A molecular marker for diagnosis and treatment of osteoporosis

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

Active Publication Date: 2018-10-02
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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  • A molecular marker for diagnosis and treatment of osteoporosis
  • A molecular marker for diagnosis and treatment of osteoporosis
  • A molecular marker for diagnosis and treatment of osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0063] 1. Research objects:

[0064] Randomly select patients who are about to undergo marrow joint replacement and test their bone density. Select 8 patients with osteoporosis and 8 patients with normal bone density control group (traumatic fractures, no osteoporosis tested), aged 52-68 years. Questionnaire surveys 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 "Chinese Osteoporosis Recommended Diagnostic Criteria (Second Draft); (2) All patients have informed consent.

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

[0067] 2. Obtaining RNA from bone tissue

[0068] From the human femoral head removed from the marrow joint replacement, 1g of hard bone was taken and quickly stored in liquid nitrogen. Trizol one-step method wa...

Embodiment 2

[0082] Example 2 QPCR experiment to verify the differential expression of KPNA3 gene

[0083] 1. Research objects: randomly selected patients about to undergo marrow joint replacement surgery, tested bone density, selected 50 patients with osteoporosis, normal bone density control group (traumatic fracture, no osteoporosis detected) 40 patients, age 50-70 years old. The inclusion criteria and exclusion criteria were the same as in Example 1.

[0084] 2. Extraction of RNA from bone tissue

[0085] The steps are the same as in Example 1.

[0086] 3. Reverse transcription

[0087] Use reverse transcription buffer to reverse transcribe 1 μg of total RNA to synthesize cDNA. A 25 μl reaction system was used, and 1 μg total RNA was used as template RNA for each sample. The following components were added to the PCR tube: DEPC water, 5× reverse transcription buffer, 10 mmol / L dNTP, 0.1 mmol / l DTT, 30 μmmol / l Oligo dT, 200 U / μl M-MLV, template RNA. Incubate at 42°C for 1 h, 72°C for 10 min,...

Embodiment 3

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

[0093] 1. Research object Same as Example 2.

[0094] 2. Collection and culture of human osteoblasts

[0095] Cancellous bone is taken from the human femoral head removed from the arthroplasty, soft tissue is removed, and the bone is repeatedly washed with normal saline. After the washing solution is clear, rinse with PBS solution and shake 3 times, cut into 1mm3 fragments, and adopt enzyme digestion method Isolate and purify osteoblasts. Inoculate it in a 30ml culture flask (add appropriate amount of DMEM-F12 (1:1) medium and 10% fetal bovine serum to the culture flask), and place it in a 37°C, 5% CO2 constant temperature incubator. Change the medium after 2 days and remove the non-adherent cells, then change the medium once every 3 days and observe under an inverted microscope. After the primary cells were fused into a monolayer, they were digested and passaged with 2.5 g / L trypsin.

[0096] 3. Extraction ...

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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 present invention relates to the field of biotechnology, in particular to the use of KPNA3 gene in the diagnosis and treatment of osteoporosis. Background technique [0002] Osteoporosis, or osteoporosis, is a group of bone diseases caused by various reasons. Bone tissue has normal calcification, calcium salt and matrix are in a normal ratio, and a metabolic bone disease characterized by a decrease in the amount of bone tissue per unit volume. . In most osteoporosis, the reduction of bone tissue is mainly due to increased bone resorption. It is characterized by bone pain and easy fracture. [0003] Epidemiological data show that about 8 million people in the United States suffer from osteoporosis, 20 million people have reduced bone mass, and 1.5 million people suffer fractures due to osteoporosis each year. The incidence of osteoporosis in white women after menopause is 17%, and that in black women is 6%. Among people older than 50 years old, nearly ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68A61K45/00A61K48/00A61K38/17A61P19/10
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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