阅读说明：本技术 基于血液滤过方式清除体外循环突触核蛋白的方法 (Method for clearing extracorporeal circulation synuclein based on hemofiltration mode ) 是由 莫明树 郭文苑 陈国云 陈祥 卢林 毛珩旭 罗阳赋 徐评议 于 2021-06-24 设计创作，主要内容包括：本发明公开了基于血液滤过方式清除体外循环突触核蛋白的方法,属于医学领域。具体公开：获取血液中突触核蛋白二聚体的比例S；根据大量的临床试验数据或者文献历史数据,获取血液滤过的血流速度和交换膜面积；根据上述确定的血流速度和交换膜面积进行血液滤过,采用线性回归法构建不同突触核蛋白二聚体的比例S与血液滤过透析膜孔径数值D的计算模型；利用上述计算模型设置血液滤过参数,即能预期体外循环突触核蛋白清除率。本发明首次发现了血液滤过有助于降低体外循环α-突触核蛋白水平,因此开发一种基于α-突触核蛋白二聚体比例构建设置血液滤过参数的计算模型,以为临床医学上缓解α-突触核蛋白相关毒性症状提供了科学数据和新方案。(The invention discloses a method for clearing extracorporeal circulation synuclein based on a hemofiltration mode, belonging to the field of medicine. Specifically disclosed is: obtaining the proportion S of synuclein dimers in blood; obtaining the blood flow velocity of hemofiltration and the area of an exchange membrane according to a large amount of clinical test data or literature historical data; performing hemofiltration according to the determined blood flow velocity and the area of the exchange membrane, and constructing a calculation model of the ratio S of different synuclein dimers and the aperture numerical value D of the hemofiltration dialysis membrane by adopting a linear regression method; the extracorporeal circulation synuclein clearance can be expected by setting the blood filtration parameters by using the calculation model. The invention discovers for the first time that hemofiltration is helpful for reducing the level of alpha-synuclein in extracorporeal circulation, and therefore develops a calculation model for setting hemofiltration parameters based on the alpha-synuclein dimer proportion, thereby providing scientific data and a new scheme for relieving alpha-synuclein related toxicity symptoms in clinical medicine.)

1. A method for constructing an extracorporeal circulation synuclein clearance calculation model based on a hemofiltration mode is characterized by comprising the following steps:

obtaining the proportion S of synuclein dimers in blood;

obtaining the blood flow velocity of hemofiltration and the area of an exchange membrane according to a large amount of clinical test data or literature historical data;

performing hemofiltration according to the determined blood flow velocity and the area of the exchange membrane, analyzing the correlation between the proportion S of different synuclein dimers and the aperture value D of the hemofiltration dialysis membrane by adopting a linear regression method, and constructing a calculation model between the S and the D;

and (3) performing hemofiltration by adopting the proportion S of the synuclein dimer and the dialysis membrane aperture determined by the calculation model, namely expecting the clearance rate of the extracorporeal circulation synuclein.

2. The method of claim 1, wherein the computational model is formulated as:

D＝5.44S+1.7376，R²＝0.7671；

wherein D is dialysis membrane pore diameter, S is synuclein dimer proportion, R²Is the correlation coefficient.

3. The method of claim 1, wherein the ratio of synuclein dimers in the blood S is obtained by an enzyme-linked immunosorbent assay, a biological activity assay, an enzyme-linked immunospot assay, and other molecular biological assays.

4. Use of the method of claim 1 for clearing of in vitro circulating synuclein.

5. The use according to claim 4 for clearance of synuclein in α -synuclein synucleinopathies.

6. A method for clearing extracorporeal circulation synuclein based on a hemofiltration mode is characterized by comprising the following steps:

obtaining the proportion S of synuclein dimers in blood;

obtaining the pore diameter D of an exchange membrane required for hemofiltration through the calculation model in claim 1;

and (3) selecting a dialysis membrane according to the pore size of the exchange membrane for hemofiltration, and then detecting the level of the synuclein in blood to evaluate the clearance of the synuclein in extracorporeal circulation.

Technical Field

The invention relates to the field of medicine, in particular to a method for removing extracorporeal circulation synuclein based on a hemofiltration mode.

Background

The alpha-synuclein clearance disorder can cause the development of Parkinson's Disease (PD), Multiple System Atrophy (MSA) and other alpha-synuclein spectrum diseases, and can cause immune disorder and neuroinflammatory reaction of the organism. Blood purification can effectively remove toxic substances in blood, such as inflammatory factors, autoantibodies, toxic protein molecules and the like; for example, plasma or peritoneal dialysis has been reported to be effective in clearing the body of toxic a β proteins and inflammatory factors. Similar pathogenesis exists in alpha-synuclein diseases such as PD, MSA and the like and AD, and the alpha-synuclein diseases similar to A beta-like diffusion property have similar clearance obstacle, so that neuroimmune disorder and inflammatory reaction are caused. At present, no effective method for eliminating alpha-synuclein to inhibit immune inflammation exists.

The hemofiltration technique is characterized in that a certain amount of replacement fluid is continuously supplemented in a blood vessel passage without using dialysate in the blood purification process, the replacement fluid is fully mixed with blood, and ultrafiltration is carried out at the same speed so as to achieve the aim of removing excessive water and toxin in the body. Compared with hemodialysis, hemofiltration has the advantages of small influence on hemodynamics, high removal rate of medium molecular substances and the like. Alpha-synuclein exhibits a variety of morphologies including an extended state, a pre-globular solubilized state, an alpha-helical state, a beta-lamellar state, a dimeric state, an oligomeric state, and an insoluble amorphous and fibrous state, among others. The alpha-synuclein with different forms has different molecular weights, and the toxic effect of the alpha-synuclein in the states of dimer and oligomer is particularly obvious. The molecular weight of the toxic alpha-synuclein is just in the hemofiltration adaptation range, and the regulation of the hemofiltration mode according to individual conditions can help to eliminate the peripheral circulating toxic alpha-synuclein and play a role in improving the alpha-synuclein related toxicity symptoms. At present, filtration parameters are set according to the molecular weight of substances needing to be filtered in hemofiltration, and alpha-synuclein has a dimer structure, so that the molecular weight distribution of an alpha-synuclein pedigree of each patient is different, and no reference study is set for the hemofiltration parameters of protein dimers or polymers.

Disclosure of Invention

The invention aims to provide a method for clearing extracorporeal circulation synuclein based on a hemofiltration mode, which aims to solve the problems in the prior art, and by designing a calculation model for clearing the extracorporeal circulation synuclein, different hemofiltration parameters can be designed according to different synuclein dimer contents, so that the efficiency of clearing the synuclein can be expected, and scientific data and a new scheme are provided for relieving alpha-synuclein related toxicity symptoms in clinical medicine.

In order to achieve the purpose, the invention provides the following scheme:

the invention provides a method for constructing an extracorporeal circulation synuclein clearance calculation model based on a hemofiltration mode, which comprises the following steps:

obtaining the proportion S of synuclein dimers in blood;

obtaining the blood flow velocity of hemofiltration and the area of an exchange membrane according to a large amount of clinical test data or literature historical data;

performing hemofiltration according to the determined blood flow velocity and the area of the exchange membrane, analyzing the correlation between the proportion S of different synuclein dimers and the aperture numerical value D of the exchange membrane for hemofiltration by adopting a linear regression method, and constructing a calculation model between the S and the D;

and (3) performing hemofiltration by adopting the proportion S of the synuclein dimer and the pore diameter of the exchange membrane determined by the calculation model, namely expecting the clearance rate of the extracorporeal circulation synuclein.

Preferably, the formula of the calculation model is as follows:

D＝5.44S+1.7376，R²＝0.7671；

wherein D is the exchange membrane pore diameter, S is the proportion of synuclein dimer, R²Is the correlation coefficient.

Preferably, the proportion S of the synuclein dimers in the blood is obtained by an enzyme-linked immunosorbent assay method, a biological activity assay method, an enzyme-linked immunospot assay, and other molecular biological assay methods.

The invention also provides application of the method in clearing the in vitro circulating synuclein.

Preferably, the method is applied to the clearance of synuclein in alpha-synuclein synucleinopathic diseases.

The invention also provides a method for clearing the extracorporeal circulation synuclein based on a hemofiltration mode, which comprises the following steps:

obtaining the proportion S of synuclein dimers in blood;

acquiring the pore diameter D of an exchange membrane required by hemofiltration through the calculation model;

and (3) selecting a dialysis membrane according to the pore size of the exchange membrane for hemofiltration, and then detecting the level of the synuclein in blood to evaluate the clearance of the synuclein in extracorporeal circulation.

Furthermore, the hemofiltration mode comprises parameters such as the aperture of an exchange membrane, the blood flow velocity, the area of the exchange membrane and the like, and the parameters are set according to the proportion of the individual alpha-synuclein dimer.

Further, methods for determining the proportion of individuals carrying alpha-synuclein dimers include the use of ELISA, biological activity assays, enzyme-linked immunospot assays and other molecular biological assays.

Further, the antibody specific to the alpha-synuclein includes monoclonal antibodies and polyclonal antibodies. Antibodies specific for said alpha-synuclein include intact antibody molecules, any fragment or modified fragment of an antibody, and the like. As long as the fragment retains the ability to bind to alpha-synuclein. The preparation of antibodies for use at the protein level is well known to those skilled in the art and any method may be used in the present invention to prepare such antibodies.

In the context of the present invention, the expression product of the alpha-synuclein gene includes human alpha-synuclein as well as partial peptides of human alpha-synuclein. The partial peptide of the alpha-synuclein contains functional domains of diseases related to the alpha-synuclein.

"alpha-synuclein" includes human alpha-synuclein and any functional equivalent of human alpha-synuclein. The functional equivalents include human alpha-synuclein conservative variant proteins, or active fragments thereof, or active derivatives thereof, allelic variants, natural mutants, induced mutants, proteins encoded by DNA that hybridizes to DNA of human alpha-synuclein under high or low stringency conditions.

Alpha-synuclein is a protein having the following amino acid sequence:

(1) a protein consisting of an amino acid sequence shown as SEQ ID No.1 below;

MDVFMKGLSK AKEGVVAAAE KTKQGVAEAA GKTKEGVLYV GSKTKEGVVH GVATVAEKTK EQVTNVGGAV VTGVTAVAQK TVEGAGSIAA ATGFVKKDQL GKNEEGAPQE GILEDMPVDP DNEAYEMPSE EGYQDYEPEA

(2) a protein which is derived from the amino acid sequence shown in SEQ ID NO.1, has the same function with the amino acid sequence shown in SEQ ID NO.1 and is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO. 1. The number of amino acids to be substituted, deleted or added is usually 1 to 50, preferably 1 to 30, more preferably 1 to 20, most preferably 1 to 10.

(3) A polypeptide consisting of an amino acid sequence having at least 80% homology (also referred to as sequence identity) with the amino acid sequence shown in SEQ ID NO.1, more preferably at least about 90% to 95% homology, often 96%, 97%, 98%, 99% homology with the amino acid sequence shown in SEQ ID NO. 1.

In a specific embodiment of the invention, the alpha-synuclein is a protein having the amino acid sequence shown in SEQ ID No. 1.

In general, it is known that modification of one or more amino acids in a protein does not affect the function of the protein. One skilled in the art will recognize that individual amino acid changes or small percentage amino acids or individual additions, deletions, insertions, substitutions to an amino acid sequence are conservative modifications, wherein a change in a protein results in a protein with a similar function. Conservative substitution tables providing functionally similar amino acids are well known in the art.

An example of a protein modified by the addition of an amino acid or amino acid residues is a fusion protein of alpha-synuclein. There is no limitation on the peptide or protein fused with α -synuclein, as long as the resulting fusion protein retains the biological activity of α -synuclein.

The alpha-synuclein of the invention also comprises non-conservative modifications of the amino acid sequence shown in SEQ ID No.1, as long as the modified protein still can retain the biological activity of the alpha-synuclein. The number of amino acids mutated in such modified proteins is typically 10 or less, such as 6 or less, such as 3 or less.

In the context of the present invention, "a-synuclein-associated toxicity condition" includes both determining whether a subject has, and determining whether a subject is at risk of having, a synuclein-associated toxicity condition.

In the context of the present invention, "alleviation of a-synuclein-related toxicity symptoms" is divided from a change in the state of the symptoms, and may include alleviation of symptoms, complete cure of the disease.

Since alpha-synuclein is considered to be a pathogenic agent of PD. Alpha-synuclein has strong self-polymerization capability and forms oligomers and fibers with stronger toxicity than alpha-synuclein monomers. Therefore, determining whether a method has the function of improving the toxicity of alpha-synuclein can be determined by determining whether the method has the effects of inhibiting and eliminating the alpha-synuclein level in vitro and blocking the toxicity of alpha-synuclein.

In a specific embodiment of the invention, total protein in peripheral blood is extracted for studies of alpha-synuclein level detection. As known to those skilled in the art, alpha-synuclein expressed by cells in brain tissue is shed into peripheral blood for circulation, i.e., alpha-synuclein is soluble, so that the level of alpha-synuclein in blood can indirectly reflect the expression of alpha-synuclein gene in brain tissue. According to the research results of the invention, the level and composition of alpha-synuclein water detected by extracting body fluid, especially peripheral blood can be used for judging whether the subjects have alpha-synuclein related toxicity symptoms.

The invention discloses the following technical effects:

the invention discovers for the first time that hemofiltration is helpful for reducing the level of alpha-synuclein in extracorporeal circulation, so that on the premise of determining the blood flow speed and the area of the exchange membrane based on clinical experience, the correlation between the alpha-synuclein and the aperture of the exchange membrane is researched, a calculation model of the alpha-synuclein and the aperture of the exchange membrane is constructed, and on the basis, the aim of clearing rate of the alpha-synuclein in extracorporeal circulation of 20 +/-2% can be fulfilled. Therefore, the invention fills the gap of the conventional toxic alpha-synuclein clearing method, adjusts the hemofiltration mode according to individual conditions, has timeliness, specificity and effectiveness for clearing the peripheral circulating toxic alpha-synuclein, plays a role in improving the toxic symptoms related to the alpha-synuclein, and can provide scientific basis and a new scheme for clinical application.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 shows the results of UPDRS and HY grading scoring of Parkinson's disease subjects before and after dialysis;

FIG. 2 is the HAMA, HAMD, MMSE, MoCA, non-motor scale and daily Living Capacity questionnaire scores for Parkinson's disease subjects before and after dialysis;

FIG. 3 is a graph showing the pore size distribution of exchange membrane and the proportion of synuclein.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.

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

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

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

As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.

Example 1

1. In vitro verification that hemofiltration can reduce the level of alpha-synuclein in extracorporeal circulation

An HF-80S high-flux dialyzer produced by the Ferwens medical apparatus and instruments Limited company is adopted to construct an extracorporeal blood filtration model, and different filtration speeds, filtration areas and exchange membrane pore diameters are set according to experimental requirements. Adopting 3L of fresh bovine whole blood, adjusting the hematocrit to be (31 +/-0.5)%, and storing in a water bath at constant temperature of (36.7 +/-0.5) ° C; different alpha-synuclein dimer combinations are added according to the experimental requirements to simulate the human body circulation blood of the patient. The blood of the simulated patient passes through an extracorporeal blood filtration model, the synuclein levels before and after filtration are collected and tested, and the clearance efficiency is calculated.

2. The presence of α -synuclein dimers in different ratios was determined by the above assay, and the following relationship between dialysis speed, dialysis membrane area and α -synuclein dimer clearance efficiency was found in table 1.

TABLE 1

As can be seen from Table 1, the different ratios of α -synuclein dimer, different dialysis speeds and dialysis membrane areas, have small differences in the clearance efficiency, which are all around 20%, that is, the dialysis speeds and the dialysis membrane areas are not the key factors affecting the clearance efficiency of α -synuclein dimer.

Therefore, the temperature of the molten metal is controlled,for the purpose of achieving an alpha-synuclein clearance efficiency of 20 + -2%, the flow rate of the dialysate was set at 200ml/min and the filtration area was 1.35m²Next, the pore size distribution of the corresponding exchange membranes was used to study the different alpha-synuclein dimer ratios, as shown in FIG. 3.

From fig. 3, the formula of the calculation model of the correlation between the alpha-synuclein ratio and the pore diameter of the exchange membrane is as follows: 5.44S +1.7376, R²0.7671; wherein D is the exchange membrane pore diameter, S is the proportion of synuclein dimer, R²Is the correlation coefficient.

Example 2

Method for clinical determination of alpha-synuclein

1. Peripheral blood and sample collection

PD patients were from the first hospital affiliated with Guangzhou medical university, 50 cases aged 57-66 years, and all cases were diagnosed as PD with reference to UK brain bank clinical diagnosis of parkinson's disease. The control population is 40, and is selected from conventional physical examination population of the first hospital affiliated to Guangzhou medical university, and all the subjects have the age of 55-68 years and exclude diseases such as blood lipid metabolism and nervous system chronic degenerative diseases. All subjects signed an informed consent for this test item, made the UPDRS score, and provided peripheral blood for the detection of alpha-synuclein.

2. Detection kit for constructing and measuring alpha-synuclein

(1) To quantitatively determine the content of α -synuclein, two different monoclonal anti- α -synuclein antibodies to human synuclein were used, including the use of an anti- α -synuclein monoclonal antibody as the capture antibody and a biotinylated mouse anti-human α -synuclein monoclonal antibody as the detection antibody.

(2) The capture antibody was embedded overnight at room temperature in PBS buffer at a final concentration of 4.0. mu.g/ml on a 96-well assay plate.

(3) After washing 3 times with Wash Buffer (PBS Buffer containing 0.05% Tween20, PH 7.2), the plates were blocked with Reagent reagents (1% BSA in PBS) for 1h at 25 ℃.

3. Assay for serum and soluble alpha-synuclein

(1) Mu.l (or 0.25g) of whole blood was applied to RNase-Free filtration column, centrifuged at 13000rpm for 2 minutes, and the supernatant was collected.

(2) Soluble human TRME2-FC and human a-synuclein-FC were used as standards in all experiments.

(3) 100 mul of fresh and rewarming serum and a standard substance are added into a coated plate to be tested and incubated for 2h at room temperature, and after washing with Wash Buffer, 100 mul of detection antibody (final concentration: 35.0ng/ml) with the final concentration of soluble alpha-synuclein and a-synuclein detection antibody (final concentration: 25.0ng/ml) are added and incubated for 2h at room temperature.

(4) After washing with Wash Buffer, 100. mu.l of streptavidin labeled with horseradish peroxidase was added and incubated for 20min at room temperature in the dark.

(5) After washing with Wash Buffer, 100. mu.l of a color developer (R & D Catalog # DY999) was added and incubated at room temperature for 20min in the dark.

(6) The color development was stopped by the addition of 50. mu.l of 2N sulfuric acid (R & D Catalog # DY 994).

(7) The OD measured at 450 nm was determined for each well and was corrected using the OD measured at 540 nm.

4. Statistical method

The experiments were performed in 3 replicates, the results were represented as mean ± sd, and were statistically analyzed using SPSS21.0 statistical software, with the difference between the two using the t-test, and considered statistically significant when P < 0.05.

Example 3

A method for constructing an extracorporeal circulation synuclein clearance calculation model based on a hemofiltration mode is specifically applied as follows:

1. specific cases

Female, 81 years old, married, admitted 13 years into hospital because of tremor, and the patient found tremor of the left upper limb before 13 years, which often appeared at rest, and disappeared during exercise and tension without obvious aggravation or relief factor, the Zhongshan university was affiliated to the first hospital to see a doctor, and was given "Meiduoba 0.125g BID + coenzyme Q1010 mg QD", and tremor did not further aggravate. Before 3 years, the symptoms are aggravated, and the patient can hardly fall asleep at night, and can fall asleep by taking 'clonazepam 2 mg'. The tremor also appears on the right upper limb 2 years ago, the tremor is static, the frequency is the same as that on the left side, the tremor amplitude is slightly lighter than that on the left side, constipation occurs, 3-4 days/times of defecation, the dry and hard to solve can be solved by using paraffin oil or glycerine enema sometimes, the oral medicine is adjusted to be Meiduoba 0.25g TID + Senfuluo 0.25mg TID + Kedan 0.2g TID, the walking instability appears 1 year ago, the starting difficulty appears, the stride is smaller and smaller in the walking process, the turning is easy to fall down, the mandibular tremor appears, the tremor is obvious in the rest, the neck movement is hard, and the appetite is reduced. The actions of button buckling, slipper wearing and the like are not flexible and have melancholy mood, and the symptom is not obviously improved by adding 'Beiaixin 1 QD', the feeling of weakness is aggravated before 1 week, the breathing difficulty is consciously seen, the symptom is not improved after oxygen inhalation, the symptom appears at 3 pm mostly, and the symptom can disappear by oneself at 8-9 night. It is good for mental disorder, poor sleep and stool (as above) and little for patients. Admission physical examination: body temperature 36.5 deg.C, pulse rate 78 times/min, respiration 20 times/min, blood pressure 117/60 mmHg. No special heart, chest and abdomen exist. Clear mind, vague speech, answering and answering, physical examination and cooperation, face mask, dismissal and gait. Memory decline in short time, normal directional and visual space functions, and decline of executive force. Hyposmia, no defect in the lateral visual field, flexible eyeball movement, equilarge pupils on both sides, and d of 3mm, and is sensitive to light reflex and convergence reflex. Coarse binaural hearing decreased, weber test centered, AC > BC. Jaw tremor. The cranial nerve was found to be abnormal. The neck muscle tension is 3 grades, the height is increased like a gear, the mental distance between the chest and the transverse fingers is 2, and the left alternation test is relatively inflexible. The left-hand swing is smaller than the right side when walking. Pull back test (+). The muscle tension of the two upper limbs is increased like a gear, the muscle strength is 5 grade, and the tendon reflex is +++. Normal lower limb muscle tone, 5-grade muscle strength, tendon reflex +. Left lower limb straight leg elevation test (+). No obvious abnormalities were seen in superficial sensation, deep sensation and cortical complex sensation. No pathological signs are elicited. The meningeal stimulation sign was negative. Admission diagnosis: parkinson's disease; sick sinus syndrome, implanted cardiac pacemaker; coronary atherosclerotic heart disease.

Perfection check and signing informed consent: the patients have intermediate and advanced Parkinson's disease and have no obvious improvement after taking the medicine. Recent symptoms are aggravated, walking is unstable, mandibular tremor occurs, and dyspnea and chest tightness occur in the afternoon to the evening. Considering that the Parkinson disease is alpha-synuclein synucleinopathic disease, the existing body immune disorder and neuroinflammatory reaction blood purification can effectively remove toxic substances in blood plasma, such as inflammatory factors, autoantibodies and toxic protein molecules; for example, plasma or peritoneal dialysis has been reported to be effective in clearing A β toxic proteins and inflammatory factors. Similar pathogenesis exists in parkinson's disease and alzheimer's disease, due to the clearance of alpha-synucleinopathic proteins with similar abeta-like spreading properties, resulting in neuroimmune disorders and inflammatory responses. At present, no method for effectively eliminating the immune inflammation exists in the clinical treatment of the Parkinson's disease, and the blood purification technology can help to eliminate peripheral nerve immune inflammation molecules and plays an important role in the clinical prevention and treatment of the Parkinson's disease. Although the patients are older, the clinical trials of blood purification technology which can tolerate the alpha-synuclein-synucleinopathic peripheral blood toxic protein and the immune inflammatory factor cannot necessarily achieve the expectation of the patients and the families (son-daughter) through informing the risks existing in the blood purification treatment and the treatment outcome, and the patients and the families show the understanding and sign of the situations after being carefully considered. And (5) simulating the next week of blood purification.

2. Detailed description of the preferred embodiments

The method of example 2 was used to obtain the ratio of alpha-synuclein in the blood of the subjects, the results showed that the cerebrospinal fluid synuclein dimer proportion of the subjects was around 0.035, and according to the computational model formula of the present invention, the pore size of the exchange membrane was 1.928 nm. After which a treatment plan is developed.

Current blood purification treatments: selecting blood perfusion parameters as follows, the aperture of the exchange membrane is 2nm, the blood flow speed is 200ml/min, and the area of the exchange membrane is 1.35m²(ii) a The blood perfusion treatment was performed 3 times each for 2 hours.

Patient symptoms change before and after treatment: the patient is better in tremor and bradykinesia than when the patient is admitted. The physical examination is clear in mind, vague in speech, answering and answering, physical examination and cooperation, face mask, and dismissal and gait. Memory decline in short time, normal directional and visual space functions, and decline of executive force. Hyposmia, no defect in the lateral visual field, flexible movement of the eyeball, equilarge pupils and the like, and sensitive to light reflex and convergence reflex, wherein d is 3 mm. Coarse binaural hearing decreased, weber test centered, AC > BC. Jaw tremor. The cranial nerve was found to be abnormal. The neck muscle tension is 2 grade, the height is increased like a gear, the mental chest distance is 2 horizontal fingers, and the left alternation test is relatively inflexible. The left-hand swing is smaller than the right side when walking. Back pull test (-). Double upper limb muscle tension gear-like heightening, muscle strength 5 grade, tendon reflex +++. Lower limb musculature was normal, muscle strength 5 grade, tendon reflex +. Left lower limb straight leg elevation test (+). No obvious abnormalities were seen in superficial sensation, deep sensation and cortical complex sensation. No pathological signs are elicited. The meningeal stimulation sign was negative.

The cerebrospinal synuclein concentration was retested after treatment and the clearance efficiency was about 0.232. Meanwhile, the comparison of multiple scores of symptoms of Parkinson disease patients before and after dialysis is compared, and the symptoms of the Parkinson disease patients after dialysis are obviously improved as shown in the figure 1 and the figure 2.

The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.

Sequence listing

<110> the first hospital affiliated to Guangzhou medical university (Guangzhou respiratory center) Jiangxi province Leyun health technology Co Ltd

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