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Method for extracting high-purity polydeoxyribonucleotide from salmon testis

A polydeoxyribonucleotide, high-purity technology, used in cell dissociation methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems not introduced, and achieve cost-effective and economically feasible results

Pending Publication Date: 2022-03-08
BIOMEDICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the patent only mentions that the process is also applicable to other tissues of other animals and does not describe any cases

Method used

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  • Method for extracting high-purity polydeoxyribonucleotide from salmon testis
  • Method for extracting high-purity polydeoxyribonucleotide from salmon testis
  • Method for extracting high-purity polydeoxyribonucleotide from salmon testis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Sperm motility (%) in different regions of salmon testis before and after hCG treatment

[0062] Harvest the immature regions by cutting the testes of male salmon (except for region D where semen leaks, see Figure 4 ). Before treatment with hCG, sperm motility from testicular regions A and B was almost zero, sperm motility was as low as 1% in testicular region C, and sperm motility was 100% in mature semen. After hCG treatment, sperm motility in testicular region A increased to about 3%, while sperm motility in testicular region B and C increased to 35% and 89%, respectively. These results lead to the conclusion that the use of testicular region C, which accounts for 45% of the total testis weight, to produce mature spermatozoa allows the preparation of high-purity PDRN with few impurities (see Figure 4 and Table 1).

[0063] The motility of sperm in different areas of salmon testis and the weight ratio (%) of testis area before and after the hCG of table...

Embodiment 2

[0076] The sperm motility (%) of salmon testis region C under different hCG treatment conditions of embodiment 2

[0077] The testis region C of male river salmon was diluted with buffer solution and treated with different concentrations of hCG. Sperm motility was very low (2%) when zone C was untreated, steadily increased with hCG concentrations up to 50 IU / g, and ≥82% when treated with hCG concentrations of 50-200 IU / g (Table 6).

[0078] Table 6 Sperm motility (%) in testis region C of river salmon caught 2 km upstream of the estuary in October when treated with different concentrations of hCG (IU / g testis)

[0079]

[0080] *Measured 10 minutes after hCG treatment

[0081] The changes in sperm motility over time after treatment with the concentrations of hCG shown in Table 6 were compared. There was no difference in sperm motility over time in the untreated control group. Sperm motility of the group treated with 25 IU / g testis increased to a maximum of 73% 1 hour afte...

Embodiment 3

[0084] Example 3 Treat the PDRN extraction rate (%) of testis region C with hCG under different conditions

[0085] In this example, 10 g of testicular region C of male river salmon was used. For comparison, 30 mL of semen typically extracted from male river salmon was used (Table 8). The testicular area was treated with hCG at concentrations of 50 and 100 IU / g testis. After hCG treatment, ≥80% of the top layer was collected and defined as the supernatant.

[0086] The dry weight of the untreated group was 5.10 g, which was smaller than that of the hCG-treated group and larger than that of semen (2.89 g). The dry weight of the untreated group was greater than that of the semen, which is thought to be due to the presence of heavier trophoblasts. The dry weight of the hCG-treated group was greater than that of the untreated group, which is thought to be because hCG treatment stimulates meiosis of spermatogonia into spermatids, resulting in an increase in cell number. The low...

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Abstract

The present disclosure relates to a method of extracting high purity polydeoxyribonucleotide (PDRN) from a salmon testis, the method comprising: 1) separating seminal fluid and an immature testis region from the salmon testis, 2) mildly grinding the immature testis region and then diluting with artificial seminal plasma, 3) treating the diluent with a predetermined concentration of human chorionic gonadotropin (hCG), and 4) extracting the high purity polydeoxyribonucleotide (PDRN) from the salmon testis. The method comprises the following steps: (1) extracting sperm from a sperm diluent to induce artificial maturation of the sperm cells into sperms, (4) centrifuging the sperm diluent and collecting potential active sperms after hCG treatment for a preset time, and (5) extracting PDRN from the collected sperms. According to the method, 100-200 times of PDRN raw materials with higher purity can be extracted from 20 mL (maximum 50 mL) of sperms (seminal fluid) collected by salmons at a time. In addition, the method enables the extraction of PDRN in high yield in a cost-effective and economically feasible manner.

Description

technical field [0001] The present invention relates to a method for extracting high-purity polydeoxyribonucleotide (PDRN) from salmon testis, more particularly to a method for extracting high-purity PDRN by hormone treatment of immature testis, which is currently used for in other applications. Background technique [0002] Polydeoxyribonucleotides (PDRN) are present in active substances for tissue regeneration (evolution) in living organisms. PDRN is a mixture of chain-like deoxyribonucleotide polymers consisting of 50 to 2000 base pairs. PDRN is a cell growth activator that has specific effects on the regeneration of tissues (such as ligaments, tendons and skin) and the relief of inflammation in the human body. [0003] Only very small amounts of PDRN are present in the human placenta. PDRN involves ethical issues and presents difficulties in pharmaceutical production. For these reasons, PDRN in fish has been proposed as an alternative to human PDRN. The semen and te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/076C12N15/10
CPCC12N5/061C12N15/1003C12N2501/31C12N2509/00C12N2509/10Y02A40/81A61K35/60A61K31/711A61K8/987A61K8/606A61Q19/00C12N15/10C12P19/34
Inventor 权五男金兌炫金善姬
Owner BIOMEDICS INC
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