Application of CDR1as to prenatal screening of neural tube defects, and product and method for performing prenatal screening of neural tube defects

A neural tube defect and prenatal screening technology, applied in the field of clinical diagnosis, can solve problems such as inability to diagnose, and achieve the effects of low professional requirements, wide applicability, and low detection cost

Active Publication Date: 2019-07-16
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, on the one hand, this technique is too dependent on the operator's skill, and on the other hand, it cannot be distinguished before obvious morphological changes occur, so it cannot be diagnosed in early pregnancy

Method used

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  • Application of CDR1as to prenatal screening of neural tube defects, and product and method for performing prenatal screening of neural tube defects
  • Application of CDR1as to prenatal screening of neural tube defects, and product and method for performing prenatal screening of neural tube defects
  • Application of CDR1as to prenatal screening of neural tube defects, and product and method for performing prenatal screening of neural tube defects

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Screening of markers

[0052] The neural tube tissue samples (test samples) of 10 groups of children with NTDs and 10 groups of normal fresh neural tube tissue samples (control samples) were collected, ground and crushed with liquid nitrogen, and the total RNA was extracted with the ThermoFisher Tissue RNA Isolation Kit. The circular RNA chip was sequenced and microarray analysis was performed to select the five circular RNAs with the largest fold difference, including circRNA-CDR1as, Circ-TTBL C2, Circ-MYLK, CZNF292 and CircRNA100876.

[0053] For the test samples and control samples, the extracted total RNA was reverse-transcribed with random primers in a reverse transcription kit, circRNA-CDR1as primers were designed, and the expression levels of CDR1as in the test samples and control samples were detected by RT-PCR, and the statistics and clarification were made Elevation of CDR1as in test samples. The result is as figure 1 Among them, the expression dif...

Embodiment 2

[0059] Example 2 Effect of circRNA-CDR1as on cell proliferation

[0060]Using pcDNA3.1 to construct a plasmid expressing circRNA-CDR1as, using lipo2000 to transiently transfect embryonic stem cells, and then using 5uM RA to induce, successfully induce differentiation into neural stem cells, and construct circRNA-CDR1as overexpressed neural stem cells. The following experiments were carried out on the induced and differentiated neural stem cells: the neural stem cells were incubated with CCK8, and the absorbance at 450nm of the cells was detected on the 1st, 2nd, 3rd, 4th, 5th, and 6th days respectively, and the cell proliferation and growth curves were made to reflect the proliferation. The result is as figure 2 As shown, the neural stem cells overexpressing circRNA-CDR1as proliferated faster than the neural stem cells emptying the pcDNA3.1 plasmid, indicating that circRNA-CDR1as could promote cell proliferation.

[0061] Design and purchase siRNA of circRNA-CDR1as from Ruib...

Embodiment 3

[0063] Example 3 The effect of circRNA-CDR1as on the expression level of TRIM-71

[0064] Referring to the experiment in Example 2, neural stem cells with overexpression of circRNA-CDR1as and neural stem cells with empty pcDNA3.1 plasmid were obtained, and the two kinds of cells were cultured and treated with or without Dactinomycin. The total protein in the cells was extracted using the whole protein extraction kit, and after the protein concentration was measured by the BCA protein concentration assay kit, each group was diluted to the same concentration and added to the sample buffer for denaturation, and detected by western blot. GAPDH was used as an internal reference in the experiment. The differences in the expression of TRIM-71 in the cells were compared. The result is as Figure 4 As shown, the amount of TRIM71 protein in neural stem cells with circRNA-CDR1as overexpression is less than that of neural stem cells with empty pcDNA3.1 plasmid. of expression.

[0065] ...

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Abstract

The invention provides an application of CDR1as to prenatal screening of neural tube defects, and a product and method for performing prenatal screening of neural tube defects, and relates to the technical field of clinical diagnosis. The invention provides an application of circRNA-CDR1as as a marker for prenatal screening of neural tube defects. Through research, the inventor finds out that the,circRNA-CDR1as is notably highly expressed in the tissue of NTDs child patients, the circRNA-CDR1as can be separately or can be united with other markers to be used for conventional screening of congenital malformation at the early pregnancy, and has great significance to families and society. According to the product and method for performing prenatal screening of neural tube defects, through detecting the expression number of the circRNA-CDR1as, NTDs is diagnosed, the detection cost is low, the detection is accurate and quick, and the NTDs can be diagnosed at the early pregnancy.

Description

technical field [0001] The invention relates to the technical field of clinical diagnosis, in particular to an application of CDR1as in prenatal screening of neural tube defects, a product and a method for prenatal screening of neural tube defects. Background technique [0002] Neural tube defects (NTDs) are one of the most common congenital birth defects. It is a series of neural tube defects affecting the brain and spinal cord caused by the failure of the neural tube to close normally during early embryonic development. The most common NTDs are anencephaly and spinal deformity. Anencephaly usually dies in the womb before birth, forming a "stillbirth" or "stillbirth". Even if it is born, it will die within a short time. Once diagnosed , almost none survived. Those with spinal deformity have severe permanent disabilities, which bring huge mental and economic burdens to the family. Preventing the occurrence of NTDs through preventive intervention, or timely termination of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178Y02A50/30
Inventor 李方成金必莲李军亮许新科陈伟陈程林锦荣袁宏耀李杨潘君平王方宇赵灵凤
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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