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Molecular marker for diagnosis and treatment of osteoporosis

An osteoporosis, one-to-one technology, applied to the KPNA3 gene in the field of diagnosis and treatment of osteoporosis, can solve problems such as inability to give reliable judgments

Active Publication Date: 2015-11-25
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main limitation of the above method is that it cannot give a reliable judgment in the early stage of osteoporosis

Method used

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  • Molecular marker for diagnosis and treatment of osteoporosis
  • Molecular marker for diagnosis and treatment of osteoporosis
  • Molecular marker for diagnosis and treatment of osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening for differentially expressed genes related to osteoporosis

[0063] 1. Research object:

[0064] Randomly select patients who are about to undergo marrow joint replacement and measure bone density. Eight patients with osteoporosis and 8 patients with normal bone density (trauma fractures, without osteoporosis) were selected, aged 52-68 years. Questionnaires were used to investigate the subjects' lifestyle and health status.

[0065] Inclusion criteria for patients with osteoporosis: (1) Those who meet the diagnostic criteria for osteoporosis, refer to the "Recommended Diagnostic Criteria for Osteoporosis in China (Second Draft); (2) All patients have given informed consent.

[0066] Exclusion criteria for patients with osteoporosis: those with secondary osteoporosis.

[0067] 2. Obtaining bone tissue RNA

[0068] Take 1g of hard bone from the human femoral head removed by the arthroplasty, and put it in liquid nitrogen for preservation immediately...

Embodiment 2

[0082] Embodiment 2 QPCR experiment verification KPNA3 gene differential expression

[0083] 1. Research objects: randomly select patients who are about to undergo marrow joint replacement, and detect bone density, select 50 cases of osteoporosis patients, and 40 cases of normal bone density control group (trauma fractures, no osteoporosis detected), age 50-70 years old. The inclusion and exclusion criteria are the same as in Example 1.

[0084] 2. Extraction of bone tissue RNA

[0085] Step is with embodiment 1.

[0086] 3. Reverse transcription

[0087] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to the PCR tube: DEPC water, 5× reverse transcription buffer, 10mmol / LdNTP, 0.1mmol / lDTT, 30μmmol / lOligodT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes. ...

Embodiment 3

[0092] Example 3 Differential expression of KPNA3 protein in osteoblasts

[0093] 1. The research object is the same as in Example 2.

[0094] 2. Preparation and cultivation of human osteoblasts

[0095] Cancellous bone was taken from the human femoral head removed by the arthroplasty, the soft tissue was removed, the bone was washed repeatedly with normal saline, after the washing solution was clarified, it was rinsed and shaken 3 times with PBS solution, and cut into 1mm 3 Fragments of the osteoblasts were isolated and purified by enzymatic digestion. Inoculate it in a 30ml culture flask (add appropriate amount of DMEM-F12 (1:1) medium and 10% fetal calf serum to the culture flask), place it at 37°C, 5% CO 2 cultured in a constant temperature incubator. After 2 days, the medium was changed and unadhered cells were removed, and the medium was changed every 3 days thereafter, and observed under an inverted microscope. After the primary cells fused into a monolayer, they we...

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Abstract

The invention discloses a molecular marker for diagnosis and treatment of osteoporosis, wherein the marker for diagnosis and treatment of osteoporosis is KPNA3 gene and expression products thereof. The high-throughput sequencing and QPCR method show that the KPNA3 gene and expression products thereof have differences in expression in bone tissues between normal people and patients with osteoporosis, and according to the correlation between the expression differences, the KPNA3 gene and expression products thereof can be used for serving as the molecular marker for diagnosis and treatment of osteoporosis. In addition, in vivo experiments show that over-expression of the KPNA3 gene can promote the proliferation of bone cells, therefore the KPNA3 gene and the expression products thereof can also be used for development of drugs for treating osteoporosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of KPNA3 gene in the diagnosis and treatment of osteoporosis. Background technique [0002] Osteoporosis, or osteoporosis, is a group of bone diseases caused by a variety of reasons. Bone tissue has normal calcification, and the ratio of calcium salt to matrix is ​​normal. It is a metabolic bone lesion characterized by a decrease in the amount of bone tissue per unit volume. . In most cases of osteoporosis, the loss of bone tissue is mainly due to increased bone resorption. Characterized by bone pain and susceptibility to fractures. [0003] Epidemiological data show that about 8 million people in the United States suffer from osteoporosis, 20 million people suffer from osteopenia, and 1.5 million patients suffer from fractures due to osteoporosis every year. The incidence of osteoporosis in postmenopausal white women is 17%, and the incidence rate in black women i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68A61K45/00A61K48/00A61K38/17A61P19/10
CPCA61K38/17A61K45/00A61K48/00C12Q1/6883C12Q2600/158G01N33/6893G01N2800/108
Inventor 李曙光杨承刚
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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