CN105567862B - CDK18 is preparing the purposes in diagnosis of coronary heart disease product - Google Patents
CDK18 is preparing the purposes in diagnosis of coronary heart disease product Download PDFInfo
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Abstract
The invention discloses the molecular markers that CDK18 gene can be used as diagnosis of coronary heart disease.Research of the present invention has shown that be compared with normal people, and the mRNA expression of CDK18 gene is remarkably decreased in patients with coronary heart disease blood.According to the existing correlation between CDK18 gene and coronary heart disease, the kit of diagnosis of coronary heart disease can be prepared, the kit can be by the mRNA level in-site of CDK18 gene in detection subject's blood to determine whether coronary heart disease occurs, which can clinically be widely applied.
Description
Technical field
The invention belongs to molecular diagnosis fields, are related to a kind of molecular marker for diagnosis of coronary heart disease, and in particular to blood
Application of the molecular marker-CDK18 gene in the product for preparing diagnosis of coronary heart disease in liquid.
Background technique
Atherosclerotic Cardio-Cerebrovascular Diseases have become the major health concern of world wide concern.The world in 2004
Health organization (WHO) report display, the annual cardiovascular disease in the whole world caused death toll based on coronary heart disease and cerebral apoplexy
Up to up to 17,200,000, the one third of all death tolls is accounted for.It is expected that this number of the year two thousand twenty will be further increased 50%, up to
25000000, cardiovascular disease is global human " number one killer ".It is also shown in the extensive perspective study that China carries out
Show, heart disease has become the major causes of death of Chinese population, point column male, the 2nd of women die reason and the 1st.
500,000 people of myocardial infarction is newly sent out in China every year, as living-pattern preservation and the relevant risk factor of atherosclerosis are held
Continuous to increase, coronary heart disease and myocardial infarction morbidity will yet be in continue ascendant trend.
A large amount of research data shows that coronary heart disease is a kind of complex disease, is long by multiple minor genes and environmental factor
Caused by phase interaction.Therefore tumor susceptibility gene relevant to coronary heart disease or Disease-causing gene are identified, is further screened in crowd
The tumor susceptibility gene for increasing disease risks determines susceptible individual, it will help the onset risk prediction of coronary heart disease, new drug development, diagnosis
And individualized treatment.
Summary of the invention
In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a kind of coronary disease disease early diagnosis of can be used for
Molecular marker.Compared to the diagnostic method of traditional coronary heart disease, carry out diagnosis of coronary heart disease with timeliness, spy using gene marker
Anisotropic and sensitivity, for risk height, takes corresponding prevention to make patient that can know disease risks in disease early stage
And remedy measures.
To achieve the goals above, the present invention adopts the following technical scheme:
The present invention provides application of the product of detection CDK18 gene expression in the tool for preparing diagnosis of coronary heart disease.
Further, the product of detection CDK18 gene expression mentioned above includes: by RT-PCR, real-time quantitative
PCR, immune detection, in situ hybridization, chip or high-flux sequence detection of platform CDK18 gene expression dose are with diagnosis of coronary heart disease
Product.
Further, the product with RT-PCR diagnosis of coronary heart disease includes at least drawing for a pair of of specific amplified CDK18 gene
Object；The product with real-time quantitative PCR diagnosis of coronary heart disease includes at least the primer of a pair of of specific amplified CDK18 gene；It is described
Product with immune detection diagnosis of coronary heart disease includes: the antibody in conjunction with CDK18 protein-specific；It is described to be diagnosed in situ hybridization
The product of coronary heart disease includes: the probe with the nucleic acid array hybridizing of CDK18 gene；The product packet with chip diagnosis of coronary heart disease
It includes: protein chip and genetic chip；Wherein, protein chip includes the antibody in conjunction with CDK18 protein-specific, genetic chip packet
Include the probe with the nucleic acid array hybridizing of CDK18 gene.
In specific embodiments of the present invention, the product with real-time quantitative PCR diagnosis of coronary heart disease includes at least one
To the sequence of the primer of specific amplified CDK18 gene as shown in SEQ ID NO.3 and SEQ ID NO.4.
Preferably, the diagnostic tool includes chip, kit, test paper or high-flux sequence platform.Wherein, high pass measures
Sequence platform is a kind of special diagnostic tool, and the product of detection CDK18 gene expression can be applied to the platform and realize to CDK18
The detection of the expression of gene.It, will be to the building of the gene expression profile of a people with the development of high throughput sequencing technologies
For very easily work.Which by comparing the gene expression profile of Disease and normal population, it is easy that gene analyzed
It is abnormal related to disease.Therefore, know that the exception of CDK18 gene is related to coronary heart disease in high-flux sequence and also belong to CDK18
The purposes of gene, equally within protection scope of the present invention.
The present invention also provides a kind of tool of diagnosis of coronary heart disease, the diagnostic tool include chip, kit, test paper,
Or high-flux sequence platform.
Wherein, the chip includes genetic chip, protein-chip；The genetic chip includes solid phase carrier and fixation
In the oligonucleotide probe of solid phase carrier, the oligonucleotide probe includes for detecting being directed to for CDK18 gene transcription level
The oligonucleotide probe of CDK18 gene；The protein-chip includes solid phase carrier and the CDK18 egg for being fixed on solid phase carrier
White specific antibody；The genetic chip can be used for detecting multiple genes including CDK18 gene (for example, and coronary disease
The relevant multiple genes of disease) expression.The protein-chip can be used for detecting multiple eggs including CDK18 albumen
The expression of white matter (such as multiple protein relevant to coronary heart disease).By the way that multiple markers with coronary heart disease are examined simultaneously
It surveys, is greatly improved the accuracy rate of diagnosis of coronary heart disease.
Wherein, the kit includes gene detecting kit and protein immunization detection kit；The genetic test examination
Agent box includes the reagent for detecting CDK18 gene transcription level；The protein immunization detection kit includes CDK18 albumen
Specific antibody.Further, the reagent includes using RT-PCR, real-time quantitative PCR, immune detection, in situ hybridization or chip
Method detects reagent needed for CDK18 gene expression dose process.Preference, the reagent include for CDK18 gene
Primer and/or probe.It is easy to design according to the nucleotide sequence information of CDK18 gene and can be used for detecting CDK18 gene table
Up to horizontal primer and probe.
It can be DNA, RNA, DNA-RNA chimera, PNA or other with the probe of the nucleic acid array hybridizing of CDK18 gene
Derivative.There is no limit for the length of the probe, as long as completing specific hybrid, being specifically bound with purpose nucleotide sequence,
Any length is ok.The length of the probe can be as short as 25,20,15,13 or 10 bases longs.Equally, the probe
Length can be grown to 60,80,100,150,300 base-pairs or longer or even whole genes.Since different probe lengths is to miscellaneous
Efficiency, signal specificity is handed over to have different influences, the length of the probe is typically at least 14 base-pairs, and longest does not surpass generally
30 base-pairs are crossed, the length complementary with purpose nucleotide sequence is best with 15-25 base-pair.The probe self-complementary sequence
Column are most preferably less than 4 base-pairs, in order to avoid influence hybridization efficiency.
The high-flux sequence platform includes the reagent for detecting CDK18 gene expression dose.
The test paper includes test paper carrier and the oligonucleotides that is fixed on test paper carrier, and the oligonucleotides is able to detect
The transcriptional level of CDK18 gene.
Further, the specific antibody of the CDK18 albumen includes monoclonal antibody, polyclonal antibody.The CDK18 egg
White specific antibody include complete antibody molecule, antibody any segment or modification (for example, chimeric antibody, scFv,
Fab, F (ab ') 2, Fv etc..As long as the segment can retain the binding ability with CDK18 albumen.For protein level
Antibody preparation when well known to a person skilled in the art and the present invention may use any method to prepare the antibody.
In specific embodiments of the present invention, the primer sequence for CDK18 gene is as follows: forward primer sequence
As shown in SEQ ID NO.3, reverse primer is as shown in SEQ ID NO.4.
It include but is not limited to blood, tissue fluid, urine for the CDK18 gene of diagnosis of coronary heart disease and its source of expression product
Liquid, saliva, spinal fluid etc. can obtain the body fluid of genomic DNA.In specific embodiments of the present invention, for diagnosing coronary disease
The CDK18 gene of disease and its source of expression product are blood.
In the context of the present invention, " CDK18 gene " includes any function etc. of CDK18 gene and CDK18 gene
The polynucleotides of jljl.CDK18 gene includes and CDK18 gene in the public GenBank GeneBank in the current world
(NC_000001.11) DNA sequence dna has 70% or more homology, and encodes the DNA sequence dna of identical function protein；
Preferably, the coded sequence of CDK18 gene includes following any DNA molecular:
(1) DNA sequence dna shown in SEQ ID NO.1 in sequence table；
(2) under strict conditions with 1) DNA sequence dna that limits hybridizes and encodes the DNA sequence dna of identical function protein；
(3) DNA sequence dna limited with (1) or (2) has 70%, preferably, 90% or more homology, and encodes identical function
The DNA molecular of energy protein.
In specific embodiments of the present invention, the coded sequence of the CDK18 gene is shown in SEQ ID NO.1
DNA sequence dna.
In the context of the present invention, CDK18 gene expression product includes the part of CDK18 albumen and CDK18 albumen
Peptide.The partial peptide of the CDK18 albumen contains functional domain relevant to coronary heart disease.
" CDK18 albumen " includes any functional equivalent of CDK18 albumen and CDK18 albumen.The functional equivalent
Including CDK18 albumen conservative variation protein or its active fragment or its reactive derivative, allelic variant, natural mutation
Body, induced mutants, can be with the encoded protein of DNA of the DNA hybridization of CDK18 under high or low stringent condition.
Preferably, CDK18 albumen is the protein with following amino acid sequences:
(1) protein that the amino acid sequence shown in SEQ ID NO.2 in sequence table forms；
(2) amino acid sequence shown in SEQ ID NO.2 by the substitution of one or several amino acid residues and/or is lacked
Lose and/or addition and with the ammonia with the same function as shown in SEQ ID NO.2 of amino acid sequence shown in SEQ ID NO.2
Protein derived from base acid sequence.The number for the amino acid for replacing, lacking or adding is usually 1-50, preferably 1-30
It is a, more preferably 1-20, most preferably 1-10.
(3) there is at least 80% homology (also known as sequence identity) with amino acid sequence shown in SEQ ID NO.2,
It is highly preferred that the homology with amino acid sequence at least about 90% to 95% shown in SEQ ID NO.2, be often 96%, 97%,
98%, the polypeptide that the amino acid sequence of 99% homology is constituted.
In specific embodiments of the present invention, the CDK18 albumen is with amino acid sequence shown in SEQ ID NO.2
The protein of column.
It is known that, conventionally, the modification of one or more amino acid will not influence the function of protein in a protein.
Those skilled in the art can approve the amino acid for changing single amino acids or small percentage or to amino acid sequence it is individual add,
Missing, insertion, replacement are conservative modifications, and wherein the change of protein generates the protein with identity function.Function phase is provided
As the Conservative substitution tables of amino acid be well known in the art.
Example by one amino acid of addition or the protein of more amino acid modification is the fusion of CDK18 albumen
Albumen.For the peptide or protein with CDK18 protein fusion, there is no limit as long as resulting fusion protein retains CDK18 egg
White biological activity.
CDK18 albumen of the invention also includes the non-conservative modification to amino acid sequence shown in SEQ ID NO.2, as long as
Protein by modification still is able to retain the biological activity of CDK18 albumen.It is mutated in such modification protein
Amino acid number be usually 10 perhaps less such as 6 perhaps less such as 3 or less.
In the context of the present invention, " diagnosis of coronary heart disease " both include judge subject whether suffered from coronary heart disease or
Including judging that subject whether there is the risk with coronary heart disease.
The advantages of the present invention:
Present invention firstly discovers that CDK18 gene expression is related to coronary heart disease, pass through the table of CDK18 in detection subject
It reaches, it can be determined that whether subject suffers from coronary heart disease or judge that subject whether there is the risk with coronary heart disease, to refer to
It leads clinician and provides prevention scheme or therapeutic scheme to subject.
Present invention finds a kind of new molecular marked compound-CDK18 genes, compared to traditional detection means, gene diagnosis
More in time, more special, sensitiveer, it can be realized the early diagnosis of coronary heart disease, to reduce the death rate of coronary heart disease.
Detailed description of the invention
Fig. 1 shows the differential expression using QPCR detection CDK18 gene in patients with coronary heart disease and normal person.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.Following embodiment is merely to illustrate this
It invents rather than limits the scope of the invention.Test method without specific conditions in embodiment, usually according to conventional strip
Part, such as Sambrook et al., molecular cloning: laboratory manual (New York:Cold Spring HarborLaboratory
Press, 1989) condition described in, or according to the normal condition proposed by manufacturer.
Embodiment 1 screens the gene of differential expression in patients with coronary heart disease and normal person
1, research object:
Collect patients with coronary heart disease 5, normal person 5.
CHD group is included in standard: according to the diagnostic criteria for the ischemic heart disease that the World Health Organization formulates, selection
Underwent coronary radiography confirms the patients with coronary heart disease for having one or more hemadostewnosis degree >=50%.The inclusion criteria of control group
Are as follows: 1) screened in epidemiological survey the age, gender, it is national match with CAD group, by questionnaire survey, physical examination,
Cardiac ultrasound and ECG examination and laboratory check the not clinical manifestation of coronary heart disease and Major Risk Factors
Examinee；2) the parallel coronary artery revasualization of row health examination of being hospitalized excludes coronary heart disease and age, gender, national and CAD group
Matcher；Meet one person of any of the above to enter to be selected as control group.Two groups are signed informed consent form before being included in research.
Rejecting standard: the infull person of CAD group-clinical data and the one for merging following disease simultaneously are rejected.
(such as: Congenital Heart patient, dissection of aorta, multiple organ failure, rheumatic heart disease and have spirit
Obstacle is not able to cooperate person).Control group: having mentally disturbed or checking through Doppler confirms there is carotid plaques or narrow person, gives
To reject.
2, in blood total serum IgE extraction
The extraction of blood total serum IgE is carried out using hundred Tyke blood rna extracts kits
(1) research object at least 12h on an empty stomach is required, at room temperature in next morning 7:00~8:00, venous blood samples take complete
Into RNase-Free Filter column, 13000rpm is centrifuged 2 minutes 250 μ l (or 0.25g) of blood, collects lower liquid, 0.75ml is added and splits
Solve liquid RLS.
(2) homogenised sample is acutely shaken to mixing, 5 minutes are incubated under the conditions of 15-30 DEG C so that ribosome divides completely
Solution.
(3) optional step: 12,000rpm is centrifuged 10 minutes under conditions of 4 DEG C, and supernatant is carefully taken to be transferred to a new nothing
In the centrifuge tube of RNA enzyme.
(4) every 1ml RLS adds 0.2ml chloroform.Sample tube cover is covered tightly, acutely vibrate 15 seconds and is incubated at room temperature 3
Minute.
(5) be centrifuged 10 minutes in 4 DEG C of 12,000rpm, sample can be divided into three layers: lower layer's organic phase, middle layer and upper layer without
The water phase of color, RNA are present in water phase.The capacity of aqueous layer is about the 60% of added RLS volume, and water phase is transferred to new pipe
In, carry out next step operation.
(6) 1 times of 70% ethyl alcohol of volume is added, is mixed by inversion (at this time it is possible that precipitating), obtained solution and possibility
Precipitating is transferred in adsorption column RA (absorption column sleeve is in collecting pipe) together.
(7) 10,000rpm are centrifuged 45 seconds, discard waste liquid, adsorption column is recovered collecting pipe again.
(8) plus 500 μ l protein liquid removal RE, 12,000rpm centrifugations 45 seconds discard waste liquid.
(9) 700 μ l rinsing liquid RW are added, 12,000rpm centrifugations 60 seconds discard waste liquid.
(10) 500 μ l rinsing liquid RW are added, 12,000rpm centrifugations 60 seconds discard waste liquid.
(11) adsorption column RA is put back in sky collecting pipe, 12,000rpm centrifugations 2 minutes, as far as possible removing rinsing liquid, in order to avoid drift
Residual ethanol inhibits downstream reaction in washing lotion.
(12) adsorption column RA is taken out, is put into the centrifuge tube without RNA enzyme, according to expected RNA yield in adsorbed film
Intermediate position adds 50-80 μ l without the water of RNA enzyme, is placed at room temperature for 2 minutes, and eluent is collected in 12,000rpm centrifugations 1 minute.
3, the purity analysis (NanoDrop1000 spectrophotometer) of RNA sample
NanoDrop1000 spectrophotometer detects RNA sample, OD260/OD280 1.8-2.2.
4, the quality analysis (Agilent Technologies 2100Bioanalyzer) of RNA sample
Agilent Technologies 2100Bioanalyzer detects RNA sample quality, observes 28S rRNA and 18S
RRNA master tape is obvious, cDNA library structure is sequenced without degradation, the RNA-seq that meets that RNA Perfection Index is qualified, concentration reaches requirement
The requirement built can be used for library construction and sequencing.
5, high-throughput transcript profile sequencing
(1) RNA-seq read positions
Low-quality read is removed to obtain cleaning read first, then using TopHat v1.3.1 will clean segment and
UCSC H.sapiens is matched with reference to genome (hg19), the H.sapiens UCSC hg19 editions indexes constructed in advance
It is downloaded from TopHat homepage, and as reference genome, when matching using TopHat with genome, allows each read (default
To 20) having multiple matching sites, most 2 mispairing.TopHat establishes possible according to exon region and GT-AG shear signal
Shearing site library navigates to the read for not navigating to genome on genome according to these shearing site libraries.We use
The system default parameter of TopHat method.
(2) transcript abundance is assessed
The read file matched is handled by Cufflinks v1.0.3, and Cufflinks v1.0.3 is by RNA-seq piece
Number of segment mesh is standardized the relative abundance for calculating transcript.FPKM value refers to being matched in every 1,000,000 sequencing fragment specific
The segment number of the exon region of gene 1kb long.The confidence interval of FPKM estimated value is calculated by Bayesian inference method.
The GTF comment file for the reference that Cufflinks is used downloads (Homo_ from Ensembl database
sapiens.GRCh37.63.gtf)。
(3) detection of difference expression gene
It is transferred to Cuffdiff by the Ensembl GTF file of downloading and by the matched original document of TopHat,
Cuffdiff re-evaluates the gene expression abundance for the transcript listed in GTF file using original matching files, detects difference table
It reaches.The only q value < 0.01 in Cuffidff output, test display is more just considered as successfully differential expression.
6, result
RNA-seq is the results show that be compared with normal people, and the mRNA level in-site of CDK18 gene is significant in patients with coronary heart disease blood
Decline, difference have statistical significance (P < 0.05).
The gene of differential expression in embodiment 2QPCR experimental verification patients with coronary heart disease and normal person
1, research object:
Screening criteria is with embodiment 1, patients with coronary heart disease and normal person each 50.
2, in blood total serum IgE extraction
Step is the same as embodiment 1.
3, reverse transcription
The synthesis of cDNA uses TaqMan reverse transcription reagent box, in 10 μ l reverse transcription buffers (including 5.5mmol/L
MgCl2, random 6 nucleotide primer of 2.5mmol/L, 4U RNase inhibitor and 31.25U MultiScribe reverse transcriptase) and right
The total serum IgE of 200ng carries out reverse transcription, operates according to the following steps: 25 DEG C of 10min, 37 DEG C of 60min, 95 DEG C of 5min.
4、QPCR
MRNA abundance is quantified using real-time fluorescence detection method.If the cDNA of above-mentioned acquisition is in ABI Prism
It is expanded on 7700 sequential detectors, CDK18 gene primer, 18S ribosomal RNA gene design of primers are as follows:
CDK18:
Upstream primer: 5 '-ACATTGGCTTTGGGAAAC-3 ' (SEQ ID NO.3)；
Downstream primer: 5 '-CCTATGCCACAGTCTTCA-3 ' (SEQ ID NO.4),
18S rRNA:
Upstream primer: 5 '-AATCAGGGTTCGATTCCGGA-3 ' (SEQ ID NO.5)；
Downstream primer: 5 '-CCAAGATCCAACTACGAGCT-3 ' (SEQ ID NO.6),
The expression of CDK18 gene makees internal reference using 18S rRNA.With 2 times of amount SYBR Green PCR of 12.5 μ l
Master mix (upstream primer and 5 μm of ol/L downstream primers including 5 μm of ol/L) carries out PCR, and final volume is 25 μ l.PCR item
Part are as follows: 95 DEG C of 10min, 50 circulations (95 DEG C of 30s, 56 DEG C of 30s, 72 DEG C of 30s).Use 7700 type Sequence Detection of ABIprism
The fluorescence that instrument emits reporter fluorescence dyestuff carries out real-time monitoring.Fluorescent emission amount reflects recurring number, and by Sequence Detector
Software read, provide the recurring number threshold value significant to PCR amplification, value and the genomic DNA logarithm of recurring number linearly close
System carries out relative quantification using Δ Δ CT method.
5, statistical method
Result data is indicated in a manner of mean+SD, is united using SPSS13.0 statistical software
Meter analysis, difference between the two is examined using t, it is believed that has statistical significance as P < 0.05.
6, result
As a result as shown in Figure 1, being compared with normal people, the mRNA level in-site of CDK18 gene significantly drops in patients with coronary heart disease blood
Low, difference has statistical significance (P < 0.05), as a result tests with RNA-Seq.
The explanation of above-described embodiment is only intended to understand method and its core concept of the invention.It should be pointed out that for this
For the those of ordinary skill in field, without departing from the principle of the present invention, several improvement can also be carried out to the present invention
And modification, these improvement and modification will also be fallen into the protection scope of the claims in the present invention.
Claims (10)
1. detecting application of the product of CDK18 gene expression in the tool for preparing diagnosis of coronary heart disease.
2. application according to claim 1, which is characterized in that the product include: by RT-PCR, real-time quantitative PCR,
Immune detection, in situ hybridization, chip or high-flux sequence detection of platform CDK18 gene expression dose are with the production of diagnosis of coronary heart disease
Product.
3. application according to claim 2, which is characterized in that the product with RT-PCR diagnosis of coronary heart disease includes at least
The primer of a pair of of specific amplified CDK18 gene；The product with real-time quantitative PCR diagnosis of coronary heart disease includes at least a pair of special
Expand the primer of CDK18 gene；The product with immune detection diagnosis of coronary heart disease includes: in conjunction with CDK18 protein-specific
Antibody；The product in situ hybridization diagnosis of coronary heart disease includes: the probe with the nucleic acid array hybridizing of CDK18 gene；Institute
Stating with the product of chip diagnosis of coronary heart disease includes: protein chip and genetic chip；Wherein, protein chip includes and CDK18 albumen
The antibody of specific binding, genetic chip include the probe with the nucleic acid array hybridizing of CDK18 gene.
4. application according to claim 3, which is characterized in that the product with real-time quantitative PCR diagnosis of coronary heart disease is extremely
The primer for a pair of of the specific amplified CDK18 gene for including less is as shown in SEQ ID NO.3 and SEQ ID NO.4.
5. application according to claim 1, which is characterized in that the tool can pass through CDK18 gene in detection sample
Expression carry out diagnosis of coronary heart disease.
6. application according to claim 5, which is characterized in that the tool includes chip, kit, test paper or high throughput
Microarray dataset.
7. application according to claim 6, which is characterized in that the chip includes genetic chip, protein-chip；It is described
Genetic chip includes solid phase carrier and the oligonucleotide probe for being fixed on solid phase carrier, and the oligonucleotide probe includes being used for
Detect the oligonucleotide probe for CDK18 gene of CDK18 gene transcription level；The protein-chip includes solid phase carrier
And it is fixed on the specific antibody of the CDK18 albumen of solid phase carrier；The kit includes gene detecting kit and albumen
Immunity detection reagent；The gene detecting kit includes the reagent for detecting CDK18 gene transcription level；The albumen
Immunity detection reagent includes the specific antibody of CDK18 albumen；The test paper includes for detecting CDK18 gene transcription level
Reagent；The high-flux sequence platform includes the reagent for detecting CDK18 gene transcription level.
8. application according to claim 7, which is characterized in that it is described detection CDK18 gene transcription level reagent include
For the primer and/or probe of CDK18 gene.
9. application according to claim 8, spy are characterized in that, the primer sequence for CDK18 gene is as follows: just
To primer sequence as shown in SEQ ID NO.3, reverse primer is as shown in SEQ ID NO.4.
10. the application according to any one of claim 5-9, which is characterized in that the sample is blood.
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| CN105861735A (en) * | 2016-06-17 | 2016-08-17 | 北京泱深生物信息技术有限公司 | Application of RAP1B in coronary heart disease diagnosis |
| CN106801095A (en) * | 2017-02-14 | 2017-06-06 | 徐州市中心医院 | Application of the PRRT1 genes in diagnosis of coronary heart disease product is prepared |
| CN108796069A (en) * | 2018-07-03 | 2018-11-13 | 北京泱深生物信息技术有限公司 | Diagnosis marker-ING1 the genes of myocardial infarction |
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| WO2003083096A2 (en) * | 2002-03-28 | 2003-10-09 | Qlt Inc. | Cancer associated protein kinases and their uses |
| CN102690878A (en) * | 2011-03-23 | 2012-09-26 | 上海人类基因组研究中心 | Application of CDK18 gene and expression product thereof |
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