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Auxiliary detection kit for scoliosis in early stage and application thereof

A scoliosis and auxiliary detection technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, bone diseases, etc., to achieve rapid and effective early diagnosis and reduce the damage of X-ray radiation

Inactive Publication Date: 2017-11-07
北京赛尔维康生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of molecular biology research on AIS, some biomarkers related to it have been found, such as calmodulin, leptin in peripheral blood, serum osteopontin, soluble CD44, SH3GL1, etc., but the effects and conclusions still need to be further studied verify

Method used

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  • Auxiliary detection kit for scoliosis in early stage and application thereof
  • Auxiliary detection kit for scoliosis in early stage and application thereof
  • Auxiliary detection kit for scoliosis in early stage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Screening of differentially expressed genes by high-throughput sequencing

[0055] 1. Sampling

[0056] A total of 20 patients with scoliosis who were treated in the Department of Orthopedics of Peking Union Medical College Hospital from October 2012 to December 2015 were selected. A total of 20 cases were collected in the case group. All patients had typical clinical manifestations and were confirmed by X-ray examination. Controls were collected from patients with other diseases who were hospitalized in the Orthopedics Department during the same period, and a total of 15 cases were collected. The patients were adolescents between the ages of 10 and 18. Blood samples from all subjects were collected and stored in a -80°C low-temperature refrigerator after being numbered. All clinical samples in this study were informed to the patients and approved by the ethics committee of the hospital.

[0057] 2. Total RNA extraction from blood samples

[0058] use LS...

Embodiment 2

[0069] Example 2 qRT-PCR to verify the expression of HMGN1, P3H2 and NOLC1 genes in scoliosis patients

[0070] 1. Materials

[0071] Fifteen patients with scoliosis who were treated in the Department of Orthopedics of Peking Union Medical College Hospital from October 2012 to December 2015 were selected. All patients had typical clinical manifestations and were diagnosed by X-ray examination. Controls were collected from patients with other diseases who were hospitalized in the orthopaedic department during the same period, and a total of 8 cases were collected. The patients were adolescents between the ages of 10 and 18. Blood samples from all subjects were collected and stored in a -80°C low-temperature refrigerator after being numbered. All clinical samples in this study were informed to the patients and approved by the ethics committee of the hospital.

[0072] 2. Method

[0073] 2.1 Extract the total RNA from the blood samples, the same as the extraction method in Ex...

Embodiment 3

[0084] Example 3 RNAi interferes with the above-mentioned gene expression in chondrocytes of patients with scoliosis

[0085] 1. Cell Culture

[0086] (1) Wash the obtained iliac cartilage from patients with scoliosis three times with D-HANKS (Huamaike, product number HMK03230) solution under sterile conditions.

[0087] (2) Cut the iliac cartilage with scissors (about 1mm 3 size) in a sterile centrifuge tube.

[0088] (3) Digest with 0.25% trypsin at 37°C for 20 min, shaking every 5 min.

[0089] (4) Centrifuge at 800 rpm for 5 min, and discard the supernatant.

[0090] (5) 0.2% type II collagenase was digested at 37°C for 4 hours, and filtered through a 200-mesh sieve.

[0091] (6) The filtrate was centrifuged at 800 rpm for 5 min, and the supernatant was discarded.

[0092] (7) Rinse with DMEM medium, centrifuge at 800 rpm for 5 min, and repeat 3 times.

[0093] (8) After the cells were counted, inoculated into culture flasks, and DMEM medium containing 10% volume of ...

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Abstract

The invention discloses an auxiliary detection kit for scoliosis in early stage. The invention also discloses an siRNA molecule for restraining HMGN1, P3H2 and NOLC1 and an application of the siRNA molecule in preparing a drug compound for treating scoliosis. The utilization of HMGN1, P3H2 and NOLC1 genes for detecting scoliosis not only can quickly and effectively realize early diagnosis but also can supply a therapeutic target and an important basis for forecast and diagnosis of genesis and development of AIS and even for the treatment of AIS at biological level in future.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an early auxiliary detection kit for scoliosis and its application. Background technique [0002] Scoliosis, also known as scoliosis, is a complex three-dimensional spinal deformity with lateral curvature of the spine and rotation of the vertebral bodies. According to the etiology, it can be divided into 3 categories: congenital scoliosis, syndromic scoliosis and idiopathic scoliosis. Congenital scoliosis is caused by deformities of spinal development; syndromic scoliosis is caused by neuromuscular diseases, skeletal diseases, connective tissue diseases and neurofibromatosis; the exact cause of idiopathic scoliosis is not known Clearly, relevant studies suggest that genetic factors, growth and development and hormones, connective tissue lesions, abnormalities of the muscular system, abnormalities of the central nervous system, and the role of melatonin may all be related to the pathog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113A61K31/713A61P19/00
Inventor 王耀贾冰寒李睿
Owner 北京赛尔维康生物医学科技有限公司
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