CN110850086A - Application of serum diagnosis marker ACLY for ischemic biliary tract lesion after liver transplantation - Google Patents
Application of serum diagnosis marker ACLY for ischemic biliary tract lesion after liver transplantation Download PDFInfo
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Abstract
The invention discloses an application of a serum polypeptide molecular diagnostic marker ACLY for ischemic biliary tract lesion after liver transplantation, wherein the amino acid sequence of the serum polypeptide molecular diagnostic marker ACLY is shown in SEQ.ID.NO. 1. This molecule is called ACLY, an ATP citrate lyase, with an exact molecular weight of 1949.9 daltons. The ACLY presents specific low expression in the serum detection of patients with ischemic biliary tract diseases after liver transplantation, and the expression level of the ACLY is detected by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) or an ELISA method, so that the method can be used as the detection method of the serum of patients with ischemic biliary tract diseases after liver transplantation.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and relates to application of a serum polypeptide molecular diagnostic marker ACLY for ischemic biliary tract lesion after liver transplantation.
Background
Since the first application of Liver Transplantation (LT) technology to clinic by Starzl in 1963, liver transplantation has become the only effective means for treating end-stage liver diseases at present after half a century of development. With the development of new immunosuppressive agents and the maturation of surgical techniques, rejection and surgical-technique-related complications have been significantly reduced, and 1 year postoperative mortality has fallen below 5%. The ischemic biliary tract disease (ITBL) refers to the condition that non-anastomotic segmental stenosis, dilatation and intrahepatic bile duct disappearance appear on a transplanted hepatic bile duct tree due to the limited or diffuse bile duct damage caused by the blood supply disorder of the bile duct, and finally mechanical obstruction and secondary biliary tract infection of the bile duct are caused, and the incidence rate is 2-19%; because the etiology and pathogenesis are not clear, early diagnosis is difficult, clinical treatment is troublesome, and the method becomes one of the main reasons influencing the long-term survival of liver transplantation patients and causing graft loss. Therefore, the research on the precise molecular mechanism of the occurrence and development of the ITBL is carried out, and the search for a novel specific diagnostic marker has important clinical significance for the early diagnosis and treatment of ITBL, the improvement of the curative effect of liver transplantation and the improvement of the life quality of patients.
Before any pathological change occurs in any disease, the intracellular proteins are altered in composition and quantity and are reflected by the pattern of proteins in the serum. Therefore, by comparing the expression of different proteins in the serum of different disease populations, it is possible to screen out disease-related marker molecules. Serum proteomics refers to the study of all proteins expressed in the serum of a selected target population, and on the basis of establishing a normal Protein Expression Map (PEM), differential protein spots of the proteins are searched, and disease-related proteins are identified, so that the structure and function of the proteins are further studied, and a new way is developed for studying major disease pathophysiological mechanisms, specific markers for early diagnosis, drug action targets and the like. A large amount of proteins and polypeptides exist in human serum, and the existence, deletion and expression of partial proteins and polypeptides are closely related to the health degree of human beings, so that the human serum becomes a biomarker for disease diagnosis.
Serum diagnosis is considered to be the most recent and effective method for early diagnosis of cancer. The method judges the occurrence and development of the tumor by searching tumor markers in blood, particularly protein markers in the blood, thereby realizing early diagnosis of the tumor. A large amount of proteins and polypeptides exist in human serum, and the existence, deletion and expression of partial proteins and polypeptides are closely related to the health degree of human beings, so that the human serum becomes a biomarker for disease diagnosis.
Currently, screening for ischemic biliary lesions after liver transplantation lacks a sensitive and specific diagnostic method.
Disclosure of Invention
The invention aims to provide a serum polypeptide molecular diagnostic marker for ischemic biliary tract lesion after liver transplantation and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention discloses an application of a serum polypeptide molecular diagnostic marker ACLY in preparing a serum diagnostic medicine for ischemic biliary tract lesion after liver transplantation, wherein the amino acid sequence of the serum polypeptide molecular diagnostic marker is shown in SEQ.ID.NO. 1.
Preferably, the serum polypeptide molecular diagnostic marker ACLY is ATP citrate lyase and has a molecular weight of 1949.9 daltons.
Preferably, the detection parameter of the serum polypeptide molecular diagnostic marker ACLY in serum is 456.07-802.50 pg/mL.
Preferably, the serum diagnosis drug for ischemic biliary tract diseases after liver transplantation is a serum polypeptide molecular drug for detecting ischemic biliary tract diseases after liver transplantation by ELISA.
The invention also discloses the application of the molecule combined with the serum polypeptide molecular diagnostic marker ACLY in preparing a serum diagnostic medicine for ischemic biliary tract lesion after liver transplantation.
The invention also discloses the application of the molecule combined with the ACLY protein in the preparation of the serum diagnosis medicament for ischemic biliary tract lesion after liver transplantation.
Compared with the prior art, the invention has the following beneficial effects:
the invention discloses a serum polypeptide molecule of ischemic biliary tract lesion after liver transplantation, the amino acid sequence of the serum polypeptide molecule is shown in SEQ.ID.NO.1, and the molecule is called ACLY. ACLY is ATP citrate lyase, with an exact molecular weight of 1949.9 daltons. ACLY exhibits specifically low expression in serum detection in patients with ischemic biliary lesions after liver transplantation: the range of expression in serum in normal healthy people is: 863.21-1291.79 pg/mL; the range of expression in sera of patients with good postoperative recovery was: 631.07-1006.79 pg/mL; the expression range in the serum of patients with ischemic biliary tract lesion after liver transplantation is as follows: 456.07-802.50pg/mL, and the expression between different groups has very significant difference (p < 0.01).
In view of the specific low expression of ACLY in serum of ischemic biliary tract lesion after liver transplantation, ACLY can be used as a serum diagnostic marker of ischemic biliary tract lesion after liver transplantation; the method for detecting the expression level of the ACLY by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) or an ELISA method can be used as a method for detecting patients with ischemic biliary tract lesions after liver transplantation. For the serum diagnosis of ischemic biliary tract lesion after liver transplantation by ELISA, ACLY can be used as a new target point of ELISA detection medicines.
Drawings
FIG. 1 is a diagram showing the results of Flex analysis of specifically low-expressed 7 protein polypeptides in serum of patients with ischemic biliary tract diseases: through the expression comparison of M/Z:1949.9 in patients with ischemic biliary tract diseases (red), patients with good postoperative recovery (green) and normal healthy people (blue), the protein polypeptide peak pattern of M/Z:1949.9 is found to be remarkably low expressed in the serum of patients with ischemic biliary tract diseases, so that the sequence identification is carried out on the protein polypeptide peak pattern and the protein polypeptide peak pattern is preferably further identified as a marker.
FIG. 2 shows the result of gel chromatography separation of serum protein polypeptides from patients with ischemic biliary tract diseases: the abscissa in the chromatogram represents the sample outflow time, the ordinate represents the relative abundance of the polypeptide, the set time of the chromatogram is 78min, fractions are collected from 5min, the polypeptide components are mainly separated after 6min, a gradient elution mode is adopted, the elution efficiency is improved, and the capture time is set to collect the fractions: 25 fractions of the peptide fragment were collected and the peak for the 1.9K Da peptide of interest flowed out at about 62 min.
FIG. 3 is a MS/MS mass spectrometric identification profile of ACLY.
FIG. 4 is the expression levels of ACLY protein in different groups: the results of enzyme-linked immunosorbent assay analysis show that the expression levels of ACLY in different detection groups are normal healthy people, good postoperative recovery patients, patients with ischemic biliary tract lesion after liver transplantation, and the three groups have significant difference.
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It is noted that the terms "comprises" and "comprising," and any variations thereof, as used in the description and claims of this invention, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the accompanying drawings:
the serum polypeptide molecule for ischemic biliary tract lesion after liver transplantation is a newly screened serum diagnosis marker for ischemic biliary tract lesion after liver transplantation, has specificity in expression, and can be applied to diagnosis of ischemic biliary tract lesion after liver transplantation.
The specific screening of the serum diagnosis marker for ischemic biliary tract lesion after liver transplantation comprises the following steps:
firstly, separating and extracting serum protein polypeptides of an ischemic biliary tract pathological change patient after liver transplantation, a good postoperative recovery patient and a normal healthy population by using a liquid protein chip technology, capturing the serum protein polypeptide spectra of the ischemic biliary tract pathological change patient after liver transplantation by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology, the serum protein polypeptide spectra of the good postoperative recovery patient and the normal healthy population, comparing and analyzing the serum protein polypeptide spectra of the ischemic biliary tract pathological change patient after liver transplantation by using ClinProTools2.1 software, finding out protein polypeptide molecules which are remarkably and differentially expressed among groups, and screening out the serum marker of the ischemic biliary tract pathological change after liver transplantation from the protein polypeptide peak which is remarkably and lowly expressed in the serum of the ischemic biliary tract pathological change patient after liver transplantation.
The screened polypeptide serum diagnostic markers for the ischemic biliary tract lesion after liver transplantation are verified as follows:
the protein polypeptide mixture separated from the serum of the ischemic biliary tract lesion patient after the liver transplantation is divided into 20-30 components by HPLC, secondary mass spectrum identification is carried out on the protein polypeptide mixture, an enzyme-linked immunosorbent assay is carried out on the identified protein polypeptide, and serum regression verification results prove that the protein polypeptide mixture is remarkably low in expression and specific in the serum of the ischemic biliary tract lesion patient after the liver transplantation and can be used as a biomarker for screening the serum of the ischemic biliary tract lesion patient after the liver transplantation.
1. Collecting and processing a sample:
9 patients with ischemic biliary lesions after cholangiography (7 men; 2 women; mean age 53 years) collected from hepatobiliary surgery (3 months to 12 months at 2016) of the first subsidiary hospital of the university of transport, west-ampere, 10 patients with good postoperative recovery (8 men; 2 women; mean age 51 years) and 10 normal healthy populations (7 men; 3 women; mean age 50 years). The sample considers factors such as age, sex, collection time, whether the storage conditions are consistent, whether basic diseases exist and the like. Collecting blood of the collected person with fasting state, collecting 5mL whole blood with vacuum blood collection tube (yellow cap, with isolation gel), and standing at room temperature for 30 min; centrifuging at room temperature for 5min (3000g), subpackaging the upper layer serum into 100 μ L/tube, immediately storing at-80 deg.C, and avoiding repeated freeze thawing.
Reagents and instrumentation:
the serum proteins were extracted using the magnetic bead kit "weak cationic" (MB-WCX) from Bruker, Germany, and spectrally pure (HPLC grade) acetonitrile, trifluoroacetic acid (Merck, Germany), α -cyano-4-hydroxycinnamic acid (HCCA) (Sigma, USA).
2. Preparation of serum protein samples
Capturing serum protein polypeptide by weak cation (MB-WCX) magnetic beads, which comprises the following specific operation steps:
① mixing the magnetic bead suspension completely for 1 min;
② adding 10 μ L MB-WCX binding solution and 10 μ L MB-WCX magnetic bead into PCR tube, mixing, adding 5 μ L serum, mixing for at least 5 times, and standing for 5 min;
③ placing PCR tube into magnetic column separator to make magnetic beads adhere to wall for 1min, and removing supernatant after the liquid is clear;
④ adding 100 mu LMB-WCX washing liquid, moving the PCR tube on the magnetic column separator back and forth for 10 times, removing the supernatant after the magnetic beads are attached to the wall, repeating steps ③ and ④ twice;
⑤ washing adherent magnetic beads with 5 μ LMB-WCX eluent, repeatedly blowing for 10 times to make the magnetic beads adhere to the wall for 2min, and transferring the supernatant into a clean centrifuge tube;
⑥ adding 5 μ LMB-WCX stable solution into centrifuge tube and mixing, the extracted protein polypeptide can be used for direct MALDI-TOF-MS detection or frozen in refrigerator at-20 deg.C for 24h mass spectrum analysis.
Mass spectrometry analysis:
mixing 1 μ L of protein sample obtained by separation and collection with 10 μ L of substrate α -cyano-4-hydroxycinnamic acid, placing 1 μ L of the mixture on an Anchorchip target plate (Bruker, Germany), placing each sample with three targets for three times, drying at room temperature, putting the target plate into a mass spectrometer for flight time mass spectrometry, correcting standard products by FlexControl 2.0 software, starting sample detection, generating a mass spectrogram after each sample is subjected to laser targeting for 300 times (5 times of point targeting and 2 × 30 times of targeting each time), obtaining a protein polypeptide spectrogram consisting of different mass-to-nuclear ratios (m/z), analyzing protein polypeptide spectrums of two groups of serum samples by ClinProTools2.1 software in combination with a genetic algorithm and other biological statistics and bioinformatics methods, carrying out normalization smoothing treatment on the total ion flow chart, eliminating chemical and electrical physical noise, analyzing proteins with difference between groups, calculating the difference size, arranging the proteins from large to small according to find out protein peak expression with difference (P < 0.001).
After blood serum samples of ischemic biliary tract lesion patients, postoperative recovery patients and normal healthy people after liver transplantation are processed by a magnetic bead separation system and analyzed by MALDI-TOF-MS, protein polypeptide maps of each sample of ischemic biliary tract lesion patients, postoperative recovery patients and normal healthy people after liver transplantation are drawn, 76 protein polypeptide peak maps are detected in a molecular weight range of 1000 Da-10000 Da, and the three-time repeated stability of each sample is high.
Analyzing serum protein polypeptide spectra of ischemic biliary tract pathological patients, postoperative recovery good patients and normal healthy people after liver transplantation captured by mass spectrometry by adopting ClinProTools2.1 software, and comparing and analyzing the serum polypeptide spectra of ischemic biliary tract pathological patients and postoperative recovery good patients with normal healthy people after liver transplantation to detect 9 protein polypeptide peak maps with significant differences (P <0.05), wherein 4 protein polypeptide peak maps have significant differences among three groups (P <0.001), wherein 7 protein polypeptides are significantly reduced in expression of ischemic biliary tract pathological patients after liver transplantation, and the expression of the rest 2 protein polypeptides is significantly increased in ischemic biliary tract pathological patients after liver transplantation, which is specifically shown in Table 1:
when 7 specific low-expression protein polypeptides in the serum of the ischemic biliary tract lesion patient in table 1 are subjected to Flexalysis software analysis, the results are shown in fig. 1, and the expression comparison of M/Z:1949.9 in the serum of the ischemic biliary tract lesion patient (red), the serum of a patient with good postoperative recovery (green) and the serum of a normal healthy population (blue) shows that the protein polypeptide peak diagram of M/Z:1949.9 is remarkably low-expression in the serum of the ischemic biliary tract lesion patient, so that the sequence identification is carried out on the protein polypeptide peak diagram and the first-choice further identification of the protein polypeptide as a marker is carried out.
3. Sequence identification of serum potential marker of ischemic biliary tract lesion after liver transplantation
Specifically, a liquid chromatography separation and mass spectrometry combined technology is adopted to identify a serum polypeptide marker M/Z:1949.9 of a patient with ischemic biliary tract lesion after liver transplantation, and two-dimensional gel chromatography separation is carried out on the serum protein polypeptide remaining after the magnetic bead separation and collection of mass spectrometry sample loading by using Nano Acity UPLC of Waters company, and 25 peptide fragment fractions are collected: the 1.9kDa peak was shed at about 62min and detected in the collection; and then, the sequence identification is carried out on protein polypeptide M/Z:1949.9 which is expressed and reduced in the serum of the patient with the ischemic biliary tract lesion after the liver transplantation by using an LTQ Orbitrap XL mass spectrum system of Thermo Fisher company.
3.1 sample Pre-treatment
Mixing the extracted protein samples, converting the protein samples to 13300 revolutions for 5 minutes, taking supernatant, drying the supernatant by a freeze dryer to ensure that the final volume is 50ul to obtain liquid A, extracting the liquid A by an Agilent zip extraction column, and concentrating the liquid A. the treatment method comprises the steps of blowing and beating ① zip columns for 5 times by 100% acetonitrile to activate the columns, repeatedly blowing and sucking ② activated zip columns in the liquid 1 for 10 times to avoid bubbles as much as possible, washing 3 times of the zip columns by ③ 40% ACN 0.2% TFA aqueous solution, repeatedly blowing and sucking ④ and zip columns in 0.2% TFA to elute a sample eluent, repeating the steps 1-4 and 30 times by ⑤, merging 30 times of eluent 2 by ⑥, freezing and drying the eluent to 10ul for mass spectrum identification.
3.2 chromatographic separation
The original sample was added with 10ul of mobile phase A and transferred to a sample vial for a total of 20 ul.
One-dimensional ultra-high performance liquid phase system: nano Aquity UPLC (Waters Corporation, Milford, USA). A chromatographic column:
a trapping column:
C18,3μm,0.10×20mm,nanoAcquityTMColumn
and (3) analyzing the column:
C18,1.9μm，0.15×120mm,nanoAcquityTMColumn
mobile phase A: 5% acetonitrile, 0.1% formic acid in water
The mobile phase B is 95 percent of acetonitrile and 0.1 percent of aqueous solution of formic acid; all solutions were HPLC grade.
The collecting flow rate is 600nl/min, the collecting time is 3min, and the analysis flow rate is 400 nl/min; the analysis time is 60min, and the temperature of a chromatographic column is 35 ℃; the Partial Loop mode was injected in a volume of 18. mu.l.
The gradient elution procedure is shown in table 2 below:
TABLE 2
| Time (min) | A | B | Flow rate (nL/min) |
| 0 | 95％ | 5％ | 600 |
| 16 | 90％ | 10％ | 600 |
| 51 | 78％ | 22％ | 600 |
| 71 | 70％ | 30％ | 600 |
| 72 | 5％ | 95％ | 600 |
| 78 | 5％ | 95％ | 600 |
The results of gel chromatography are shown in FIG. 2. The abscissa in the chromatogram represents the sample outflow time, the ordinate represents the relative abundance of the polypeptide, the set time of the chromatogram is 78min, fractions are collected from 5min, the polypeptide components are mainly separated after 6min, a gradient elution mode is adopted, the elution efficiency is improved, and the capture time is set to collect the fractions: 25 fractions of the peptide fragment were collected and the peak for the 1.9K Da peptide of interest flowed out at about 62 min.
3.3 LTQ-Orbitrap XL Mass Spectrometry
Q-exact HF mass spectrometry system (Thermo Fisher Co., Ltd.) was used. Nano liter flow rate HPLC liquid phase system Easy nLC 1000, spray voltage 1.8 kV; the mass spectrum scanning time is 78 min; the experimental modes are data dependence (DataDependency) and Dynamic Exclusion (Dynamic Exclusion), and parent ions are added into an Exclusion list for 80 seconds after being cascaded for 2 times within 60 seconds; the scanning range is 300-1400 m/z; the first-order scan (MS) uses obirrap with a resolution set to 70000; CID and secondary scanning use LTQ; selecting single isotope of 20 ions with the strongest intensity as parent ion in MS spectrogram to carry out MS/MS (single charge exclusion without being used as parent ion).
And (3) data analysis: database searching identification is carried out by using data analysis software mascot incorporation PD 2.0. The error of parent ion is set to be +/-15 ppm, the error of fragment ion is set to be 20mmu, the enzyme cutting mode is non-enzyme cutting, and the variable modification is M (Methionine) methionine oxidation. The search result parameter is set to be equal to or less than 0.01.
The retrieval result is as follows: 1950.106 for m/z; uniprot, P53396; gene Symbol ═ ACLY; the sequence is as follows: iliiggsanftnvaatfk.g.
1949.9 is called ACLY, which is an ATP citrate lyase with an exact molecular weight of 1949.9 daltons and the amino acid sequence: iliiggsanftnvaatfk.g (as shown in seq. id. No. 1).
Therefore, the method for detecting the expression level of the ACLY by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) or an ELISA method can be used as a method for detecting patients with ischemic biliary tract lesions after liver transplantation.
It is suggested that ACLY is a protein specifically associated with ischemic biliary lesions after liver transplantation, and further verified by ELISA detection.
4. ELISA serum verification analysis of ischemic biliary tract lesion serum ACLY expression after liver transplantation
1) Serum samples: serum verification analysis by ELISA was performed by collecting 9 cases (8 cases in males, 1 case in females; mean age 52) of serum from patients with ischemic biliary tract disease after liver transplantation, 10 cases (8 cases in males, 2 cases in females; mean age 48) of serum from patients with good postoperative recovery, and 10 cases (7 cases in males, 3 cases in females; mean age 55.3) of serum from normal healthy population. All serum samples come from Xian traffic university's first subsidiary college, and the collection time is 1 month in 2017 to 9 months in 2017;
2) the detection method comprises the following steps: the expression level of serum ACLY of patients with ischemic biliary tract diseases, patients with good postoperative recovery and normal healthy people after liver transplantation is detected by adopting an enzyme-linked immunosorbent assay (ELISA), and the kit is purchased from American R & D company. The kit adopts a double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA): to the coated microwells previously coated with anti-human ACLY protein (ACLY) antibody, the specimen, standard, HRP-labeled detection antibody were added in sequence, incubated and washed thoroughly. The color is developed with the substrate TMB, which is converted to blue by the catalysis of peroxidase and to the final yellow color by the action of an acid. The shade of color is positively correlated with the ACLY protein (ACLY) in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated. The specific experimental steps refer to the kit specification, and the positive judgment standard is defined according to the kit specification;
3) the statistical method comprises the following steps: one-way analysis of variance (ANOVA) and T-test of independent samples were performed using graphpad. prism. v5.01 software;
4) and (4) analyzing results: the results of the analysis by the enzyme-linked immunosorbent assay show that the expression levels of ACLY in different detection groups are normal healthy people, good postoperative recovery patients, and patients with ischemic biliary tract lesions after liver transplantation, and the specific results are shown in table 3 and fig. 4, wherein the differences between two groups in the three groups are significant.
TABLE 3
The result of ELISA detection of ACLY in serum of patients with ischemic biliary tract disease after liver transplantation, patients with good postoperative recovery and normal healthy people shows that the expression of ACLY has specificity: the range of expression in serum in patients with ischemic biliary tract lesions after liver transplantation was: 456.07-802.50 pg/mL; the range of expression in sera of patients with good postoperative recovery was: 631.07-1006.79 pg/mL; the expression range in the serum of patients with normal healthy population is as follows: 863.21-1291.79 pg/mL, and the expression between different groups is significantly different (p < 0.01). This indicates that: ACLY is a protein closely related to a patient with ischemic biliary tract lesion after liver transplantation, and can be used as a primary detection index of ischemic biliary tract lesion after liver transplantation. Therefore, the ACLY expression of the serum sample to be detected can be preliminarily judged to be a patient with ischemic biliary tract lesion after liver transplantation (456.07-802.50pg/mL), a patient with good postoperative recovery (631.07-1006.79 pg/mL) or a normal healthy population (863.21-1291.79 pg/mL) through an ELISA experiment.
The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.
Sequence listing
<110> university of west ampere traffic
Application of serum polypeptide molecular diagnostic marker ACLY for ischemic biliary tract lesion after liver transplantation
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>1
Ser Pro Pro Met Gly Leu Val Val Ala Pro Thr Gly Leu
1 5 10
Claims (6)
1. The application of a serum polypeptide molecular diagnostic marker ACLY in preparing a serum diagnostic medicine for ischemic biliary tract lesion after liver transplantation is characterized in that the amino acid sequence of the serum polypeptide molecular diagnostic marker is shown in SEQ ID No. 1.
2. The use of claim 1, wherein the serum polypeptide molecular diagnostic marker ACLY is ATP citrate lyase and has a molecular weight of 1949.9 daltons.
3. The use of claim 1, wherein the serum polypeptide molecular diagnostic marker ACLY has a detection parameter in serum of 456.07-802.50 pg/mL.
4. The use of claim 1, wherein the serum diagnostic agent for ischemic biliary tract diseases after liver transplantation is a serum polypeptide molecule agent for ELISA detection of ischemic biliary tract diseases after liver transplantation.
5. Use of a molecule that binds to the serum polypeptide molecular diagnostic marker ACLY of claim 1 for the preparation of a serum diagnostic medicament for ischemic biliary lesions after liver transplantation.
6. The application of the molecule combined with the ACLY protein in preparing a serum diagnosis medicament for ischemic biliary tract lesion after liver transplantation.
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