Use Of Biomarker Or Biomarkers Combinations In Preparing A Diagnostic Reagent For Fulminant Myocarditis And A Drug For Fulminant Myocarditis

Abstract

The invention “use of biomarker or biomarkers combination in preparing a diagnostic reagent for fulminant myocarditis and a drug for fulminant myocarditis” belongs to the field of disease diagnostic reagents and drugs. The biomarker comprises Siglec-5. The biomarkers combination is further selected from the group consisting of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4. The invention finds that the levels of the biomarker in plasma of patients with fulminant myocarditis is increased, and proves that the biomarker plays a role in diagnosing fulminant myocarditis and a role as a drug target in relieving and/or improving fulminant myocarditis. The level of the biomarker can be detected to forecast or auxiliary diagnose fulminant myocarditis or estimate prognosis of fulminant myocarditis. Meanwhile, level of the biomarkers is controlled through drugs, more intensified diagnosis and follow-up treatment can be carried out on diseases, and the biomarkers have huge clinical application value.

Claims

1. Use of biomarker or biomarkers combination in preparing diagnostic reagents for fulminant myocarditis, wherein the biomarker comprises: Siglec-5.

2. The use of biomarker or biomarkers combination in preparing diagnostic reagents for fulminant myocarditis of claim 1, wherein the biomarkers combination is selected from the group consisting of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

3. The use of biomarker or biomarkers combination in preparing diagnostic reagents for fulminant myocarditis of claim 1, wherein the diagnostic reagents for fulminant myocarditis includes the biomarkers combination and other diagnostically acceptable auxiliary reagents; the diagnostically acceptable auxiliary reagents include: ELISA test reagents, or flow detection reagents.

4. The use of biomarker or biomarkers combination in preparing diagnostic reagents for fulminant myocarditis of claim 1, wherein the diagnosis includes determining whether suffer from fulminant myocarditis, or assessing prognosis of fulminant myocarditis; and/or the diagnosis refers to whether suffer from fulminant myocarditis, or prognosis of fulminant myocarditis is assessed based on expression level of biomarker in the biomarkers combination in subject; and/or when Siglec-5 expression level≥89.56 pg/mL, and/or sST2 expression level≥21.39 ng/mL, and/or PAI-1expression level≥41.29 ng/mL, and/or CD163 expression level≥133.8 ng/mL, and/or CD40 expression level≥67.43 pg/mL, and/or P-Cadherin expression level≥2634 pg/mL, and/or CD14 expression level≥262.4 ng/mL, and/or CTLA4 expression level≥11.19 pg/mL, fulminant myocarditis is diagnosed; and/or when Siglec-5 expression level<89.56 pg/mL, and/or sST2 expression level<21.39 ng/mL, and/or CD163 expression level<133.8 ng/mL, the prognosis of fulminant myocarditis is good.

5. The use of biomarker or biomarkers combination in preparing diagnostic reagents for fulminant myocarditis of claim 1, wherein the fulminant myocarditis is selected from fulminant myocarditis caused or triggered by viruses, bacteria, allergies, drugs, upper respiratory tract infections, and/or intestinal infection.

6. Use of biomarker or biomarkers combination in preparing drug to prevent and cure fulminant myocarditis, wherein the biomarker comprises: Siglec-5.

7. The use of biomarker or biomarkers combination in preparing drug to prevent and cure fulminant myocarditis of claim 6, wherein the biomarkers combination is selected from the group consisting of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

8. The use of biomarker or biomarkers combination in preparing drug to prevent and cure fulminant myocarditis of claim 6, wherein drug target of the drug to prevent and cure fulminant myocarditis is biomarker in the biomarkers combination; and/or the drug to prevent and cure fulminant myocarditis includes an active ingredient that reduces, downregulates, or inhibits the biomarker; and/or the active ingredient that reduces, downregulates or inhibits the biomarker is selected from inhibitor, neutralizing antibody or polypeptide of the biomarker.

9. The use of biomarker or biomarkers combination in preparing drug to prevent and cure fulminant myocarditis of claim 6, wherein the drug to prevent and cure fulminant myocarditis takes the biomarker in the biomarkers combination as the drug target, and achieves the effect of preventing and curing fulminant myocarditis by inhibiting expression level of the biomarker; and/or the drug to prevent and cure fulminant myocarditis achieves effect of preventing and curing fulminant myocarditis by reducing expression level of biomarker in plasma of patients with fulminant myocarditis by inhibitor, neutralizing antibody or peptide against the biomarker; and/or the fulminant myocarditis is defined as fulminant myocarditis caused or triggered by viruses, bacteria, allergies, drugs, upper respiratory tract infections, and/or intestinal infections.

10. The use of biomarker or biomarkers combination in preparing drug to prevent and cure fulminant myocarditis of claim 6, wherein the drug to prevent and cure fulminant myocarditis also includes pharmaceutically acceptable excipients.

Description

DESCRIPTION OF DRAWINGS

[0057] In order to more clearly state the technical solution in embodiments of this invention, a brief description of drawings required for descriptions of the embodiments is given below, clearly, drawings referring to description below are just some examples of the present invention. For a person skilled in the art, other relevant drawings can be obtained from these drawings without creative works.

[0058] FIG. 1 shows expression profile of inflammatory factors in peripheral blood of control population and patients with fulminant myocarditis at admission. In the figure, control represents the control population, FM represents patients with fulminant myocarditis; True indicates that value is greater than threshold and difference is significant. False indicates that value is less than threshold and difference is not significant.

[0059] FIG. 2 shows change profile of inflammatory factors in peripheral blood of patients with fulminant myocarditis before and after treatment. In the figure, FM-admission represents blood test results of patients with fulminant myocarditis when admitted without untreated. FM-discharge represents result of a blood sample test performed on patient with fulminant myocarditis after treatment and before discharge; True indicates that value is greater than threshold and difference is significant. False indicates that value is less than threshold and difference is not significant.

[0060] FIG. 3 shows expression levels of specific inflammatory factors in patients with fulminant myocarditis. In the figure, FM-admission represents expression levels of inflammatory factors in blood samples of patients with fulminant myocarditis when admitted without treatment. FM-discharge represents expression level of inflammatory cytokines in blood sample of a patient with fulminant myocarditis after treatment and before discharge; control represents expression level of inflammatory factors in blood samples of control population; folds refers to a fold change value of expression level. Cytokines refer to inflammatory factors.

[0061] FIG. 4 shows chart of content analysis on 8 inflammatory factors in blood samples of control population and patients with fulminant myocarditis, indicating that 8 inflammatory factors can be used in diagnosis and evaluation of fulminant myocarditis. In the figure, con represents expression level of inflammatory factors in blood samples of control population. FM represents expression level of inflammatory factors in blood samples from patients with fulminant myocarditis.

[0062] FIG. 5 shows analysis of correlation between 9 inflammatory factors and cardiac ejection fraction, among which 3 inflammatory factors can be used to evaluate prognosis of fulminant myocarditis. In the figure, Ejection Fraction represents Ejection Fraction, and FM represents expression level of inflammatory factors in blood samples of patients with fulminant myocarditis; hs-CRP represents positive control.

[0063] FIG. 6 shows ROC curve of diagnostic efficacy of peripheral blood levels of specific inflammatory factors for fulminant myocarditis; in the figure, hs-CRP represents positive control.

[0064] FIG. 7 shows bar chart of levels of inflammatory factors in four groups of mice in experiment example 4, indicating that levels of various inflammatory factors in peripheral blood of fulminant myocarditis A/J mice are increased. In the figure, C57 represents C57 mice, A/J represents A/J mice, Balb/c represents Balb/c mice, C3H represents C3H mice, Control represents blank control group, and CVB3 represents CVB3 myocarditis model group.

[0065] FIG. 8 shows numerical bar graphs of maximum velocity of increase of left ventricular pressure (−dP/dt.sub.min) and maximum velocity of decrease of left ventricular pressure(−dP/dt.sub.min) of three groups of mice in experiment example 5, indicating that sST2 can induce cardiac dysfunction in A/J mice.

[0066] FIG. 9 shows TUNEL staining of cardiomyocytes and bar chart of TUNEL positive cell rate per 100 cardiomyocytes (TUNEL positive rate) of 3 groups of mice in experiment example 5, indicating that sST2 can induce apoptosis of cardiomyocytes in A/J mice. In FIG. 7 and FIG. 8, control represents blank control group; PBS represents negative control group; sST2 represents experimental group.

[0067] FIG. 10 shows chart of analysis on content of sST2 in blood samples of mice in 4 groups of experimental example 6, indicating that sST2 neutralizing antibody can inhibit the increased sST2 in peripheral blood of mice with A/J fulminant myocarditis induced by CVB3.

[0068] FIG. 11 shows chart of analysis on content of sST2 in myocardial tissue of 4 groups of mice in experiment example 6, indicating that sST2 neutralizing antibody can inhibit the increased sST2 in heart of mice with A/J fulminant myocarditis induced by CVB3.

[0069] FIG. 12 shows chart of analysis on survival rate of 4 groups of mice in experiment example 6, indicating that sST2 neutralizing antibody can improve survival rate of A/J fulminant myocarditis mice induced by CVB3.

[0070] FIG. 13 shows numerical bar graphs of maximum velocity of decrease of left ventricular pressure (−dP/dt.sub.min) and maximum velocity of increase of left ventricular pressure(−dP/dt.sub.max) of 4 groups of mice in experiment example 6, indicating that sST2 neutralizing antibody can improve the damaged cardiac function of mice with A/J fulminant myocarditis induced by CVB3.

[0071] FIG. 14 shows staining of myocardial tissue sections and histogram of proportion of inflammatory regions in myocardial tissue of 4 groups of mice in experiment example 6, indicating that sST2 neutralizing antibody can reduce inflammatory cell infiltration in heart of mice with A/J fulminant myocarditis induced by CVB3.

[0072] FIG. 15 shows TUNEL staining of cardiomyocytes and bar chart of TUNEL positive cell rate per 100 cardiomyocytes (TUNEL positive rate) of 4 groups of mice in experiment example 6, indicating that sST2 neutralizing antibody can reduce cardiomyocyte apoptosis in heart of mice with A/J fulminant myocarditis induced by CVB3.

[0073] In FIGS. 10-15, control represents blank control group; PBS+goat IgG represented negative control group. CVB3+PBS represented CVB3 fulminant myocarditis model group; CVB3+ anti-sST2 or anti-sST2 represented treatment group.

[0074] FIG. 16 shows expression levels of inflammatory factors in peripheral blood of fulminant myocarditis mice in experimental example 7, suggesting that anti-CD163 neutralizing antibodies can inhibit increase of CD163. In the figure, ordinate is expression level of CD163 inflammatory factor, and on abscissa, control represents blank control group, PBS+IgG represents negative control group, CVB3 represents CVB3 fulminant myocarditis model group, and CVB3+anti-CD163 represents treatment group.

[0075] FIG. 17 shows cardiac function of mice in experimental example 7, indicating that anti-CD163 neutralizing antibody can improve cardiac dysfunction of A/J fulminant myocarditis mice induced by CVB3. In the figure, on the left ordinate, cardiac output represents the measured value of cardiac output, and on the right ordinate, stroke volume represents stroke output, control represented blank control group, PBS+IgG represented negative control group, CVB3 represented CVB3 fulminant myocarditis model group, and CVB3+ anti-CD163 represented treatment group.

EMBODIMENTS

[0076] Technical solution of this present invention will be clearly and completely described in the light of the following specific embodiments. Clearly, the described embodiments are only a part of embodiments of this invention and not all of them. Based on embodiments of the present invention, all other embodiments obtained by a person skilled in the art without creative works will fall within the scope of protection of the present invention.

[0077] Sources of Biological Materials

[0078] Mice used in the experimental examples of this invention can be purchased commercially.

[0079] Examples Group 1, Use of the Biomarker Combination of the Invention

[0080] This group of examples provides use of biomarker or biomarkers combination in preparing diagnostic reagents for fulminant myocarditis, characterized in that, the biomarker comprises: Siglec-5.

[0081] The biomarkers combination is selected from the group consisting of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

[0082] Specifically, the biomarkers combination is selected from one or more of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

[0083] More specifically, the biomarkers combinations is selected from one, two, three, four, five, six, seven, or eight of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

[0084] The diagnostic reagents for fulminant myocarditis include the biomarkers combination and other diagnostically acceptable auxiliary reagents; the diagnostically acceptable auxiliary reagents include: ELISA test reagents, or flow detection reagents.

[0085] The diagnosis includes determining whether suffer from fulminant myocarditis, or assessing prognosis of fulminant myocarditis;

[0086] preferably, the diagnosis refers to whether suffer from fulminant myocarditis, or prognosis of fulminant myocarditis is assessed based on expression level of biomarker in the biomarkers combination in subject;

[0087] preferably, when Siglec-5 expression level≥89.56 pg/mL,

[0088] and/or sST2 expression level≥21.39 ng/mL,

[0089] and/or PAI-1expression level≥41.29 ng/mL,

[0090] and/or CD163 expression level≥133.8 ng/mL,

[0091] and/or CD40 expression level≥67.43 pg/mL,

[0092] and/or P-Cadherin expression level≥2634 pg/mL,

[0093] and/or CD14 expression level≥262.4 ng/mL,

[0094] and/or CTLA4 expression level≥11.19 pg/mL, fulminant myocarditis is diagnosed;

[0095] preferably, when Siglec-5 expression level<89.56 pg/mL,

[0096] and/or sST2 expression level<21.39 ng/mL,

[0097] and/or CD163 expression level<133.8 ng/mL, prognosis of fulminant myocarditis is good.

[0098] The fulminant myocarditis is selected from fulminant myocarditis caused or triggered by viruses, bacteria, allergies, drugs, upper respiratory tract infections, and/or intestinal infection.

[0099] All above markers in this disclosure have conventional technical meanings commonly understood by a person skilled in the art, such as:

[0100] Siglec-5 can be Siglec-5 as described in Patent Application No. 201980062880.6;

[0101] sST2 can be the sST2 recorded in “Chinese Guidelines for Diagnosis and Treatment of Heart Failure”;

[0102] PAI-1 can be the PAI-1 described in an article of “Research on Application of PAI-1 Gene Testing for Ischemic stroke Prevention”;

[0103] CD163 can be the CD163 described in an article of “Research Progress of M2 macrophage marker CD163 and tumor”;

[0104] CD40 can be the CD40 described in an article of “Research on correlation between serum soluble CD40 ligand levels and Disease Activity in Crohn's Disease”;

[0105] P-Cadherin can be the P-Cadherin described in “Ultrasensitive Detection of Fn14, P-Cadherin and N-Cadherin Based on Single Molecule Array Technology”;

[0106] CD14 can be the CD14 recorded in an article of “Research on correlation between CT signs of AIDS complicated with pulmonary tuberculosis and plasma soluble CD14”;

[0107] CTLA4 can be the CTLA4 described in an article of “Correlation between serum GP73 and CTLA4 levels and degree of liver fibrosis in patients with chronic hepatitis B”;

[0108] hs-CRP can be the hs-CRP described in an article of “Study on Relationship between serum PTX3, CAV-1, hs-CRP and condition and prognosis of patients with acute cerebral infarction”.

[0109] Examples Group 2, Pharmaceutical Use of Biomarkers Combination of this Invention

[0110] This group of examples provides use of biomarker or biomarkers combination in preparing drug to prevent and cure fulminant myocarditis, characterized in that, the biomarker comprises: Siglec-5.

[0111] The biomarkers combination is selected from the group consisting of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

[0112] Specifically, the biomarkers combination is selected from one or more of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4;

[0113] More specifically, the biomarkers combination is selected from one, two, three, four, five, six, seven, or eight of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4.

[0114] Drug target of the drug to prevent and cure fulminant myocarditis is biomarker in the biomarkers combination.

[0115] Preferably, the drug to prevent and cure fulminant myocarditis includes an active ingredient that reduces, downregulates, or inhibits the biomarker.

[0116] Preferably, the active ingredient that reduces, downregulates or inhibits the biomarker is selected from inhibitor, neutralizing antibody or polypeptide of the biomarker.

[0117] The drug to prevent and cure fulminant myocarditis takes the biomarker in the biomarkers combination as the drug target, and achieves the effect of preventing and curing fulminant myocarditis by inhibiting expression level of the biomarker.

[0118] Preferably, the drug to prevent and cure fulminant myocarditis achieves effect of preventing and curing fulminant myocarditis by reducing expression level of biomarker in plasma of patients with fulminant myocarditis by inhibitor, neutralizing antibody or peptide against the biomarker.

[0119] Preferably, the fulminant myocarditis is defined as fulminant myocarditis caused or triggered by viruses, bacteria, allergies, drugs, upper respiratory tract infections, and/or intestinal infections.

[0120] The drug to prevent and cure fulminant myocarditis also includes pharmaceutically acceptable excipients.

[0121] Preferably, the pharmaceutically acceptable excipients include: solvent, propellant, solubilizer, cosolvent, emulsifying agent, colorant, adhesive, disintegrant, filling agent, lubricant, wetting agent, osmotic pressure regulator, stabilizer, flow aid, flavoring agent, preservative, suspending agent, coating material, aromatic, anti-adhesive, integrator, osmosis enhancer, pH regulator, buffering agent, plasticizer, surface active agent, foaming agent, elimination foaming agent, thickening agent, inclusion agent, humectant, absorbent, diluent, flocculant, deflocculant, filter aid, release blocker.

THE EXPERIMENTAL EXAMPLE 1

[0122] Expression Levels of Multiple Inflammatory Factors in Peripheral Blood of Patients with Fulminant Myocarditis are Changed

[0123] Study population and study design: From April 2017 to March 2019, consecutive inpatients with fulminant myocarditis were enrolled in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province. The inclusion criteria included: {circle around (1)} older than 18 years; {circle around (2)} Diagnosis of fulminant myocarditis: myocardial biopsy suggests myocarditis or clinical diagnosis (rapid onset of severe hemodynamic disturbance symptoms, increased cTnI, cardiac ultrasound suggested rapid reduction of left ventricular EF value or less than 40%). Exclusion criteria included: {circle around (1)} acute myocardial infarction; {circle around (2)} common acute myocarditis; {circle around (3)} severe neurological diseases (Alzheimer's disease, progressive stage of Parkinson's syndrome), lower limb disability or deaf-mute patients.

[0124] Detection of inflammatory factors expression profile: fasting peripheral blood was collected into EDTA anticoagulant tube, and serum was isolated immediately by centrifugation at 3000 g for 8 minutes and then frozen at −80° C. until analysis. Plasma inflammatory factors profile was detected by Human Cytokine Arrays (RayBiotech Company).

[0125] Experimental results: 4 patients with fulminant myocarditis were enrolled in this study, and 4 people with matched baseline data including gender, age and etc., were enrolled in control group. The results showed that compared with the control group, levels of 58 inflammatory factors in peripheral blood of patients with fulminant myocarditis were significantly changed (FIG. 1). These results suggested that changed levels of numerous inflammatory factors in plasma were associated with fulminant myocarditis. In the figure, control represents the control group population, FM represents patients with fulminant myocarditis, and red dots represent inflammatory factors with significant differences.

THE EXPERIMENTAL EXAMPLE 2

[0126] Expression Level of Specific Inflammatory Factors in Peripheral Blood of Patients with Fulminant Myocarditis were Back to Normal Level After Treatment and Improved

[0127] Study population and study design: From April 2017 to March 2019, consecutive inpatients with fulminant myocarditis were enrolled in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province. The inclusion criteria included: {circle around (1)} older than 18 years; {circle around (2)} Diagnosis of fulminant myocarditis: myocardial biopsy suggested myocarditis or clinical diagnosis (rapid onset of severe hemodynamic disturbance symptoms, increased cTnI, cardiac ultrasound suggested rapid reduction of left ventricular EF value or less than 40%). Exclusion criteria included: {circle around (1)} acute myocardial infarction; {circle around (2)} common acute myocarditis; {circle around (3)} severe neurological diseases (Alzheimer's disease, progressive stage of Parkinson's syndrome), lower limb disability or deaf-mute patients.

[0128] Detection of inflammatory factor expression profile: fasting peripheral blood was collected into EDTA anticoagulant tube, and serum was isolated immediately by centrifugation at 3000 g for 8 minutes and then frozen at −80° C. until analysis. Plasma inflammatory factor expression profile was detected by Human Cytokine Arrays (RayBiotech Company).

[0129] Experimental results: 4 patients with fulminant myocarditis were enrolled in this study, blood samples were collected at admission without treatment and after treatment as well as improved before discharge. The results showed that after effective treatment, content of some changed inflammatory factors significantly returned (FIG. 2). These results suggested that the changed levels of specific inflammatory factors in peripheral blood were associated with fulminant myocarditis, namely sST2, PAI-1, Siglec-5, CD163, IL-17B, IL-4, VEGF-C, CD40, P-Cadherin, CD14, Angiostatin and CTLA4 (FIG. 3). * means p<0.05.

THE EXPERIMENTAL EXAMPLE 3

[0130] Levels of Specific Inflammatory Factors in Peripheral Blood for Diagnosis of Fulminant Myocarditis and Prognostic Evaluation of Fulminant Myocarditis

[0131] Study population and study design: From April 2017 to March 2019, consecutive inpatients with fulminant myocarditis were enrolled in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province. The inclusion criteria included: {circle around (1)} older than 18 years; {circle around (2)} Diagnosis of fulminant myocarditis: myocardial biopsy suggested myocarditis or clinical diagnosis (rapid onset of severe hemodynamic disturbance symptoms, increased cTnI, cardiac ultrasound suggested rapid reduction of left ventricular EF value or less than 40%). Exclusion criteria included: {circle around (1)} acute myocardial infarction; {circle around (2)} common acute myocarditis; {circle around (3)} severe neurological diseases (Alzheimer's disease, progressive stage of Parkinson's syndrome), lower limb disability or deaf-mute patients.

[0132] Detection of inflammatory factor levels: fasting peripheral blood was collected into EDTA anticoagulant tube, and serum was isolated immediately by centrifugation at 3000 g for 8 minutes and then frozen at −80° C. until analysis. Plasma inflammatory factor level was detected by ELISA kit (R&D Company).

[0133] Experimental results: 32 patients with fulminant myocarditis were enrolled in this study, and 16 patients whose matching baseline data like gender, age and etc., has no difference with 4 patients were enrolled in control group. The results showed that levels of 8 inflammatory factors (sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14 and CTLA4) in peripheral blood of patients with fulminant myocarditis were significantly changed, which had diagnostic value (FIG. 4). Among them, levels of 3 inflammatory factors (sST2, Siglec-5 and CD163) were correlated with EF values of cardiac function, which had prognostic value (FIG. 5). Moreover, levels of specific inflammatory factors (sST2, Siglec-5 and CD163) in peripheral blood of patients had better diagnostic efficacy than CRP (FIG. 6). * represents p<0.05.

THE EXPERIMENTAL EXAMPLE 4

[0134] Levels of Inflammatory Factors in Peripheral Blood of Fulminant Myocarditis Mice Increased

[0135] Mouse model preparation: 8-week-old male C57, A/J, Balb/c and C3H mice were purchased from Model Animal Center of Nanjing University, and were raised in SPF animal room of Tongji Medical College, Huazhong University of Science and Technology. Mice with different backgrounds were intraperitoneally injected with CVB3 virus (10.sup.4 dissolved in 0.1 ml PBS), respectively, after one week of adaptation in the animal house, and used as model group of CVB3 myocarditis.

[0136] Level of inflammatory factors was detected by ELISA Kit of R&D company.

[0137] Experimental results: cardiac catheter detection was performed 8 days after intraperitoneal injection of CVB3 virus, and then animals were sacrificed to collect tissue samples for above detection. The results showed that sST2, C163, P-Cadherin, CD14 and CTLA4 were most significantly increased in peripheral blood of mice with A/J myocarditis after CVB3 infection compared with control group (FIG. 7). * represents p<0.05.

THE EXPERIMENTAL EXAMPLE 5

[0138] Taking sST2 as an Example, Effects of Certain Inflammatory Factors on Cardiac Function of A/J Mice were Observed

[0139] Mouse model preparation: 8-week-old male A/J mice were purchased from the Model Animal Center of Nanjing University, and were raised in SPF animal room of Tongji Medical College, Huazhong University of Science and Technology. Mice were intraperitoneally injected with recombinant sST2 protein (5 ug dissolved in 0.1 ml PBS) one week after adaptation to the animal room, as intervention group (experimental group). Another group of mice were treated with PBS as negative control group, and the third group was blank control group without any treatment (control).

[0140] Cardiac hemodynamic assays in mice were performed by using the Millar Pressure-volume system of Millar Instrument PowerLab Company. After animals were anesthetized to a proper depth, a median incision was made in their neck, the right common carotid artery was separated, its distal end was ligated, and the proximal end was clipped. A V-shaped incision was made in artery with microscissors, and a microcatheter was inserted until the left ventricle, and signals were recorded through conduction system. Hemodynamic data such as heart rate (HR), left ventricular end-diastolic pressure (PED), left ventricular end-systolic pressure (PES), maximum rate of decline of left ventricular pressure (−dP/dt.sub.min) and maximum rate of rise of left ventricular pressure (+dP/dt.sub.max) were obtained.

[0141] Histological detection: myocardial tissue was immersed and fixed in neutral formaldehyde solution in the embedding frame, dehydrated, embedded in paraffin to obtain paraffin block. The paraffin block was cut into sections with a thickness of 4 μm on microtome. HE staining was used to observe gross appearance of myocardium, and TUNEL staining was detected by test kit of Roche Company.

[0142] Experimental results: cardiac catheter detection was performed 10 days after intraperitoneal injection of recombinant sST2 protein, and then the animals were sacrificed and tissue samples were collected for above detection. The results showed that intraperitoneal injection of recombinant sST2 protein significantly decreased cardiac function of mice compared with control group (FIG. 8). Moreover, intraperitoneal injection of recombinant sST2 protein increased TUNEL positive rate in cardiomyocytes, suggesting increased apoptosis of cardiomyocytes (FIG. 9). * represents p<0.05.

THE EXPERIMENTAL EXAMPLE 6

[0143] Anti-sST2 Neutralizing Antibody Can Inhibit Myocardial Injury Induced by CVB3 in A/J Fulminant Myocarditis Mice

[0144] Mouse model preparation: 8-week-old male A/J mice were purchased from the Model Animal Center of Nanjing University, and were raised in SPF animal room of Tongji Medical College, Huazhong University of Science and Technology. Mice were given intraperitoneal injection of CVB3 virus (10.sup.4 dissolved in 0.1 ml PBS) one week after acclimation to the animal room as model group of fulminant myocarditis of CVB3. Another group of fulminant myocarditis model mice were intraperitoneally injected with anti-sST2 neutralizing antibody (10 μg dissolved in 0.1 ml PBS) as treatment group. The third group was blank control group without any treatment. The fourth group was negative control group which received intraperitoneal injection of goat IgG antibody (10 μg dissolved in 0.1 ml PBS).

[0145] sST2 levels were detected by Mouse ST2/IL-33R Quantikine ELISA Kit of R&D company.

[0146] Cardiac hemodynamic assays in mice were performed using the Millar Pressure-volume system from Millar Instrument PowerLab Company. After animals were anesthetized to a proper depth, a median incision was made in their neck, the right common carotid artery was separated, its distal end was ligated, and the proximal end was clipped. A V-shaped incision was made in artery with microscissors, and a microcatheter was inserted until the left ventricle, and signals were recorded through conduction system. Hemodynamic data such as heart rate (HR), left ventricular end-diastolic pressure (PED), left ventricular end-systolic pressure (PES), maximum rate of decline of left ventricular pressure (−dP/dt.sub.min) and maximum rate of rise of left ventricular pressure (+dP/dt.sub.max) were obtained.

[0147] Histological detection: myocardial tissue was immersed and fixed in neutral formaldehyde solution in the embedding frame, dehydrated, embedded in paraffin to obtain paraffin block. The paraffin block was cut into sections with a thickness of 4 μm on microtome. HE staining was used to observe gross appearance of myocardium, and TUNEL staining was detected by test kit of Roche Company.

[0148] Experimental results: cardiac catheter detection was performed 8 days after intraperitoneal injection of anti-sST2 neutralizing antibody, and then the animals were sacrificed and tissue samples were collected for above detection. The results showed that sST2 in peripheral blood of mice with fulminant myocarditis after CVB3 infection increased compared with control group, while anti-sST2 neutralizing antibody could reduce it (FIG. 10). sST2 was increased in heart of mice with fulminant myocarditis after CVB3 infection, which was decreased by anti-sST2 neutralizing antibodies (FIG. 11). Survival rate of fulminant myocarditis mice after CVB3 infection was significantly decreased, which was increased by anti-sST2 neutralizing antibody (FIG. 12). Cardiac function of fulminant myocarditis mice after CVB3 infection was significantly decreased, which was enhanced by anti-sST2 neutralizing antibodies (FIG. 13). Inflammatory cell infiltration was significantly increased in heart of fulminant myocarditis mice after CVB3 infection, which was reduced by anti-sST2 neutralizing antibody (FIG. 14). TUNEL positive rate in cardiomyocytes of mice with fulminant myocarditis after CVB3 infection was significantly increased, which was reduced by anti-sST2 neutralizing antibody (FIG. 15). * represents p<0.05.

THE EXPERIMENTAL EXAMPLE 7

[0149] Anti-CD163 Neutralizing Antibody Can Improve Cardiac Dysfunction in A/J Fulminant Myocarditis Mice Induced by CVB3

[0150] Mouse model preparation: 8-week-old male A/J mice were purchased from the Model Animal Center of Nanjing University, and were raised in SPF animal room of Tongji Medical College, Huazhong University of Science and Technology. Mice were given intraperitoneal injection of CVB3 virus (10.sup.4 dissolved in 0.1 ml PBS) one week after acclimation to the animal room as model group of fulminant myocarditis of CVB3. Mice without any treatment were used as blank control group, and mice with goat IgG antibody (10 μg dissolved in 0.1 ml PBS) injected intraperitoneally were used as negative control group. In addition, mice in fulminant myocarditis model group were intraperitoneally injected with anti-CD163 neutralizing antibody (10 μg dissolved in 0.1 ml PBS) as treatment group.

[0151] Level of inflammatory factors was detected by ELISA Kit of R&D company.

[0152] Mouse cardiac ultrasound testing: cardiac ultrasound was performed by using Vevo 2100 Small animal ultrasound of Visual Sonics Inc. Main measuring indicators included heart rate (HR), left ventricular ejection fraction (LVEF), left ventricular short-axis shortening rate (LVFS), left ventricular septal thickness during diastole or systole (LVIDd and LVIDs), and left ventricular posterior wall thickness (LVPWd and LVPWs).

[0153] Experimental results: cardiac catheter detection was performed 8 days after intraperitoneal injection of anti-CD163 neutralizing antibody. The animals were sacrificed and tissue samples were collected for above detection. The results showed that CD163 level in peripheral blood of fulminant myocarditis mice after CVB3 infection was increased compared with control group. However, treatment with anti-CD163 neutralizing antibodies decreased CD163 levels (FIG. 16). Cardiac function was significantly decreased in fulminant myocarditis mice after CVB3 infection, which was enhanced by anti-CD163 neutralizing antibody treatment (FIG. 17). * represents p<0.05.

[0154] The aforementioned are merely examples of the present invention. For those skilled in the art, this application can still modify the technical solutions described in the aforementioned embodiments, or equivalently replace some of the technical features. However, these modifications or substitutions do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of various examples of the present invention.