CN114414806A - Marker for diagnosing gallbladder cancer and application thereof - Google Patents
Marker for diagnosing gallbladder cancer and application thereof Download PDFInfo
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- CN114414806A CN114414806A CN202210116583.9A CN202210116583A CN114414806A CN 114414806 A CN114414806 A CN 114414806A CN 202210116583 A CN202210116583 A CN 202210116583A CN 114414806 A CN114414806 A CN 114414806A
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- gallbladder cancer
- glyceraldehyde
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- phosphate dehydrogenase
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- 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
- G01N33/57488—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 involving compounds identifable in body fluids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/90203—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/904—Oxidoreductases (1.) acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
Abstract
The invention discloses a marker for diagnosing gallbladder cancer and application thereof, wherein the marker is selected from at least one of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase. The invention discovers that glyceraldehyde-3-phosphate dehydrogenase, particularly the combination of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase can be used as a primary diagnosis marker for diagnosing gallbladder cancer for the first time, and has higher detection specificity.
Description
Technical Field
The invention belongs to the field of medical detection, in particular to a marker for diagnosing gallbladder cancer and application thereof, which is used for constructing a reagent or a kit for diagnosing gallbladder cancer, belongs to the field of medical examination and inspection service, and particularly relates to the fields of reagents, test paper, matched equipment and consumables for in-vitro diagnosis in-vitro diagnosis and detection.
Background
The gallbladder is an important digestive organ in human body, and not only has the functions of storage, concentration and excretion (contraction), but also has the important function of regulating the pressure of the internal and external biliary tracts of the liver and the important functions of secretion and immunity. Gallbladder cancer (GBC) refers to a malignant tumor of gallbladder (including gallbladder bottom, body, neck and cystic duct), and accounts for 80% -95% of the malignant tumor of biliary tract. Gallbladder cancer has the characteristics of invisibility, easy invasion and metastasis, difficult early diagnosis, poor prognosis and the like. The incidence rate of gallbladder cancer in China accounts for 0.4-3.8% of that of biliary tract diseases at the same stage, and the gallbladder cancer occupies the 6 th position of digestive tract tumors. In the past decades, gallbladder cancer treatment strategies have not made substantial progress, and due to the lack of typical symptoms, the early diagnosis rate is only 19.1%, most patients are already in the middle and late stages when they are clearly diagnosed, and lymph node metastasis has occurred when 50% of patients are found. Surgical resection remains by far the only treatment for patients with gallbladder cancer. Surgical resection, however, is of little therapeutic interest for patients with advanced stage disease. The gallbladder cancer is not sensitive to the treatment measures such as radiotherapy, chemotherapy and the like, so the integral prognosis is very poor, and the total survival rate of the gallbladder cancer patients in 5 years is only 5 percent. In addition, the tendency to relapse or metastasize after surgery largely determines the poor prognosis of advanced gallbladder cancer. The pathogenesis of gallbladder cancer is mainly related to various genes and environmental factors, and chronic biliary tract system infection, special chemical substances, heavy metal environmental exposure, dietary factors and the like are related to the formation of the gallbladder cancer. Estrogen, cholesterol circulation and Salmonella infections are among the major causes of high gallbladder cancer incidence in developing countries. Although remarkable progress is made in the aspect of molecular biological pathogenesis of the gallbladder cancer, the functions of signal pathways and genes such as PI3K-AKT-mTOR, Notch, ErbB, MAPK-ERK and the like in the gallbladder cancer are basically clarified, however, the research cannot change the treatment and cure pathways of the gallbladder cancer, and the research on the biomarker for early detection for diagnosing the gallbladder cancer still has a crucial position in the treatment of the gallbladder cancer.
Chinese granted patent CN108931633B discloses marker PIM1 for diagnosing and prognosing gallbladder cancer, and also discloses a kit for diagnosing and prognosing gallbladder cancer, which can be used for diagnosing, predicting, detecting or screening human gallbladder cancer cell spreading, gallbladder cancer clinical stage or gallbladder cancer patient prognosis. The patent also discloses the application of the PIM1 protein inhibitor in preparing anti-gallbladder cancer drugs. The research result of the patent shows that the PIM1 has very important significance in clinical practice, can possibly become a new generation of tumor biological diagnosis marker and a molecular target for tumor treatment, opens up the research of a brand new target of gallbladder cancer diseases mainly based on the PIM1, and also opens up a new way for developing new anti-tumor drugs.
Chinese patent application CN111455049A discloses a diagnostic marker for gallbladder cancer, which is lncrnameng 3 and MALAT 1. The patent application screens the markers LncRNA MEG3 and MALAT1(LncRNA MEG3 is low in expression in gallbladder cancer, MALAT1 is high in expression in gallbladder cancer) from the gallbladder cancer tissues by the technical means of bioinformatics and existing molecular biology, and provides guidance and help for the treatment of the gallbladder cancer.
However, there is no data disclosing the effect of the above markers in the diagnosis of gallbladder cancer. How to discover more markers capable of being used for gallbladder cancer targeting remains one of the problems to be solved urgently by the technical personnel in the field.
Disclosure of Invention
The invention aims to provide a marker for diagnosing gallbladder cancer, which solves the problems in the prior art.
Glyceraldehyde-3-phosphate dehydrogenase (GAPD) is widely distributed in various tissues and organs of the human body, is known to catalyze the oxidation (dehydrogenation) and phosphorylation of Glyceraldehyde-3-phosphate to generate 1, 3-diphosphoglycerate, is a central link in sugar metabolism, and is used for the relief and treatment of patients with sugar metabolism problems.
Human acetyl-CoA dehydrogenase has been commercialized for detection, but its use in the diagnostic test of gallbladder cancer has not been found.
Without being bound to any theory, the present inventors have unexpectedly found that glyceraldehyde-3-phosphate dehydrogenase and acetyl-coa dehydrogenase exist characteristically in gallbladder cancer patients, and thus have completed the present invention. The embodiment of the invention provides the following technical scheme:
the present invention relates, in one aspect, to a marker for the diagnosis of gallbladder cancer, the marker being selected from at least one of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-coa dehydrogenase.
In a preferred embodiment of the present invention, the marker is a combination of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase.
In a preferred embodiment of the present invention, the glyceraldehyde-3-phosphate dehydrogenase and the human acetyl-CoA dehydrogenase refer to glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase in serum.
In another aspect, the invention also relates to a kit for diagnosing gallbladder cancer, which comprises a reagent for detecting the concentrations of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase in human serum.
In a preferred embodiment of the present invention, the reagents for measuring the concentrations of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase in human serum are a glyceraldehyde-3-phosphate dehydrogenase ELISA reagent and a human acetyl-CoA dehydrogenase ELISA reagent, respectively.
The invention also relates to application of the marker and the kit in preparation of a reagent for diagnosing gallbladder cancer.
Advantageous effects
Compared with the prior art, the invention at least realizes the following beneficial effects:
the invention discovers that glyceraldehyde-3-phosphate dehydrogenase, particularly the combination of glyceraldehyde-3-phosphate dehydrogenase and human acetyl coenzyme A dehydrogenase can be used as a preliminary diagnosis marker for diagnosing the gallbladder cancer for the first time, has higher detection specificity, and provides a quick and effective new choice for early diagnosis of the gallbladder cancer.
Drawings
FIG. 1 is a graph showing a comparison of the concentration of glyceraldehyde-3-phosphate dehydrogenase in the serum of gallbladder cancer patients with that of healthy people.
FIG. 2 is a graph showing a comparison of the concentration of human acetyl-CoA dehydrogenase in the serum of gallbladder cancer patients with that of healthy people.
FIG. 3 is a ROC diagram showing glyceraldehyde-3-phosphate dehydrogenase combined with human acetyl-CoA dehydrogenase as a diagnosis of gallbladder cancer.
Detailed Description
In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to 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 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.
Unless otherwise specified, the reagents involved in the examples of the present invention are all commercially available products, and all of them are commercially available.
Example 1
Sample collection
The diagnosis time of 40 patients with gallbladder cancer is 6-12 months in 2020 in Henan province, the patients are 40-50 years old, the average age is 46.8 years old, 22 men and 18 women. A whole blood sample was collected from 18 healthy people, with a mean age of 46.3 years, 10 men, and 8 women.
Taking a whole blood sample of each subject, placing the whole blood sample at room temperature for 2 hours, centrifuging the whole blood sample for 5 minutes at 3000g, sucking supernatant, transferring the supernatant to a sterile EP tube, subpackaging, and storing at-80 ℃ until the whole blood sample is detected.
Test methods and results
The ELISA kit method is adopted to detect the content of glyceraldehyde-3-phosphate dehydrogenase and human acetyl coenzyme A dehydrogenase in the serum of a subject. The ELISA detection kit for glyceraldehyde-3-phosphate dehydrogenase is from Yaji Biotechnology Ltd of Shanghai, and the operation is carried out according to the kit instructions. The human acetyl-coa dehydrogenase ELISA kit was from shanghai caryopsis laboratory equipment ltd, and its operation was performed according to the kit instructions.
The detection results are shown in fig. 1 and fig. 2, and the results are: the average relative concentration of glyceraldehyde-3-phosphate dehydrogenase in the serum of the healthy group is 100 +/-27.4%, the average relative concentration of glyceraldehyde-3-phosphate dehydrogenase in the serum of the gallbladder cancer patient group is 176.6 +/-22.7%, and the concentrations of glyceraldehyde-3-phosphate dehydrogenase in the serum of the healthy group and the serum of the gallbladder cancer patient group have significant difference. Similarly, the average relative concentration of human acetyl-CoA dehydrogenase in the serum of the healthy group is 100 + -22.3%, the average relative concentration of human acetyl-CoA dehydrogenase in the serum of the gallbladder cancer patient group is 150.6 + -21.8%, and the concentrations of human acetyl-CoA dehydrogenase in the serum of the healthy group and the gallbladder cancer patient group have significant differences.
The experimental results show that the glyceraldehyde-3-phosphate dehydrogenase and the human acetyl-CoA dehydrogenase are both significantly higher than those of healthy people, and that the glyceraldehyde-3-phosphate dehydrogenase and the human acetyl-CoA dehydrogenase can be used as diagnostic markers in gallbladder cancer patients.
Example 2
Another 100 patients with gallbladder cancer were selected from the gallbladder cancer patients diagnosed in tumor hospitals of Henan province from 6-12 months at 2021, and AUC values were determined based on the content of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase in serum, as shown in Table 1 below, wherein the ROC curve of the combination of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase as diagnostic markers is shown in FIG. 3.
Table 1: AUC value for diagnosis of gallbladder cancer patient
| Marker substance | AUC value |
| Glyceraldehyde-3-phosphate dehydrogenase | 0.635 |
| Human acetyl-CoA dehydrogenase | 0.574 |
| Glyceraldehyde-3-phosphate dehydrogenase + human acetyl-CoA dehydrogenase | 0.763 |
The results show that the AUC value of glyceraldehyde-3-phosphate dehydrogenase alone as a diagnostic marker for gallbladder cancer is 0.635, and the AUC value of human acetyl-CoA dehydrogenase alone as a diagnostic marker for gallbladder cancer is 0.574. Whereas the AUC value of the combined diagnosis of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase in combination as a marker was 0.763. Therefore, the combination of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase as a marker has higher accuracy in the diagnosis of gallbladder cancer.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (7)
1. A marker for diagnosing gallbladder cancer, wherein the marker is selected from at least one of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-coa dehydrogenase.
2. The marker according to claim 1, wherein the marker is a combination of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-coa dehydrogenase.
3. The marker according to claim 1, wherein the glyceraldehyde-3-phosphate dehydrogenase and the human acetyl-coa dehydrogenase are glyceraldehyde-3-phosphate dehydrogenase and human acetyl-coa dehydrogenase in serum.
4. Use of a marker according to any one of claims 1 to 3 in the preparation of a reagent for the diagnosis of gallbladder cancer.
5. A kit for diagnosing gallbladder cancer, which comprises a reagent for detecting the concentrations of glyceraldehyde-3-phosphate dehydrogenase and human acetyl-CoA dehydrogenase in serum.
6. The kit of claim 5, wherein the reagents are glyceraldehyde-3-phosphate dehydrogenase ELISA reagent and human acetyl-CoA dehydrogenase ELISA reagent, respectively.
7. Use of the kit according to claim 5 or 6 for the preparation of a reagent for diagnosing gallbladder cancer.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210116583.9A CN114414806A (en) | 2022-02-07 | 2022-02-07 | Marker for diagnosing gallbladder cancer and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210116583.9A CN114414806A (en) | 2022-02-07 | 2022-02-07 | Marker for diagnosing gallbladder cancer and application thereof |
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2002033062A1 (en) * | 2000-10-16 | 2002-04-25 | Bayer Aktiengesellschaft | REGULATION OF HUMAN ACYL-CoA DEHYDROGENASE |
| US20130338027A1 (en) * | 2012-06-15 | 2013-12-19 | Nuclea Biotechnologies, Inc. | Predictive Markers For Cancer and Metabolic Syndrome |
| US20160069889A1 (en) * | 2012-12-19 | 2016-03-10 | Caris Science, Inc. | Compositions and methods for aptamer screening |
| CN107022635A (en) * | 2017-06-02 | 2017-08-08 | 北京泱深生物信息技术有限公司 | The application of ACADL genes and its expression product in abdominal aneurvsm diagnosis and treatment product is prepared |
| TW201816399A (en) * | 2016-10-27 | 2018-05-01 | 百威研發股份有限公司 | Method for diagnosis of Alzheimer's disease |
| US20190022191A1 (en) * | 2016-01-20 | 2019-01-24 | The Johns Hopkins University | Mutant glyceraldehyde-3-phosphate dehydrogenase (gapdh) compositions and methods of treating cancer |
| US20190083436A1 (en) * | 2016-03-08 | 2019-03-21 | Agency For Science, Technology And Research | Methods of diagnosing cancer |
| US20200150125A1 (en) * | 2017-03-12 | 2020-05-14 | Yeda Research And Development Co., Ltd. | Methods of diagnosing and prognosing cancer |
| CN111198271A (en) * | 2018-11-16 | 2020-05-26 | 山东泽济生物科技有限公司 | Chemiluminescent enzyme-linked immunosorbent assay kit for glyceraldehyde-3-phosphate dehydrogenase detection |
-
2022
- 2022-02-07 CN CN202210116583.9A patent/CN114414806A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002033062A1 (en) * | 2000-10-16 | 2002-04-25 | Bayer Aktiengesellschaft | REGULATION OF HUMAN ACYL-CoA DEHYDROGENASE |
| US20130338027A1 (en) * | 2012-06-15 | 2013-12-19 | Nuclea Biotechnologies, Inc. | Predictive Markers For Cancer and Metabolic Syndrome |
| US20160069889A1 (en) * | 2012-12-19 | 2016-03-10 | Caris Science, Inc. | Compositions and methods for aptamer screening |
| US20190022191A1 (en) * | 2016-01-20 | 2019-01-24 | The Johns Hopkins University | Mutant glyceraldehyde-3-phosphate dehydrogenase (gapdh) compositions and methods of treating cancer |
| US20190083436A1 (en) * | 2016-03-08 | 2019-03-21 | Agency For Science, Technology And Research | Methods of diagnosing cancer |
| TW201816399A (en) * | 2016-10-27 | 2018-05-01 | 百威研發股份有限公司 | Method for diagnosis of Alzheimer's disease |
| US20200150125A1 (en) * | 2017-03-12 | 2020-05-14 | Yeda Research And Development Co., Ltd. | Methods of diagnosing and prognosing cancer |
| CN107022635A (en) * | 2017-06-02 | 2017-08-08 | 北京泱深生物信息技术有限公司 | The application of ACADL genes and its expression product in abdominal aneurvsm diagnosis and treatment product is prepared |
| CN111198271A (en) * | 2018-11-16 | 2020-05-26 | 山东泽济生物科技有限公司 | Chemiluminescent enzyme-linked immunosorbent assay kit for glyceraldehyde-3-phosphate dehydrogenase detection |
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