Application of IGFBP3 (insulin-like growth factor-binding protein 3) in preparation of products to diagnose type I neurofibromatosis with spinal deformity

A technique for neurofibromas, spinal deformities, used in biotechnology and medical testing

Active Publication Date: 2018-08-17
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the fact that there is no product for early diagnosis of spinal deformity combined with type I neurofibroma, if the corresponding markers can be found and the corresponding diagnostic kits can be

Method used

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  • Application of IGFBP3 (insulin-like growth factor-binding protein 3) in preparation of products to diagnose type I neurofibromatosis with spinal deformity
  • Application of IGFBP3 (insulin-like growth factor-binding protein 3) in preparation of products to diagnose type I neurofibromatosis with spinal deformity
  • Application of IGFBP3 (insulin-like growth factor-binding protein 3) in preparation of products to diagnose type I neurofibromatosis with spinal deformity

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Experimental program
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Embodiment 1

[0030] The collection of embodiment 1 sample and the arrangement of sample data

[0031] From January 2017 to December 2017, peripheral blood samples from 5 patients with neurofibroma combined with spinal deformity and 5 relatively normal peripheral blood samples were obtained. All patients did not receive chemotherapy or radiotherapy before the operation, and the informed consent of the patients was obtained before the operation, which was reviewed by the ethics committee.

[0032] Sample processing: EDTA anticoagulated whole blood was mixed with Trizol at a ratio of 1:1, mixed thoroughly and placed in a 1.8ml cell cryopreservation tube, cooled rapidly in liquid nitrogen for 30 seconds and stored in a -80°C refrigerator.

Embodiment 2

[0033] Example 2 Peripheral blood RNA extraction

[0034] 1. Homogenization treatment

[0035] Take fresh blood, add 3 times the volume of erythrocyte lysate, mix well, place at room temperature for 10 minutes, and centrifuge at 10,000 rpm for 1 minute. Discard the supernatant and collect the white blood cell pellet. Add 1ml Trizol to every 100-200ul blood collected leukocyte pellet.

[0036] 2. Layering

[0037] After the sample was added to Trizol, it was left at room temperature for 5 minutes to fully lyse the sample.

[0038] Add 200ul of chloroform to every 1ml of Trizol, vortex vigorously and mix well, then place at room temperature for 3-5min to allow natural phase separation.

[0039] 3. RNA precipitation

[0040] Centrifuge at 12000rpm at 4°C for 10-15min. The sample will be divided into 3 layers: yellow organic phase, intermediate layer and colorless aqueous phase. RNA is mainly in the aqueous phase. Transfer the aqueous phase (usually absorbing 550-600ul) to a n...

Embodiment 3

[0051] Example 3 RNA-seq sequencing library construction and quality inspection of peripheral blood RNA

[0052] 1. Data quality assessment

[0053] The construction and sequencing of the cDNA library was entrusted to Beijing Novogene Technology Co., Ltd. to complete. The sequencing data of 10 samples were checked for sequencing error rate, GC content distribution and raw data filtering to obtain clean reads for subsequent analysis. The data summary is shown in Table 2.1. It is generally considered that RNA-seq data is used for gene differential expression analysis between different libraries, the total number of reads in the library is at least 10M, the overall GC content of the data is kept at 40%-60%, and the Q30 is more than 80%. In the data obtained by this sequencing, clean bases accounted for more than 7.2G, Q2 bases accounted for more than 95.18%, Q3 bases accounted for more than 89.15%, and the GC content remained stable between samples, ranging from 54.29% to 56.80%...

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Abstract

The invention discloses application of IGFBP3 (insulin-like growth factor-binding protein 3) in the preparation of products to diagnose type I neurofibromatosis with spinal deformity. Related markersIGFBP3 for NF1 with spinal deformity are acquired by screening via transcriptome sequencing; the role of IGFBP3 in genesis and development of bone healing defects in NF1 with spinal deformity is explored, and the acting mechanism of IGFBP3 is given to play; IGFBP3 as a novel biomarker of NF1 with spinal deformity provides important theoretical basis for early clinical intervention and targeted treatment of NF1 with spinal deformity.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and medical detection, in particular to the application of IGFBP3 in the preparation and diagnosis of type I neurofibroma combined with spinal deformity. Background technique [0002] Neurofibromatosis type 1 (NF1) is a relatively common autosomal dominant genetic disease; regardless of sex or race, the global incidence is 1 / 3000. In 1882, Frederich von Recklinghausen first described NF1, In 1987, the NIH published formal diagnostic criteria. Genetic studies have found that neurofibromatosis type I is caused by mutations in the NF1 tumor suppressor gene. The NF1 gene is located at 17q11.2, the long arm of chromosome 17, and encodes a cytoplasmic protein with a molecular weight of 220kDa—neurofibromin. Part of the protein’s role is to negatively regulate the Ras proto-oncogene, and Ras is a regulator of cell growth. important signaling molecules. In patients with genetic mutations, each c...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61P35/00A61P19/08
CPCA61K45/00A61P19/08A61P35/00C12Q1/6886C12Q2600/158
Inventor 蔡思逸邱贵兴吴志宏
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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