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The use of apol6 gene as a molecular marker for the diagnosis of heavy ion radiation exposure

A molecular marker, heavy ion radiation technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of small dose, inability to observe the molecular mechanism and microscopic changes of radiation damage, and difficult molecular markers, etc.

Active Publication Date: 2021-10-22
CHINA INST FOR RADIATION PROTECTION
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

For the damage caused by heavy ion radiation to the human body, the existing detection methods mainly include MTT colorimetry, flow cytometry, and chromosome and micronucleus detection methods, but the disadvantage of these methods is that they can only detect the degree of radiation damage from the cellular level , unable to observe the molecular mechanism and microscopic changes of radiation damage
[0003] Using the principle of molecular changes after heavy ion radiation to select molecular markers for diagnosis / detection of heavy ion radiation is a promising diagnostic method for heavy ion radiation that can overcome the above-mentioned defects to a large extent, but the selection of specific detection molecules (Molecular markers) need to pay attention to the following two issues: first, the sensitivity and specificity of detecting molecular changes, and the dose required to induce molecular changes should be small; It is necessary to reflect the entire radiation exposure dose in a timely manner
Therefore, it is difficult to select all molecular markers that meet the above conditions to evaluate the damage of previous radiation exposure or low dose rate radiation exposure

Method used

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  • The use of apol6 gene as a molecular marker for the diagnosis of heavy ion radiation exposure
  • The use of apol6 gene as a molecular marker for the diagnosis of heavy ion radiation exposure
  • The use of apol6 gene as a molecular marker for the diagnosis of heavy ion radiation exposure

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1: Preliminary Screening of Gene Molecular Markers for Heavy Ion Radiation Exposure Diagnosis

[0024] Using gene chip technology, by detecting the differential expression of different genes in human lymphocytes after heavy ion radiation exposure, the gene molecular markers for the diagnosis of heavy ion radiation exposure were initially screened.

[0025] 1) Extraction, quantification, quality inspection and purification of total cellular RNA

[0026] Human lymphocytes were subjected to 0.1, 0.5, 2.0Gy 12 C 6+ Cultivate for 24 hours after ion radiation irradiation (these three groups are the experimental group; non-irradiation is the control group). Total RNA was extracted from the four groups of cells according to the Trizol reagent extraction instructions. Concentration and purity were measured by spectrophotometer, and 5 g of total RNA was taken for electrophoresis inspection in 1% formaldehyde-agarose gel. According to QIAGEN Kit (QIAGEN, Germany) was...

Embodiment 2

[0038] Example 2: PCR verification confirms the feasibility of APOL6 gene as a molecular marker for the diagnosis of heavy ion radiation exposure

[0039] Extract the total RNA of the radiation-irradiated group and the non-irradiated group with Trizol reagent, measure the concentration with an ultraviolet spectrophotometer, use the total RNA as a template, and reverse-transcribe cDNA according to the instructions of the reverse transcription kit. The reverse transcription system is 20 μl.

[0040] The appropriate amount of cDNA samples from the radiation exposure group and the control group were taken to verify the differentially expressed gene of APOL6 by PCR. The primer sequences and PCR amplification conditions are shown in Table 2.

[0041] Table 2 Primer sequences and amplification conditions for RT-PCR amplified genes

[0042]

[0043]Result analysis: Compared with the cells of the control group, there were 504 differentially expressed genes in the three dose groups,...

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Abstract

The invention belongs to the application field of gene molecular markers, and relates to the use of APOL6 gene as a molecular marker in preparing reagents for heavy ion radiation exposure diagnosis. Using the APOL6 gene as a molecular marker in the detection of heavy ion radiation exposure can be more convenient and accurate, and can determine the radiation dose.

Description

technical field [0001] The present invention generally relates to the use of molecular markers in the preparation of reagents for the diagnosis of diseases or health conditions, and in particular to the use of molecular markers in the preparation of reagents / reagent sets for the diagnosis of radiation exposure. Background technique [0002] Ionizing radiation is radiation that can cause the ionization of matter. Heavy ions are a kind of dense ionizing radiation with high energy transfer linear density, which can cause serious damage to the human body. At the cellular level, heavy ion radiation damage mainly includes decreased cell survival, cell cycle arrest, and chromosomal aberrations. For the damage caused by heavy ion radiation to the human body, the existing detection methods mainly include MTT colorimetry, flow cytometry, and chromosome and micronucleus detection methods, but the disadvantage of these methods is that they can only detect the degree of radiation damage...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158
Inventor 张睿凤董娟聪林海鹏党旭红张忠新王超原雅艺任越刘建功左雅慧段志凯
Owner CHINA INST FOR RADIATION PROTECTION
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