CN105606818A - IDH1-R132H and ATRX expression based glioma prognostic system - Google Patents
IDH1-R132H and ATRX expression based glioma prognostic system Download PDFInfo
- Publication number
- CN105606818A CN105606818A CN201610024304.0A CN201610024304A CN105606818A CN 105606818 A CN105606818 A CN 105606818A CN 201610024304 A CN201610024304 A CN 201610024304A CN 105606818 A CN105606818 A CN 105606818A
- Authority
- CN
- China
- Prior art keywords
- idh1
- atrx
- patients
- glioma
- kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 206010018338 Glioma Diseases 0.000 title claims abstract description 64
- 102000056014 X-linked Nuclear Human genes 0.000 title claims abstract description 59
- 108700042462 X-linked Nuclear Proteins 0.000 title claims abstract description 59
- 101150020330 ATRX gene Proteins 0.000 title claims abstract description 54
- 102200069690 rs121913500 Human genes 0.000 title claims abstract description 53
- 208000032612 Glial tumor Diseases 0.000 title claims abstract description 38
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 18
- 230000004083 survival effect Effects 0.000 claims abstract description 18
- 238000004393 prognosis Methods 0.000 claims description 14
- 238000004043 dyeing Methods 0.000 claims description 12
- 238000011156 evaluation Methods 0.000 claims description 7
- 210000005170 neoplastic cell Anatomy 0.000 claims description 7
- 210000004940 nucleus Anatomy 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical group Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
- 210000004698 lymphocyte Anatomy 0.000 claims description 3
- 210000002540 macrophage Anatomy 0.000 claims description 3
- 210000000274 microglia Anatomy 0.000 claims description 3
- 210000001130 astrocyte Anatomy 0.000 claims description 2
- 210000004881 tumor cell Anatomy 0.000 claims description 2
- 208000005017 glioblastoma Diseases 0.000 abstract description 12
- 238000012360 testing method Methods 0.000 abstract description 4
- 238000011022 operating instruction Methods 0.000 abstract 3
- 208000020372 Infective dermatitis associated with HTLV-1 Diseases 0.000 abstract 1
- 206010003571 Astrocytoma Diseases 0.000 description 22
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 15
- 239000012188 paraffin wax Substances 0.000 description 15
- 238000000034 method Methods 0.000 description 14
- 239000001993 wax Substances 0.000 description 14
- 238000003745 diagnosis Methods 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 8
- 102000007999 Nuclear Proteins Human genes 0.000 description 6
- 108010089610 Nuclear Proteins Proteins 0.000 description 6
- 201000010133 Oligodendroglioma Diseases 0.000 description 6
- 235000019441 ethanol Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 208000030883 malignant astrocytoma Diseases 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000006059 cover glass Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 238000011532 immunohistochemical staining Methods 0.000 description 3
- 230000007170 pathology Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000009966 trimming Methods 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 239000005662 Paraffin oil Substances 0.000 description 2
- 239000000084 colloidal system Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 235000015110 jellies Nutrition 0.000 description 2
- 239000008274 jelly Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 229920001206 natural gum Polymers 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YULFFYZCWUUTNC-YWPNNVDBSA-N (1r)-1-[(4r,4ar,8as)-2,6-bis(4-chlorophenyl)-4-methyl-8,8a-dihydro-4ah-[1,3]dioxino[5,4-d][1,3]dioxin-4-yl]ethane-1,2-diol Chemical compound C([C@@H]1OC(O[C@]([C@@H]1O1)(C)[C@H](O)CO)C=2C=CC(Cl)=CC=2)OC1C1=CC=C(Cl)C=C1 YULFFYZCWUUTNC-YWPNNVDBSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 101001042041 Bos taurus Isocitrate dehydrogenase [NAD] subunit beta, mitochondrial Proteins 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 101000960234 Homo sapiens Isocitrate dehydrogenase [NADP] cytoplasmic Proteins 0.000 description 1
- 108010075869 Isocitrate Dehydrogenase Proteins 0.000 description 1
- 102000012011 Isocitrate Dehydrogenase Human genes 0.000 description 1
- 102100039905 Isocitrate dehydrogenase [NADP] cytoplasmic Human genes 0.000 description 1
- 101710088105 Isocitrate dehydrogenase [NAD] subunit 1, mitochondrial Proteins 0.000 description 1
- 101710086399 Isocitrate dehydrogenase [NAD] subunit 2, mitochondrial Proteins 0.000 description 1
- 102100021332 Isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial Human genes 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000003140 astrocytic effect Effects 0.000 description 1
- 238000007622 bioinformatic analysis Methods 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 201000000053 blastoma Diseases 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 239000000104 diagnostic biomarker Substances 0.000 description 1
- -1 dimethylbenzeneOn dimethylbenzene Chemical compound 0.000 description 1
- XPPKVPWEQAFLFU-UHFFFAOYSA-N diphosphoric acid Chemical compound OP(O)(=O)OP(O)(O)=O XPPKVPWEQAFLFU-UHFFFAOYSA-N 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 201000008184 embryoma Diseases 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 208000029824 high grade glioma Diseases 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- 230000002962 histologic effect Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 201000011614 malignant glioma Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229940005654 nitrite ion Drugs 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000000505 pernicious effect Effects 0.000 description 1
- 210000002706 plastid Anatomy 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 229940005657 pyrophosphoric acid Drugs 0.000 description 1
- 238000012175 pyrosequencing Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Classifications
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6827—Total protein determination, e.g. albumin in urine
- G01N33/6839—Total protein determination, e.g. albumin in urine involving dyes, e.g. Coomassie blue, bromcresol green
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Bioinformatics & Computational Biology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to an IDH1-R132H and ATRX expression based glioma prognostic system which comprises (1), a tumor paraffin-embedding kit for glioma patients and an operating instruction thereof if necessary; (2), a kit for immunohistochemically testing ATRX protein expression level of tumor of glioma patients and an operating instruction thereof if necessary; and (3), a kit for immunohistochemically testing IDH1-R132H protein expression of tumor of glioma patients and an operating instruction thereof if necessary. Progression free survival (PFS) of WHO II-level and III-level of glioma is typed according to loss states of IDH1-R132H and ATRX; patients with both loss of IDH1-R132H and ATRX has longer PFS than patients with both expression of IDH1-R132H and ATRX, and PFS of patients with IDH-WT is close to PFS of patients with glioblastoma.
Description
Technical field
The present invention relates to disease prognosis field, particularly relate to based on IDH1-R132H and ATRXGlioma prognosis system.
Background technology
According to the classification of WHO, dispersivity glioma is divided into II level, III level and glioblastThe Astrocytic glioma of knurl, prominent astroglioma and mesoglioma less. Compared with low level colloidThe patient of knurl (LGGs, II level) has more and has than the patient of High Grade Gliomas (III level and IV level)The prognosis of profit, in the low level patients with gliomas of 50-75%, tumour continued growth and developing intoFor higher level, cause sacred disease and final dead.
Astrocytoma is modal histological type in brain tumor, very easily recurrence or malignant progression,Even be also difficult to cure through operation, radiation and chemotherapy. Pernicious astrocytoma is as colloid motherCytoma is the most fatal ICT. Astrocytoma patient's prognosis depend on some clinical because ofElement, as age of onset, KPS scoring, Operative Range, histological type, tumour rank andMolecular marked compound feature. Current, diagnosis of glioma only, taking morphology as basis, is difficult to clear and definite visitorSee ground diagnosis glioma pathology type, can not reflect the biological behaviour of tumour comprehensively, can notAccurately judge patient's prognosis and instruct postoperative clinical treatment.
IDH (isocitratedehydrogenase, isocitric dehydrogenase) and ATRX (X-linkedAlphathalassaemiamentalretardation) the early stage appearance that sudden change occurs in gliomaWith the specific hypotype that characterizes glioma. Most carcinogenic IDH1 sudden changes are heterozygosis missense mutation,395 positions (G395A) G becomes A, has caused at enzyme active sites codon 132(IDH1-R132H) arginine is substituted by histidine. ATRX sudden change or loss affect star glueThe biological behaviour of cell plastid, relevant with astrocytic tumor patient's favourable existence.
Summary of the invention
The object of the invention is to provide a kind of glioma based on IDH1-R132H and ATRX expressionPrognosis system.
In one embodiment, the glioma based on IDH1-R132H and ATRX expression is pre-Rear system, it comprises: (1) detects ATRX protein expression level in patients with gliomas tumourKit and optionally described kit operation instructions; (2) detect patients with gliomas tumourThe kit of middle IDH1-R132H protein expression and optionally described kit operation instructions.
In one embodiment, the glioma based on IDH1-R132H and ATRX expression is pre-Rear system, described system comprises: (1) patients with gliomas tumour FFPE kit and optionalThe described kit operation instructions in ground; (2) SABC detects ATRX in patients with gliomas tumourThe kit of protein expression level and optionally described kit operation instructions; (3) immunityGroupization detects the kit of IDH1-R132H protein expression in patients with gliomas tumour and optionallyDescribed kit operation instructions.
In one embodiment, the evaluation criteria of IDH1-R312H dyeing: (1) is swollen widelyThe strong dyeing of oncocyte matter is defined as the positive; (2) the weak positive being dispersed in or macrophage dyeing quiltBe defined as feminine gender.
In one embodiment, in ATRX core, express the evaluation criteria of loss: if tumour is thinKaryon is not dyed to brown, but not neoplastic cell nuclei, microglia, lymphocyte and starCell shows as strong positive, is defined as so in ATRX core and expresses and lose.
In one embodiment, described SABC detects ATRX in patients with gliomas tumourIt is 2211-2413 that the kit of protein expression level detects ATRX Argine Monohydrochloride sequence site.
Except by the method for SABC, IDH1-R132H also can be by the pyrophosphoric acid skill that checks orderArt (pyro-sequencing) detects and realizes, and ATRX sudden change can be by the extron detection of checking order, itsExpressing loss also can detect by transcribing group order-checking (rna-seq). The current technology of these detection methodsComplexity, and price is higher, and popularization is poor, contrast therewith, IDH1R132H and ATRX coreInterior loss of proteins can detect by the method for SABC, and method uniformity is strong, and technical requirement is notHeight, price is more economical, and Clinical practicability and popular strong, is applicable to applying.
Other side of the present invention is due to disclosure herein, to those skilled in the art andSpeech is apparent. In should be understood that within the scope of the present invention, above-mentioned each technology of the present inventionFeature and in below (eg embodiment) group mutually between specifically described each technical charactericticClose, thereby form new or preferred technical scheme. As space is limited, this is no longer going to repeat them.
Below in conjunction with specific embodiment, further set forth the present invention. Should be understood that these embodimentOnly be not used in and limit the scope of the invention for the present invention is described. Unreceipted in the following exampleThe experimental technique of actual conditions, carries out according to the normal condition in affiliated field conventionally.
Brief description of the drawings
Fig. 1 is the immunohistochemical staining result figure of IDH1-R132H albumen in samples of human glioma:Disperse astrocytoma, less prominent astroglioma and mesoglioma (A/B/C) showIDH1-R132H protein positive expression; Between become astrocytoma, the prominent astrocytoma that tails off andBetween become mesoglioma (D/E/F) and also show the IDH1-R132H positive; And former hair jelly matter motherCytoma and recurrence glioblastoma are IDH1-R132H feminine gender (G/H); Secondary colloid motherCytoma is the IDH1-R132H positive (I).
Fig. 2 is the immunohistochemical staining result figure of ATRX nuclear protein in samples of human glioma:Disperse astrocytoma (WHOII, A), become astrocytoma (WHOIII, C) andSecondary glioblastoma (WHOIII, E) shows ATRX in obvious neoplastic cell nucleiProtein expression loss; Oligodendroglioma (WHOII, B), a change oligodendrogliaKnurl (WHOIII, D) and former glioblastoma (WHOIV, F) show significantlyATRX protein positive expression in neoplastic cell nuclei; Present positive expression with vascular endothelial cellATRX nuclear protein, can be used as positive control.
Fig. 3 is that IDH1-R132H and ATRX nuclear protein lose as glioma pathology typeThe ROC evaluation graph of diagnosis: use IDH1-R132H as differentiating pGBM (former hair jelly matter motherCytoma) and the diagnosis marker of II, III level glioma and sGBM (secondary glioblastoma)AUC be 0.7414 (sensitiveness 63.19%, specificity 85.09%, A); ATRX damagesLose as distinguishing pGBM, oligodendroglioma (WHO classification II/III), and sGBM,The AUC of astrocytoma (WHO classification II/III) diagnostic biomarkers is 0.8241 (quickPerception 76.67%, specificity 88.15%, Fig. 3 B); And when associating IDH1-R132H withATRX loss is distinguished pGBM and oligodendroglioma as diagnosis marker, and (WHO dividesLevel II/III), when astrocytoma (WHO classification II/III), sGBM, AUC is 0.7407(sensitiveness 53.33%, specificity 94.815%, Fig. 3 C), wherein specificity is obviously highIn IDH1-R132H or these two single labelled things of ATRX loss situation.
Fig. 4 is the patients with gliomas Progression free survival phase of IDH1-R132H protein expression and lossAnalysis chart: in WHOII level patients with gliomas, IDH1-R132H is indicating that patient is longer(IDH1-R132H-meta PFS=893 days, IDH1-WT is (wild for PFS (Progression free survival phase)Type)-meta PFS=580 days, p=0.0009, A); At change glioma and a glioblastIn knurl patient, IDH1-R132H type patient is becoming with longer PFS than IDH1-WT type patientGesture (p=0.0702, p=0.0914, B/C).
Fig. 5 is that ATRX nuclear protein is expressed and the patients with gliomas Progression free survival phase of loss dividesAnalyse figure: for WHOII level (A) or III level (B) patients with gliomas, ATRX tableReaching loss is indicating the better Progression free survival phase of patient; ATRX expresses loss and the star of dashing forward lessCytoma patient (C) or the Progression free survival phase longer compared with low level patients with gliomas (D) haveClose, express astrocytoma patient's Progression free survival phase longer (E) of loss with ATRX;In addition, associating IDH1-R132H and ATRX loss state can be WHOII, III level glueThe Progression free survival phase of matter knurl is carried out somatotype (p < 0.0001), IDH1-R132H and ATRXThe patient that the patient of loss expresses than IDH1-R132H and ATRX simultaneously is simultaneously with longerThe trend of PFS, the patient PFS of IDH-WT is close to glioblastoma patient's PFS (F).
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail.
1. patient's data and Specimen origin
Amount to 211 routine gliomas and enter this research of group, all from Beijing Tiantan Hospital's gliomaTherapeutic community. All patients in year March in January, 2008 to 2015 undergo surgery excision, putTreat and alkylating agent treatment, Pathological diagnoses is glioma (according to WHO maincenter god in 2007Through system tumor criteria for classification).
The male sex's 112 examples in all patients, women's 99 examples, age 16-69 year, average (40)Year. Patient's Informed Consent Form is ratified and is signed in this research by Ethics Committee of units concerned.
2. tumor tissues FFPE
2.1 key instruments and reagent
Paraffin wax embedding: manufacturer: Hao Silin; Production code member: TEC2800
Milli-Q type ultra-pure water system: manufacturer: Millipore
Ethanol, dimethylbenzene and paraffin all come from Beijing Chemical Plant.
2.2 experimental procedure
2.2.1 dehydration
Table 1: dewatering process flow
2.2.2 transparent
Table 2: transparent flow process
| Solution | Time (min) |
| 1/2 ethanol+1/2 dimethylbenzene | 60min |
| Dimethylbenzene I | 60min |
| Dimethylbenzene II | 60min |
2.2.3 infiltration
Table 3: infiltration flow process
| Solution | Time (min) | Temperature (DEG C) |
| 1/2 dimethylbenzene+1/2 paraffin | 90min | 62 |
| 1/2 paraffin | 90min | 62 |
| Paraffin I | 120min | 62 |
| Paraffin II | 120min | 62 |
2.2.4 embedding
Tissue is put into the mould that fills paraffin, set position, in cold of paraffin wax embeddingUpper cooling.
3. paraffin section
3.1 key instrument and reagent
Paraffin slicing machine: manufacturer: state SLEE; Production code member: CUT4062;
Slicer: manufacturer: state SLEE;
Milli-Q type ultra-pure water system: manufacturer: Millipore;
Water-bath heater: manufacturer: Beijing Chang Feng instrument company; Production code member:HHSY11-N1；
Slide: manufacturer: Sang Ge bio tech ltd, Shanghai
Insulating box: manufacturer: Thermo company
The set of 3.2 paraffin masses and trimming
After embedding, just can cut into slices. Embedded paraffin mass is loaded onto slicer and is cutBefore sheet, also must carry out set and trimming.
Set: on general rotary microtom, all have can set paraffin mass metal shallow bid, thisAlso can substitute with onesize platform wood. Shovel embedded block be pasted on set thing with the wax of heating,And make tissue block outwardly, after being convenient to, cut out rapidly required slice, thin piece.
Trimming: the wax shovel of use heating or blade, by the embedded block surrounding equating of set, make upper and lower twoFace is accomplished parallel surface, and normal reservation organized the paraffin that adheres to wide 2～3mm around, and the wax of fixingPiece is rectangle. Also can prune one jiao so that identification section on wax band.
3.3 section
Slicer is the custom-designed precision optical machinery of one that is used for various histotomies, conventionalBe rotary microtom, its folder thing part move up and down before and after advance, and folder cutter portionDivide and maintain static. Critical piece is the cutter platform that slicer is installed, the object sland that embedded block is installedWith the inching gear of controlling slice thickness. When section, slicer maintains static, rotating wheel, markThis TV station moves up and down and pushes ahead certain distance by the slice thickness mixing up, and tissue block is upper and lowerMotion once, just obtains the section of an opening and closing thickness requirement on blade.
Slicer is directly related with the quality of section. Before section, must whet a knife, method is: will cutSheet cutter is loaded onto handle of a knife, knife back folder, is dripped a little paraffin oil on level and smooth whetstone, and cutter is being pastedWhetstone, with the mill of edge of a knife direction dorsad, should be cleaned paraffin oil with dimethylbenzene after finishing using in time.
Before section, the edge of a knife is put to large Microscopic observation, select the part that the smooth nothing of the edge of a knife is incisedCut. The embedded block that will cut is fixed on object sland, makes the outer tangent plane of embedded block and markThis folder cross section is parallel, and allows embedded block slightly expose one section. After being pushed into outer rim, cutter platform unclamps bladeThe spiral of folder, first-class blade, makes slicer plane and organizes the angle that is 15 ° of left and right between tangent plane,Parallel with the edge of a knife below on embedded block. The thickness that regulates section to require on inching gear, regulatesTime should notice that pointer can not be between two scales, otherwise easily damage slicer, Jiang Daotai movesTo nearly object sland place, allow the edge of a knife contact slightly with organizing tangent plane, at this moment just can start to have cut into slices.
Dicing method: right hand rotating wheel, left hand is held writing brush and slightly descended termination Cui to cut at the edge of a knifeSlice, thin piece, and hold the wax band cutting, after wax band forms certain length, the right hand stops operating,Hold another writing brush and gently wax band is provoked, lie against in the carton that is lined with black paper, note sectionSpeed should not be too fast, and shake runner firmly should be even, prevents that slicer vibrations severity from causing sectionBecame uneven is even, should also be noted that the direction of rotation, cuts less than slice, thin piece in case move after object sland.Cut into slices complete, should with chloroform, the relative section of slicer be cleaned in time.
3.4 pasters are baked sheet
The section cutting must be attached on slide and just can be for further processing, but section is normalThere is tiny band, must after flattening, could attach, otherwise impact dyeing and observation. AdoptDrag for sheet method and carry out paster. After wax disk(-sc) is cut into, the right hand is taken the photograph wax disk(-sc) with pincet, and left hand is used writing brush edgeThe knife edge separates wax disk(-sc) gently, and the tepidarium of 48 DEG C put into section down by tangent plane, shakeouts rear useTweezers separate continuous wax slice gently, then pick up with slide, and wax disk(-sc) should drag for 1/3 place at slide,Wax disk(-sc) thickness is generally at 3～5 μ m. Wax disk(-sc) is put on roasting horse, more than putting into 62 DEG C of insulating box 6h.
4. immunohistochemical staining
4.1 key instruments and reagent
The desk-top mould release of trace: manufacturer: Eppendorf; Model: 541D;
Turbine mixer: manufacturer: TonIDa; Model: TDX-1;
Milli-Q type ultra-pure water system: manufacturer: Millipore
Water-bath heater: manufacturer: Beijing Chang Feng instrument company; Production code member:HHSY11-N1；
Cover glass: manufacturer: Sang Ge bio tech ltd, Shanghai
Microscope: manufacturer: Japanese Olympus company; Model: IX51;
Magnetic force thermostatic mixer: manufacturer: Shanghai Nanhui Telecommunication Apparatus Factory; Model: CHJ-1;
Ethanol, dimethylbenzene, H2O2With citrate from Beijing Chemical Plant;
Anti-ATRX antibody: manufacturer: Abcam company; Model: ab97508
Anti-IDH1-R132H antibody: manufacturer: German Dianova company; Model: H09
Two anti-HRP polymers: manufacturer: Beijing Quanshijin Biotechnology Co., Ltd; TypeNumber: PV-6000
DAB developer: manufacturer: Beijing Quanshijin Biotechnology Co., Ltd;
4.2 experimental procedure
4.2.1 aquation as follows dewaxes
4.2.2 flow process dewaxes
Table 4: dewaxing flow process
4.2.3 distilled water washing 5min.
4.2.4 antigen retrieval is put into slide the antigen repairing box that fills citrate buffer. MicrowaveHigh fire screen 3min in stove, makes temperature reach 95 DEG C of left and right, is adjusted to low fire screen, heating 15min. GetGo out antigen repairing box, naturally cool to room temperature. Take out slide, the washing of PBST buffer solution, 5min/Inferior x3 time.
4.2.53%H2O2 room temperature lucifuge sealing 8-10min.
4.2.6PBS buffer solution washing, 5min/ time x3 time.
4.2.7 add primary antibodie (anti-ATRX antibody, 1:800; Or anti-IDH1-R132H antibody, 1: 60),Hatch 12h for 4 DEG C.
4.2.8 get rid of primary antibodie, the washing of PBS buffer solution, 5min/ time x3 time.
4.2.9 hatch two and resist, 37 DEG C, 30min, PBS washing 3 times, each 5min.
4.2.10DAB colour developing. Press kit explanation configuration DAB nitrite ion, drip DAB dye liquorIn tissue, micro-Microscopic observation, develops the color and rinses and stop dyeing with running water in time when suitable.
4.2.11 bush uniformly dyeing core 2min.
4.2.12 unhurried current water rinses 10min.
4.2.131% hydrochloride alcohol color separation 5s, flowing water rinses running water and returns blue 10min.
4.2.14 dewater transparent, step is as follows:
The transparent flow process 4.2.15 dewater
Table 5: the transparent flow process of dewatering
| Solution | Time (min) |
| 95% ethanol | 5min |
| Absolute ethyl alcohol I | 5min |
| Absolute ethyl alcohol II | 5min |
| Dimethylbenzene I | 5min |
| Dimethylbenzene II | 5min |
| Dimethylbenzene III | 5min |
4.2.16 mounting: slide after sucking-off, is sucked to unnecessary dimethylbenzene, at sample from dimethylbenzeneOn dimethylbenzene add a natural gum before not yet parching, carefully cover covered, avoid producingBubble, also must not drag cover glass, in order to avoid sample is destroyed. Natural gum amount look cover glass size andFixed, do not make too much very few. Labelled, indicate material, decoration method, put in vent cabinet and dry,Hide after agent is solidified and just become a successful paraffin section until envelope. Microscopic examination, photographic analysis.
5. coloration result criterion
The evaluation criteria of IDH1-R312H dyeing: dye by force by fixed (1) widely tumour cell matterJustice is positive; (2) the weak positive being dispersed in or macrophage dyeing are defined as feminine gender.
In ATRX core, express the evaluation criteria of loss: brown if neoplastic cell nuclei is not dyed to,But not neoplastic cell nuclei is as endothelial cell, microglia, lymphocyte and astrocyte performanceFor strong positive, be defined as so in ATRX core and express and lose.
6. statistics and bioinformatic analysis
6.1ROC (Receiveroperatingcharacteristic) curve is used to assess ATRX and damagesLose specificity and the sensitiveness of diagnosis glioma.
6.2Kaplan-Meier curve is used to assess the time of patient's prognosis between different grouping,Log-rank is used to check prognosis difference.
6.3 Progression free survival phases were defined as from diagnosing to the Patients on Recurrence time.
It is meaningful that 6.4p < 0.05 is considered to difference.
6.5 all analyses are all to use software GraphPadPrism (GraphPadSoftware, LaJolla, CA) and SPSSversion16.0 (SPSS, Chicago, IL, USA).
7. result
The frequency that 7.1IDH1-R132H and the internal loss of ATRX pyrenoids occur in glioma and examiningDisconnected value
In order to study the correlation of the internal loss of ATRX pyrenoids and the diagnosis of glioma pathology, weEnter to have organized 211 routine patients with gliomas, continuous sampling, comprise astrocytoma 103 examples (A,AA), 25 routine oligodendrogliomas (O, AO), astrocytoma 123 examples of dashing forward less (OA,AOA) and 181 routine glioblastomas (former glioblastoma (pGBM), secondary glueMatter blastoma (sGBM)). Wherein 30 examples are organized in and approach whole tissue or specific pattern does not haveThere is immune response, or have necrotic area, therefore do not consider statistical appraisal.
IDH1-R132H stained positive and feminine gender are clear and definite. Under IDH1-R132H stained positiveShow strong cytoplasm dyeing weak nuclear staining (Fig. 1) of while. At our 64 routine II level starsIn shape cytoma, 37 examples positive (57.81%, Figure 1A). II level is prominent astroglioma (40/ less49,81.63%, Figure 1B) and mesoglioma (9/12,75%, Fig. 1 C) have one moreHigh positive rate. Between become astrocytoma (13/27,48.15%, Fig. 1 D), prominent star tails offShape cytoma (38/68,55.88%%, Fig. 1 E) and a change mesoglioma (5/9,55.56%,Fig. 1 F) IDH1-R132H incidence is also very high. In 114pGBM, detect positive 17 examples(14.91%, Fig. 1 G/H), and 40 examples positive (67.8%, Fig. 1 I) in 59 routine sGBMIn conjunction with IDH1-R132H (table 6, p=0.009, Fisher Precision Test).
In 174 (402) example tissues of the ATRX SABC positive, it is obvious that immune response isNuclear staining (Fig. 2, table 6). In the obvious negative tumor tissues of 228 example, neoplastic cell nucleiBe ATRX loss, and endothelial cell and Inflammatory infiltrating cell and residual neuron are ATRXExpress positive. Frequency higher in astrocytoma (A, 49/64,76.56% of ATRX loss;AA, 21/27,77.78%; SGBM, 45/59,76.27%, Fig. 2 A/C/E),OA, AOA lower (OA, 23/49,46.94%; AOA, 20/68,29.41%),O, frequency of loss minimum (O, 1/12,8.33% in AO and pGBM; AO, 1/9,11.11%;PGBM, 14/114,12.28%, Fig. 2 B/D/F, P=0.009, Fisher Precision Test).
Table 6IDH1-R132H and the occurrence frequency of ATRX core internal loss in glioma
We have also carried out sensitiveness that recipient's operating characteristics (ROC) analysis delimit and specialProperty IDH1-R132H and (or) ATRX loss histologic classification. As shown in Figure 3, useIDH1-R132H is as the diagnostic markers of differentiating pGBM and II, III level glioma and sGBMThe AUC of thing is 0.7414 (sensitiveness 63.19%, specificity 85.09%, Fig. 3 A). ATRXPGBM is distinguished in loss conduct, oligodendroglioma (WHO classification II/III), and sGBM,The AUC of the biomarker of astrocytoma (WHO classification II/III) diagnosis is 0.8241(sensitiveness 76.67%, specificity 88.15%, Fig. 3 B). When associating IDH1-R132H andATRX loss is distinguished pGBM and oligodendroglioma (WHO classification as diagnosis markerII/III),, when astrocytoma (WHO classification II/III), sGBM, AUC is 0.7407(sensitiveness 53.33%, specificity 94.815%, Fig. 3 C), wherein specificity is obviously highIn IDH1-R132H or these two single labelled things of ATRX loss situation. These results show,IDH1-R132H associating ATRX loss situation has the histodiagnosis of good glioma pathologyPredictive ability.
7.2IDH1-R132H and the prognostic value assessment of ATRX pyrenoids internal loss to patients with gliomas
We have analyzed IDH1-R132H and ATRX egg in the cohort study of patients with gliomasWhite nucleus internal loss state is as the biological molecule mark of the Progression free survival phase of assessment patients with gliomasNote thing. In our research, we are evaluated at WHOII-IV level patient's Progression free survivalPhase, there were significant differences (meta PFS (low level glioma)=830 days, a meta PFS (changeProperty glioma)=495 days, meta PFS (glioblastoma)=394 day, P < 0.0001,Table 7).
The Progression free survival phase between table 7, patients with gliomas different pathological types is analyzed
In WHOII level patients with gliomas, IDH1-R132H is indicating the PFS that patient is longer(IDH1-R132H-meta PFS=893 days, IDH1-WT-meta PFS=580 days, P=0.0009, Fig. 4 A). In a change glioma and glioblastoma patient, IDH1-R132HType patient is trend (p=0.0702, the p=with longer PFS than IDH1-WT type patient0.0914, Fig. 4 B/C).
The life although the II level that ATRX loss and ATRX express or III level tumor patient get nowhereThe phase of depositing is without remarkable significant difference (Fig. 5 A/B), but ATRX loss II/III level prominent star lessOr the II/III level patients with gliomas Progression free survival phase longer (Fig. 5 C/D). At astrocytic tumorIn patient, the tumor patient of losing ATRX expression has the longer Progression free survival phase (Fig. 5 E).
Associating IDH1-R132H and ATRX loss state can be WHOII, III level colloidThe Progression free survival phase of knurl is carried out somatotype (p < 0.0001), and IDH1-R132H and ATRX are simultaneouslyThe patient that the patient of loss expresses than IDH1-R132H and ATRX is simultaneously with longer PFS'sTrend, the patient PFS of IDH-WT is close to glioblastoma patient's PFS (Fig. 5 F).The WHOII/III level patients with gliomas that IDH1-R132H is positive and ATRX nuclear protein lacksWHOII/III level glioma positive with IDH1-R132H and that ATRX nuclear protein is expressed is suffered fromPerson compares, and has the trend (p=0.0845) of longer Progression free survival phase. IDH1-R132H the moonThe WHOII/III level patients with gliomas of property and the WHOII/III level glue of the IDH1-R132H positiveMatter knurl patient compares, the Progression free survival time shorter (p=0.0242). IDH1-R132H feminine genderProgression free survival time and the glioblastoma patient of WHOII/m level patients with gliomas connectClosely (p=0.1288).
All documents of mentioning in the present invention are all quoted as a reference in this application, just as oftenOne section of document is quoted separately as a reference. In addition should be understood that read of the present inventionAfter foregoing, those skilled in the art can carry out to the present invention the change of various unsubstantialitiesOr amendment, these equivalent form of values fall within the protection that the application's appended claims limits equallyIn scope.
Claims (6)
1. the glioma prognosis system based on IDH1-R132H and ATRX expression, its feature existsComprise in described system: (1) detects ATRX protein expression level in patients with gliomas tumourKit and optionally described kit operation instructions; (2) detect patients with gliomas tumourThe kit of middle IDH1-R132H protein expression and optionally described kit operation instructions.
2. glioma prognosis system as claimed in claim 1, is characterized in that described system bagDraw together: (1) patients with gliomas tumour FFPE kit and optionally described kit use to be saidBright book; (2) SABC detects the examination of ATRX protein expression level in patients with gliomas tumourAgent box and optionally described kit operation instructions; (3) SABC detects glioma troubleIn person's tumour the kit of IDH1-R132H protein expression and optionally described kit use and sayBright book.
3. glioma prognosis system as claimed in claim 2, is characterized in that:The evaluation criteria of IDH1-R312H dyeing: (1) strong dyeing of tumour cell matter is widely defined asPositive; (2) the weak positive being dispersed in or macrophage dyeing are defined as feminine gender.
4. glioma prognosis system as claimed in claim 2, is characterized in that: ATRX coreThe evaluation criteria of interior expression loss: brown if neoplastic cell nuclei is not dyed to, but not tumour is thinKaryon, microglia, lymphocyte and astrocyte show as strong positive, are defined as soIn ATRX core, express loss.
5. glioma prognosis system as claimed in claim 2, is characterized in that: described systemFor the Progression free survival phase somatotype of WHOII, III level glioma, IDH1-R132H and ATRXThe patient that the patient of loss expresses than IDH1-R132H and ATRX simultaneously is simultaneously with longer nothingProgress life cycle trend.
6. glioma prognosis system as claimed in claim 2, is characterized in that: described immunityThe kit that groupization detects ATRX protein expression level in patients with gliomas tumour detects ATRXArgine Monohydrochloride sequence site is 2211-2413.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610024304.0A CN105606818A (en) | 2016-01-15 | 2016-01-15 | IDH1-R132H and ATRX expression based glioma prognostic system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610024304.0A CN105606818A (en) | 2016-01-15 | 2016-01-15 | IDH1-R132H and ATRX expression based glioma prognostic system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105606818A true CN105606818A (en) | 2016-05-25 |
Family
ID=55986910
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610024304.0A Pending CN105606818A (en) | 2016-01-15 | 2016-01-15 | IDH1-R132H and ATRX expression based glioma prognostic system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105606818A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108727500A (en) * | 2018-05-03 | 2018-11-02 | 南京基诺米医疗科技有限公司 | The anti-human IDH1 R132H mutain monoclonal antibodies of mouse prepare and its immunohistochemistry purposes |
| CN112940133A (en) * | 2021-04-09 | 2021-06-11 | 福州迈新生物技术开发有限公司 | Monoclonal antibody of anti-ATRX protein, cell strain, preparation method and application thereof |
| CN114381525A (en) * | 2022-01-20 | 2022-04-22 | 上海交通大学医学院附属第九人民医院 | A set of molecular markers for prognostic typing of glioma and their typing methods and applications |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2253716A1 (en) * | 2009-05-15 | 2010-11-24 | Deutsches Krebsforschungszentrum, Stiftung des öffentlichen Rechts | Diagnostic methods for the prognosis of a brain tumor |
| WO2015116868A2 (en) * | 2014-01-29 | 2015-08-06 | Caris Mpi, Inc. | Molecular profiling of immune modulators |
-
2016
- 2016-01-15 CN CN201610024304.0A patent/CN105606818A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2253716A1 (en) * | 2009-05-15 | 2010-11-24 | Deutsches Krebsforschungszentrum, Stiftung des öffentlichen Rechts | Diagnostic methods for the prognosis of a brain tumor |
| WO2015116868A2 (en) * | 2014-01-29 | 2015-08-06 | Caris Mpi, Inc. | Molecular profiling of immune modulators |
Non-Patent Citations (4)
| Title |
|---|
| BENEDIKT WIESTLER等: "ATRX loss refines the classification of anaplastic gliomas and identifies a subgroup of IDH mutant astrocytic tumors with better prognosis", 《ACTA NEUROPATHOL》 * |
| DAVID CAPPER等: "Characterization of R132H Mutation-specific IDH1 Antibody Binding in Brain Tumors", 《BRAIN PATHOLOGY》 * |
| DAVID E. REUSS等: "ATRX and IDH1-R132H immunohistochemistry with subsequent copy number analysis and IDH sequencing as a basis for an "integrated" diagnostic approach for adult astrocytoma, oligodendroglioma and glioblastoma", 《ACTA NEUROPATHOL》 * |
| 孙红军等: "胶质瘤基于组学方法的分子标记物的研究进展", 《国际神经病学神经外科学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108727500A (en) * | 2018-05-03 | 2018-11-02 | 南京基诺米医疗科技有限公司 | The anti-human IDH1 R132H mutain monoclonal antibodies of mouse prepare and its immunohistochemistry purposes |
| CN112940133A (en) * | 2021-04-09 | 2021-06-11 | 福州迈新生物技术开发有限公司 | Monoclonal antibody of anti-ATRX protein, cell strain, preparation method and application thereof |
| CN112940133B (en) * | 2021-04-09 | 2022-04-22 | 福州迈新生物技术开发有限公司 | Monoclonal antibody of anti-ATRX protein, cell strain, preparation method and application thereof |
| CN114381525A (en) * | 2022-01-20 | 2022-04-22 | 上海交通大学医学院附属第九人民医院 | A set of molecular markers for prognostic typing of glioma and their typing methods and applications |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Otali et al. | A standard tissue as a control for histochemical and immunohistochemical staining | |
| Bryan et al. | Ductal carcinoma in situ with basal-like phenotype: a possible precursor to invasive basal-like breast cancer | |
| Dhir et al. | Early identification of individuals with prostate cancer in negative biopsies | |
| Babic et al. | The impact of pre-analytical processing on staining quality for H&E, dual hapten, dual color in situ hybridization and fluorescent in situ hybridization assays | |
| EP2046971A2 (en) | Quantifiable internal reference standards for immunohistochemistry and uses thereof | |
| Onozato et al. | Evaluation of a completely automated tissue-sectioning machine for paraffin blocks | |
| CN112326961A (en) | Analysis method and storage device for proportion of PD-L1 positive tumor cells in non-small cell lung cancer | |
| CN105606818A (en) | IDH1-R132H and ATRX expression based glioma prognostic system | |
| Houcine et al. | Neuroendocrine differentiation in basal cell carcinoma | |
| Phillips et al. | Increased p16 expression in high-grade serous and undifferentiated carcinoma compared with other morphologic types of ovarian carcinoma | |
| CN105675868A (en) | Glioma typing system based on IDH1-R132H and ATRX expression | |
| CN113281516A (en) | Application of CUL9 as marker in colorectal cancer prognosis evaluation | |
| Sun et al. | An improved processing method for breast whole-mount serial sections for three-dimensional histopathology imaging | |
| CN111948395A (en) | Quadruple marker for diagnosing immune regulation subtype of triple negative breast cancer and application thereof | |
| WO2010007509A1 (en) | Method for preparing cell standard | |
| CN115165511A (en) | PD-L1 immunohistochemical detection quality control product and reference product | |
| Salih et al. | Double immunohistochemical labelling of PRAME and Melan A in slow mohs biopsy margin assessment of lentigo maligna and lentigo maligna melanoma | |
| Mohanty et al. | Does Immunohistochemistry Add to Morphology in Differentiating Trichoepithelioma, Desmoplastic Trichoepithelioma, Morpheaform Basal Cell Carcinoma, and Microcystic Adnexal Carcinoma? | |
| Seidl et al. | Critical assessment of staining properties of a new visualization technology: a novel, rapid and powerful immunohistochemical detection approach | |
| CN113834941A (en) | Prognostic and diagnostic markers of colon cancer based on B cell expression and their uses | |
| Kahveci et al. | Detection of spermatogonial stem/progenitor cells in prepubertal mouse testis with deep learning | |
| Jasani et al. | Measurement of estrogen receptor status by immunocytochemistry in paraffin wax sections | |
| Kerger et al. | Microscopic assessment of fresh prostate tumour specimens yields significantly increased rates of correctly annotated samples for downstream analysis | |
| Ghasemi et al. | Immunohistochemical investigation of mutant BRAF V600E in common pigmented skin neoplasms, study on a sample of Iranian patients | |
| Chowers et al. | MIB-1 and PC-10 immunostaining for the assessment of proliferative activity in primary acquired melanosis without and with atypia |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160525 |