver function-related indicators in patients with chronic hepatitis B-related end-stage liver disease according to the present invention. Detailed Implementation

[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0024] This invention addresses the shortcomings of existing methods for predicting the progression of chronic hepatitis B-related end-stage liver disease, including the use of single indicators, limited clinical model adaptability, difficulty in identifying different disease stages, and poor practicality of some detection methods. It proposes a solution using the plasminogen lysate (PLG) / prothrombin time (PT) ratio as the core predictive indicator. By collecting sodium citrate-anticoagulated venous blood and properly separating and preserving plasma, PLG activity and PT values ​​are detected using a conventional fully automated coagulation analyzer via chromogenic substrate and coagulation methods. The ratio is calculated and combined with critical thresholds determined by ROC curves to assess the progression of chronic hepatitis B, including hepatitis, cirrhosis, and liver failure. This method can be further validated by combining liver function indicators, HBV viral DNA quantification, or the MELD scoring system. This approach achieves a multi-dimensional integrated assessment of the imbalance between liver synthetic function and the coagulation and fibrinolytic system. The predictive AUC for liver failure reaches 0.9458, while the predictive AUCs for hepatitis and cirrhosis are 0.821 and 0.6139, respectively. It covers the entire disease course, is convenient to operate, and cost-effective, providing a reliable basis for precision clinical diagnosis and treatment.

[0025] See instruction manual attached Figures 1 to 5 As shown, in order to solve the above-mentioned technical problems, this invention proposes a method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio, comprising the following steps: S1. Sample collection: Collect sodium citrate anticoagulated venous blood from the subjects to be predicted, and separate the plasma after centrifugation at 3000 r / min for 15 min; S2. Indicator detection: Plasminogen levels in plasma were detected using the chromogenic substrate method, and prothrombin time in plasma was detected using the coagulation method. All tests were performed on the STGAO fully automated coagulation analyzer. S3. Ratio Calculation: Calculate the ratio of PLG to PT, which serves as a core predictive indicator for the progression of chronic hepatitis B-related end-stage liver disease. S4. Progress Judgment: Based on the PLG / PT ratio and a preset threshold, determine the disease progression stage of the subject to be predicted. The progression stages include chronic hepatitis B, chronic hepatitis B-related cirrhosis, and chronic hepatitis B-related liver failure.

[0026] In this embodiment, the preset threshold in step S4 is determined as follows: Clinical samples were collected from healthy individuals, patients with chronic hepatitis B, patients with chronic hepatitis B-related cirrhosis, and patients with chronic hepatitis B-related liver failure. A total of 513 cases were included in this embodiment, comprising 95 healthy controls, 84 patients with chronic hepatitis B, 108 patients with chronic hepatitis B-related cirrhosis, and 81 patients with chronic hepatitis B-related liver failure. Sample inclusion followed the 2022 edition of the "Guidelines for the Prevention and Treatment of Chronic Hepatitis B," excluding cases under 18 years of age, those with other viral hepatitis, rheumatic immune system diseases, pregnancy, and non-liver tumors. PLG and PT were measured in each group of samples, and the PLG / PT ratio was calculated. ROC curve analysis was performed using GraphpadPrism and SPSS software to determine the critical thresholds corresponding to each stage of the process, namely the first threshold, the second threshold, the third threshold, and the fourth threshold. Analysis showed that PLG / PT had a predictive AUC of 0.821 for chronic hepatitis B, 0.6139 for chronic hepatitis B-related cirrhosis, and 0.9458 for chronic hepatitis B-related liver failure, all of which were superior to the predictive efficacy of single indicators such as PLG and PLG / ALB.

[0027] In this embodiment, the preset threshold in step S4 is determined in the following way: Clinical samples were collected from healthy individuals, patients with chronic hepatitis B, patients with chronic hepatitis B-related cirrhosis, and patients with chronic hepatitis B-related liver failure. PLG and PT were measured and the PLG / PT ratio was calculated. ROC curve analysis was used to determine the critical thresholds corresponding to each stage of the process.

[0028] In this embodiment, the criteria for judging the PLG / PT ratio corresponding to each process stage are as follows: When the PLG / PT ratio is between the first threshold and the second threshold, the object to be predicted is determined to be in the chronic hepatitis B stage. When the PLG / PT ratio is between the second and third thresholds, the subject to be predicted is determined to be in the stage of chronic hepatitis B-related cirrhosis. When the PLG / PT ratio falls between the third and fourth thresholds, the subject of the prediction is determined to be in the stage of chronic hepatitis B-related liver failure.

[0029] It is important to note that the PLG / PT ratio criteria for each of the above-mentioned stages of the disease process cannot be used to independently diagnose chronic hepatitis B / cirrhosis / liver failure. Rather, they are used to determine the stage of disease progression in patients already diagnosed with chronic hepatitis B.

[0030] Among them, the expression level of ALB in the healthy control group, the chronic hepatitis B control group and the end-stage liver disease group showed significant differences (p<0.05), and the expression level in the chronic hepatitis B-related cirrhosis and liver failure group was significantly lower than that in the chronic hepatitis B group (p<0.05). The PLT count was significantly different in all groups of patients with chronic hepatitis B-related end-stage liver disease compared with the healthy control group and the chronic hepatitis B group, and showed a significant decreasing trend with disease progression (p<0.05), which can be used as an important auxiliary judgment basis. PLG and ALB showed a significant positive correlation (p < 0.001), R< 0.001. 2 =0.678, further supporting the scientific validity of PLG as an indicator related to liver synthetic function.

[0031] In this embodiment, in step S1, the amount of sodium citrate anticoagulated venous blood collected is 2-3 ml, and the plasma is separated and stored at 2-8°C for no more than 4 hours.

[0032] In this embodiment, in step S3, PLG is expressed as an activity level in U / ml, PT is expressed as time in seconds, and the PLG / PT ratio is expressed as "U·s". -1 The results are presented in units of measurement.

[0033] In this embodiment, in step S2, the reaction temperature for detecting PLG using the chromogenic substrate method is controlled at 37±0.5℃, and the reagent activation time for detecting PT using the coagulation method is 1-2 minutes.

[0034] Specific application examples of the embodiments of the present invention: The subject of the prediction was a 52-year-old male. 2 ml of his sodium citrate-anticoagulated venous blood was collected, centrifuged at 3000 rpm for 15 min to separate the plasma, and stored at 4°C. Testing was completed within 3 hours. Using the STGAO fully automated coagulation analyzer, the PLG activity level was measured at 37°C using the chromogenic substrate method, and was found to be 0.85 U / ml. After reagent activation for 1.5 minutes, the PT value was measured using the coagulation method, and was found to be 16.2 s. The calculated PLG / PT ratio was 0.052 U·s. -1 ; Based on the preset thresholds, this ratio falls between the third and fourth thresholds, initially indicating chronic hepatitis B-related liver failure. Further testing of auxiliary indicators showed a significant decrease in ALB levels and a significant increase in TBIL levels, consistent with the PLG / PT ratio assessment, ultimately confirming the patient's diagnosis of chronic hepatitis B-related liver failure.

[0035] The above are merely specific embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio, characterized in that: Includes the following steps: S1. Sample collection: Collect sodium citrate anticoagulated venous blood from the subjects to be predicted, and separate the plasma after centrifugation at 3000 r / min for 15 min; S2. Indicator detection: Plasminogen levels in plasma were detected using the chromogenic substrate method, and prothrombin time in plasma was detected using the coagulation method. All tests were performed on the STGAO fully automated coagulation analyzer. S3. Ratio Calculation: Calculate the ratio of PLG to PT, which serves as a core predictive indicator for the progression of chronic hepatitis B-related end-stage liver disease. S4. Progress Judgment: Based on the PLG / PT ratio and a preset threshold, determine the disease progression stage of the subject to be predicted. The progression stages include chronic hepatitis B, chronic hepatitis B-related cirrhosis, and chronic hepatitis B-related liver failure.

2. The method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio as described in claim 1, characterized in that: The preset threshold in step S4 is determined in the following way: We collected data from healthy individuals, patients with chronic hepatitis B, patients with chronic hepatitis B-related cirrhosis, and patients with chronic hepatitis B-related liver failure. We measured PLG and PT and calculated the PLG / PT ratio. We then used ROC curve analysis to determine the critical thresholds for each stage of the process.

3. The method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio according to claim 2, characterized in that: The criteria for judging the PLG / PT ratio for each process stage are as follows: When the PLG / PT ratio is between the first threshold and the second threshold, the object to be predicted is determined to be in the chronic hepatitis B stage. When the PLG / PT ratio is between the second and third thresholds, the subject to be predicted is determined to be in the stage of chronic hepatitis B-related cirrhosis. When the PLG / PT ratio is between the third and fourth thresholds, the subject to be predicted is determined to be in the stage of chronic hepatitis B-related liver failure.

4. The method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio according to claim 1, characterized in that: In step S1, the amount of sodium citrate anticoagulated venous blood collected is 2-3 ml. After plasma separation, it is refrigerated at 2-8℃ and the storage time does not exceed 4 hours.

5. The method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio according to claim 1, characterized in that: In step S3, PLG is expressed as activity level in U / ml, PT is expressed as time in seconds, and the PLG / PT ratio is expressed as U·s. -1 The results are presented in units of measurement.

6. The method for predicting the progression of chronic hepatitis B-related diseases based on the PLG to PT ratio according to claim 1, characterized in that: In step S2, the reaction temperature for detecting PLG using the chromogenic substrate method is controlled at 37±0.5℃, and the reagent activation time for detecting PT using the coagulation method is 1-2 minutes.

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