CN117310165A - Application of renin as biomarker in prediction, diagnosis or monitoring of sepsis and septic lung injury in children - Google Patents

Abstract

The invention discloses application of renin serving as a biomarker in prediction, diagnosis or monitoring of sepsis and septic lung injury in children, and belongs to the technical field of biological medicines, wherein the ages of the children are 28 days to 18 years. The invention discovers that the renin can be used as a diagnosis marker of the sepsis of children, has better prospect, can specifically detect the serum renin level in the patients suffering from the infection, is used for prompting whether the pediatric patients suffer from the sepsis or not and provides a basis for the prognosis of the patients; the invention has strong specificity, can predict the occurrence of sepsis by detecting the expression condition of the peripheral renin of a patient through ELISA means, can be directly applied to clinic, and has practical clinical use value. The invention has the advantages of high efficiency, high specificity, high sensitivity, high reliability, intuitiveness and high accuracy.

Description

Application of renin as biomarker in prediction, diagnosis or monitoring of sepsis and septic lung injury in children

Technical Field

The invention relates to the technical field of biological medicines, in particular to application of renin as a biomarker in prediction, diagnosis or monitoring of sepsis and septic lung injury in children.

Background

Sepsis is an infectious disease with high morbidity and mortality, and is one of the leading causes of death in children. The pathological features of sepsis are excessive inflammation and immunosuppression, and multiple organ failure caused by deregulation of the host's response to infection is the leading cause of sepsis mortality. About 120 tens of thousands of children are diagnosed with sepsis each year worldwide. The disease progress of the sepsis in children is fast, the short-term death rate is high, and most deaths occur within 3-7 days of admission to the hospital. It was counted that 64% of dead children 760 thousands of ages less than 5 in 2010 die from sepsis or septic shock caused by severe infections. Sepsis often accumulates systemic multiple organs, with lung injury being the most common type of complication and progressing very easily to respiratory distress syndrome (Acute Respiratory Distress Syndrome, ARDS), with high mortality and very poor prognosis.

Immune dysregulation plays a critical role in the development and progression of sepsis. Immunosuppression and immunoparalysis result in ineffective anti-sepsis infections and make it easier for sepsis patients to combine secondary and latent reactivation. The children population has specificity, and all viscera and function indexes of the children population are in continuous development and perfection, including an immune system playing a decisive role in sepsis. It has been reported that the distribution and proportion of cell subsets of innate immunity and adaptive immunity of children of different ages are greatly different from those of adults from fetal development, neonatal stage to childhood and adolescence. Thus, the results of studies of adult sepsis and biomarkers may not necessarily be applicable in pediatric sepsis patients.

Renin has been found since 1898 to have played a vital role in the regulation of cardiovascular disease and hypertension for more than 120 years. Renin is released by the kidneys, and its regulatory mechanisms include stimulation of capillary postoutput during renal perfusion and adrenergic stimulation of beta receptors. However, the current clinical studies on renin and infectious diseases are mostly focused on the adult field, the change characteristics and rules of renin in pediatric sepsis infections are currently unclear, and the current clinical studies on renin and pediatric sepsis and septic lung injury are reported to be relatively few. Because the early warning specific marker for the sepsis of children is lacking, a unified diagnosis and treatment guideline cannot be formulated so far for the sepsis, and standardized scientific researches are also influenced, the early warning specific marker for the sepsis of children is discovered, and is a key for accurately tracking and displaying the development change of the illness state, evaluating prognosis and formulating an individualized treatment scheme in the future.

Disclosure of Invention

The invention aims to provide application of renin as a biomarker in prediction, diagnosis or monitoring of sepsis and septic lung injury in children so as to solve the problems in the prior art.

In order to achieve the above object, the present invention provides the following solutions:

the present invention provides the use of renin as a biomarker in the manufacture of a product for diagnosing, prognosticating or monitoring or staging the progression of sepsis or septic lung injury in children aged 28 days to 18 years.

Preferably, the product is used to stage the progression of sepsis or septic lung injury, wherein the sepsis is staged as normal infection, sepsis or septic shock.

Preferably, the method of diagnosing, predicting or monitoring or staging the progression of sepsis or septic lung injury is a qualitative, quantitative detection of renin from a biological sample obtained from the child.

Preferably, the biological sample comprises plasma, serum or blood.

Preferably, the biological sample is analyzed using an enzyme-linked immunosorbent assay.

Preferably, an elevated level of renin in the biological sample as compared to a control sample or a reference sample indicates an increased severity of sepsis or septic lung injury as it progresses to various stages.

The invention also provides a detection reagent for children sepsis and septic lung injury, which takes renin as a detection object.

The invention also provides a detection kit for children sepsis and septic lung injury, which comprises the detection reagent.

The invention also provides the use of renin as a biomarker in the manufacture of a product for determining the severity of sepsis or septic lung injury in a child by qualitatively and quantitatively detecting renin in a biological sample obtained from the child and comparing the level of renin to a control sample or a reference sample to derive a score.

The invention also provides the use of renin as a biomarker in the manufacture of a product for determining childhood organ dysfunction or childhood sequential organ failure by qualitatively and quantitatively detecting renin in a biological sample obtained from the child and comparing the level of renin with a control sample or a reference sample.

The invention discloses the following technical effects:

the invention is found by the study of 108 clinically diagnosed children sepsis patients: there was a marked increase in renin expression in pediatric sepsis patients. In addition, renin was found to be of significant value for the diagnosis of pediatric sepsis by study of patient clinical and prognostic data in combination with gold standards now used for sepsis diagnosis. In addition, the expression condition has higher value for predicting the prognosis of patients. The invention has the advantages that: 1. the invention discovers that the renin can be used as a diagnosis marker of the sepsis of children, has better prospect, can specifically detect the serum renin level in the patients suffering from the infection, is used for prompting whether the pediatric patients suffer from the sepsis or not and provides a basis for the prognosis of the patients; 2. the invention has strong specificity, can predict the occurrence of sepsis by detecting the expression condition of the peripheral renin of a patient through ELISA means, can be directly applied to clinic, and has practical clinical use value. 3. The invention has the advantages of high efficiency, high specificity, high sensitivity, high reliability, intuitiveness and high accuracy.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is an experimental operational flow;

fig. 2 shows differences and characteristics of the expression levels of renin in different groups of different cohorts, wherein a is the pioneer cohort (sepsis n=58, normal infection group n=14, healthy group n=37), and the differences of the expression levels of renin different groups are compared; b is a comparison of differences in expression levels of renin in different groups in the validation cohort (sepsis group n=50, normal infection group n=37, healthy group n=46); c is the comparison of the differences in the expression levels of renin in the different groups in the total cohort (sepsis n=108, normal infection group n=51, healthy group n=83); d is a comparison of the differential levels of expression of renin at different time points of sepsis in healthy groups (sepsis group n=32, healthy group n=83);

fig. 3 shows differences and characteristics of renin expression levels in different subgroups of sepsis, wherein a is a comparison of differences in renin expression levels in two subgroups of simple sepsis group (n=76) and sepsis shock (n=32); b is a comparison of the difference in expression levels of renin in the 28-day survival group (n=95) and the 28-day death group (n=13);

FIG. 4 is a correlation analysis of various indexes of the severity of the condition of the infant with sepsis, wherein A is the correlation analysis of the expression of renin and the lactic acid level within 24 hours of hospitalization of the infant with sepsis; b is a correlation analysis of renin expression and a childhood organ dysfunction score 2 (PELOD-2) within 24 hours of hospitalization of the sepsis infant; c is a correlation analysis of renin expression with the sequential organ failure score (pSOFA) of children within 24 hours of hospitalization of the sepsis infant;

fig. 5 is a comparison of mechanical ventilation characteristics of renin in pediatric sepsis, wherein a is a comparison of differences in expression levels of renin in the mechanical ventilation free group (n=25) and mechanical ventilation in the mechanical ventilation free group (n=83) of 108 sepsis; b is a comparison of differences in renin expression levels within 1 week (including 1 week) of mechanical ventilation (n=46) and above 1 week (n=37) of mechanical ventilation in the sepsis mechanical ventilation group (n=83); c is a comparison of the difference in expression levels in 108 sepsis, with an oxygenation index (partial pressure of oxygen/concentration of inhaled oxygen) above 300 (n=21) and within 300 (containing 300) (n=87) in two different subgroups;

FIG. 6 is a ROC curve of renin in a pediatric sepsis model and its predicted potency versus Area Under Curve (AUC) compared to Procalcitonin (PCT);

fig. 7 shows the survival curves and cut-off values (cutoff values) associated with renin and 108 cases of sepsis infants.

Detailed Description

Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples are exemplary only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.

The technical scheme of the invention is conventional in the field, and the reagents or raw materials are purchased from commercial sources or are disclosed.

Example 1

All patients who were included in the study were from the major medical disciplines of the affiliated child hospitals of the university of Chongqing medical science, and the general condition of the patients, including patient name, sex, age, critical illness score, organ function index, ventilator parameters, etc., were collected immediately after the patients were admitted. The routine collection after admission is complete and comprises clinical examination indexes such as blood routine, liver and kidney functions, blood coagulation routine, blood lactic acid and the like. On the day of sepsis diagnosis (D1), on the third day (D3), and on the seventh day (D7), 2mL of patient peripheral blood was again collected at different time points, and the frozen serum was isolated, frozen at-80 ℃ and tested within 1 week.

In order to ensure the design rigor and the data reliability, the pioneer queue and the verification queue are designed, samples collected at different times are detected by adopting the same detection method, and the expression stability and consistency of the renin level in the sepsis of children are tested.

Before collection of the enrolled patients, the present example establishes strict inclusion criteria (see inclusion criteria and exclusion criteria for details below) in accordance with the international guidelines for pediatric sepsis. The normal infection group is the infant who enters PICU at the same time and has the infection but does not reach the sepsis diagnosis standard. The health study population is a child population for carrying out health examination by a physical examination center of an affiliated child hospital of the university of Chongqing medical science, and inquires about the past medical history condition of the child population and immediately collects peripheral blood after the child population is informed of the consent of the child population and/or family members. For the patients who do not have diseases in the past and all the examination indexes of physical examination are normal, the patients are brought into a healthy control group, 2mL of peripheral blood is collected, frozen serum is also separated at-80 ℃, and the next experiment is carried out within 1 week. The experimental operation flow is shown in fig. 1, and the specific experimental details are as follows:

(1) nano-row standard

Experimental group:

inclusion criteria: a. meets the international consensus on sepsis for children in 2005: (1) Reaching two or more systemic inflammatory response syndrome standards; (2) a confirmed or suspected invasive infection; (3) There is cardiovascular dysfunction, acute respiratory distress syndrome, or two or more organ dysfunctions; b. age range 29 days to 18 years; c. informed consent and complete study data were available.

Exclusion criteria: a. is not in accordance with the international consensus on sepsis for children in 2005; b. age ranges from 28 days to 18 years of age are not satisfied; c. informed consent and complete study data were not available.

Control group:

inclusion criteria: a1, collecting infants suffering from PICU common infection (with etiology evidence) but not sepsis in the same-period income of the auxiliary child hospitals of Chongqing medical university at the sample collecting stage; a2, a child patient is detected by a health physical examination center of a child affiliated hospital of Chongqing medical university (a 1 and a2 can meet one of the requirements); b. age range 28 days to 18 years; c. informed consent and complete study data were available.

Exclusion criteria: a. age ranges from 28 days to 18 years of age are not satisfied; b. informed consent and complete study data were not available.

(2) Sample collection and processing: the blood samples of the infants who are incorporated into the standard experiment group on the 1 st, 3 rd and 7 th days after admission are 2mL each, the children in the control group collect 2mL of the blood samples on the PICU or physical examination day, and the blood samples are subjected to centrifugal treatment (3500 rpm, 8 minutes) within 1 hour to separate serum, and 200 mu L/tube are packaged and then stored at the ultralow temperature of-80 ℃.

(3) Sample detection: and detecting the levels of RAS related factors such as renin and the like on the collected samples of the experimental group and the control group by adopting an enzyme-linked immunosorbent assay (ELISA), and collecting related clinical data of the two groups.

(4) And (3) data quality control: double rechecking is carried out on the data of the experimental group and the control group, and statistical analysis is carried out by adopting SPSS17.0 software. The data for comparison were evaluated using a normalization test and the metering data conforming to the normal distribution were expressed as mean ± standard deviation. Correlation analysis was performed using the Spearman scale test. All statistical tests were double-sided tests, all performed at a significant level of p=0.05. A subject operating characteristic (ROC) curve is constructed, and an optimal cut-off point is determined from the ROC curve. All charts were created by GraphPad Prism 8.0.

Experimental results:

the differences in the expression levels of renin in different groups of different queues and their characteristics are shown in FIG. 2. Data are expressed as mean ± standard deviation, P <0.01; * P < 0.0001. Fig. 2 a shows that the expression level of renin in the pioneer cohort in the pediatric sepsis group is significantly higher than in the normal infected group, n=37, the healthy group, n=14, the sepsis group, n=58; in fig. 2B, the expression level of renin in the cohort in the pediatric sepsis group is significantly higher than in the healthy group, n=46, normal infection group n=37, sepsis group n=50; fig. 2C shows that the expression level of renin in the total cohort in the pediatric sepsis group is significantly higher than in the normal infected group, n=83, the normal infected group n=51, and the sepsis group n=108; in fig. 2, D shows that the expression level of renin in the total cohort was significantly higher on day 1, day 3 and day 7 of the pediatric sepsis group than in the healthy group, there was no significant difference in renin expression levels in the different disease day groups of the sepsis group, the number of sepsis samples of D1/3/7 n=32, and the healthy group=83.

The differences in the expression levels of renin in different subgroups of sepsis are shown in figure 3. Data are expressed as mean ± standard deviation, P <0.001; * P < 0.0001. Fig. 3 a shows that renin is expressed at significantly higher levels in the septic shock group than in the normal sepsis group; in fig. 3B shows that renin was expressed at significantly higher levels in the 28-day-death group than in the 28-day-survival group with sepsis.

The correlation analysis of various indexes of the severity of the renin and the sepsis infant is shown in figure 4. Fig. 4 a shows that renin levels in sepsis infants are positively correlated with blood lactate levels (r=0.2396; p < 0.05); FIG. 4B shows that renin levels in sepsis patients are positively correlated with childhood organ dysfunction score 2 (PELOD-2) (r=0.3849; P < 0.0001); in fig. 4C shows that renin levels in infants with sepsis are positively correlated with the children sequential organ failure score (pSOFA) (r=0.3577; p=0.0001).

Comparison of mechanical ventilation characteristics of renin in pediatric sepsis is shown in figure 5. Data are expressed as mean ± standard deviation, P <0.05. Fig. 5 a shows that renin expression levels in the sepsis-combined mechanical ventilation group were significantly higher than in the sepsis-non-mechanical ventilation group (< 0.05, < P); fig. 5B shows that the expression level of renin in the sepsis subgroup with mechanical ventilation time exceeding 1 week is higher than in the sepsis subgroup with mechanical ventilation time less than 1 week (< 0.05); FIG. 5C shows that renin is expressed at higher levels in the sepsis subgroup than P/F > 300 in the subgroup with the ratio of arterial blood oxygen partial pressure to inhaled oxygen saturation (P/F). Ltoreq.300, but without significant statistical differences.

ROC curves of renin in pediatric sepsis model versus predicted potency of Procalcitonin (PCT) -Area Under Curve (AUC) are shown in fig. 6. As shown in fig. 6: renin has good predicted efficacy, with an area under the curve (AUC) of 0.8074, which is higher than PCT (0.7536).

The relevant survival curve (Kaplan-Meier survival curve) of renin and 108 cases of sepsis infants is shown in FIG. 7. The cut-off value (Cutoff value) of renin is calculated to be 842.4pg/mL according to the Johnson index, the cut-off value is divided into two groups, survival curve correlation analysis is carried out according to the 28-day survival rate, the difference has statistical significance (P < 0.05), and the fact that the serum renin level is higher than 842.4pg/mL has larger prediction value for children sepsis is suggested.

The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims (10)

1. Use of renin as a biomarker for the manufacture of a product for diagnosing, prognosticating or monitoring or staging the progression of sepsis or septic lung injury in children aged 28 days to 18 years.

2. The use of claim 1, wherein the product is for staging the progression of sepsis or septic lung injury, wherein the sepsis is staged as normal infection, sepsis or septic shock.

3. The use according to any one of claims 1-2, wherein the method of diagnosing, predicting or monitoring or staging the progression of sepsis or septic lung injury is qualitative, quantitative detection of renin from a biological sample obtained from the child.

4. The use according to claim 3, wherein the biological sample comprises plasma, serum or blood.

5. The use according to any one of claims 3 to 4, wherein the biological sample is analysed by enzyme-linked immunosorbent assay.

6. The use of claim 5, wherein an increase in the level of renin in the biological sample as compared to a control sample or a reference sample indicates an increase in the severity of sepsis or septic lung injury as it progresses to the various stages.

7. A detection reagent for children sepsis and septic lung injury is characterized by taking renin as a detection object.

8. A test kit for pediatric sepsis and septic lung injury comprising the test reagent of claim 7.

9. Use of renin as a biomarker in the manufacture of a product for determining the score of the severity of sepsis or septic lung injury in a child by qualitatively and quantitatively detecting renin in a biological sample obtained from the child and comparing the level of renin to a control sample or a reference sample to derive a score.

10. Use of renin as a biomarker in the manufacture of a product for determining childhood organ dysfunction or childhood sequential organ failure by qualitatively and quantitatively detecting renin in a biological sample obtained from the child and comparing the level of renin with a control sample or a reference sample.