Application of serglycin as a specific marker for Kawasaki disease in men

Abstract

The present invention discloses the application of serglycan protein as a specific marker for male Kawasaki disease. The present invention finds that serglycan protein (SRGN) is significantly highly expressed in male Kawasaki disease patients and male Kawasaki disease mouse models by measuring the serum of patients and mice and coronary artery tissues, while there is no significant difference in female Kawasaki disease patients and female Kawasaki disease group mice, indicating that SRGN can be used as a specific biomarker for diagnosing male Kawasaki disease patients, as a basis for diagnosing male Kawasaki disease patients, or used to prepare Kawasaki disease-related reagents or kits for auxiliary diagnosis of Kawasaki disease.

Description

Technical Field

[0001] The present invention relates to the field of disease diagnosis, and in particular to the application of serglycin protein as a specific marker for male Kawasaki disease. Background Art

[0002] Kawasaki disease (KD) is an acute febrile rash disease with unknown etiology and pathogenic mechanism, with systemic vasculitis as the main lesion. KD is common in infants and young children under 5 years old, among which coronary artery injury (CAL) is the most common. Its main harm is to cause coronary artery dilatation or coronary artery aneurysm formation. In the late stage, coronary artery stenosis or thrombosis may occur, and even myocardial infarction may occur. It is currently the leading cause of acquired heart disease in children. More and more studies have shown that KD is a new risk factor for atherosclerosis and ischemic heart disease, and CAL sequelae are an important cause of myocardial infarction and sudden death in adolescents and adults. Epidemiological and clinical data from various countries show that there are obvious gender differences in the incidence of KD. Among KD children, the incidence rate of male children is significantly higher than that of female children. The incidence rate of coronary artery injury and coronary artery aneurysm in male children is also significantly higher than that in female children.

[0003] In 1967, Japanese scholar Tomisaku Kawasaki first reported KD, and it was subsequently reported all over the world, with the highest incidence in Asia. According to reports from 2011 to 2012 in Japan, the male-to-female ratio was 1.33:1 to 1.49:1, and the peak age of KD was under 1 year old. Studies have also found that the gender distribution is more obvious in younger age groups, that men are more likely to develop coronary artery damage, and that boys have more common heart diseases than girls. [1] According to reports from South Korea, the male-to-female ratio was slightly higher than that of Japan from 2007 to 2014, ranging from 1.37:1 to 1.52:1. [2] Epidemiological surveys in some regions have shown that the male-to-female ratio of children under 5 years old increases with age, and coronary artery damage in boys is significantly higher than in girls. Males are an independent high-risk factor for KD-induced coronary artery aneurysms. [3-5] , and 95% of patients with giant aneurysms are boys [6] In addition, male KD patients treated with IVIG or steroids have a higher relapse rate than female KD patients. [7] So far, only one article has reported that FCGR2A gene polymorphism is associated with male children with Kawasaki disease. [8] .

[0004] So far, the studies on the significantly higher incidence and coronary artery abnormalities in male KD patients than in female patients at home and abroad have mainly focused on the epidemiological statistical analysis of clinical data, and no research on its molecular mechanism has been reported. The pathogenic mechanism of why the pathological damage of the coronary arteries in male KD patients is more serious than that in female KD patients has not been reported. Therefore, exploring the molecular mechanism of higher coronary artery damage in male KD patients than in female KD patients, finding potential drug targets to reduce coronary artery damage in male KD patients for targeted prevention and treatment, and reducing the coronary artery damage rate in male KD patients have become clinical problems that need to be solved urgently. It has important theoretical significance for the prevention and treatment of KD, and can provide clues for the targeted and precise treatment of male KD patients in clinical practice.

[0005] Chinese patent application with publication number 201280070975.0 discloses the use of PDFGC gene expression as a diagnostic biomarker for Kawasaki disease; Chinese patent application with publication number 201711452354.X discloses that eight genes and proteins, including CD247, TNFRSF1A, SMAD2, MMP9, BMP2, ACVR2B, CXCL14 and CD14, can be used as biomarkers in a detection kit for diagnosing the risk of coronary artery lesions in Kawasaki disease; Chinese patent applications with publication numbers 201811602267.2 and 201610194943.1 disclose that miRNA molecules can be used as biomarkers in a detection kit for diagnosing Kawasaki disease or the risk of complications.

[0006] Serglycin (SRGN) belongs to the family of small molecule proteoglycans and is composed of a smaller core protein and various polysaccharide side chains such as GAG chains, chondroitin sulfate, heparin, etc. [9] SRGN protein has three main functional domains: a signal peptide region, encoded by exon 1; the N-terminal structure is encoded by exon 2; the glycosylation domain is encoded by exon 3. It is the only intracellular proteoglycan to date and is mainly involved in the storage and release of peptide hydrolases, inflammatory chemokines or other cytokines.

[0007] SRGN has been shown to be a marker for acute myeloid leukemia (AML) and to be used to differentiate acute lymphoblastic leukemia from Philadelphia chromosome-negative chronic myeloproliferative disease.

[10] In addition, SRGN can inhibit the classical pathway and lectin pathway of complement through the chondroitin sulfate chains in its own structure, thereby protecting tumor cells from attack by the complement system.

[11] SRGN also helps myeloma cells adhere to the extracellular matrix of the bone marrow and plays an important role in tumor invasion and migration.

[12] SRGN expression was found to be upregulated in hematopoietic-derived drug-resistant tumor cells, indicating that SRGN may be associated with chemotherapy resistance in tumor cells

[13] . Among them, high levels of SRGN can independently serve as a prognostic indicator for disease-free survival and distant metastasis-free survival of primary nasopharyngeal carcinoma, and are related to the recurrence rate and survival rate of patients. In the extracellular matrix, SRGN can play an important role in the occurrence of inflammation, wound repair, and cell invasion by activating MMPs.

[14] These studies have shown that SRGN may promote the malignancy of tumors, but the potential related mechanisms still need further exploration. However, so far, there have been no reports on the use of SRGN in the diagnosis of male children with a high incidence of Kawasaki disease coronary artery aneurysms and as a biomarker for the assessment and monitoring of coronary artery damage. Summary of the invention

[0008] The primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide the use of serglycin protein as a specific marker for male Kawasaki disease.

[0009] Another object of the present invention is to provide the use of serglycin protein as a specific marker in the preparation of a diagnostic tool for male Kawasaki disease.

[0010] Another object of the present invention is to provide the use of serglycin protein as a specific marker in the preparation of drugs for preventing and treating male Kawasaki disease.

[0011] The purpose of the present invention is achieved through the following technical solutions:

[0012] Application of serglycin protein as a specific marker for Kawasaki disease in men.

[0013] The serglycin (SRGN) is composed of a core protein, a serine repeat sequence and various glycosaminoglycan (GAG) chains, and is mainly involved in the storage and release of peptide hydrolases, inflammatory chemokines or other cytokines; the structure of SRGN is composed of a core peptide chain and a polysaccharide chain connected to the N or O position, and is mainly expressed in embryonic stem cells, hematopoietic cells and endothelial cells.

[0014] The nucleotide sequence of the serglycin protein is shown in SEQ ID NO.1; serglycin is highly expressed in male Kawasaki disease patients.

[0015] The male is a male child; preferably a male infant under 5 years old.

[0016] Application of serglycan protein as a specific marker in the preparation of diagnostic tools (products) for male Kawasaki disease.

[0017] The diagnostic tool includes a diagnostic reagent or a kit, a serglycan protein (SRGN)-specific antibody, an antibody set-specific indicator reagent, and the like.

[0018] The male is a male child; preferably a male infant under 5 years old.

[0019] The application is to detect and / or analyze the serglycin protein in a biological sample from a subject.

[0020] The biological sample is serum or coronary artery tissue.

[0021] The coronary artery tissue is heart coronary artery tissue.

[0022] The detection of the serglycin protein can be performed by at least one of a mass spectrometry quantitative method, a Western-Blot quantitative method, ELISA, immunomagnetic beads and immunohistochemistry.

[0023] Application of serglycan protein as a specific marker or drug target in the preparation of drugs for preventing and treating Kawasaki disease in men.

[0024] The male is a male child; preferably a male infant under 5 years old.

[0025] The drug includes a pharmaceutically acceptable carrier; the carrier is a diluent, a buffer, a suspension, an emulsion, a granule, an encapsulation, an excipient, a filler, an adhesive, a spray, a transdermal absorbent, a wetting agent, a disintegrant, an absorption accelerator, a surfactant, a colorant, a flavoring agent or an adsorption carrier, etc.

[0026] Application of serglycan protein as a specific marker in screening drugs for the treatment of Kawasaki disease in men.

[0027] The male is a male child; preferably a male infant under 5 years old.

[0028] The screening is performed in an in vitro environment.

[0029] Use of a reagent for detecting the expression level of serglycin protein in serum or coronary artery tissue, and / or for determining the content of serglycin protein in serum or coronary artery tissue in the preparation of a reagent or kit for diagnosing male Kawasaki disease.

[0030] The coronary artery tissue is heart coronary artery tissue.

[0031] The male is a male child; preferably a male infant under 5 years old.

[0032] Application of serglycan protein inhibitor in the preparation of medicine for preventing and treating Kawasaki disease in men.

[0033] The male is a male child; preferably a male infant under 5 years old.

[0034] Compared with the prior art, the present invention has the following advantages and effects:

[0035] (1) The present invention provides a SRGN protein marker specifically for the diagnosis of KD in male children with Kawasaki disease and a detection method thereof. This marker has not been reported before, and this is the first time that a detection marker method for the diagnosis of Kawasaki disease with gender specificity has been proposed. The marker SRGN protein mainly includes mass spectrometry quantitative method and Western-Blot quantitative method, and other methods such as ELISA, immunomagnetic beads and immunohistochemistry. The expression level of SRGN protein in serum (such as serum of children and mouse serum) and coronary artery tissue of the heart is measured to assist in the diagnosis of Kawasaki disease.

[0036] (2) The present invention relates to the use of clinical specimens and mouse models of Kawasaki disease patients in combination with mass spectrometry analysis technology and Western-Blot analysis technology to obtain protein markers with gender-specific differences: proteomics and Western blotting analysis were performed in serum samples of male and female children with Kawasaki disease, and the results showed that SRGN was significantly more highly expressed in male children with Kawasaki disease than in female children with Kawasaki disease; in addition, the expression of SRGN in male mice with Kawasaki disease was also consistent with clinical samples. The marker was highly expressed in male children with Kawasaki disease and male Kawasaki disease mouse models. These phenomena indicate that SRGN can be used as a specific biomarker for diagnosing male children with Kawasaki disease, and can be used as a basis for the diagnosis of male children with Kawasaki disease, or to prepare Kawasaki disease-related reagents or kits (including SRGN-specific antibodies and antibody set-specific indicator reagents) for auxiliary diagnosis of Kawasaki disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The figure shows the expression of differential proteins between male and female patients with Kawasaki disease analyzed by protein mass spectrometry. A shows the significantly differential proteins between male patients with Kawasaki disease and normal controls (a total of 115 proteins); B shows the significantly differential proteins between female patients with Kawasaki disease and normal controls (a total of 100 proteins).

[0038] Figure 2 yes Figure 1 Venn diagram of differentially expressed proteins (DEPs) between male and female children with Kawasaki disease (37 proteins are differentially expressed proteins specific to male children with Kawasaki disease, among which SRGN expression difference is the most significant).

[0039] Figure 3 This is a graph of the expression verification results of SRGN protein in the serum of clinical patients with Kawasaki disease and the normal control group (C) ("*" and "ns" are used to indicate statistical significance, "***" p<0.001; "**" p<0.01; "*" p<0.05; "ns" p>=0.05); wherein, A is the expression level of SRGN protein in the serum of male normal control group and male children with Kawasaki disease; B is the expression level of SRGN protein in the serum of female normal control group and female children with Kawasaki disease; C is the quantitative result analysis of A; D is the quantitative result analysis of B.

[0040] Figure 4 This is a graph of the expression verification results of SRGN protein in the serum of Kawasaki disease mice and normal control groups ("*" and "ns" are used to indicate statistical significance, "***" p<0.001; "**" p<0.01; "*" p<0.05; "ns" p>=0.05); wherein, A is the expression level of SRGN protein in the serum of male normal control group and male Kawasaki disease mouse group; B is the expression level of SRGN protein in the serum of female normal control group and female Kawasaki disease mouse group; C is the quantitative result analysis of A; D is the quantitative result analysis of B.

[0041] Figure 5 This is a graph of the verification results of SRGN protein expression in the coronary artery tissue of the heart of Kawasaki disease mice and normal control group ("*" and "ns" are used to indicate statistical significance, "***" p<0.001; "**" p<0.01; "*" p<0.05; "ns" p>=0.05); wherein, A is the expression level of SRGN protein in the coronary artery tissue of the male normal control group and the male Kawasaki disease mouse group; B is the expression level of SRGN protein in the coronary artery tissue of the female normal control group and the female Kawasaki disease mouse group; C is the quantitative result analysis of A; D is the quantitative result analysis of B. DETAILED DESCRIPTION

[0042] The present invention is further described in detail below in conjunction with examples, but embodiments of the present invention are not limited thereto. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, reagents and raw materials used in the present invention can be obtained commercially.

[0043] The protein sequence of the serglycin protein (SRGN) (UniProt ID: P10124) in the present invention is as follows (SEQ ID NO.1):

[0044] MMQKLLKCSRLVLALALILVLESSVQGYPTRRARYQWVRCNPDSNSANCLEEKGPMFE LLPGESNKIPRLRTDLFPKTRIQDLNRIFPLSEDYSGSGFGSGSGSGSGSGFLTEMEQDYQL VDESDAFHDNLRSLDRNLPSDSQDLGQHGLEEDFML.

[0045] Example 1 Identification of sex-specific protein biomarkers

[0046] Identification of sex-specific protein biomarkers in the serum of male and female children with Kawasaki disease:

[0047] (1) Clinical sample collection and processing: Five healthy male and female children matched in age and gender (equal numbers of males and females, both Kawasaki disease children and healthy children were under 5 years old and in similar age groups) were randomly selected as the healthy control group and 10 Kawasaki disease children (five male Kawasaki disease children and five female Kawasaki disease children). All Kawasaki disease children were diagnosed according to the Kawasaki disease diagnostic guidelines published by the American Heart Association in 2017. [6] The blood of these clinical samples was centrifuged at 1,000 × g for 10 min to separate serum samples, and the serum components were collected and stored in a -80°C freezer.

[0048] (2) The serum of the five male Kawasaki disease patients, five male healthy children, five female Kawasaki disease patients, and five female healthy children was depleted of albumin and immunoglobulin G (IgG) using the ProteoPrep Blue Albumin & IgG Depletion Kit (Sigma, St. Louis, MO, USA); the serum proteins were then extracted using a lysis buffer (Beyotime Institute of Biotechnology, Haimen, China); and the concentrations of these proteins were then determined using a Bradford protein assay kit (Bio-Rad Laboratories Inc., Hercules, CA, USA). An equal amount of total protein obtained in the above steps was taken from each sample, and finally the protein expression of the above samples was analyzed by a non-label (Label-Free) protein quantification method. The specific steps are as follows:

[0049] ① Take 100 μg of total protein from each sample, use a final concentration of 5 mM Tris (2-carboxyethyl) -phosphine (TCEP, Sciex) reducing agent at 60 ° C to reduce the protein disulfide bonds and then alkylate with methylthiosulfonate cysteine ​​blocking agent iodoacetamide (Sciex) at room temperature for 30 minutes.

[0050] ② The protein was digested overnight at 37°C with 2% Promega Corporation (Madison, Wi, USA) at a ratio of 1:50 (the protease in the kit and the total protein of the sample were mixed in a mass ratio of 1:50); the enzymatic peptide solution of each sample was centrifuged at 6000×g at 25°C for 4 hours in a vacuum centrifuge.

[0051] ③Then, the enzymatic peptide sample was diluted to 100 μl with H2O buffer (NH3·H2O, pH=10), desalted by C-18 Stagetips adsorption column, and then peptide separation was performed in Proxeon EASY-nLC 1200U HPLC liquid phase separation system (Thermo Scientific) coupled to Qbitrap Fusion Lumos Mass mass spectrometer (ThermoScientific): mobile phase acetonitrile (ACN), linear gradient of 6-30% (containing 0.1% (v / v) formic acid); separation elution rate was 270 nL / min, separation time was 100-130 min, and ACN concentration was linearly increased to 90% in the last 20 min of elution.

[0052] ④ MS analysis was performed on a Qbitrap Fusion Lumos Mass spectrometer (Thermo Scientific) with the following parameters: DDA scan mode; source operating condition 2.0 kV; MS spectra were acquired in the high scan range of 400-1500 m / z; resolution was 60,000 FWHM, AGC was set to 4E5 (maximum injection time was 50 ms); followed by MS / MS scan with resolution set to 15,000 FWHM, AGC set to 4E5 (maximum injection time was 30 ms); data collection correlation mode was set to the highest speed; interval window was set to 1.6, dynamic exclusion was set to 90 s; by MaxQuant (ver.1.5.8.3) The MS raw data were compared and searched against the human protein database, with the precursor ion tolerance set to 20 ppm and the fragmentation ion tolerance window set to 0.5 m / z. A maximum of two missed cleavage sites were allowed for the peptide segment, and formyl methylation of cysteine ​​residues and oxidative modification of methionyl were allowed. At the same time, the human proteome sequence reverse library was used to calculate the false discovery rate (FDR), and FDR was set to < 0.01.

[0053] (3) Protein spectrum data analysis results:

[0054] Using mass spectrometry protein identification and quantification technology, we identified 115 differentially expressed proteins between the healthy control group and male Kawasaki disease patients, and 100 differentially expressed proteins between the healthy control group and female Kawasaki disease patients (defined as the protein expression fold change rate between Kawasaki disease and normal control group Fold Change>1.50 or Fold Change<0.67). The relative expression levels of these proteins are shown in the heat map ( Figure 1 ).

[0055] In this result ( Figure 2 and Table 1), we determined that serglycin (SRGN) was highly expressed in male children with Kawasaki disease (Fold change = 7.844, q value < 0.01), but there was no significant difference in female children with Kawasaki disease.

[0056] Table 1 Mass spectrometry results of differentially expressed proteins (DEPs) in male children with Kawasaki disease

[0057]

[0058]

[0059]

[0060] Example 2 Western-Blot detection of SRGN protein expression in serum of children with Kawasaki disease

[0061] In order to further confirm that SRGN protein can be used as a specific diagnostic marker for male children with Kawasaki disease, this example uses Western-Blot technology for verification:

[0062] (1) 30 male healthy children and 30 female healthy children were randomly selected as the control group, and 30 male Kawasaki disease children and 30 female Kawasaki disease children were randomly selected as the experimental group (the healthy children and Kawasaki disease children were all under 5 years old). According to the method of Example 1, the protein in the serum of these children was extracted, and the protein concentration was determined using lysis buffer (Beyotime Institute of Biotechnology, Haimen, China) and Bradford protein assay kit (Bio-Rad Laboratories, Inc).

[0063] (2) 25 μg of protein was taken from each sample and then separated by 10-15% SDS-PAGE electrophoresis. The separated proteins were transferred to polyvinylidene fluoride (PVDF) membranes and blocked with 5% (w / v) skim milk powder at room temperature for 2 hours. Then, they were incubated with primary anti-Serglycin antibody (abcam, catalog number ab156991) at a dilution ratio of 1:1000 in a 4°C refrigerator for 12 hours; then, horseradish peroxidase goat anti-rabbit secondary antibody (abcam, catalog number ab6721) was diluted at a ratio of 1:2,000 at room temperature and incubated at room temperature for 2 hours, then developed by ECL method, and finally, the protein band intensity was quantitatively analyzed using Image J software (version 2.0), and statistical analysis was performed using GraphPad Prism (version 5.04).

[0064] (3) Western-Blot analysis results of SRGN protein in the serum of children with Kawasaki disease Figure 3 Shown: In Figure 3 A and the corresponding quantitative analysis results ( Figure 3 C) shows that the expression of SRGN protein in the serum of male Kawasaki disease patients is higher than that of the male normal control group. Figure 3 B and Figure 3D shows that there is no significant difference in the expression of SRGN protein in the serum of the female Kawasaki disease group compared with the female normal control group (the figure uses "*" and "ns" to indicate the statistical significance of the difference between different groups, "***" p<0.001; "**" p<0.01; "*" p<0.05; "ns" p>=0.05). The above examples show that the high expression of SRGN protein can be used as a specific protein marker for the diagnosis of male Kawasaki disease.

[0065] Example 3 Western-Blot detection of SRGN protein expression in Kawasaki disease mouse samples

[0066] (1) According to the method reported in the literature (Exp Ther Med. 2017Jun; 13(6): 3438-3442. doi: 10.3892 / etm.2017.4434.) to induce KD in mice using Lactobacillus casei cell wall extract (LCWE), the male and female Kawasaki disease mouse models required for this project were established. The specific experimental steps are as follows:

[0067] ① Lactobacillus casei (Cat. GDMCC 1.80, Guangdong Microbial Culture Collection Center) was cultured in lactic acid bacteria broth medium (Cat. 022010, Guangdong Huankai Microbiological Technology Co., Ltd.), and the bacteria in the logarithmic growth phase were collected. 4% SDS was added to the Lactobacillus casei for lysis overnight, and RNAse, DNAse and pancreatin were added respectively for incubation at 37°C, and the bacterial fragments were collected and ultrasonically lysed. The supernatant was taken as LCWE.

[0068] ② Use phenol / sulfuric acid colorimetry to detect the concentration of rhamnose in the supernatant to determine the content of LCWE. Dilute LCWE to 1 mg / ml with PBS, filter and store at -80℃ for later use.

[0069] ③BALB / C mice (Experimental Animal Center of Southern Medical University, 4 weeks old, 16 g) were intraperitoneally injected with 0.5 ml LCWE and PBS buffer, respectively, and orbital blood was collected (orbital blood was collected 10 days later), serum samples were separated, and the coronary artery tissues of the mice were taken for HE staining and EVG staining to examine their pathological changes. The mouse serum was taken and TNF-a, IL-1B and IL-6 were used to detect its inflammation. It was confirmed that the Kawasaki disease mouse model was successfully established, and male Kawasaki disease mouse models and female Kawasaki disease mouse models were obtained.

[0070] (2) Referring to the steps of Example 2 above, changes in SRGN protein were detected in the serum and coronary artery tissues of 30 male healthy mice (control), 30 female healthy mice (control), 30 male Kawasaki disease mice and 30 female Kawasaki mice.

[0071] (3) The results are as follows Figure 4 and Figure 5 (In the figure, "*" and "ns" are used to indicate the statistical significance of the differences between different groups, "***" p<0.001; "**" p<0.01; "*" p<0.05; "ns" p>=0.05) as shown:

[0072] exist Figure 4 A and the corresponding quantitative analysis results Figure 4 In C, the expression of SRGN protein in the serum of male Kawasaki disease mice was significantly higher than that of male normal control mice. Figure 4 B and Figure 4 D shows that there is no significant difference in the expression of SRGN protein in the serum of female Kawasaki disease group mice compared with that of female normal control group mice.

[0073] exist Figure 5 A and the corresponding quantitative analysis results Figure 5 In C, the expression of SRGN protein in the coronary artery tissue of male Kawasaki disease mice was higher than that of male normal control mice. Figure 5 B and Figure 5 D shows that there is no significant difference in the expression of SRGN protein in the coronary artery tissue of female Kawasaki disease group mice compared with that of female normal control group mice.

[0074] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

[0075] References:

[0076] [1]Makino, N. et al. Descriptive epidemiology of Kawasaki disease in Japan, 2011-2012: from the results of the 22nd nationwide survey. J Epidemiol 25, 239-245, doi: 10.2188 / jea.JE20140089(2015).

[0077] [2]Ha,S.,Seo,G.H.,Kim,K.Y.&Kim,D.S.Epidemiologic Study on KawasakiDisease in Korea,2007-2014: Based on Health Insurance Review&AssessmentService Claims.Journal of Korean medical science 31, 1445-1449,doi:10.3346 / jkms.2016.31.9.1445(2016).

[0078] [3]Du,Z.D.et al.Epidemiologic study on Kawasaki disease in Beijingfrom 2000through 2004.The Pediatric infectious disease journal 26,449-451,doi:10.1097 / 01.inf.0000261196.79223.18(2007).

[0079] [4]Chen,J.J.et al.Epidemiologic Features of Kawasaki Disease inShanghai From 2008Through 2012.The Pediatric infectious disease journal 35,7-12,doi:10.1097 / INF.0000000000000914(2016).

[0080] [5]Tang,Y.,Gao,X.,Shen,J.,Sun,L.&Yan,W.Epidemiological and ClinicalCharacteristics of Kawasaki Disease and Factors Associated with CoronaryArtery Abnormalities in East China:Nine Years Experience.J Trop Pediatr 62,86-93,doi:10.1093 / tropej / fmv077(2016).

[0081] [6]Dietz,S.M.et al.Giant aneurysms:A gender-specific complication ofKawasaki disease?Journal of Cardiology 70,359(2017).

[0082] [7]Sudo,D.&Nakamura,Y.Nationwide surveys show that the incidence ofrecurrent Kawasaki disease in Japan has hardly changed over the last30years.Acta paediatrica 106,796-800(2017).

[0083] [8]Kwon,Y.C.et al.Male-specific association of the FCGR2A His167Argpolymorphism with Kawasaki disease.PLoS One 12,e0184248,doi:10.1371 / journal.pone.0184248(2017).

[0084] [9]Kolset,S.O.et al.Serglycin and secretion in humanmonocytes.Glycoconjugate Journal,2008,25(4):305-311.

[0085]

[10] Niemann,C.U.et al.Serglycin proteoglycan in hematologicmalignancies:a marker of acute myeloid leukemia.Leukemia,2007,21(12):2406-2410.

[0086]

[11] Angeliki,K.et al.Serglycin is implicated in the promotion ofaggressive phenotype of breast cancer cells.Plos one,2013,8(10):e78157.

[0087]

[12] Scully,OJet al.Serglycin in health and diseases.AnatomicalRecord Advances in integrative Anatomy&Evolutionary Biology,2012,295(9):1415.

[0088]

[13] Beyer-Sehlmeyer, G. et al. Suppressive subtractive hybridization reveals differential expression of serglycin, sorcin, bone marrow proteoglycan and prostate-tumour-inducing gene I (PTI-1) in drug-resistant and sensitive tumor cell lines of haematopoetic origin. European Journal of Cancer, 1999, 35(12): 1735-1742.

[0089]

[14] Lundequist, A. et al. Mast cell-dependent activation of pro matrixmetalloprotease 2: A role for serglycin proteoglycan-dependent mast cellproteases. Biological Chemistry, 2006, 118 (10 / 11): 1513-1519. Sequence Listing <110> Jinan University <120> Application of serglycin as a specific marker for Kawasaki disease in men <160> 1 <170> SIPOSequenceListing 1.0 <210> 1 <211> 158 <212> PRT <213> Artificial Sequence <220> <223> Serglycin <400> 1 Met Met Gln Lys Leu Leu Lys Cys Ser Arg Leu Val Leu Ala Leu Ala 1 5 10 15 Leu Ile Leu Val Leu Glu Ser Ser Val Gln Gly Tyr Pro Thr Arg Arg 20 25 30 Ala Arg Tyr Gln Trp Val Arg Cys Asn Pro Asp Ser Asn Ser Ala Asn 35 40 45 Cys Leu Glu Glu Lys Gly Pro Met Phe Glu Leu Leu Pro Gly Glu Ser 50 55 60 Asn Lys Ile Pro Arg Leu Arg Thr Asp Leu Phe Pro Lys Thr Arg Ile 65 70 75 80 Gln Asp Leu Asn Arg Ile Phe Pro Leu Ser Glu Asp Tyr Ser Gly Ser 85 90 95 Gly Phe Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Phe 100 105 110 Leu Thr Glu Met Glu Gln Asp Tyr Gln Leu Val Asp Glu Ser Asp Ala 115 120 125 Phe His Asp Asn Leu Arg Ser Leu Asp Arg Asn Leu Pro Ser Asp Ser 130 135 140 Gln Asp Leu Gly Gln His Gly Leu Glu Glu Asp Phe Met Leu 145 150 155

Claims

1. Use of serglycin protein as a specific marker in the preparation of a diagnostic tool for male Kawasaki disease, characterized in that: The diagnostic tool includes a diagnostic reagent or a kit.

2. The use according to claim 1, characterized in that: The diagnostic tool comprises a serglycan protein-specific antibody or an antibody set-specific indicator reagent.

3. The use according to claim 1, characterized in that: The application is to detect and / or analyze the serglycin protein in a biological sample from a subject; The biological sample is serum or coronary artery tissue.

4. The use according to claim 3, characterized in that: The detection of serglycan protein is carried out by at least one method selected from the group consisting of mass spectrometry quantification method, Western-Blot quantification method, ELISA, immunomagnetic beads and immunohistochemistry.

5. Use of a reagent for detecting the expression level of serglycin protein in serum or coronary artery tissue, and / or for determining the content of serglycin protein in serum or coronary artery tissue in the preparation of a reagent or kit for diagnosing Kawasaki disease in men.

6. The use according to any one of claims 1 to 5, characterized in that: The male mentioned is a male infant under 5 years old; The nucleotide sequence of the serglycin protein is shown in SEQ ID NO.1.

Images