Application of soluble factor in paralytic dementia detection

Abstract

The invention relates to application of soluble factors in paralytic dementia detection. Specifically, the invention relates to an application of a soluble factor, NF-L and / or neopterin in preparation of the product, and the soluble factor is selected from any one or a combination of any two or more of 19 soluble factors such as LIF, IL-10 and the like. The invention also relates to an assessment system for assessing the patient with PD based on the cerebrospinal fluid concentration of said soluble factor, NF-L and / or neopterin. According to the method, the PD can be distinguished from other types of syphilis, so that early recognition of the PD is realized, and then targeted diagnosis and treatment can be performed on the PD as soon as possible.

Description

[0001] This application requests the priority of the invention patent whose applicant is "Beijing Ditan Hospital Affiliated to Capital Medical University", whose invention name is "Application of soluble factors in the detection of paralytic dementia" and whose application number is "202311735576.8". Technical Field

[0002] The invention relates to the technical field of biomedicine, and in particular to the application of soluble factors in the detection of paralytic dementia. Background Art

[0003] Syphilis, as a sexually transmitted disease, has had a huge impact on global public health and is one of the most common sexually transmitted diseases. Neurosyphilis is the most common, most serious, and most disabling complication of syphilis, and its incidence is increasing year by year.

[0004] Neurosyphilis is a chronic infectious disease that causes damage to the brain, spinal cord, and peripheral nerves caused by Treponema pallidum invading the nervous system. It can occur in all stages of syphilis. With the annual increase in syphilis cases, the incidence of neurosyphilis has also increased. The clinical manifestations of neurosyphilis are similar to other nervous system diseases, making the diagnosis of neurosyphilis more complicated and difficult. Many patients are diagnosed only after surgery or autopsy. Misdiagnosis and mistreatment often lead to adverse consequences such as paralysis, aphasia, paraplegia, epilepsy, mania, and brain herniation. Therefore, early and accurate diagnosis of neurosyphilis is particularly important for the patient's clinical cure and improvement of their quality of life.

[0005] In the early stage of syphilis, neurosyphilis most often affects the cerebrospinal fluid (CSF), meninges, and vascular system, manifesting as asymptomatic neurosyphilis, meningeal neurosyphilis, and meningeal vascular neurosyphilis; in the late stage of the disease, it most often affects the brain and spinal cord parenchyma, manifesting as paralytic dementia and tabes dorsalis. There is overlap between these manifestations, and even multiple types of overlap.

[0006] Paralytic dementia (PD) belongs to late neurosyphilis, which is caused by Treponema pallidum invading the brain parenchyma. It is the most common, most prevalent and most serious type of neurosyphilis, and its disability rate is extremely high. PD has nerve paralysis, progressive dementia, personality changes, mental abnormalities and other symptoms as the first symptoms. However, due to the complex and changeable clinical manifestations of PD, and the lack of clarity on the sensitivity and specificity of laboratory tests, the recognition rate of early PD is low, and it is difficult to distinguish it from Alzheimer's disease, Lewy body dementia, frontotemporal dementia, vascular dementia, chronic alcoholism, Parkinson's disease, various mental illnesses, depression, autoimmune encephalitis, and syphilis, especially other types of neurosyphilis, which makes it difficult to make a diagnosis, delays the best treatment time, and leads to worsening of the disease and even life-threatening.

[0007] Due to the complex and changeable clinical manifestations of PD and the atypical early symptoms, the recognition rate of PD is low in the early stages, which makes it difficult to make a good diagnosis of PD and delays the timing of early treatment.

[0008] Currently, there is no method to distinguish PD from other types of syphilis, especially neurosyphilis, so accurate detection of PD cannot be achieved, and timely targeted treatment cannot be achieved. Summary of the invention

[0009] In a first aspect, the present invention provides a product for detecting PD, which comprises a first detection reagent for detecting the cerebrospinal fluid concentration of a soluble factor, and the detection of PD is achieved based on the cerebrospinal fluid concentration of the soluble factor, wherein the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

[0010] The second aspect of the present invention provides the use of soluble factors, NF-L (neurofilament light chain) and / or neopterin in the preparation of the product described in the first aspect of the present invention, wherein the soluble factors are selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

[0011] In a third aspect, the present invention provides an evaluation system for determining the risk of a patient suffering from PD, the evaluation system comprising a detection module and a determination module; the detection module comprises at least one detection module selected from a first detection module, a second detection module, a third detection module and a fourth detection module, the first detection module being a factor detection module for detecting the cerebrospinal fluid concentration of a soluble factor, the second detection module being an NF-L detection module for detecting the cerebrospinal fluid concentration of NF-L, the third detection module being a neopterin detection module for detecting the cerebrospinal fluid concentration of neopterin, the fourth detection module being a protein detection module for detecting the cerebrospinal fluid concentration of a protein; the evaluation module The block includes a readable carrier storing an evaluation rule, wherein the evaluation rule is: when the detected cerebrospinal fluid concentration is higher than the corresponding cutoff value, the syphilis patient is evaluated as having a high risk of suffering from PD; otherwise, the syphilis patient is evaluated as having a low risk of suffering from PD; the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

[0012] Compared with the prior art, the present invention has the following beneficial technical effects: the present invention can realize accurate and timely detection of PD, especially can distinguish PD from other types of syphilis, thereby realizing early identification of PD, and then can carry out targeted diagnosis and treatment of PD as soon as possible. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The differential expression of soluble factors LIF, IL-10, Eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18, and IL-17A between the PD patient group and the non-PD patient group is shown. Among them, the number of patients in the PD patient group was 32, and the number of patients in the non-PD patient group was 80. Each box plot includes the lower quartile (lower horizontal line), median (middle horizontal line), and upper quartile (upper horizontal line). Student's T-Test was used to compare the normally distributed data between the two groups, and the Mann-Whitney Utest test was used to compare the non-normally distributed data. p value ≤ 0.05 was considered statistically significant.

[0014] Figure 2The differential expression of soluble factors TNF-β, IFN-α, IL-1β, IL-22, MIP-1β, MCP-1, IL-23, IL-27, IL-5, IL-15, IL-13, IL-9, IL-1α, IL-21, and IL-31 between the PD patient group and the non-PD patient group is shown. Among them, the number of patients in the PD patient group was 32, and the number of patients in the non-PD patient group was 80. Each box plot includes the lower quartile (lower horizontal line), the median (middle horizontal line), and the upper quartile (upper horizontal line). Student's T-Test was used to compare the normally distributed data between the two groups, and the Mann-Whitney U test was used to compare the non-normally distributed data. p value ≤ 0.05 was considered statistically significant.

[0015] Figure 3 The differential expression of neopterin and neurofilament light chain (NF-L) between the PD patient group and the non-PD patient group is shown. The number of patients in the PD patient group was 32, and the number of patients in the non-PD patient group was 80. Each box plot includes the lower quartile (lower horizontal line), median (middle horizontal line), and upper quartile (upper horizontal line). Student's T-Test was used to compare the normally distributed data between the two groups, and Mann-Whitney U test was used to compare the non-normally distributed data. p value ≤ 0.05 was considered statistically significant.

[0016] Figure 4 The differential expression of clinical parameters, cerebrospinal fluid leukocyte count and cerebrospinal fluid protein level, between the PD patient group and the non-PD patient group is shown. The number of patients in the PD patient group was 32, and the number of patients in the non-PD patient group was 80. Each box plot included the lower quartile (lower horizontal line), median (middle horizontal line), and upper quartile (upper horizontal line). Student's T-Test was used to compare the normally distributed data between the two groups, and Mann-Whitney U test was used to compare the non-normally distributed data. p values ​​≤ 0.05 were considered statistically significant. DETAILED DESCRIPTION

[0017] definition

[0018] In the present invention, the term "cerebrospinal fluid concentration" refers to the concentration of the substances mentioned (eg, soluble factors, NF-L, neopterin, proteins, etc.) in a cerebrospinal fluid sample.

[0019] In the present invention, the term "two or more" or "multiple" refers to more than one situation (i.e. more than one). Wherein "more than" includes the number, for example, "two or more" includes two situations and more than two situations.

[0020] In the present invention, the terms "first", "second", "third" or "fourth" do not mean order or importance, but are only for the convenience of describing the multiple parallel objects mentioned. For example, the second detection reagent refers to a detection reagent for detecting the cerebrospinal fluid concentration of NF-L, and the third detection reagent refers to a detection reagent for detecting the cerebrospinal fluid neopterin concentration. It does not mean that the NF-L concentration measurement needs to be performed before the neopterin concentration measurement, nor does it mean that the cerebrospinal fluid NF-L concentration measurement is more important than the cerebrospinal fluid neopterin concentration measurement. It is only for the convenience of describing the detection reagent for detecting the cerebrospinal fluid concentration of NF-L and the detection reagent for detecting the cerebrospinal fluid neopterin concentration and preventing confusion.

[0021] In the present invention, the abbreviation "NF-L" means neurofilament light chain.

[0022] In the present invention, the abbreviation "CSF" means cerebrospinal fluid.

[0023] In the present invention, the names of soluble factors such as IL-7, TNF-α and IFN-γ have the meanings commonly understood by those skilled in the art. For example, IL-7 means interleukin-7.

[0024] The present invention will be further explained below by describing some specific implementation methods, but these specific implementation methods are only partial technical solutions of the present invention, and the protection scope of the present invention is not limited to these mentioned specific implementation methods.

[0025] As described above, the present invention provides a product for detecting PD in the first aspect, the product includes a first detection reagent for detecting the cerebrospinal fluid concentration of a soluble factor, and the detection of PD is achieved based on the cerebrospinal fluid concentration of the soluble factor, wherein the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A. The inventors found that the area under the curve (AUC) of the operating characteristic curve (ROC) depicted according to the cerebrospinal fluid concentration of these soluble factors is greater than 0.7 and the corresponding p-value is less than 0.001, which can be used to distinguish PD patients from other types of syphilis patients. It is generally believed that if the AUC of a factor is greater than 0.7 and the p-value is less than 0.05, it can be considered to have acceptable sensitivity and specificity, and thus has diagnostic value. The inventors found through experiments that the p-values ​​of the above factors are not only less than 0.05, but also less than 0.001, thus having significant specificity.

[0026] In some preferred embodiments, the soluble factors are selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α and RANTES. The inventors found that the area under the curve (AUC) of the operating characteristic curve (ROC) drawn according to the cerebrospinal fluid concentration of these soluble factors is greater than 0.75 and the corresponding p value is less than 0.001, which is suitable for distinguishing PD patients from other types of syphilis patients.

[0027] In some more preferred embodiments, the soluble factors are selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, and IFN-γ. The inventors found that the area under the curve (AUC) of the operating characteristic curve (ROC) drawn according to the cerebrospinal fluid concentration of these soluble factors is greater than 0.80 and the corresponding p-value is less than 0.05, which is particularly suitable for distinguishing PD patients from other types of syphilis patients.

[0028] In some more preferred embodiments, the soluble factors are selected from one or both of LIF and IL-10. The inventors found that the area under the curve (AUC) of the operating characteristic curve (ROC) drawn according to the cerebrospinal fluid concentration of these soluble factors is greater than 0.85 and the corresponding p value is less than 0.001. Compared with the other soluble factors mentioned above, only from the sensitivity and specificity of the detection, it is more suitable for distinguishing PD patients from other types of syphilis patients.

[0029] In some preferred embodiments, the product further comprises one or more selected from a second detection reagent, a third detection reagent and a fourth detection reagent, wherein the second detection reagent is a detection reagent for detecting NF-L in cerebrospinal fluid, the third detection reagent is a detection reagent for detecting neopterin in cerebrospinal fluid, and the fourth detection reagent is a detection reagent for detecting protein in cerebrospinal fluid. The inventors found that the area under the curve (AUC) of the operating characteristic curve (ROC) drawn according to these NF-L cerebrospinal fluid concentrations, neopterin cerebrospinal fluid concentrations and protein cerebrospinal fluid concentrations is greater than 0.7 and the corresponding p values ​​are less than 0.001, so it can be used in combination with the above-mentioned soluble factors for the detection of PD, especially for distinguishing PD patients from other types of syphilis patients.

[0030] For example, the product may include the first detection reagent and the second detection reagent, thereby detecting PD by jointly determining the cerebrospinal fluid concentration of the soluble factor and NF-L. Alternatively, the product may include the first detection reagent and the third detection reagent, thereby detecting PD by jointly determining the cerebrospinal fluid concentration of the soluble factor and neopterin. Alternatively, the product may include the first detection reagent and the fourth detection reagent, thereby detecting PD by jointly determining the cerebrospinal fluid concentration of the soluble factor and the concentration of cerebrospinal fluid protein. Alternatively, the product may include the first detection reagent, the second detection reagent, and the third detection reagent, thereby detecting PD by jointly determining the cerebrospinal fluid concentration of the soluble factor, NF-L, and neopterin. Alternatively, the product may include the first detection reagent, the second detection reagent, the third detection reagent, and the fourth detection reagent, thereby detecting PD by jointly determining the cerebrospinal fluid concentration of the soluble factor, NF-L, neopterin, and cerebrospinal fluid protein.

[0031] In addition, given that the areas under the curves (AUCs) of the operating characteristic curves (ROC) drawn based on the cerebrospinal fluid concentration of NF-L, neopterin and protein were all greater than 0.7 and the corresponding p values ​​were all less than 0.001, NF-L, neopterin and cerebrospinal fluid protein can also be used independently to detect PD, especially to distinguish PD patients from patients with other types of syphilis.

[0032] Therefore, the present invention provides a product for detecting PD in some aspects, which comprises a second detection reagent for detecting the cerebrospinal fluid concentration of NF-L, and detects PD based on the cerebrospinal fluid concentration of NF-L.

[0033] In other aspects, the present invention provides a product for detecting PD, which includes a third detection reagent for detecting the cerebrospinal fluid concentration of neopterin, and detects PD based on the cerebrospinal fluid concentration of neopterin.

[0034] In other aspects, the present invention provides a product for detecting PD, which includes a fourth detection reagent for detecting the concentration of cerebrospinal fluid protein, and detects PD based on the concentration of cerebrospinal fluid protein.

[0035] In yet other aspects, the present invention provides a product for detecting PD, which comprises two or all three of the second detection reagent, the third detection reagent and the fourth detection reagent, and detects PD based on the cerebrospinal fluid concentration of at least two of the cerebrospinal fluid NF-L, cerebrospinal fluid neopterin and cerebrospinal fluid protein corresponding to the selected detection reagent combination.

[0036] For example, the product may include the second detection reagent and the third detection reagent, and the product realizes the detection of PD based on the cerebrospinal fluid concentration of NF-L and neopterin. Alternatively, the product may include the second detection reagent and the fourth detection reagent, and the product realizes the detection of PD based on the cerebrospinal fluid concentration of NF-L and cerebrospinal fluid protein. Alternatively, the product may include the third detection reagent and the fourth detection reagent, and the product realizes the detection of PD based on the cerebrospinal fluid concentration of neopterin and cerebrospinal fluid protein. Alternatively, the product includes the second detection reagent, the third detection reagent and the fourth detection reagent, and realizes the detection of PD based on the concentration of cerebrospinal fluid NF-L, cerebrospinal fluid neopterin and cerebrospinal fluid protein.

[0037] The present invention has no particular limitation on the form of the product, as long as it can detect PD based on the cerebrospinal fluid concentration of the soluble factor, especially can distinguish PD (PD) from other types of syphilis such as latent syphilis (LS), asymptomatic neurosyphilis (ANS), ocular syphilis (OS), early neurosyphilis such as meningeal neurosyphilis (MNS) and meningovascular neurosyphilis (MVNS).

[0038] In some embodiments, the product is any one of a detection composition, a detection kit, a detection chip, and a detection array.

[0039] In some embodiments, the first detection reagent, the second detection reagent, the third detection reagent and the fourth detection reagent are independently antibodies or antibody pairs. Preferably, the antibodies are monoclonal antibodies. Also preferably, the antibody pair is a combination of a coating antibody and a detection antibody.

[0040] In a second aspect, the present invention provides the use of soluble factors, NF-L and / or neopterin in the preparation of the products described in the above aspects of the present invention, wherein the soluble factors are selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

[0041] In some preferred embodiments, the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α and RANTES.

[0042] In some more preferred embodiments, the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ. In some further preferred embodiments, the soluble factor is selected from one or both of LIF and IL-10.

[0043] In a third aspect, the present invention provides an evaluation system for determining a patient's risk of suffering from PD, the evaluation system comprising a detection module and a determination module; the detection module comprises at least one detection module selected from a first detection module, a second detection module, a third detection module and a fourth detection module, the first detection module being a factor detection module for detecting the cerebrospinal fluid concentration of a soluble factor, the second detection module being a factor detection module for detecting the cerebrospinal fluid concentration of NF-L, the third detection module being a factor detection module for detecting the cerebrospinal fluid concentration of neopterin, and the fourth detection module being a factor detection module for detecting the cerebrospinal fluid concentration of a protein; the evaluation module It includes a readable carrier storing an evaluation rule, wherein the evaluation rule is: when the detected cerebrospinal fluid concentration is higher than the corresponding cutoff value, the syphilis patient is evaluated as having a high risk of suffering from PD; otherwise, the syphilis patient is evaluated as having a low risk of suffering from PD; the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

[0044] In some preferred embodiments, the detection module includes the first detection module.

[0045] In some preferred embodiments, the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α and RANTES. More preferably, the soluble factor is selected from one or more of LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ. Further preferably, the soluble factor is selected from one or both of LIF and IL-10.

[0046] Example 1

[0047] The present invention will be illustrated below by examples, but the protection scope of the present invention is not limited to these examples.

[0048] 1. Subject profile

[0049] The subjects mentioned in the embodiments of the present invention are all patients recruited by the inventors in the past few years at Beijing Ditan Hospital affiliated to Capital Medical University, a total of 112. It was confirmed by diagnosis that none of these patients had AIDS, and the diagnosis of syphilis was confirmed by the syphilis-specific antibody test TPPA test. The inventors grouped these patients into 32 PD patients and 80 other types of syphilis patients (including 29 latent syphilis (LS) patients, 30 asymptomatic neurosyphilis (ANS) patients and 21 non-PD symptomatic neurosyphilis patients according to the 2018 Health Industry Standards of the People's Republic of China.

[0050] Among them, the criteria for latent syphilis, asymptomatic neurosyphilis, symptomatic neurosyphilis and paralytic dementia are as follows:

[0051] A. Criteria for latent syphilis (LS): (1) history of syphilis infection; (2) positive serum non-treponemal test; (3) positive serum treponemal test; (4) no symptoms or signs of neurological damage; (5) no abnormalities in CSF.

[0052] B. Criteria for asymptomatic neurosyphilis (ANS): (1) history of syphilis infection; (2) positive serum Treponema pallidum test; (3) no symptoms and signs of nervous system damage; (4) positive CSF VDRL (TRUST) test; or negative CSF VDRL (TRUST) test and positive CSF Treponema pallidum test or CSF white blood cell count>5 / μl or CSF protein>45mg / dl.

[0053] C. Criteria for determining symptomatic neurosyphilis (SNS): (1) history of syphilis infection; (2) positive serum Treponema pallidum test; (3) symptoms and signs of nervous system damage; (4) positive CSF VDRL (TRUST) test; or negative CSF VDRL (TRUST) test with positive CSF Treponema pallidum test or CSF white blood cell count >5 / μl or CSF protein >45mg / dl.

[0054] D. Criteria for diagnosis of paralytic dementia (PD)

[0055] The patient meets the criteria for symptomatic neurosyphilis (SNS) in C and has clinical manifestations of mental and behavioral abnormalities, including inattention, forgetfulness, impaired judgment and memory, cognitive impairment, dementia, mood changes, depression, personality changes, delusions, mania or mental confusion. The patient may also have pupil abnormalities, dysarthria, hypotonia of the face and limbs, involuntary movements of the face, tongue and hands, epileptic seizures, stroke symptoms and nutritional disorders.

[0056] The examples of the present invention were approved by the Ethics Committee of Beijing Ditan Hospital Affiliated to Capital Medical University, and all participants provided signed informed consent.

[0057] 2. CSF Sample Collection

[0058] CSF samples were collected as part of routine diagnostic workup. After lumbar puncture, 2 mL CSF samples were collected. The supernatant of CSF was extracted by centrifugation at 1500 rpm for 10 minutes and stored at -80°C until analysis.

[0059] 3. Determination of CSF concentrations of neopterin and neurofilament light chain (NF-L)

[0060] The concentrations of CSF neopterin and NF-L were measured using commercial ELISA kits (IBL, Hamburg, Germany) according to the manufacturer's instructions. Optical density was measured at 450 nm using a microplate reader (Spectra Max M2, England). CSF neopterin levels were expressed in nmol / L and NF-L levels were expressed in pg / mL.

[0061] 4. CSF soluble factor concentration determination

[0062] In order to compare the concentrations of soluble factors, the present inventors used the Luminex xMAP method (ProcartaPlex Human Cytokine / Chemokine / Growth Factor Panel 1 (45plex) 96 tests, AffymetrixeBioscience Ltd., Hatfield, United Kingdom) to detect 45 soluble factors in the CSF of the two groups of patients according to the manufacturer's instructions. These soluble factors include:

[0063] (1) 29 cytokines: IL-1RA, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8 (or CXCL8), IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17A, IL-18, IL-21, IL-22, IL-23, IL-27, IL-31, TNF-α, TNF-β, IFN-α, IFN-γ, brain-derived neurotrophic factor (BDNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), stem cell factor (SCF), and leukemia inhibitory factor (LIF);

[0064] (2) 8 chemokines: growth-regulated oncogene α (GRO-α, also known as CXCL1), MCP-1 (also known as CCL2), macrophage inflammatory protein 1α (MIP-1α, also known as CCL3), MIP-1β (also known as CCL4), regulatory factor expressed and secreted on activated normal T cells (RANTES, also known as CCL5), IFN-γ-induced protein 10 (IP-10, also known as CXCL10), eotaxin (also known as CCL11), and stromal cell-derived factor 1α (SDF-1α, also known as CXCL12);

[0065] (3) 8 growth factors: epidermal growth factor (EGF), fibroblast growth factor (FGF-2), hepatocyte growth factor (HGF), β-nerve growth factor (β-NGF), platelet-derived growth factor BB (PDGF-BB), placental growth factor (PLGF), vascular endothelial growth factor A (VEGF-A) and vascular endothelial growth factor (VEGF-D).

[0066] 5. Data Statistical Analysis

[0067] Statistical analyses of all data were performed using the R program (https: / / cran.r-project.org / ), SPSS (IBM Corporation, New York, NY, USA), or GraphPad Prism 7 (GraphPad Software, La Jolla, CA, USA). Data were presented as mean ± SD for normally distributed data and as median (interquartile range) for non-normally distributed data. Student's T-Test was used to compare normally distributed data between two groups, and Mann-Whitney U test was used to compare non-normally distributed data. A p value ≤ 0.05 was considered statistically significant.

[0068] 6. Data processing

[0069] (1) Detection of intergroup differences

[0070] GraphPad Prism 7 software was used to perform statistical analysis of the differences in different parameters between the two groups. For normally distributed data, the data were expressed as mean ± standard deviation, while for non-normally distributed data, the data were expressed as median (interquartile range). Student's T-Test was used to compare the normally distributed data between the two groups, and Mann-Whitney Utest test was used to compare the non-normally distributed data. p value ≤ 0.05 was considered statistically significant.

[0071] (2) ROC analysis

[0072] SPSS software was used to perform ROC analysis to evaluate the diagnostic accuracy of each parameter. AUC was used to evaluate the discriminatory ability of each parameter.

[0073] 7. Results Analysis

[0074] Figure 1 The expression of soluble factors LIF, IL-10, Eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18, and IL-17A in the PD patient group and the non-PD patient group was compared. Among them, the number of patients in the PD patient group was 32; the number of patients in the non-PD patient group was 80, including 8 MNS patients, 4 MVNS patients, 29 LS patients, 30 ANS patients, and 9 OS patients. Figure 1 It can be seen that the expressions of these detected soluble factors were significantly different between the PD patient group and the non-PD patient group.

[0075] Figure 2 The expression of soluble factors TNF-β, IFN-α, IL-1β, IL-22, MIP-1β, MCP-1, IL-23, IL-27, IL-5, IL-15, IL-13, IL-9, IL-1α, IL-21, and IL-31 in the PD patient group and the non-PD patient group was compared. Among them, the number of patients in the PD patient group was 32; the number of patients in the non-PD patient group was 80, including 8 MNS patients, 4 MVNS patients, 29 LS patients, 30 ANS patients, and 9 OS patients. Figure 2 It can be seen that the expression of some soluble factors (IFN-α, IL-1β, IL-22, MIP-1β, MCP-1, IL-23, IL-27, IL-5) between the PD patient group and the non-PD patient group was significantly different, while the expression of other soluble factors (TNF-β, IL-15, IL-13, IL-9, IL-1α, IL-21, IL-31) between the PD patient group and the non-PD patient group was not significantly different.

[0076] Figure 3 The expression of neopterin and NF-L in PD patients and non-PD patients was compared. There were 32 patients in the PD group and 80 patients in the non-PD group, including 8 MNS patients, 4 MVNS patients, 29 LS patients, 30 ANS patients and 9 OS patients. Figure 3 It can be seen that there are significant differences in the expression of neopterin and neurofilament light chain between the PD patient group and the non-PD patient group.

[0077] Figure 4 The expression comparison of clinical parameters cerebrospinal fluid white blood cell count (CSF wbc) and cerebrospinal fluid protein (CSF protein) levels in PD patient group and non-PD patient group was shown. Among them, the number of patients in PD patient group was 32; the number of patients in non-PD patient group was 80, including 8 MNS patients, 4 MVNS patients, 29 LS patients, 30 ANS patients and 9 OS patients. Figure 4 It can be seen that the expression of clinical parameters CSF leukocyte count and CSF protein level in the PD patient group and the non-PD patient group was significantly different.

[0078] In order to confirm whether the soluble factors and clinical parameters of neopterin, NF-L, clinical parameters cerebrospinal fluid white blood cell count (CSF wbc) and cerebrospinal fluid protein (CSF protein) levels and the expression of these soluble factors detected in the PD patient group and the non-PD patient group have significant differences in can be used for the detection of PD patients, for example, whether they can be used to distinguish PD patients from non-PD patients, the present inventors also performed ROC analysis based on the cerebrospinal fluid concentrations of neopterin, NF-L and these soluble factors and the clinical parameters, and the results are shown in Table 1 below.

[0079] Table 1. Receiver operating characteristic curve analysis data of CSF concentrations of some soluble factors, NF-L and neopterin, and CSF clinical parameters (wbc, protein).

[0080]

[0081]

[0082] As can be seen from Table 1, soluble factors selected from LIF, IL-10, eotaxin, GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A, NF-L and neopterin can be used to detect PD, and are particularly suitable for distinguishing PD from other types of neurosyphilis, while other soluble factors detected cannot. In addition, the clinical parameter cerebrospinal fluid protein level can be used to detect PD, while the cerebrospinal fluid white blood cell count cannot distinguish PD from other types of neurosyphilis.

[0083] The inventors further randomly selected 78 patient sample measurement data from the above 112 patient sample measurement data as the test group, and the remaining 34 patient sample data as the verification group (the ratio of the number of samples in the test group and the verification group is 7:3). The Cut-off values ​​of the corresponding indicators (the first three factors in Table 1 and CSF protein and CSFwbc) were calculated using the 78 sample measurement data of the test group, and then the Cut-off values ​​were used to judge the remaining 34 patient sample measurement data (the random grouping operation was repeated 5 times), and the sensitivity, specificity and accuracy results are shown in Table 2 below.

[0084] Table 2. Sensitivity, specificity, and accuracy of partial factorial group validation.

[0085]

[0086] As can be seen from the data in Table 2, the group verification results are consistent with those in Table 1, with high sensitivity, specificity and accuracy, proving that soluble factors such as LIF, IL-10 and eotaxin can be used to detect PD, and are particularly suitable for distinguishing PD from other types of neurosyphilis, while cerebrospinal fluid leukocyte count cannot distinguish PD from other types of neurosyphilis, especially the specificity is obviously too low. Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some of the technical features therein by equivalents; and these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of soluble factors, NF-L and / or neopterin in the preparation of a product for detecting paralytic dementia, wherein the soluble factors are selected from one or more of the soluble factor group comprising LIF, IL-10 and eotaxin.

2. The use according to claim 1, characterized in that: The soluble factor group further comprises one or more of GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

3. The use according to claim 1, characterized in that: In addition to the first detection reagent for detecting the cerebrospinal fluid concentration of the soluble factor, the product also includes one or more selected from a second detection reagent, a third detection reagent and a fourth detection reagent, wherein the second detection reagent is a detection reagent for detecting cerebrospinal fluid NF-L, the third detection reagent is a detection reagent for detecting cerebrospinal fluid neopterin, and the fourth detection reagent is a detection reagent for detecting cerebrospinal fluid protein.

4. The use according to claim 1, characterized in that: The product is any one of a detection composition, a detection kit, a detection chip and a detection array.

5. The use according to claim 4, characterized in that: The first detection reagent, the second detection reagent, the third detection reagent and the fourth detection reagent are independently antibodies or antibody pairs.

6. The use according to claim 5, characterized in that: The antibody is a monoclonal antibody, and the antibody pair is a combination of a coating antibody and a detection antibody.

7. An assessment system for determining a patient's risk of suffering from paralytic dementia, characterized in that: The evaluation system includes a detection module and a judgment module; The detection module includes at least one detection module selected from a first detection module, a second detection module, a third detection module and a fourth detection module, wherein the first detection module is a factor detection module for detecting the cerebrospinal fluid concentration of a soluble factor, the second detection module is a NF-L detection module for detecting the cerebrospinal fluid concentration of NF-L, the third detection module is a neopterin detection module for detecting the cerebrospinal fluid concentration of neopterin, and the fourth detection module is a protein detection module for detecting the cerebrospinal fluid concentration of a protein; The evaluation module includes a readable carrier storing an evaluation rule, wherein the evaluation rule is: when the detected cerebrospinal fluid concentration is higher than the corresponding cutoff value, the patient is evaluated as having a high risk of suffering from paralytic dementia; otherwise, the patient is evaluated as having a low risk of suffering from paralytic dementia; The soluble factors are selected from one or more of the soluble factor group consisting of LIF, IL-10 and eotaxin.

8. The evaluation system according to claim 7, characterized in that: The detection module at least includes the first detection module.

9. The evaluation system according to claim 7 or 8, characterized in that: The soluble factor group also includes one or more of GRO-α, IP-10, SDF-1α, IL-6, IL-8, MIP-1α, IFN-γ, IL-12p70, IL-7, IL-4, IL-2, TNF-α, RANTES, IL-1RA, IL-18 and IL-17A.

Images