CN106596916A - Correlation analysis method between peripheral blood lymphocyte connexin 43 and IL-2 and IL-6 - Google Patents
Correlation analysis method between peripheral blood lymphocyte connexin 43 and IL-2 and IL-6 Download PDFInfo
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Abstract
The invention discloses a correlation analysis method of peripheral blood lymphocyte connexin 43 and IL-2 and IL-6. The method comprises the steps of selecting and taking 20 SHR mice and 20 WKY big mice wherein the WKY big mice have normal blood pressure; collecting abdominal aorta peripheral blood to be subjected to centrifugation, wherein blood plasma is used for detecting expression levels of IL-2 and IL-6 through an ELISA method; lower bottom haemocytes are used for separating peripheral blood lymphocyte; collecting 5*105 lymphocytes, and adopting a flow cytometry to detect a positive expression rate of CD4+CD8+T cell C*34; diving the lymphocytes into a blank control group, a concanavalin group and a Gap27+ConA group; extracting a lymphocyte RNA, and adopting real-time fluorescence quantification RCR to detect relative expression quantity of IL-2mRNA and IL-6mRNA in each group of cells.
Description
Technical field
The invention belongs to biological technical field, is related to a kind of peripheral blood lymphocyte connection protein 43 and IL-2 and IL-6 tables
The correlation analysis for reaching, specifically, are related to a kind of spontaneous hypertensive rat peripheral blood lymphocyte connection protein 43
The correlation analysis expressed with IL-2 and IL-6.
Background technology
Hypertension is used as the multifactor chronic disease for causing, and its cause of disease and pathogenesis are sufficiently complex.The research such as ARIMA is sent out
Existing, the cardiovascular disease such as hypertension, atherosclerosiss is close with body immune system functional relationship.Immune system, especially
Expression connection albumen (connexin, Cx) in periphery blood T cell, B cell and natural killer cell.Cx is the base of gap connection
This composition unit, the transmission information material between flanking cell constitutes gap connection cell-cell communication.The research discovery such as MATSUE,
In the case where lipopolysaccharide and tumor necrosis factor ɑ stimulate, Cx43 expression in mononuclear cell can be raised significantly.Therefore, gap connection and machine
Body Immune inflammatory reaction is related, participates in being communicated between immune system cell.CD4+T cells and CD8+T cells are used as periphery blood T cell
Major subpopulations cell, participate in adjusting immunity of organism inflammatory reaction, play important immunomodulating in infection, anti-tumor aspect
Effect.
The content of the invention
It is an object of the invention to overcome defect present in prior art, there is provided a kind of peripheral blood lymphocyte connects egg
White 43 correlation analysis expressed with IL-2 and IL-6, the pass between Primary Study Cx43 and Hypertensive Rats inflammatory reaction
System, theoretical foundation is provided to illustrate Cx43 to the mechanism of action of hyperpietic's inflammatory reaction.
Its concrete technical scheme is：
The correlation analysis that a kind of peripheral blood lymphocyte connection protein 43 is expressed with IL-2 and IL-6, including it is following
Step：
Step 1, rat blood pressure are determined and measure rat tail artery blood pressure using BP-6 non-invasive blood pressure measuring instruments, it is to avoid rat
Restless, the continuous measurement 3 times under the clear-headed rest state of rat, per minor tick at least 1min, takes 3 meansigma methodss of measurement and is rat
Systolic pressure.
Step 2, the pentobarbital sodium anesthetized rat of sample collection 3%, ethylenediaminetetraacetic acid (EDTA) anticoagulant tube collection abdomen master
Periarterial blood 5ml, 1 500r/min are centrifuged 5min, take blood plasma in -20 DEG C of preservations, and the lower floor's hemocyte after centrifugation is used to separate
Peripheral blood lymphocyte.
Step 3, inflammatory factor IL-2 and IL-6 expression take frozen in -20 DEG C of plasma sample, ELISA method detection IL-2
With IL-6 expressions, operate is carried out in strict accordance with kit specification.
Step 4, separation of lymphocytes are by hemocyte and 0.9% sodium chloride solution 1:1 equal-volume dilutes, and piping and druming is mixed.Take
Aseptic 15ml centrifuge tubes, add lymphocyte separation medium, by cell suspension slowly according to 1:1 ratio is added to lymphocyte separation medium
Upper strata, it is ensured that blood is placed in lymphocyte separation medium upper strata.Lymphocyte, low speed centrifuge are separated according to density－gradient centrifuga－tion method
Horizontal centrifugal 30min (1 500r/min delays and rises slow drop).By detached white opacity buffy coat suction out to aseptic 15ml from
In heart pipe, 3 times of 0.9% sodium chloride solution washing is added, washed 2 times with 1 800r/min centrifugation 6min, abandon supernatant, precipitation is drenched
Bar cell.Add 1ml culture medium resuspended, draw 50 μ l to 1.5ml centrifuge tubes, plus 450 μ l phosphate buffers (PBS) dilutions 10
Times, counted under microscope is simultaneously active with Trypan Blue observation of cell.Cytoactive is more than 95%, and adjustment cell concentration is 1
×106Individual/ml, cell is seeded in 24 orifice plates.
Step 5, Flow cytometry CD4 +And CD8 +The expression of T cell Cx43 collects 5 × 105Individual cell is interior in EP pipes,
100 μ l PBS re-suspended cells, are separately added into corresponding streaming antibody (CD4- APC, APC Isotype control；CD8- PE, PE Isotype control) 1
μ l, and set up negative control.125 μ l fixatives fix lymphocyte, 4 DEG C of incubation 20min, the anti-1 μ l antibody of Cx43 mono- and 99 μ l
1 × rupture of membranes agent rupture of membranes, 37 DEG C of incubation 20min；FITC bis- resists 1 μ l, 37 DEG C of incubation 20min；The agent of 1 × ruptures of membranes of 1ml is washed, and is abandoned
Clear liquid, PBS re-suspended cells, flow cytomery, streaming CELLQuest software is analyzed.
Step 6, real-time fluorescence quantitative PCR detection IL-2mRNA and IL-6mRNA expression count the lymphocyte isolated,
Adjustment culture medium to cell concentration is 1 × 106Individual/ml.Lymphocyte is divided into into blank control group, canavaline
(Concanavalin A, ConA) group (adding 5 μ g/ml ConA after culture 48h) and Gap27+ConA groups (add 5 before culture
00 μm of ol/L Gap275, after culture 48h 5 μ g/ml ConA are added).Add 10% autoserum (serum process step：56℃
Inactivation 30min, with 4 000r/min 30min, 4 DEG C of preservations are centrifuged), in 37 DEG C, 5%CO248h is cultivated in incubator.
TRIzol methods extract the lymphocyte mRNA of above-mentioned culture, and nucleic acid-protein content detection instrument determines A260/A280Value,
1% denaturing formaldehyde gel electrophoresis detection extracts quality.From GADPH as reference gene, according to rat IL-2, IL-6 gene
CDNA sequence, setting primer sequence (being shown in Table 1).MRNA samples, random primer, Rtmix, dNTP mixed liquor, the coke that will be extracted
Diethyl phthalate (DEPC) water prepares reaction system according to reverse transcription system, according to 37 DEG C of reverse transcription 60min, 90 DEG C of inactivation reverse transcriptions
The response procedures of enzyme 5min carry out reverse transcription synthesis cDNA.By 2 × QuantiNova SYBR Green PCR Master Mix,
10 × miScript Nucleics Mix universal primers, template cDNA and RNase-Free water are placed on ice, and gently mix
It is even, reaction system is prepared, according to 95 DEG C of denaturations 2min, 1 circulation；95 DEG C of degeneration 5s, 60 DEG C of annealing plus extension 10s, 40
It is circularly set Bio-Rad quantitative real time PCR Instruments, record IL-2mRNA and IL-6mRNA relative expression quantities.
Compared with prior art, beneficial effects of the present invention：
The study find that, SHR rats increase with inflammatory reaction, proinflammatory factor release, while constitute gap between T cell connecting
The Cx43 expression for connecting road is significantly raised, using proinflammatory factor after information communication between gap junction blocker blocking lymphocyte
Release is remarkably decreased, and points out gap connection to participate in hypertension inflammatory reaction.
Specific embodiment
Technical scheme is described in more detail with reference to specific embodiment.
1 materials and methods
1.1 material
1.1.1 laboratory animal chooses SHR and normotensive WKY rats each 20 in April, 2015-2016 year April
Only, 16～18 week old, male, 150～220g of weight, from the purchase of Beijing laboratory animal Co., Ltd of dimension tonneau China
(licence numbering SCXK capital 2007-0001), has reached cleaning grade standard, and animal quality is primary standard.Feeding environment is required：
Good ventilation, air filtering system, environment quiet, room temperature keep 20 DEG C, humid control 45% or so, it is daily change bedding and padding and
Drinking water, freely absorbs water and food.
1.1.2 instrument and reagent flow cytometer (E6) are purchased from Mairui Biological Medical Electronic Co., Ltd., Shenzhen, water
Bath (H216) is purchased from Shanghai Bo Xun Industrial Co., Ltd.s, and low speed large capacity centrifuge (LC-4016) is purchased from good in section in Anhui
Scientific instrument company limited, difference inverted microscope (XD30) is purchased from Ningbo Shun Yu Co., Ltds, Biohazard Safety Equipment (BHC-
1300 II A2) it is purchased from Altay laboratory equlpment (Beijing) company limited, CO2Constant incubator (HF151) is purchased from Shanghai power Shen
Scientific instrument company limited, gel imaging instrument (BioDoc Imaging) is purchased from UVP companies of the U.S., grads PCR instrument (TC-96/G/
H (b) C) Hangzhou BIOER Technology Co., Ltd is purchased from, microplate reader, micro-spectrophotometer (K5500) are sent out purchased from the triumphant AudioCodes skill in Beijing
Exhibition company limited, quantitative real time PCR Instrument (TIB-8000) is purchased from Thymopetidum Injection thing science and technology (China) company limited.
The little anti-rat CD of other phycocyanin (allophycocyanin, APC) labelling4Antibody (B159112), algae red egg
The little anti-rat CD of (phycoerythrin, PE) labelling in vain8aAntibody (B166485), PE labelling IgG1, к Isotype Ctrl are same
Type control (B179044) is purchased from BioLegend companies, anti-rat Anti-Connexin 43 (ab-79010) purchase in mice source
From Abcam companies, goat anti-mouse IgG/FITC labellings two anti-(ZF-0312) are purchased from the limited public affairs of Beijing Zhong Shan Golden Bridge biotechnology
Department, it is public that rat IL-2 (230240922), IL-6 (230641023) ELISA kit are purchased from Hangzhou connection section biology Limited Liability
Department, lymphocyte separation medium (LTS10770125) is purchased from Tianjin Hao oceans science and technology limited Company, the culture medium of RPMI 1640
(1279443) purchased from gibco companies of the U.S., Concanavalin A (C-2010) are purchased from Sigma Co., USA, gap connection resistance
Disconnected agent Gap27 (A1045) is purchased from Apexbio companies, real-time quantitative PCR (real-time quantitative PCR, qRT-
PCR) purchased from Beijing Zhuan Meng biologies company limited, TRIzol is purchased from life techologies companies to reagent, and primer sequence is by giving birth to
The design synthesis of work biological engineering limited company.
1.2 method
1.2.1 rat blood pressure is determined and measures rat tail artery blood pressure using BP-6 non-invasive blood pressure measuring instruments, it is to avoid rat is hot-tempered
Dynamic, the continuous measurement 3 times under the clear-headed rest state of rat, per minor tick at least 1min, takes 3 meansigma methodss of measurement and is rat receipts
Contractive pressure.
1.2.2 the pentobarbital sodium anesthetized rat of sample collection 3%, ethylenediaminetetraacetic acid (EDTA) anticoagulant tube collection abdomen master
Periarterial blood 5ml, 1 500r/min are centrifuged 5min, take blood plasma in -20 DEG C of preservations, and the lower floor's hemocyte after centrifugation is used to separate
Peripheral blood lymphocyte.
1.2.3 inflammatory factor IL-2 and IL-6 expression take it is frozen in -20 DEG C of plasma sample, ELISA method detection IL-2 and
IL-6 expressions, operation is carried out in strict accordance with kit specification.
1.2.4 separation of lymphocytes is by hemocyte and 0.9% sodium chloride solution 1:1 equal-volume dilutes, and piping and druming is mixed.Take
Aseptic 15ml centrifuge tubes, add lymphocyte separation medium, by cell suspension slowly according to 1:1 ratio is added to lymphocyte separation medium
Upper strata, it is ensured that blood is placed in lymphocyte separation medium upper strata.Lymphocyte, low speed centrifuge are separated according to density－gradient centrifuga－tion method
Horizontal centrifugal 30min (1 500r/min delays and rises slow drop).By detached white opacity buffy coat suction out to aseptic 15ml from
In heart pipe, 3 times of 0.9% sodium chloride solution washing is added, 6min is centrifuged with 1 800r/min.Washing 2 times, abandons supernatant, and precipitation is
Lymphocyte.Add 1ml culture medium resuspended, draw 50 μ l to 1.5ml centrifuge tubes, plus 450 μ l phosphate buffers (PBS) dilutions
10 times, counted under microscope is simultaneously active with Trypan Blue observation of cell.Cytoactive is more than 95%, and adjustment cell concentration is
1×106Individual/ml, cell is seeded in 24 orifice plates.
1.2.5 Flow cytometry CD4 +And CD8 +The expression of T cell Cx43 collects 5 × 105Individual cell is interior in EP pipes, 100 μ
L PBS re-suspended cells, are separately added into corresponding streaming antibody (CD4- APC, APC Isotype control；CD8- PE, PE Isotype control) 1 μ l,
And set up negative control.125 μ l fixatives fix lymphocyte, 4 DEG C of incubation 20min, and the anti-1 μ l antibody of Cx43 mono- and 99 μ l 1 ×
Rupture of membranes agent rupture of membranes, 37 DEG C of incubation 20min；FITC bis- resists 1 μ l, 37 DEG C of incubation 20min；The agent of 1 × ruptures of membranes of 1ml is washed, and abandons supernatant
Liquid, PBS re-suspended cells, flow cytomery, streaming CELLQuest software is analyzed.
1.2.6 real-time fluorescence quantitative PCR detection IL-2mRNA and IL-6mRNA expression counts the lymphocyte isolated, and adjusts
Whole culture medium to cell concentration is 1 × 106Individual/ml.Lymphocyte is divided into into blank control group, canavaline
(Concanavalin A, ConA) group (adding 5 μ g/ml ConA after culture 48h) and Gap27+ConA groups (add 5 before culture
00 μm of ol/L Gap275, after culture 48h 5 μ g/ml ConA are added).Add 10% autoserum (serum process step：56℃
Inactivation 30min, with 4 000r/min 30min, 4 DEG C of preservations are centrifuged), in 37 DEG C, 5%CO248h is cultivated in incubator.
TRIzol methods extract the lymphocyte mRNA of above-mentioned culture, and nucleic acid-protein content detection instrument determines A260/A280Value,
1% denaturing formaldehyde gel electrophoresis detection extracts quality.From GADPH as reference gene, according to rat IL-2, IL-6 gene
CDNA sequence, setting primer sequence (being shown in Table 1).MRNA samples, random primer, Rtmix, dNTP mixed liquor, the coke that will be extracted
Diethyl phthalate (DEPC) water prepares reaction system according to reverse transcription system, according to 37 DEG C of reverse transcription 60min, 90 DEG C of inactivation reverse transcriptions
The response procedures of enzyme 5min carry out reverse transcription synthesis cDNA.By 2 × QuantiNova SYBR Green PCR Master Mix,
10 × miScript Nucleics Mix universal primers, template cDNA and RNase-Free water are placed on ice, and gently mix
It is even, reaction system is prepared, according to 95 DEG C of denaturations 2min, 1 circulation；95 DEG C of degeneration 5s, 60 DEG C of annealing plus extension 10s, 40
It is circularly set Bio-Rad quantitative real time PCR Instruments, record IL-2mRNA and IL-6mRNA relative expression quantities.
The real-time fluorescence quantitative PCR primer sequence of table 1
1.3 statistical methods carry out statistical analysis using the softwares of SPSS 18.0, and measurement data is represented with (x ± s), and two
Compare homogeneity of variance person between group using two sample t-tests, Heteroscedasticity person is checked using approximate t；Multigroup is compared employing side
Difference is analysed, and is compared two-by-two between group and is checked using SNK-q.With P<0.05 is that difference is statistically significant.
2 results
2.1SHR and WKY rat systolic pressures compare SHR rats systolic pressure for (211 ± 15) mm Hg (1mm Hg=
0.133kPa), higher than systolic pressure (122 ± 5) the mm Hg of WKY rats, difference statistically significant (t=25.22, P<0.01).
2.2SHR and WKY rat inflammatory factor expression levels compare SHR rat IL-2 expressions for (152.40 ±
29.62) pg/ml, higher than (73.41 ± 20.16) pg/ml of WKY rats, difference statistically significant (t=7.24, P<
0.05)；SHR rat IL-6 expressions are (29.74 ± 1.90) pg/ml, higher than (22.58 ± 1.06) pg/ of WKY rats
Ml, difference statistically significant (t=14.72, P<0.05).
2.3SHR and WKY rat CD4 +And CD8 +T cell Cx43 expresses SHR rat CD4 +T cell Cx43 positive expression rate is
(3.48 ± 0.58) %, higher than Cx43 positive expression rate (1.36 ± 0.18) % of WKY rats, the statistically significant (t=of difference
15.61, P<0.05).SHR rat peripheral blood CD8 +T lymphocyte Cx43 positive expression rate is (39.02 ± 4.77) %, is higher than
Cx43 positive expression rate (27.48 ± 1.90) % of WKY rats, difference statistically significant (t=10.05, P<0.05).
2.4 each group cell IL-2mRNA, IL-6mRNA relative expression levels compare 5 μ g/ml ConA and 500 μm of ol/L
Gap27 stimulates lymphocyte, qRT-PCR detections to find that the expression of ConA active sets IL-2, IL-6 is all remarkably higher than after Gap27 blockings
Reactivation group (P<0.01, n=3), difference has statistical significance (being shown in Table 2).
The SHR rats of table 2 and WKY rat each group cell IL-2mRNA, IL-6mRNA relative expression levels compare (x ± s, n
=3)
The above, preferably specific embodiment only of the invention, protection scope of the present invention not limited to this are any ripe
Those skilled in the art are known in the technical scope of present disclosure, the letter of the technical scheme that can be become apparent to
Altered or equivalence replacement are each fallen within protection scope of the present invention.
SEQUENCE LISTING
<110>Shihezi Univ
<120>The correlation analysis that a kind of peripheral blood lymphocyte connection protein 43 is expressed with IL-2 and IL-6
<160> 6
<170> PatentIn version 3.3
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<211> 20
<212> DNA
<213>Artificial sequence
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ctctctgctc ctccctgttc 20
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<213>Artificial sequence
<400> 2
gccaaatccg ttcacaccg 19
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<213>Artificial sequence
<400> 3
acgcttgtcc tcgcta 16
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<211> 20
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gcacctgtaa gtccagcaac 20
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ttgggactga tgttgttgac 20
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<211> 19
<212> DNA
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<400> 6
tgtgggtggt atcctctgt 19
Claims (2)
1. a kind of correlation analysis that peripheral blood lymphocyte connection protein 43 is expressed with IL-2 and IL-6, its feature exists
In comprising the following steps：
Step 1, rat blood pressure are determined
Rat tail artery blood pressure is measured using BP-6 non-invasive blood pressure measuring instruments, it is to avoid rat is restless, in the clear-headed rest state of rat
Lower continuous measurement 3 times, per minor tick at least 1min, takes 3 meansigma methodss of measurement and is rat systolic pressure；
Step 2, sample collection
3% pentobarbital sodium anesthetized rat, ethylenediaminetetraacetic acid anticoagulant tube collection ventral aorta peripheral blood 5ml, 1 500r/min
Centrifugation 5min, takes blood plasma in -20 DEG C of preservations, and the lower floor's hemocyte after centrifugation is used to separate peripheral blood lymphocyte；
Step 3, inflammatory factor IL-2 and IL-6 expression
Frozen in -20 DEG C of plasma sample, ELISA method detection IL-2 and IL-6 expression is taken, is operated in strict accordance with test kit
Description is carried out；
Step 4, separation of lymphocytes
By hemocyte and 0.9% sodium chloride solution 1:1 equal-volume dilutes, and piping and druming is mixed；Aseptic 15ml centrifuge tubes are taken, lymph is added
Cell separation liquid, by cell suspension slowly according to 1:1 ratio is added to lymphocyte separation medium upper strata, it is ensured that it is thin that blood is placed in lymph
Born of the same parents' separating liquid upper strata；Lymphocyte, low speed centrifuge horizontal centrifugal 30min, 1 500r/ are separated according to density－gradient centrifuga－tion method
Min, delays and rises slow drop；Detached white opacity buffy coat is suctioned out to aseptic 15ml centrifuge tubes, 3 times of 0.9% chlorine is added
Change sodium solution washing, 6min is centrifuged with 1 800r/min；Washing 2 times, abandons supernatant, and precipitation is lymphocyte；Add 1ml culture medium
It is resuspended, 50 μ l to 1.5ml centrifuge tubes are drawn, plus 450 μ l phosphate buffers dilute 10 times, counted under microscope simultaneously uses trypan blue
Dyeing observation of cell activity；Cytoactive is more than 95%, and adjustment cell concentration is 1 × 106Individual/ml, by cell 24 are seeded in
In orifice plate；
Step 5, Flow cytometry CD4 +And CD8 +The expression of T cell Cx43 collects 5 × 105Individual cell is interior in EP pipes, 100 μ l
PBS re-suspended cells, are separately added into corresponding streaming antibody 1 μ l, CD4- APC, APC Isotype control；CD8- PE, PE Isotype control, and set
Vertical negative control；125 μ l fixatives fix lymphocyte, 4 DEG C of incubation 20min, the anti-1 μ l antibody of Cx43 mono- and 99 μ 1 × ruptures of membranes of l
Agent rupture of membranes, 37 DEG C of incubation 20min；FITC bis- resists 1 μ l, 37 DEG C of incubation 20min；The agent of 1 × ruptures of membranes of 1ml is washed, and abandons supernatant, PBS
Re-suspended cell, flow cytomery, streaming CELLQuest software is analyzed；
Step 6, real-time fluorescence quantitative PCR detection IL-2mRNA and IL-6mRNA expression count the lymphocyte isolated, adjustment
Culture medium to cell concentration is 1 × 106Individual/ml；Lymphocyte is divided into into blank control group, canavaline group and Gap27+ConA
Group；Add 10% autoserum, serum process step：56 DEG C of inactivation 30min, with 4 000r/min 30min is centrifuged, and 4 DEG C preserve,
In 37 DEG C, 5%CO248h is cultivated in incubator；
TRIzol methods extract the lymphocyte mRNA of above-mentioned culture, and nucleic acid-protein content detection instrument determines A260/A280Value, 1% first
Aldehyde denaturing gel electrophoresis Detection and Extraction quality；From GADPH as reference gene, according to rat IL-2, IL-6 gene cDNA sequence
Row, set primer sequence；By extract mRNA samples, random primer, Rtmix, dNTP mixed liquor, pyrocarbonic acid diethyl ester water according to
Reverse transcription system prepares reaction system, and according to 37 DEG C of reverse transcription 60min, the response procedures of 90 DEG C of inactivation reverse transcription 5min are carried out
Reverse transcription synthesizes cDNA；By 2 × QuantiNova SYBR Green PCR Master Mix, 10 × miScript
Nucleics Mix universal primers, template cDNA and RNase-Free water are placed on ice, and gently mix, and prepare reactant
System, according to 95 DEG C of denaturations 2min, 1 circulation；95 DEG C of degeneration 5s, 60 DEG C of annealing plus extension 10s, 40 are circularly set Bio-
Rad quantitative real time PCR Instruments, record IL-2mRNA and IL-6mRNA relative expression quantities.
2. the correlation analysiss that peripheral blood lymphocyte connection protein 43 according to claim 1 is expressed with IL-2 and IL-6
Method, it is characterised in that Gap27 makes lymphocyte inflammatory factor IL-2mRNA and IL-6mRNA down-regulated expression, points out gap to connect
Blocker is connect by changing Cx43 channel designs or suppressing intracellular Ca2+ concentration and then change passage permeability, it is thin so as to affect
The release of intracellular cytokine.
Priority Applications (1)
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| CN111208306A (en) * | 2020-03-28 | 2020-05-29 | 安徽大学 | Method for measuring expression quantity of nicotinic acetylcholine receptor in rat lymphocyte |
| CN114438017A (en) * | 2022-02-18 | 2022-05-06 | 中山大学附属第一医院 | Correlation analysis method of endothelial progenitor cell function and cardiac function of sepsis patient and application of correlation analysis method in diagnosis of sepsis cardiomyopathy |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111208306A (en) * | 2020-03-28 | 2020-05-29 | 安徽大学 | Method for measuring expression quantity of nicotinic acetylcholine receptor in rat lymphocyte |
| CN114438017A (en) * | 2022-02-18 | 2022-05-06 | 中山大学附属第一医院 | Correlation analysis method of endothelial progenitor cell function and cardiac function of sepsis patient and application of correlation analysis method in diagnosis of sepsis cardiomyopathy |
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