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Molecular marker for detecting neural tube malformation and application of marker

A neural tube defect and molecular marker technology, which is applied in the biological field, can solve the problems of functional experimental research without a single specific protein modification, and the lack of access to human NTDs molecular markers, so as to be easy to popularize and use, and realize early diagnosis and intervention. , the effect of simple production process

Active Publication Date: 2019-07-09
首都儿科研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the existing technology has found that Wnt, PCP, Notch, Hh / SHH, BMP and other signaling pathways are all related to human NTDs, there is no functional experimental study on a single specific protein modification, especially histones and related genes and NTDs Whether there are relevant reports, and there is no clear molecular marker of human NTDs, which can be used for clinical diagnosis and detection of human NTDs

Method used

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  • Molecular marker for detecting neural tube malformation and application of marker
  • Molecular marker for detecting neural tube malformation and application of marker
  • Molecular marker for detecting neural tube malformation and application of marker

Examples

Experimental program
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experiment example 1

[0056] Embryonic stem cells are mouse embryonic stem cells Sv / 129, which come from Beijing Xuanwu Hospital, and are frozen in the cell bank of Beijing Capital Institute of Pediatrics. Methotrexate (MTX) is purchased from Pfizer (Perth) Pty Limited, and the concentration of the storage solution is 500mg / 20mL.

[0057]The mouse embryonic stem cells were cultured in DMEM high-glucose medium at 37°C in a carbon dioxide incubator, and the medium formula (mouse embryonic stem cells (mESC) Sv / 129 was maintained in Dulbecco's modified Eagle medium (DMEM, Gibco, USA), the medium contains 0.1mM β-mercaptoethanol (Invitrogen, Carlsbad, USA), 0.1mM non-essential amino acid (NEAA) (Invitrogen, Carlsbad) (USA), 0.1mM glutamic acid (Invitrogen, Carlsbad, USA) , 15% fetal bovine serum (Gibco, USA) and 1000 U / ml mouse leukemia inhibitory factor (Millipore, Billerica, USA), 0.2% gelatin (Invitrogen, Carlsbad, USA)). Place the cells in a humidified incubator at 37°C, 5% CO 2 Conditioned cultu...

experiment example 2

[0079] 1. Experimental animals and materials

[0080] SPF grade adult C57BL / 6J mice, male and female, 7-8 weeks old, weighing 17-19 g. Purchased from Beijing Jinmuyang Experimental Animal Breeding Co., Ltd. Methotrexate (MTX) was purchased from Pfizer (Perth) Pty Limited, and the concentration of the stock solution was 500mg / 20mL.

[0081] 2. Preparation of mouse model

[0082] After 2 days of adaptive feeding, the purchased mice were randomized into groups. The grouped female mice were housed overnight with the same batch of male mice at a ratio of 2:1, and the vaginal suppositories were found the next morning and were determined to be 0.5 days pregnant. The experimental group was given intraperitoneal injection of different doses of MTX on the 7.5th day of pregnancy; the control group was given the same volume of normal saline. On the 13.5th day of pregnancy, the pregnant mice were taken from the eyeballs to collect blood. After blood collection, the mice were killed by...

Embodiment 1

[0090] The present embodiment provides a method for detecting neural tube defects, comprising the following steps:

[0091] 1) Extract H2AK119 and Mdm2 of the sample to be tested;

[0092] 2) Quantitatively analyze the content of H2AK119, Mdm2, specific marker genes Pax6, Nestin, Bmp4; 3) Compared with the control samples, if the levels of H2AK119 and Mdm2 are up-regulated, and the specific marker genes Pax6, Nestin, Bmp4 are all down-regulated, the neuronal Tube deformities occur.

[0093] The sample to be tested is the brain tissue of a patient with neural tube defects, and the control sample is the brain tissue of a normal human body. The above-mentioned tissues were taken from Lvliang City, Shanxi Province, and the family members knew and agreed to transport them on dry ice to Beijing Capital Institute of Pediatrics and store them in a -80 refrigerator.

[0094] The extraction method of H2AK119 is as follows: the EPIGENTEK kit EpiQuik TM Total HistoneExtraction Kit is us...

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Abstract

The invention belongs to the field of biology, in particular to a molecular marker for detecting neural tube malformation and an application of the marker. The molecular marker comprises specific marker gene and / or proteins, wherein the specific marker genes is at least one of Pax6, Nestin and Bmp4; and the proteins are H2AK119, Mdm2 and / or a gene encoding at least one of the H2AK119 and the Mdm2.The molecular marker can be used for preparing products for diagnosing or detecting neural tube defects, can quickly and conveniently detect changes of the molecular marker, and can be used for prenatal diagnosis, timely treatment and reduction of the incidence rate of neonatal neural tube malformation.

Description

technical field [0001] The invention relates to the field of biology, in particular to a molecular marker for detecting neural tube defects and an application thereof. Background technique [0002] Neural tube defects (NTDs) is a high-incidence birth defect disease with the highest morbidity and mortality among birth defects, and it is also the most common and serious birth defect disease that affects the quality of the population in my country and the world. . NTDs usually occur early in pregnancy and are malformations from the head to the spine due to a non-closed or insufficient neural tube. Deformities ranging from anencephaly to mild spina bifida: including anencephaly, deencephaly, microcephaly, macrocephaly, hydrocephalus, meningocele, craniospina bifida, and spina bifida. The deformity rate of NTDs in newborns is as high as 0.1-0.5%, and its etiology and pathogenesis are still not very clear. The development of the neural tube is a complex multi-step process, which...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883G01N33/6893C12Q2600/158G01N2800/28
Inventor 裴培王珊张霆崔小岱
Owner 首都儿科研究所
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