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Intervertebral disc degenerative change diagnosis and treatment marker

A technology for degenerative diseases and intervertebral discs, applied in disease diagnosis, gene therapy, bone diseases, etc.

Active Publication Date: 2016-03-16
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conservative treatment includes rest on a hard board bed, massage, traction, acupuncture, etc., all of which are symptomatic; surgical treatment, including nucleus pulposus excision, spinal fusion, etc., is also symptomatic, and is based on sacrificing part or the entire intervertebral disc structure However, allograft intervertebral disc transplantation, artificial nucleus pulposus replacement and artificial intervertebral disc prosthesis have clear disadvantages and defects, and it has been reported that such operations have a failure rate of 15%-40%. The success rate of the second operation is even lowered to 50%, and the fusion of the intervertebral disc will lead to early degeneration of the adjacent upper and lower normal segmental intervertebral discs

Method used

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  • Intervertebral disc degenerative change diagnosis and treatment marker
  • Intervertebral disc degenerative change diagnosis and treatment marker
  • Intervertebral disc degenerative change diagnosis and treatment marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Differences in expression of TMED2 gene in normal control tissue and intervertebral disc degenerative disease tissue

[0059] 1. Sample collection

[0060] Five normal human intervertebral disc tissues were obtained from patients with spinal burst fractures caused by trauma. The intervertebral disc tissues of 5 patients with degenerated intervertebral disc disease were collected, with an average age of 75 years. According to the Gr1es 〔5〕 scoring standard, all of them were severely degenerated. The samples were collected with the consent of the patients and their families.

[0061] 2. Extraction of nucleus pulposus tissue

[0062] Normal nucleus pulposus tissue: The normal intervertebral disc tissue is nearly completely removed during the operation, and the peripheral white fibrous ring and the jelly-like nucleus pulposus tissue in the center can be seen. Soak the intervertebral disc in normal saline containing double-antibody (penicillin-streptomycin) for ...

Embodiment 2

[0085] Example 2 Large sample verification of differentially expressed genes

[0086] 1. Sample collection and tissue processing

[0087] 40 normal human intervertebral disc tissues were obtained from patients with spinal burst fractures caused by trauma. The intervertebral disc tissues of 40 patients undergoing surgery for degenerative disc disease were collected, with an average age of 75 years. According to the Gr1es 〔5〕 scoring standard, all of them were severely degenerated. The samples were collected with the consent of the patients and their families.

[0088] The extraction of nucleus pulposus tissue was the same as in Example 1.

[0089] 2. Detection of differential expression of TMED2 gene at the transcriptional level

[0090] 2.1 Extraction of total tissue RNA

[0091] Step is with embodiment 1.

[0092] 2.2 Reverse transcription

[0093] The reverse transcription of RNA was carried out using the reverse transcription kit of TAKARA company.

[0094] 2.3 QPCR ...

Embodiment 3

[0121] Example 3 TMED2 gene overexpression

[0122] 1. Construction of TMED2 gene expression vector

[0123] Amplification primers were designed according to the coding sequence of the TMED2 gene (as shown in SEQ ID NO.1). The coding sequence of the full-length TMED2 gene was amplified from the cDNA library of adult fetal brain (clontech company, catalog number: 638831). The obtained recombinant vector pcDNA3.1-TMED2 was connected to the eukaryotic cell expression vector pcDNA3.1 double-digested with enzymes BamHI and XhoI for subsequent experiments.

[0124] 2. Cell culture

[0125] Put the intervertebral disc nucleus pulposus tissue into a 100ml beaker filled with 10ml2% type II collagenase, and stir it with a magnetic stirrer for about 60 minutes. After the tissue is completely dissolved, centrifuge at 1000r / min for 10 minutes, suck out the supernatant, gently blow off the cells with 1ml of DMEM medium containing 10% fetal bovine serum, suck it into a 50ml culture bottle...

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Abstract

The invention discloses a gene TMED2 which can serve as a molecular marker for diagnosing the intervertebral disc degenerative change. Experiments prove that in comparison with a normal person, the expression quantity of the gene TMED2 in intervertebral disc degenerative change patients is large. The invention further discloses the gene TMED2 which can be used for preparing a drug for treating the intervertebral disc degenerative change. Apoptosis experiments prove that overexpression of the gene TMED2 can promote nucleus pulposus cell apoptosis. The research result provides a new intervertebral disc degenerative change clinic diagnosis method and provides a drug new target for treating the intervertebral disc degenerative change.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of human TMED2 gene in the diagnosis and treatment of intervertebral disc degeneration. Background technique [0002] The intervertebral disc is the earliest tissue in the human body that is most likely to degenerate with age. The exact cause of the disease is still unclear, but it has a significant relationship with accumulated strain and trauma, and also has a certain relationship with living habits, occupation, nutrition, general condition or endocrine. Generally speaking, the shape of the gel-like nucleus pulposus is lost, the original clear boundary between the nucleus pulposus and the annulus fibrosus gradually disappears, the laminae of the annulus fibrosus are rough and progressively fibrosis, and finally cracks and pigmentation appear. decrease and decrease. At present, the methods for treating intervertebral disc mainly include conservative treatment and su...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68A61K45/00A61K48/00A61K38/17A61P19/08
CPCA61K38/17A61K45/00C12Q1/6883C12Q2600/158G01N33/6893G01N2800/10
Inventor 杨承刚孙耀兰
Owner GUAN BOJIAN BIOTECH CO LTD
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