CN113933229B - Platelet activation performance identification method - Google Patents
Platelet activation performance identification method Download PDFInfo
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- CN113933229B CN113933229B CN202111137109.6A CN202111137109A CN113933229B CN 113933229 B CN113933229 B CN 113933229B CN 202111137109 A CN202111137109 A CN 202111137109A CN 113933229 B CN113933229 B CN 113933229B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
Abstract
The invention discloses a platelet activation performance identification method, which uses the concentration of triglyceride and uric acid as an index to judge the activation performance of platelets. The platelet activation performance identification method provided by the invention evaluates the platelet activation level by taking the serum triglyceride and uric acid as judgment indexes, can screen more qualified platelets for clinical infusion, reduces unnecessary medical resource waste, and has important significance for clinical patient first-aid infusion.
Description
Technical Field
The invention relates to the field of blood detection, in particular to a method for identifying platelet activating performance.
Background
Platelet activation is one of the major problems faced by current clinical infusions of platelets, and surface markers of activated platelets can be detected by flow cytometry, such as CD63 and CD62p as surface markers of platelet activation are expressed in plasma membranes of platelets, and CD63 and CD62p expression in activated platelets is increased accordingly, which is a gold standard for current platelet activation detection. The prior art is restricted, the platelets have a special molecular structure, the storage time of the platelets separated from fresh blood under the oscillation condition of 22 ℃ and 60 times/min is only 5 days, and the shorter storage time greatly limits the emergency treatment of patients needing blood transfusion in clinic. Platelets are activated during storage to increase adhesion capacity, platelet activation performance determines whether the platelets can be used, and platelet activation is also affected by other biochemical indexes, but research on other biochemical indexes affecting platelet activation is not available. It is now necessary to find more biochemical indicators that lead to platelet activation and evaluate the platelet activation performance.
Disclosure of Invention
The invention aims to solve the technical problem of providing a platelet activation performance identification method aiming at the defects in the prior art.
In order to solve the technical problems, the invention adopts the following technical scheme: a method for identifying the activating performance of blood platelet features that the concentration of triglyceride and urea acid is used as the index to judge the activating performance of blood platelet.
Preferably, the method comprises the steps of:
1) Pre-constructing a corresponding relation table of triglyceride concentration, uric acid concentration and platelet activation degree, wherein in the table, triglycerides with different concentrations correspond to first index scores with different scores, uric acid with different concentrations correspond to second index scores with different scores, and the sum of the first index score and the second index score is marked as a total score, and the total score reflects the platelet activation degree;
2) Detecting the triglyceride concentration and uric acid concentration of a sample to be detected, obtaining a first index score and a second index score of the sample through the corresponding relation table obtained in the step 1), calculating a total score, and judging the platelet activation degree of the sample.
Preferably, the step 1) specifically includes:
1-1) classifying the serum triglycerides into a normal serum triglyceride group, a critical high serum triglyceride level group and a high serum triglyceride level group according to the serum triglyceride concentration, and classifying the serum triglycerides into a normal uric acid group and a high uric acid expression group according to the uric acid concentration;
1-2) detecting the CD63 expression intensity and the CD62p expression intensity in the normal serum triglyceride group, the critical high serum triglyceride level group and the elevated serum triglyceride level group respectively, and judging the platelet activation degree according to the CD63 expression intensity and the CD62p expression intensity, thereby giving different scores to the normal serum triglyceride group, the critical high serum triglyceride level group and the elevated serum triglyceride level group as A1, A2 and A3;
detecting the expression intensity of CD63 and the expression intensity of CD62p in the uric acid normal group and the uric acid high expression group respectively, judging the activation degree of platelets according to the expression intensity of CD63 and the expression intensity of CD62p, so as to endow the uric acid normal group and the uric acid high expression group with second index scores with different scores, and marking the scores as B1 and B2 in sequence;
the total score was noted as: aAi + bBj, where a and b are weights, i=1, 2,3, j=1, 2, respectively;
1-3) tabulating to obtain the corresponding relation table.
Preferably, wherein a=0.5 and b=0.5.
Preferably, wherein the higher the serum triglyceride concentration, the higher the corresponding first index score; the higher the uric acid concentration, the higher the corresponding second index score; the higher the total score, the higher the degree of platelet activation.
Preferably, wherein the serum triglyceride normal group concentration is: the critical high group concentrations of serum triglyceride levels are < 1.7 mmoL/L: 1.7-2.25mmoL/L, serum triglyceride level elevation group concentration: not less than 2.26mmoL/L.
Preferably, wherein the uric acid normal group concentration is: 154.7-357.0 mu mol/L, uric acid high expression set concentration is: > 357.0. Mu. Mol/L.
Preferably, the concentration of serum triglycerides and uric acid is measured using the Beckmann Coulter Biochemical analysis System.
Preferably, the expression intensities of the platelet surface CD62p and CD63 are detected using a flow cytometer, among others.
The beneficial effects of the invention are as follows: the platelet activation performance identification method provided by the invention evaluates the platelet activation level by taking the serum triglyceride and uric acid as judgment indexes, can screen more qualified platelets for clinical infusion, reduces unnecessary medical resource waste, and has important significance for clinical patient first-aid infusion.
Drawings
FIGS. 1a and 1b show the results of comparing platelet activities in the normal and critical TG groups;
FIGS. 2a and 2b are results of comparing platelet activities in the normal and elevated TG groups;
FIGS. 3a and 3b are results of comparing platelet activity in normal and elevated UA groups.
Detailed Description
The present invention is described in further detail below with reference to examples to enable those skilled in the art to practice the same by referring to the description.
It will be understood that terms, such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
The method for identifying platelet activation performance in this embodiment uses the concentration of triglyceride and uric acid as an index to determine the platelet activation performance, and includes the following steps:
1) Pre-constructing a corresponding relation table of triglyceride concentration, uric acid concentration and platelet activation degree, wherein in the table, triglycerides with different concentrations correspond to first index scores with different scores, uric acid with different concentrations correspond to second index scores with different scores, and the sum of the first index score and the second index score is marked as a total score, and the total score reflects the platelet activation degree;
the method specifically comprises the following steps:
1-1) classifying the serum triglycerides into a normal serum triglyceride group, a critical high serum triglyceride level group and a high serum triglyceride level group according to the serum triglyceride concentration, and classifying the serum triglycerides into a normal uric acid group and a high uric acid expression group according to the uric acid concentration;
in a preferred embodiment, the normal group concentration of serum triglycerides is the critical high group concentration of serum triglyceride levels is: 1.7-2.25mmoL/L, serum triglyceride level elevation group concentration: not less than 2.26mmoL/L. Uric acid normal group concentration is: 154.7-357.0 mu mol/L, uric acid high expression set concentration is: > 357.0. Mu. Mol/L.
1-2) detecting the CD63 expression intensity and the CD62p expression intensity in the normal serum triglyceride group, the critical high serum triglyceride level group and the elevated serum triglyceride level group respectively, and judging the platelet activation degree according to the CD63 expression intensity and the CD62p expression intensity, thereby giving different scores to the normal serum triglyceride group, the critical high serum triglyceride level group and the elevated serum triglyceride level group as A1, A2 and A3;
wherein CD62P (P selectin): the platelet lysosome membrane glycoprotein is a surface marker for platelet activation, and can accurately reflect the activation degree of the platelets; CD63: platelet granule membrane protein is an activation index of platelets;
detecting the expression intensity of CD63 and the expression intensity of CD62p in the uric acid normal group and the uric acid high expression group respectively, judging the activation degree of platelets according to the expression intensity of CD63 and CD62p, so as to endow the uric acid normal group and the uric acid high expression group with second index scores with different scores, and marking the scores as B1 and B2 in sequence;
the total score was noted as: aAi + bBj, wherein i=1, 2,3, j=1, 2; a and b are weights, respectively, and in one embodiment a=0.5 and b=0.5.
1-3) tabulating to obtain the corresponding relation table.
2) Detecting the triglyceride concentration and uric acid concentration of a sample to be detected, obtaining a first index score and a second index score of the sample through the corresponding relation table obtained in the step 1), calculating a total score, and judging the platelet activation degree of the sample.
It should be understood that the method of the present invention may be used directly for platelet activation performance identification, or may be used as an auxiliary index for platelet activation performance evaluation. For example, the method of the present invention is used as a further adjunct indicator to the conventional gold standard for platelet activation assays with activated platelets for CD63 and CD62p expression, thereby screening for more acceptable platelets for clinical infusion. That is, when the CD63 and CD62p are detected as being acceptable, the result is also acceptable as being acceptable platelets when the detection is performed by the method of the present invention.
Wherein, the higher the serum triglyceride concentration, the higher the corresponding first index score; the higher the uric acid concentration, the higher the corresponding second index score; the higher the total score, the higher the degree of platelet activation.
Wherein, the concentration of serum triglyceride and uric acid is detected by adopting a Beckmann Coulter AU biochemical analysis system, and the expression intensity of CD62p and CD63 on the surface of the blood platelet is detected by adopting a flow cytometry.
In this example, uric Acid (UA) normal group (154.7-357.0. Mu. Mol/L) and uric acid high expression group (> 357.0. Mu. Mol/L), serum Triglyceride (TG) normal group (< 1.7 mmole/L), serum triglyceride level criticality high group (1.7-2.25 mmole/L) and serum triglyceride level elevation group (. Gtoreq.2.26 mmole/L), were stored for 5 days under 60-time shaking conditions at 22℃and platelet counts were performed on days 1,2,3, 4, 5, respectively, and the expression levels of CD62P and CD63 on the platelet surface were detected using a flow cytometer, and the platelet activation levels at the different uric acid and serum triglyceride levels were observed for three groups, wherein CD62P (P selectin): the platelet lysosome membrane glycoprotein is a surface marker for platelet activation, and can accurately reflect the activation degree of the platelets; CD63: platelet granule membrane protein is an index of platelet activation.
Referring to table 1 and fig. 1a, fig. 1b are results of comparing platelet activities of normal TG group and TG critical high group:
table 1 comparison of platelet Activity in the TG Normal and TG Critical high groups
It can be seen that the serum triglyceride normal group and critical high group platelets have small differences in the expression levels of CD62P and CD63 (P > 0.05) at different shelf times, so that the assignment of A1, A2 can be approximated.
Referring to table 2 and fig. 2a, fig. 2b are comparison results of platelet activity in TG normal group and TG elevated group:
table 2 comparison of platelet Activity in the TG Normal and TG-elevated groups
It can be seen that the expression levels of CD62P and CD63 were significantly higher in the serum triglyceride elevated group platelets than in the serum triglyceride normal group (P < 0.05) at days 2,3, 4, and 5 of in vitro storage, so that A3 should be assigned a significantly greater value than A2. From the above, it can be seen that, within a certain range, higher serum triglyceride concentrations indicate higher platelet activity.
Referring to table 3 and fig. 3a, fig. 3b are comparison results of platelet activity in normal and elevated UA groups:
table 3 comparison of platelet Activity in normal and UA elevated groups of 3 UA
It can be seen that the expression levels of CD62P and CD63 were significantly higher in the uric acid-elevated group platelets than in the normal group of haematuria (P < 0.05) when stored in vitro on days 3, 4, and 5. Within a certain range, higher uric acid concentrations indicate higher platelet activity, and the assignment of B2 is greater than that of B1.
From the above results, it can be seen that the serum triglyceride concentration and uric acid concentration can reflect platelet activity within a certain range. The activation level of platelets is affected by the serum triglyceride and uric acid levels of the donor during the five-day storage period, and the platelets of the donor with normal and critical serum triglyceride levels can be kept in a static state to a certain extent, maintain the adhesion and bonding capability and reduce the transmission of blood-borne diseases when the platelets are stored under the condition of shaking at 22 ℃ for 60 times/min. Platelets from donors with higher serum triglyceride levels are activated to a higher level during storage and are unsuitable for clinical infusion. Uric acid levels in blood donors also have a significant impact on changes in platelet in vitro preservation activity, and platelets from blood donors with higher uric acid levels have higher levels of activity during preservation than normal uric acid groups. By evaluating the activation level of the mechanically-sampled platelets by taking the serum triglyceride and uric acid as judgment indexes, more qualified platelets can be screened for clinical infusion, unnecessary medical resource waste is reduced, and the method has important significance for clinical patient emergency infusion.
Although embodiments of the present invention have been disclosed above, it is not limited to the use of the description and embodiments, it is well suited to various fields of use for the invention, and further modifications may be readily apparent to those skilled in the art, and accordingly, the invention is not limited to the particular details without departing from the general concepts defined in the claims and the equivalents thereof.
Claims (3)
1. A platelet activation performance identification method is characterized in that the method judges the activation performance of platelets by taking the concentration of triglyceride and uric acid as indexes, and comprises the following steps:
1) Pre-constructing a corresponding relation table of triglyceride concentration, uric acid concentration and platelet activation degree, wherein in the corresponding relation table, triglycerides with different concentrations correspond to first index scores with different scores, uric acid with different concentrations correspond to second index scores with different scores, and the sum of the first index score and the second index score is marked as a total score, and the total score reflects the platelet activation degree;
2) Detecting the triglyceride concentration and uric acid concentration of a sample to be detected, obtaining a first index score and a second index score of the sample through the corresponding relation table obtained in the step 1), calculating a total score, and judging the platelet activation degree of the sample;
the step 1) specifically comprises the following steps:
1-1) classifying the serum triglycerides into a normal serum triglyceride group, a critical high serum triglyceride level group and a high serum triglyceride level group according to the serum triglyceride concentration, and classifying the serum triglycerides into a normal uric acid group and a high uric acid expression group according to the uric acid concentration;
1-2) detecting the CD63 expression intensity and the CD62p expression intensity in the normal serum triglyceride group, the critical high serum triglyceride level group and the elevated serum triglyceride level group respectively, and judging the platelet activation degree according to the CD63 expression intensity and the CD62p expression intensity, thereby giving different scores to the normal serum triglyceride group, the critical high serum triglyceride level group and the elevated serum triglyceride level group as A1, A2 and A3;
detecting the expression intensity of CD63 and the expression intensity of CD62p in the uric acid normal group and the uric acid high expression group respectively, judging the activation degree of platelets according to the expression intensity of CD63 and the expression intensity of CD62p, so as to endow the uric acid normal group and the uric acid high expression group with second index scores with different scores, and marking the scores as B1 and B2 in sequence;
the total score was noted as: aAi + bBj, where a and b are weights, i=1, 2,3, j=1, 2, respectively;
1-3) tabulating to obtain the corresponding relation table;
wherein a=0.5, b=0.5;
wherein, the higher the serum triglyceride concentration, the higher the corresponding first index score; the higher the uric acid concentration, the higher the corresponding second index score; the higher the total score, the higher the degree of platelet activation;
wherein, the normal group concentration of serum triglyceride is: the critical high group concentrations of serum triglyceride levels are < 1.7 mmoL/L: 1.7-2.25mmoL/L, serum triglyceride level elevation group concentration: more than or equal to 2.26 mmoL/L;
wherein, uric acid normal group concentration is: 154.7-357.0 mu mol/L, uric acid high expression set concentration is: > 357.0. Mu. Mol/L.
2. The method for identifying platelet activation performance according to claim 1, wherein the concentration of serum triglycerides and uric acid is detected using a beckmann coulter AU biochemical analysis system.
3. The method for discriminating platelet activating property according to claim 1 wherein the expression intensities of the platelet surface CD62p and CD63 are detected using a flow cytometer.
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Citations (5)
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| CN1255637A (en) * | 1998-09-03 | 2000-06-07 | 希森美康株式会社 | Antihemagglutinin and blood examining method |
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| CN101356286A (en) * | 2005-11-17 | 2009-01-28 | 英特利金遗传学有限公司 | Diagnostics and therapeutics for cardiovascular disorders |
| CN103338760A (en) * | 2010-11-15 | 2013-10-02 | 贝林格尔.英格海姆国际有限公司 | Vasoprotective and cardioprotective antidiabetic therapy |
| CN103596576A (en) * | 2011-01-20 | 2014-02-19 | 阿龙·托梅 | Platelet analysis system |
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- 2021-09-27 CN CN202111137109.6A patent/CN113933229B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1255637A (en) * | 1998-09-03 | 2000-06-07 | 希森美康株式会社 | Antihemagglutinin and blood examining method |
| JP2005503110A (en) * | 2001-01-02 | 2005-02-03 | ザ・クリーブランド・クリニック・ファンデーション | Myeloperoxidase, a risk indicator for cardiovascular disease |
| CN101356286A (en) * | 2005-11-17 | 2009-01-28 | 英特利金遗传学有限公司 | Diagnostics and therapeutics for cardiovascular disorders |
| CN103338760A (en) * | 2010-11-15 | 2013-10-02 | 贝林格尔.英格海姆国际有限公司 | Vasoprotective and cardioprotective antidiabetic therapy |
| CN103596576A (en) * | 2011-01-20 | 2014-02-19 | 阿龙·托梅 | Platelet analysis system |
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