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Novel long-chain non-coding RNA of RA marker and application of novel long-chain non-coding RNA

A long-chain non-coding and marker technology, applied in the new RA marker long-chain non-coding RNA and its application field

Active Publication Date: 2022-01-28
FIRST AFFILIATED HOSPITAL OF ANHUI UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although domestic and foreign studies on the pathogenesis of RA have achieved a series of achievements in the fields of mRNA, protein and miRNA and other epigenetics (Tsai CY, Hsieh SC, Liu CW, et al. The Expression of Non-Coding RNAs and Their Target Molecules in Rheumatoid Arthritis: A Molecular Basis for Rheumatoid Pathogenesis and Its Potential Clinical Applications[J].Int J Mol Sci,2021,22(11),doi:10.3390 / ijms22115689.), but its exact pathogenesis is still under active research Exploring

Method used

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  • Novel long-chain non-coding RNA of RA marker and application of novel long-chain non-coding RNA
  • Novel long-chain non-coding RNA of RA marker and application of novel long-chain non-coding RNA
  • Novel long-chain non-coding RNA of RA marker and application of novel long-chain non-coding RNA

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Volcano map of differentially expressed lncRNAs in RA patients and normal PBMCs

[0036] 1. Materials and methods

[0037] (1) Materials

[0038] 1. Source of cases: PBMCs of 3 RA patients were from inpatients confirmed in the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, and PBMCs of 3 normal people were from patients undergoing physical examination at the Physical Examination Center of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine during the same period.

[0039] 2. Inclusion criteria: all patients with RA were in line with the diagnostic criteria proposed by the American College of Rheumatology (ACR) and the European League Against Rheumatism (EULAR) in 2010:

[0040] A: Affected joints

[0041] 1 large joint (0 points)

[0042] 2-10 large joints (1 point)

[0043] 1-3 small joints (with or without large joints) (2 points)

[0044] 4-10 small joints (with or without large joints) (3 points)

...

Embodiment 2

[0081] RT-qPCR preliminary verification of differential expression of AC123912.4 in 45 pairs of RA patients and normal PBMCs 1. Experimental materials

[0082] The PBMCs of 45 RA patients were selected from inpatients diagnosed in the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine (see Table 1 for the specific information of 45 patients, and Table 2 for the sample characteristics), and the PBMCs of 45 normal people were obtained from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine during the same period. A patient undergoing physical examination at the physical examination center of an affiliated hospital.

[0083] Table 2 Sample Information 1

[0084]

[0085]

[0086]

[0087] Table 3 Sample Information 2

[0088]

[0089] 2. Method

[0090] 1. Extraction of PBMCs from RA patients

[0091] At room temperature, add 6mL Ficoll-Paque PLUS to a 50mL centrifuge tube; add 4mL fresh anticoagulated blood...

Embodiment 3

[0118] Application of AC123912.4 Overexpression Sequence and Small Interfering Sequence in RA-FLS

[0119]This embodiment is aimed at the full-length sequence design of AC123912.4, and synthesizes its specific overexpression sequence pcDNA3.1-AC123912.4 and small interfering sequence si-AC123912.4 (its nucleotide sequence is shown in the table below), infection RA-FLS makes AC123912.4 gene overexpression and small interference in cells, and further applies the overexpression and small interference gene carrier to regulate the inflammatory response and hypercoagulable state of RA-FLS.

[0120]

[0121]

[0122] The specific virus packaging and cell infection methods are as follows:

[0123] (1) When 293T cells are cultured in a 6cm dish until 80-90% confluent, discard the culture medium and wash the cells twice with 3mL PBS; (2) Add 1mL Trypsin-EDTA solution, mix well, and carefully suck off the trypsin solution, placed at 37°C for 3 minutes; (3) Add 2 mL of DMEM culture...

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Abstract

The invention provides novel long-chain non-coding RNA of a RA marker and application of the novel long-chain non-coding RNA, belonging to the field of molecular biology of rheumatism. The novel long-chain non-coding RNA is AC123912.4, and the nucleotide sequence of the long-chain non-coding RNA is as shown in SEQ ID NO. 1. According to the sequence of the long-chain non-coding RNA, specific real-time quantitative PCR primers and probes are designed and synthesized, and a preparation for auxiliary diagnosis or curative effect prediction of RA is prepared. A real-time quantitative PCR preparation is utilized to detect the expression level of the long-chain non-coding RNA in an RA clinical case specimen, and it is found that the expression of the long-chain non-coding RNA in the RA is obviously up-regulated; and thus, the long-chain non-coding RNA is expected to be made into a preparation for auxiliary diagnosis or curative effect prediction of the RA.

Description

technical field [0001] The invention belongs to the field of molecular biology of rheumatic diseases, and relates to the application of long-chain non-coding RNA in the preparation of auxiliary diagnosis or prognosis preparations for RA, in particular to a new RA-marked long-chain non-coding RNA and its application. Background technique [0002] Rheumatoid arthritis (RA) is a chronic, systemic, progressive, autoimmune disease with erosive, symmetrical, and polyarthritis as the main clinical manifestations. The basic pathological changes are the formation of synovitis and pannus, and Gradual destruction of articular cartilage and bone, which may eventually lead to joint deformity and loss of function (Weyand CM, Goronzy JJ. The immunology of rheumatoid arthritis[J]. Nat Immunol, 2021, 22(1), doi: 10.1038 / s41590-020-00816 -x.). The incidence of RA is mainly concentrated in 20-50 years old, accounting for about 1% of the world's population, the incidence rate of women is 2-3 t...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6883C12Q1/6851C12N15/11A61K45/00A61K31/7088A61P19/02A61P29/00
CPCC12N15/113C12Q1/6883C12Q1/6851A61K45/00A61K31/7088A61P19/02A61P29/00C12N2310/11C12Q2600/158C12Q2600/178C12Q2600/118C12Q2600/136C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 刘健文建庭王馨王杰王帆帆
Owner FIRST AFFILIATED HOSPITAL OF ANHUI UNIV OF CHINESE MEDICINE
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