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Biomarker related to occurrence and development of Parkinson's disease

A Parkinson's, product technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc.

Inactive Publication Date: 2019-11-29
潘伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, abnormally expressed lncRNAs found in diseased tissues can involve various systems of the body and are widely distributed. However, due to the large number of lncRNAs, the current research on lncRNAs in the field of Parkinson's is still in its infancy. Therefore, to explore the cause of Parkinson's The molecular basis of lncRNA is of great significance

Method used

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  • Biomarker related to occurrence and development of Parkinson's disease
  • Biomarker related to occurrence and development of Parkinson's disease
  • Biomarker related to occurrence and development of Parkinson's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Screening for gene markers associated with Parkinson's

[0051] 1. Sample collection

[0052] Blood samples from 35 normal subjects and Parkinson's patients were collected, and the patients gave informed consent, and all the above samples were obtained with the consent of the ethics committee. Five samples were taken from each group for detection and analysis of lncRNA expression profiles, screening of differentially expressed lncRNAs, and verification experiments were carried out in all 35 samples of each group.

[0053] 2. RNA sample preparation and quality analysis

[0054] Using Invitrogen's Total RNA was extracted with Reagent. For details, please refer to the instruction manual. Nanodrop2000 was used to detect the concentration and purity of the extracted RNA, agarose gel electrophoresis was used to detect RNA integrity, and Agilent2100 was used to determine the RIN value. Concentration ≥ 200ng / μl, OD260 / 280 between 1.8 and 2.2.

[0055] 3. Constru...

Embodiment 2

[0064] Example 2 QPCR sequencing verification of differential expression of LOC105373557 gene

[0065] 1. Large-scale QPCR verification of the LOC105374334 gene.

[0066] 2. RNA extraction

[0067] Using Invitrogen's Total RNA was extracted with Reagent. For details, please refer to the instruction manual.

[0068] 3. Reverse transcription

[0069] The FastQμant cDNA First Strand Synthesis Kit of TIANGEN was used for reverse transcription. For details, please refer to the instruction manual.

[0070] 4. QPCR amplification

[0071] (1) Primer design

[0072] QPCR amplification primers were designed according to the coding sequences of LOC105373557 gene and GAPDH gene in Genebank, and synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. The specific primer sequences are as follows:

[0073] LOC105373557 gene:

[0074] The forward primer is 5'-AACAATCAGGATGGAGAC-3' (SEQ ID NO.1);

[0075] The reverse primer is 5'-CGTCACTCTGCTATTTCA-3' (SEQ ID NO.2). ...

Embodiment 3

[0089] Example 3 Detection of gene silencing efficiency of LOC105373557

[0090] 1. Cell culture

[0091] PC12 cell culture medium is DMEM medium, containing 5% newborn bovine serum, 10% fetal bovine serum, 100IU / mL penicillin and 100IU / mL streptomycin, placed at 37°C, 5% CO 2 cultured in an incubator. After the cells are full, remove the supernatant, add 0.25% trypsin to digest, 1-2min, shake gently, wait for the cells to detach, remove the trypsin, add the culture medium containing serum to neutralize and terminate the digestion; blow the cells evenly, Take 1 / 4 of the cells for subculture and continue to culture at 37°C.

[0092] 2. Synthesis of siRNA

[0093] According to the sequence design siRNA of LOC105373557, synthesized by Shanghai Gemma Gene, the sequence of the siRNA against LOC105373557 is as follows:

[0094] siRNA:

[0095] The sense strand is 5'-AGUUUUUCCUGCAACUUCCCC-3' (SEQ ID NO.5),

[0096] The antisense strand is 5'-GGAAGUUGCAGGAAAAACUCC-3' (SEQ ID NO....

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Abstract

The invention discloses a biomarker related to the occurrence and development of Parkinson's disease. The biomarker is LOC105373557. The invention finds that LOC1053735757 is up-regulated in patientswith Parkinson's disease for the first time. By detecting the expression level of LOC1053735757, whether the subject has Parkinson's disease can be determined. When the level of LOC1035735757 is significantly up-regulated, the subject has Parkinson's disease; and the down-regulation of the expression level of LOC10357575 can change the proliferative activity of Parkinson's disease cells, so as toprovide a molecular treatment means for the patients with Parkinson's disease.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a biomarker related to the occurrence and development of Parkinson's, specifically the biomarker is LOC105373557. Background technique [0002] Parkinson's disease (PD) is one of the most common neurodegenerative diseases clinically. It has a high prevalence rate in middle-aged and elderly people, and seriously threatens the health of middle-aged and elderly people. The main clinical manifestation of PD is inactivity. Non-motor symptoms such as hyposmia, constipation, depression, and abnormal sleep behavior may also occur. The degeneration and loss of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies in the cell bodies of residual neurons are the main pathological features. PD is a chronic progressive disease. At present, the clinical focus is still on controlling symptoms and alleviating disease progression, and the disability rate in the later s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158
Inventor 潘伟常万生王永红
Owner 潘伟
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