CN116949168A - Application of CD73 in the diagnosis of Parkinson's disease - Google Patents

Abstract

The present invention discloses the application of CD73 in the diagnosis of Parkinson's disease. By studying the levels of CD73 in the serum of Parkinson's disease patients and healthy controls, the present invention finds that the content of CD73 in the serum of Parkinson's disease patients is significantly lower than that of healthy controls, and PD The patient's serum CD73 level is negatively correlated with H&Y stage and UPDRS-III score, which provides a new direction for the effective diagnosis of Parkinson's disease patients and has broad clinical application prospects.

Description

Application of CD73 in the diagnosis of Parkinson's disease

Technical field

The invention belongs to the field of biomedicine, and specifically relates to the application of CD73 in the diagnosis of Parkinson's disease.

Background technique

Parkinson's disease (PD) is the most common neurodegenerative disease causing movement disorders in the elderly. Its typical clinical core motor symptoms are tremor, rigidity, hypokinesia and bradykinesia, and unstable posture and gait. As the disease progresses, the motor symptoms of PD progressively worsen, becoming the main reason for the decline in the quality of life of PD patients and the increase in life dependence. Therefore, screening markers related to Parkinson's disease is of great significance for the diagnosis and subsequent treatment of Parkinson's disease.

Contents of the invention

In order to make up for the shortcomings of the existing technology, the present invention provides the application of CD73 in the diagnosis of Parkinson's disease.

In order to achieve the above objects, the present invention adopts the following technical solutions:

A first aspect of the present invention provides the use of a reagent for detecting CD73 expression levels in samples in preparing products for diagnosing and staging Parkinson's disease.

Further, the reagent is selected from a probe that specifically recognizes the CD73 gene, a primer that specifically amplifies the CD73 gene, or a binding agent that specifically binds to the protein encoded by the CD73 gene.

Further, the primer or probe is labeled with a labeling substance.

Further, the labeling substance includes fluorescent substance, radioactive isotope or enzyme.

Further, the products include chips, test strips, test kits or nucleic acid film strips.

Further, the chip includes a gene chip and a protein chip.

Further, the gene chip includes a probe targeting the CD73 gene for detecting the CD73 gene transcription level, and the protein chip includes a specific binding agent for the CD73 protein.

Further, the kits include gene detection kits and protein detection kits.

Further, the gene detection kit includes a reagent or chip for detecting the CD73 gene transcription level, and the protein detection kit includes a reagent or chip for detecting the CD73 protein expression level.

Furthermore, the kit also includes instructions.

Further, the kit also includes a buffer.

Furthermore, the kit also includes reagents for detecting CD73 gene or protein expression levels through RT-PCR, qRT-PCR, biochip detection, Southern blotting, in situ hybridization, and immunoblotting.

Further, the samples include serum, tissue, and exosomes.

Further, the sample is selected from serum.

Furthermore, when the level of CD73 in the serum shows a significant decrease, Parkinson's disease is diagnosed.

A second aspect of the present invention provides a product for diagnosing and staging Parkinson's disease, which product includes a reagent for detecting the expression level of CD73 in a sample.

Further, the products include chips, test strips, test kits or nucleic acid film strips.

Further, the samples include serum, tissue, and exosomes.

Further, the sample is selected from serum.

The third aspect of the present invention provides a method for screening candidate drugs for treating Parkinson's disease, the method comprising: treating a culture system expressing or containing the CD73 gene or the protein encoded by it with a substance to be screened; and detecting the system The expression or activity of the CD73 gene or the protein encoded by it; wherein, when the substance to be screened promotes the expression level or activity of the CD73 gene or the protein encoded by it, the substance to be screened is a candidate drug for treating Parkinson's disease.

A fourth aspect of the invention provides the use of CD73 as a target in screening candidate drugs for treating Parkinson's disease.

Further, the method of screening candidate drugs for treating Parkinson's disease includes: treating a culture system that expresses or contains the CD73 gene or its encoded protein with a substance to be screened; and detecting the expression of the CD73 gene or its encoded protein in the system. Or activity; wherein, when the substance to be screened promotes the expression level or activity of the CD73 gene or the protein encoded by it, the substance to be screened is a candidate drug for treating Parkinson's disease.

The fifth aspect of the present invention provides the application of CD73 in constructing a computational model for diagnosis and staging of Parkinson's disease.

The sixth aspect of the present invention provides the application of CD73 in constructing a system/device/computer-readable storage medium for diagnosing and staging Parkinson's disease.

Advantages and beneficial effects of the present invention: By studying the levels of CD73 in the serum of Parkinson's disease patients and healthy controls, the present invention found that the content of CD73 in the serum of Parkinson's disease patients was significantly lower than that of healthy controls, and the serum CD73 level of PD patients was significantly different from that of H&Y There is a negative correlation between stage and UPDRS-Ⅲ score (P<0.01), which provides a new direction for the effective diagnosis of Parkinson's disease patients and has broad clinical application prospects.

Description of the drawings

Figure 1 is a graph of the difference level of serum CD73 in the training set, where 1A is a graph of the difference level of serum CD73, and 1B is a ROC curve graph;

Figure 2 is the ROC curve of serum CD73 in the validation set.

Detailed ways

Definitions of some terms used in this specification are provided below. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The invention provides the application of a reagent for detecting CD73 expression levels in samples in preparing products for diagnosing and staging Parkinson's disease.

In the present invention, CD73 includes wild type, mutant type or fragments thereof. The term encompasses full-length, unprocessed CD73, as well as any form of CD73 derived from processing in the cell, as well as naturally occurring variants of CD73 (e.g., splice variants or allelic variants). The term encompasses, for example, human CD73 as well as CD73 from any other vertebrate source, including CD73 from mammals, such as primates and rodents (eg, mice and rats), Gene ID: 4907.

As used herein, diagnosis, diagnosing, making a diagnosis, and variations of these terms refer to the discovery, discovery, or condition of an individual's health or condition based on one or more signs, symptoms, data, or other information related to the individual. Judgment or cognition. An individual's health status may be diagnosed as healthy/normal (ie, the absence of a disease or disorder), or may be diagnosed as unhealthy/abnormal (ie, the assessment of a disease or disorder or characteristic is present). The above terms diagnose, diagnose, perform diagnosis, etc. include, in relation to a specific disease or disorder, the early detection of a disease; the characteristics or classification of a disease; the discovery of the progression, cure, or recurrence of a disease; the disposition or treatment of an individual, the response to the disease. Reaction discovery.

In the present invention, expression level or level refers to the absolute or relative amount of CD73 in the present invention. The expression level of CD73 in the present invention can be determined by a variety of techniques. In particular, it can be determined by using methods well known to those skilled in the art. The absolute or relative amount of CD73 in the present invention is detected.

The reagent is selected from a probe that specifically recognizes the CD73 gene, a primer that specifically amplifies the CD73 gene, or a binding agent that specifically binds to the protein encoded by the CD73 gene.

In the present invention, the probe that specifically recognizes the CD73 gene can be DNA, RNA, DNA-RNA chimera, PNA or other derivatives. There is no limit to the length of the probe. As long as it completes specific hybridization and specifically binds to the target nucleotide sequence, any length is acceptable. The probes can be as short as 10, 25, 20, 15, 13 or 10 bases in length. Likewise, the length of the probe can be 60, 80, 100, 150, 300 base pairs or more, or even the entire gene. Since different probe lengths have different effects on hybridization efficiency and signal specificity, the length of the probe is usually at least 14 base pairs, and the longest is generally no more than 30 base pairs, which is consistent with the target nucleotide sequence. The optimal complementary length is 15-25 base pairs. The self-complementary sequence of the probe is preferably less than 4 base pairs, so as not to affect the hybridization efficiency.

In the present invention, a primer refers to a single-stranded polynucleotide capable of hybridizing to a nucleic acid and allowing polymerization of complementary nucleic acid (generally by providing a free 3&apos;-OH group). The primers enable specific amplification of target sequences. Specific amplification refers to the

The primers or probes of the present invention can be chemically synthesized using the phosphorimide solid phase support method or other well-known methods. Modification can also be performed using many means known in the art. Non-limiting examples of these modifications include methylation, capping, substitution with one or more analogs of the natural nucleotide and modifications between nucleotides, e.g., modification of uncharged linkers (e.g., methyl phosphate, phosphotriester, phosphorimide, carbamate, etc.), or modified charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.).

The primer or probe is labeled with a labeling substance.

The labeling substances include, but are not limited to, fluorescent substances, radioactive isotopes or enzymes.

Among them, fluorescent substances include but are not limited to TAMRATTM, Alexa555, Alexa647, Cy3, Cy5 and fluorescein of the cyanine dye series.

Radioactive isotopes include but are not limited to 32P, 33P, and 35S.

Enzymes include, but are not limited to, alkaline phosphatase and horseradish peroxidase.

In the primer or probe according to the present invention labeled with a labeling substance, the labeling substance may be directly bound to the primer or probe, or may be bound via a linker. The linker may be any linker commonly used in this field. Specifically, for example, a nucleic acid of 1 to 3 bases is preferred, a DNA of 1 to 3 bases is more preferred, and a DNA of 2 bases is even more preferred. base DNA, particularly preferably the two bases adenine (A) and adenine (A).

The method of labeling the primers or probes according to the present invention with a fluorescent substance may be carried out according to a well-known method generally performed in this field. Specifically, for example, a method known in the art may be used. A method of incorporating fluorescein-labeled nucleotides into primers or probes.

The method of labeling the primer or probe according to the present invention with a radioactive isotope may be carried out according to a known method generally performed in this field. Specific examples include labeling with a radioactive isotope by incorporation. Methods for labeling nucleotides, etc. Specific examples include the random priming method, the nick translation method, the 5' end labeling method based on T4 polynucleotide kinase, the 3' end labeling method based on terminal deoxynucleotidyl transferase, and the like.

The method of labeling the primer or probe of the present invention with an enzyme may be carried out according to a known method generally performed in this field. Specific examples include alkaline phosphatase, horseradish, etc. Direct labeling methods in which enzyme molecules such as peroxidase are directly covalently bound to the primer or probe to be labeled, etc.

In the present invention, a binding agent refers to a naturally occurring or non-naturally occurring molecule that specifically binds to a target sequence. Examples of specific binding agents include, but are not limited to, proteins, peptides, nucleic acids, carbohydrates, and lipids.

In the present invention, binding agents that specifically bind to the protein encoded by the CD73 gene include protein CD73 receptors, lectins that bind to protein CD73, antibodies against protein CD73, peptide antibodies against protein CD73 (peptidebodies), bispecific dual binding agent or bispecific antibody format. Specific examples of specific binding agents include peptides, peptidomimetics, aptamers, spiegelmers, darpins, ankyrin repeat proteins, Kunitz-type domains, and antibodies.

The products include chips, test strips, test kits or nucleic acid film strips.

In the present invention, a chip is also called an array, which refers to a solid support containing attached nucleic acid or peptide probes. Arrays typically contain a variety of different nucleic acid or peptide probes attached to the substrate surface at different known locations. These arrays, also known as "microarrays," can generally be produced using mechanical synthesis methods or light-guided synthesis methods that incorporate a combination of photolithographic and solid-phase synthesis methods. The array may comprise a flat surface, or may be nucleic acids or peptides on beads, gels, polymeric surfaces, fibers such as optical fibers, glass or any other suitable substrate. The array can be packaged in a manner that allows diagnostics or other manipulation of the fully functional device.

The term "microarray" is an ordered arrangement of hybridization array elements, such as polynucleotide probes (eg, oligonucleotides) or binding agents (eg, antibodies), on a substrate. The substrate may be a solid substrate such as a glass or silica slide, beads, fiber optic adhesive, or a semi-solid substrate such as a nitrocellulose membrane. The nucleotide sequence can be DNA, RNA, or any arrangement thereof.

In the present invention, the chip includes a gene chip and a protein chip; the gene chip includes a solid phase carrier and a probe fixed on the solid phase carrier, and the probe includes an oligonucleotide targeting the CD73 gene for detecting the CD73 gene transcription level. The protein chip includes a solid-phase carrier and a specific antibody for the CD73 protein immobilized on the solid-phase carrier; the gene chip can be used to detect multiple genes including the human CD73 gene (for example, related to Parkinson's disease). The expression levels of multiple disease-related genes). The protein chip can be used to detect the expression levels of multiple proteins including human CD73 protein (eg, multiple proteins related to Parkinson's disease). By detecting multiple markers related to Parkinson's disease simultaneously, the accuracy of diagnosing Parkinson's disease can be greatly improved.

In the present invention, the kit may also include a fluorescent dye, and a variety of known fluorescent dyes may be used. Examples include a method using an intercalator having a labeling function, a method using a probe that binds a fluorescent substance to a nucleotide that hybridizes specifically to an amplified DNA sequence, and the like. Examples of the intercalating agent include ethidium bromide and SYBR GreenI which are unsaturated fluorescent dyes, and Resolight (manufactured by Roche) and EvaGreen (manufactured by Biotim) which are saturated fluorescent dyes. Preferred intercalating agents are SYBR Green I which is an unsaturated fluorescent dye, and EvaGreen and Resolight which are saturated fluorescent dyes, and more preferably EvaGreen and Resolight which are saturated fluorescent dyes. Regarding the dosage, follow the recommendations of the manufacturer and seller of the fluorescent pigment used.

The kit also includes instructions, which may include instructions on obtaining the sample and processing the sample.

In addition, the kit may include bacterial genomic DNA used as a positive control for PCR and sterile water used as a negative control.

In the present invention, the components of the kit may be packaged in an aqueous medium or in a lyophilized form. Suitable containers in a kit will usually include at least one vial, test tube, flask, PET bottle, syringe or other container into which one of the components can be placed and, preferably, appropriately aliquoted. Where more than one component is present in a kit, the kit will typically also contain a second, third or other additional container in which the additional components are separately located. However, different combinations of components can be contained in one vial. Kits of the present invention will also typically include a container for holding the reagents, sealed for commercial sale. Such containers may include injection molded or blow molded plastic containers in which the desired vials may be retained.

The solid support of the kit can be, for example, plastic, silicon wafer, metal, resin, glass, membrane, particle, precipitate, gel, polymer, flake, sphere, polysaccharide, capillary, film, plate or slide . The biological sample may be, for example, a cell culture, cell line, tissue, oral tissue, gastrointestinal tissue, organ, organelle, biological fluid, serum sample, urine sample or skin.

In the present invention, the nucleic acid membrane strip includes a base and a probe fixed on the base; the base can be any base suitable for fixing the probe, including but not limited to nylon membrane, nitrocellulose membrane, polypropylene membrane , glass sheets, silica gel wafers, miniature magnetic beads.

The chip, test paper, kit or nucleic acid membrane strip described in the present invention can be used to detect the expression levels of multiple genes or proteins including the CD73 gene or protein and their expression products.

In the present invention, sample or sample have the same meaning and can be used interchangeably. Sample or sample means any substance, biological fluid, tissue or cell obtained or otherwise obtained from an individual. It includes blood (including whole blood, white blood cells, peripheral blood mononuclear cells, buffy coat, plasma and serum), tissue, exosomes, sputum, tears, mucus, nasal wash, nasal aspirate, breath samples , urine, semen, saliva, meningeal fluid, amniotic fluid, glandular fluid, lymph fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate, ascites, cells, cell extracts and cerebrospinal fluid. It also includes all experimental fractions described above. For example, a blood sample may be fractionated into serum or a fraction containing specific types of blood cells, such as red blood cells or white blood cells (leukocytes). If desired, the sample may be a combination of samples from the individual, such as a combination of tissue and fluid samples. Samples also include materials containing homogeneous solid material, such as material from stool samples, tissue samples, or biopsies. Samples also include material derived from tissue culture or cell culture. Any suitable method for obtaining a biological sample may be utilized; exemplary methods include, for example, phlebotomy, swabbing (eg, buccal swabbing), and fine needle aspiration biopsy procedures. Samples may also be collected by, for example, microdissection (eg, laser capture microdissection (LCM) or laser microdissection (LMD)), smearing (eg, PAP smear), or catheter lavage. A sample obtained from or derived from an individual includes any such sample that is processed in any suitable manner after being obtained from the individual.

In embodiments of the present invention, samples include serum, tissue, and exosomes.

In a specific embodiment of the invention, said sample is selected from serum.

The present invention provides the application of CD73 in constructing a computational model for diagnosis and staging of Parkinson's disease.

In the present invention, the computational model includes the expression level of CD73. As the skilled artisan will know, the steps of correlating CD73 levels with a certain likelihood or risk can be implemented and accomplished in different ways. Preferably, the measured concentrations of CD73 and one or more other markers are mathematically combined and the combined values are correlated to the underlying diagnostic question. Determinations of marker values may be combined by any suitable prior art mathematical method.

The present invention provides the application of CD73 in constructing a system/device/computer-readable storage medium for diagnosing and staging Parkinson's disease.

In the present invention, implementation of the system may include performing or completing selected tasks manually, automatically, or a combination thereof. Furthermore, actual instruments and devices according to embodiments of the system of the present invention may use an operating system to perform a plurality of selected tasks by hardware, by software, or by firmware, or by a combination thereof.

For example, the hardware used to perform the selected task may be a chip or a circuit. As software, the selected tasks may be implemented as a plurality of software instructions executed by a computer using any suitable operating system. In the present invention, one or more tasks according to exemplary embodiments of methods and/or systems according to the present invention may be performed by a processing unit, such as a computing platform for executing a plurality of instructions. Optionally, the processing unit includes volatile memory for storing instructions and/or data, and/or non-volatile memory for storing instructions and/or data, for example, a magnetic hard disk and/or removable media. . Optionally, network connectivity is also provided. A display and/or user input device such as a keyboard or mouse may also optionally be provided.

In the present invention, computer-readable storage media, such as computer-executable code, may take many forms, including but not limited to tangible storage media, carrier wave media, or physical transmission media. Non-volatile storage media includes, for example, optical or magnetic disks, such as any storage device in any computer, and volatile storage media includes dynamic memory, such as the main memory of such a computer platform. Tangible transmission media include coaxial cable; copper wire and fiber optics, including the wires that make up the buses within computer systems. Carrier transmission media may take the form of electrical or electromagnetic signals, or acoustic or light waves, such as those generated during radio frequency and infrared data communications. Thus, common forms of computer readable media include, for example: floppy disk, floppy disk, hard drive, magnetic tape, any other magnetic media, CD-ROM, DVD or DVD-ROM, any other optical media, punched cardstock tape, tape with hole pattern any other physical storage media, RAM, ROM, PROM and EPROM, FLASH-EPROM, any other memory chip or cartridge, carrier waves that transmit data or instructions, cables or links that transmit such carrier waves, or from which a computer can read Any other medium to retrieve programming code and/or data. Many of these forms of computer-readable media may be involved in conveying one or more sequences of one or more instructions to a processor for execution.

This invention will be further described below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are presented by way of example and are not intended to be limiting of the invention. The principal features of the invention may be employed in various embodiments without departing from the scope of the invention.

Example

1 Materials and methods

1.1 Research objects

The samples of this study included PD patients hospitalized in the Geriatrics Department of the Second Hospital of Hebei Medical University from November 2019 to April 2023. Among them, 33 PD patients were included in the training set as the case group, of which 16 were female (48.5% ), 17 males (51.5%); 0 smokers; 35 healthy examination subjects received by the physical examination center during the same period were also included as healthy controls, including 20 females (57.1%) and 15 males (42.9% ); 0 smokers.

The validation set included 97 PD patients as the case group, including 51 women (52.6%), 46 men (47.4%), and 10 smokers (10.3%); healthy subjects received by the physical examination center during the same period were also included. 71 cases were healthy controls, including 31 women (43.7%), 40 men (56.3%), and 14 smokers (19.7%).

1.2 Inclusion criteria

① Meet the 2016 Chinese Parkinson's Disease Diagnosis and Preliminary Diagnosis Criteria; ② The duration of the disease is 1 year or more; ③ The patient has one or more of the following characteristics that are obvious, including postural instability, resting tremor, muscle stiffness, and non-original symptoms. ④ The clinical data are complete; ⑤ The patient and family members were informed, signed an informed consent form, and was discussed and approved by the ethics committee of our hospital (ethical review resolution number: 2022-R039).

1.3 Exclusion criteria

①Patients with secondary Parkinson's syndrome; ②Patients with Parkinson's superimposed syndrome; ③Those with severe heart, liver, kidney, and gastrointestinal dysfunction; ④Those who provide unclear detailed medical history; ⑤Infection within the past month ⑥Those with immune system diseases; ⑦Those with chronic infectious diseases; ⑧Those who have used anti-inflammatory drugs and immunomodulatory drugs in the past month; ⑨Those with a history of malignant tumors; ⑩Those who have recently suffered trauma or head surgery.

1.4 Research content and methods

The data collected on the included subjects include: ①Basic information: gender, smoking history, and disease course of the patients in the case group. ②Evaluation of the patient's condition in the case group: The unified Parkinson's disease rating scale part III (UPDRS-III) was used to evaluate the patient's motor function (all were performed after the patient temporarily stopped taking medication and was in " "off" state), this scale has 14 items in total, each item is scored from 0 to 4 according to the severity of the corresponding symptom. The lower the score, the better the quality of life; the H&Y staging is used to evaluate the severity of the patient's movement disorder. , according to the grading of patients’ motor symptoms, they were divided into mild PD group (H&Y stages I and II) and moderate-to-severe PD group (H&Y stages III-V). The above evaluations were jointly conducted by two attending neurologists. ③Serum CD73 results: The 5’-NT value obtained from the fasting venous blood drawn from all participants and sent to the Laboratory Department of the Second Hospital of Hebei Medical University for detection by a fully automatic biochemical analyzer.

1.5 Statistical methods

SPSS software version 26.0 was used for statistical analysis. If the measurement data conformed to the normal distribution, the t test was used. otherwise, the median (interquartile range) [M (Q1, Q3)] is used to represent the rank sum test; the count data is represented by the x^2 test and expressed as the number of cases (percentage) [n (%)] ; Binary logistic regression was used to analyze the correlation between each risk factor and PD; the predictive value was analyzed using the receiver operating characteristic curve to obtain the area under the curve (AUC), confidence interval, sensitivity, specificity and cutoff value. ; Pearson method (Pearson) was used to analyze the correlation between serum CD73, H&Y stage and UPDRS-Ⅲ score in the case group. One-way ANOVA was used to analyze the changes in serum CD73 levels in PD patients with different H&Y stages, and P < 0.05 was considered a statistically significant difference.

2Experimental results

2.1 General data analysis of case group and healthy control group

A total of 168 research subjects were included in the validation set of this study. Among them, there were 97 cases in the case group and 71 cases in the healthy control group. The gender distribution of the research subjects in the two groups was balanced (P>0.05), and there was no significant difference in whether they had a history of smoking (P>0.05). In addition, the disease duration of the case group was 2.0 (1.0, 4.0) years, the H&Y stage was 2.0 (2.0, 3.0), and the UPDRS-III score was 18.66±0.70 points. The expression of CD73 in the peripheral blood of PD patients was significantly reduced (P<0.01) (Table 1).

Table 1 General data analysis of validation set case group and healthy control group

Note: a is t test; b is chi-square test; c is rank sum test

2.2 Logistic regression analysis of the correlation between serum CD73 and other risk factors and Parkinson’s disease

After logistic regression analysis, after adjusting for gender and smoking history, the results showed that the lower the serum CD73 level, the greater the risk of PD (OR=0.208, P<0.01) (Table 2).

Table 2 Logistic regression analysis of the correlation between serum CD73 and other risk factors and PD in the validation set

Note: * is the healthy control group

2.3 Differential expression of serum CD73 in PD

In the training set, a ROC curve was drawn using the healthy control group as positive samples and the case group as negative samples. The results showed that the AUC of serum CD73 in predicting PD was 0.844 (Figure 1).

The AUC of serum CD73 in predicting PD in the validation set was 0.845, the best sensitivity was 90.1%, and the specificity was 67.0% (Table 3, Figure 2).

Table 3 Prediction ROC curve of serum CD73 for PD in the validation set

Note: a means the unit is U/L

2.4 General data analysis of mild PD group and moderate to severe PD group

The study subjects in the case group were divided into groups according to the H&Y stage. Stages I and II were the mild PD group (n=50); stages III-V were the moderate and severe PD group (n=47). There was no statistical difference in gender and smoking history between the two groups (P>0.05). Patients in the moderate-to-severe PD group had a longer course of disease, higher UPDRS-III scores, and significantly lower serum CD73 levels than those in the mild PD group (Table 4).

Table 4 General data analysis of the mild PD group and the moderate to severe PD group in the validation set

Note: a is t test; b is chi-square test; c is rank sum test

2.5 Correlation analysis between serum CD73 and H&Y stage and UPDRS-Ⅲ score in patients with PD

Pearson correlation analysis showed that serum CD73 levels in PD patients were negatively correlated with H&Y stage and UPDRS-Ⅲ score (P<0.01) (Table 5).

Table 5 Correlation analysis

The description of the above embodiments is only for understanding the method of the present invention and its core idea. It should be noted that those of ordinary skill in the art can make several improvements and modifications to the present invention without departing from the principles of the present invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims (10)

1. The use of a reagent for detecting the expression level of CD73 in a sample for the preparation of a product for diagnosis and staging of parkinson's disease.

2. The use according to claim 1, wherein the reagent is selected from the group consisting of a probe specifically recognizing the CD73 gene, a primer specifically amplifying the CD73 gene, or a binding agent specifically binding to a protein encoded by the CD73 gene;

preferably, the primer or probe is labeled with a labeling substance;

preferably, the labeling substance includes a fluorescent substance, a radioisotope, or an enzyme.

3. The use according to claim 1, wherein the product comprises a chip, a test strip, a kit or a nucleic acid membrane strip.

4. The use according to claim 3, wherein the chip comprises a gene chip, a protein chip;

preferably, the gene chip comprises a probe for CD73 gene for detecting the transcription level of CD73 gene, and the protein chip comprises a specific binding agent for CD73 protein;

preferably, the kit comprises a gene detection kit and a protein detection kit;

preferably, the gene detection kit comprises a reagent or chip for detecting the transcription level of the CD73 gene, and the protein detection kit comprises a reagent or chip for detecting the expression level of the CD73 protein;

preferably, the kit further comprises instructions;

preferably, the kit further comprises a buffer.

5. The use according to claim 4, wherein the kit further comprises reagents for detecting the expression level of CD73 gene or protein by RT-PCR, qRT-PCR, biochip assay, southern blotting, in situ hybridization, immunoblotting;

preferably, the sample comprises serum, tissue, exosomes;

preferably, the sample is selected from serum.

6. A product for diagnosis and staging of parkinson's disease, said product comprising an agent for detecting the level of CD73 expression in a sample;

preferably, the product comprises a chip, a test paper, a kit or a nucleic acid membrane strip;

preferably, the sample comprises serum, tissue, exosomes;

preferably, the sample is selected from serum.

7. A method of screening for a candidate agent for treating parkinson's disease, said method comprising: treating a culture system expressing or containing the CD73 gene or a protein encoded by the CD73 gene with a substance to be screened; and detecting the expression or activity of the CD73 gene or a protein encoded thereby in the system; wherein the substance to be screened is a candidate drug for treating Parkinson's disease when the substance to be screened promotes the expression level or activity of the CD73 gene or a protein encoded by the same.

Use of cd73 as a target in screening candidate drugs for the treatment of parkinson's disease;

preferably, the method for screening candidate drugs for treating parkinson's disease comprises: treating a culture system expressing or containing the CD73 gene or a protein encoded by the CD73 gene with a substance to be screened; and detecting the expression or activity of the CD73 gene or a protein encoded thereby in the system; wherein the substance to be screened is a candidate drug for treating Parkinson's disease when the substance to be screened promotes the expression level or activity of the CD73 gene or a protein encoded by the same.

Application of CD73 in constructing a calculation model for diagnosis and stage of Parkinson's disease.

Use of cd73 in constructing a system/device/computer readable storage medium for diagnosis and staging of parkinson's disease.