Non-coding RNA-LOC105376991 for diagnosing male osteoporosis
An osteoporosis, long-chain non-coding technology, applied in the field of biomedicine, can solve the problems of little help in early diagnosis of osteoporosis, low sensitivity and accuracy of osteoporosis, etc.
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Embodiment 1
[0052] Example 1 Screening of differentially expressed molecules
[0053] 1. Case collection
[0054] Five elderly male patients with osteoporosis, aged 62-79 years old, were selected; the diagnosis of osteoporosis was based on the standard set by the Osteoporosis Committee of the Chinese Geriatrics Society, that is, the BMD value at the same location was lower than the peak value in the region by two standard deviations Defined as osteoporosis. During the same period, 6 healthy elderly men (healthy control group), aged 61-78 years, were selected. Eligible subjects excluded diabetes, primary hyperparathyroidism and hyperthyroidism, hypercortisolism, liver and kidney dysfunction, alcoholism, malnutrition, rheumatoid arthritis and other diseases affecting bone metabolism; no application within six months Androgens, calcitonin, large amounts of calcium, bisphosphonates, and other drugs that affect bone metabolism. The two groups were comparable in terms of age, height, body ma...
Embodiment 2
[0095] Example 2 The differentially expressed LncRNA screened out by large sample verification
[0096] 1. Sample collection
[0097] According to the method of Example 1, 32 osteoporosis patients and 30 healthy male controls were collected.
[0098] 2. Extraction of total RNA from blood
[0099]The steps are the same as in Example 1.
[0100] 3. Validation at the RNA level
[0101] (1) Reverse transcription: According to the measured RNA concentration, 1000 ng of RNA was quantified for each sample for reverse transcription. Use Dalian Biobio's DDR037A reverse transcription kit, 20 μl of the system (see Table 1), add it to a 200 μl de-enzyme EP tube, centrifuge and place it in a PCR instrument, the temperature conditions are set as: 37 ° C for 15 min, 85 ° C for 5 sec . After the end, the cDNA was diluted 10 times and stored at -20°C for future use.
[0102] Table 1 Reverse transcription system
[0103]
[0104] (2) Real-time quantitative PCR
[0105] Shanghai Sango...
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