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Application of lncRNA AK124826 and related molecules thereof in diagnosis of polymyositis/dermatomyositis

A dermatomyositis, multiple technology, applied in the field of biotechnology and treatment, can solve the problems of complex pathogenesis, abnormal factor response, and the specific mechanism is not fully elucidated.

Inactive Publication Date: 2020-09-04
常熟市第二人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pathogenesis of PM / DM is complex, involving many factors such as genetics, infection, immune function, and abnormal expression of microRNA. In recent years, the pathogenesis of the disease has been studied in depth. However, the specific mechanism has not been fully elucidated

Method used

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  • Application of lncRNA AK124826 and related molecules thereof in diagnosis of polymyositis/dermatomyositis
  • Application of lncRNA AK124826 and related molecules thereof in diagnosis of polymyositis/dermatomyositis
  • Application of lncRNA AK124826 and related molecules thereof in diagnosis of polymyositis/dermatomyositis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Analysis of the expression of lncRNA AK124826 in PM / DM patients

[0032] In this embodiment, PM / DM patients are taken as the disease group, and healthy people with normal liver and kidney function indexes are used as the normal control group. 50 samples of PM / DM patients and 50 healthy control groups are collected from the Second People's Hospital of Wuxi City, respectively. Peripheral blood CD4+ T cells of the above subjects were isolated and serum samples were preserved.

[0033] 1. Analyze the expression of IL-27, TGF-β and creatine kinase (creatine kinase, CK) in the serum of the disease group and the control group.

[0034] The expression levels of IL-27, TGF-β and creatine kinase (Creatine Kinase, CK) in serum of healthy control group and PM / DM patients were detected by ELISA technology. The operation steps were carried out according to the ELISA kit instructions of IL-27, TGF-β and CK respectively. The microplate reader measured the absorbance of each ...

Embodiment 2

[0047] Example 2 Study on the regulatory mechanism of lncRNA AK124826 on the differentiation of Th17 / Treg cells in patients with PM / DM

[0048] (1) Construction of lncRNA AK124826 overexpression vector

[0049] Firstly, the full-length lncRNA AK124826 gene was cloned, and the cDNA chain of NcRNA AK124826 was amplified using the healthy human peripheral blood cell genome as a template. The reaction system: 1 μL of upstream primer, 1 μL of downstream primer, 1 μL of template DNA, 10 μL of 5×PBS buffer, 4 μL of dNTP Mix, HS DNA polymerase (2.5U / μL) 0.5 μL (purchased from Dalian Bao Biological Company), ddH 2 O3 2.5 μL. Reaction conditions: 98°C, 10s, 55°C (10s), 72°C (90s), 30 cycles in total. The sequence of the upstream primer is: 5'-GCAAGCTTTATGAACTTTCTGCTGTCTTG-3', and the sequence of the downstream primer is: 5'-GCTCTAGATTACCGCCTCGGCCTGTCACATC-3'. The cDNA of the amplified lncRNA AK124826 was connected between the restriction sites of the expression vector pcDNA3.1(+) di...

Embodiment 3

[0058] Example 3 Correlation between lncRNA AK124826 and IL27 gene and protein expression

[0059] One of the modes of action of lncRNA is to regulate the expression of its coding gene near the chromosomal position. We used bioinformatics to predict and analyze the cis and trans action target genes of lncRNA AK124826, and formed a network regulation map centered on lncRNA AK124826 ( Figure 6 A), it was found that IL-27 is located in its adjacent chromosome position and may be regulated by it as a target gene. We used the ELISA method to detect the level of IL-27 in the peripheral blood of PM / DM and healthy control group. The operation steps were carried out according to the instructions of the ELISA kit for IL-27. is the abscissa, the OD value of the standard is the ordinate, draws a standard curve, and calculates the concentration of each sample by the standard curve formula. The results showed that the expression level of IL-27 in peripheral blood of PM / DM patients was 13....

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Abstract

The invention discloses application of a medicine for preventing or treating polymyositis / dermatomyositis or diagnosis reagent or test kit. The invention also discloses application of the lncRNA AK124826 slow virus overexpression vector in the preparation of drugs for preventing or treating polymyositis / dermatomyositis or a diagnosis reagent or a test kit. The invention further discloses application of a RNAi interference vector of the lncRNA AK124826 in preparation of the drugs for preventing or treating polymyositis / dermatomyositis or the diagnosis reagent or the test kit.

Description

technical field [0001] The invention belongs to the field of biotechnology and treatment, and specifically relates to the application of lncRNA AK124826 and related molecules in the diagnosis of polymyositis / dermatomyositis. Background technique [0002] Polymyositis / dermatomyositis (PM / DM) is a relatively common autoimmune disease. CD4+T cells play an important role in the pathogenesis, and the number and function of Th17 / Treg cells are out of balance, but the specific pathogenesis is not yet fully understood . [0003] Polymyositis and dermatomyositis (PM / DM) is a typical autoimmune disease, which is mainly manifested by lymphocyte infiltration in muscle tissue, involving striated muscle, positive serum characteristic anti-Jo-1 antibody, In recent years, the incidence of PM / DM has been increasing year by year, and there is no effective radical treatment at present. The pathogenesis of PM / DM is complex, involving many factors such as genetics, infection, immune function, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/713A61P37/02A61P17/00A61P21/00C12Q1/6883
CPCA61K31/713A61P17/00A61P21/00A61P37/02C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 蒋廷旺薛建中韩志君龚燕萍高明珠
Owner 常熟市第二人民医院
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