circRNA AFF1 and application thereof

A circular, qrt-PCR technology, applied in the field of biomedical testing, can solve problems such as increased phosphorylation level, angiogenesis and loss of repair function

Pending Publication Date: 2021-02-26
XUHE TIANJIN YIYAO KEJI YOUXIAN GONGSI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] SAH is closely related to vascular dysfunction and vascular stress injury caused by hypoxia. Under hypoxia, circRNA AFF1 is abnormally activated in vascular endothelial cells, which leads to the up-regulation of the phosphorylation level of the final effector YAP in the Hippo / YAP signaling pathway. , resulting in partial loss of angiogenesis and repair function

Method used

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  • circRNA AFF1 and application thereof
  • circRNA AFF1 and application thereof
  • circRNA AFF1 and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0043] in CoCl 2Under the condition of induced hypoxia, the expression of CircRNA AFF1 in HUV-EC-C and HEBC-5i vascular endothelial cells abnormally increased, suggesting that CircRNA AFF1 has a certain connection with hypoxic stress injury of blood vessels. The specific operation was as follows: Genomic DNA was isolated with MiniBEST Universal Genomic DNA Extraction Kit Ver.5.0 (Takara, Japan), and total RNA was extracted from cells using TRIzol reagent (Invitrogen, USA) according to the manufacturer's instructions. For circRNA detection, the RNA was divided into two equal fractions before RNase R treatment. One of them was treated with RNase R to obtain circRNA, and the other was used to detect the expression of GAPDH. RNase R (Epicentre Technologies, USA) was used to remove linear RNA. 2 μg of total RNA was treated with or without RNase R (3 U / μg) at 37° C. for 30 minutes, and then the treated RNA was purified with RNeasy MinElute Cleanup Kit (Qiagen, Germany). Fractions...

Embodiment 2

[0048] After obtaining informed consent, the peripheral blood of 235 SAH patients was collected to establish the correlation between circAFF1 and SAH pathological changes. The clinical data of 235 SAH patients are shown in Table 1:

[0049] Table 1. Clinical data of 235 SAH patients

[0050] 0 I II III IV age 54.3±13.4 64.2±8.4 56.6±11.9 55.9±11.4 56±10.9 Male, n(%) 10(35.7) 2(40) 58(36.7) 12(37.5) 4(36.4)

[0051] The peripheral blood was collected and centrifuged within 2 hours, centrifuged at 1200g for 10 minutes, and the upper layer of plasma was separated. Add TRZOL to plasma to extract RNA, then perform reverse transcription, and finally perform real-time quantitative PCR. RNA extraction and RT-PCR experimental steps refer to Example 1. The results showed that the expression of CircRNA AFF1 was significantly increased in SAH patients compared with the expression of CircRNA AFF1 in the blood of normal people. Such as Figure ...

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Abstract

The invention belongs to the field of biomedicine examinations, and particularly relates to circRNA AFF1 used for diagnosing subarachnoid hemorrhage (SAH). The invention also relates to application ofthe circRNA AFF1 in diagnosis of the SAH, and a related kit. The nucleotide sequence of the circRNA AFF1 is disclosed by SEQ ID No.1. The RNA in peripheral blood plasma is extracted to discover thatas an important biological detection index, the circRNA AFF1 can be applied to quantitative detection so as to judge whether a patient has a SAH risk or not, early diagnosis, early prevention and early treatment of SAH patients can be realized, and the circRNA AFF1 performs an important function in the early-stage diagnosis and the differential diagnosis of the SAH.

Description

technical field [0001] The invention belongs to the field of biomedical testing, in particular, relates to a circular circRNA AFF1 for diagnosing subarachnoid hemorrhage (SAH), and also relates to the use of the circular circRNA AFF1 in diagnosing subarachnoid hemorrhage (SAH) applications and related kits. Background technique [0002] Subarachnoid hemorrhage (SAH) is a neurological emergency with a mortality rate of 50%. Except for the rupture of aneurysm, the other causes are unclear, and bleeding after onset is especially dangerous. The reasons are closely related to abnormal vascular function. SAH not only causes a devastating blow to the central nervous system, but also seriously affects multiple other organs. The most common complication is the loss of neurological function due to brain tissue ischemia and hypoxia, leaving permanent neurological dysfunction. [0003] SAH is a neurological disease with a sudden and high fatality rate. For a long time, its diagnosis ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/178C12Q2600/158
Inventor 陈双王红光闫华冯韧赖科观吴海东吕欣泽李咪咪鲍静雨杨学佳
Owner XUHE TIANJIN YIYAO KEJI YOUXIAN GONGSI
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