CN113893331A - Product for inducing female deer to grow antler, preparation method thereof, method for inducing female deer to grow antler and application thereof - Google Patents
Product for inducing female deer to grow antler, preparation method thereof, method for inducing female deer to grow antler and application thereof Download PDFInfo
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- CN113893331A CN113893331A CN202010640880.4A CN202010640880A CN113893331A CN 113893331 A CN113893331 A CN 113893331A CN 202010640880 A CN202010640880 A CN 202010640880A CN 113893331 A CN113893331 A CN 113893331A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/195—Chemokines, e.g. RANTES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
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- A—HUMAN NECESSITIES
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- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
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- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
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Abstract
The invention belongs to the technical field of agricultural production, and particularly relates to a product for inducing female deer to grow antler, a preparation method thereof, a method for inducing female deer to grow antler and application thereof. The product for inducing the antler growth of the stag comprises cytokines, wherein the cytokines comprise chemotactic factors CCL2 and/or chemotactic factors CCL3, and the mass concentration of the cytokines is 1 ng/ml-1 mg/ml. The product for inducing the stag to grow the antler provided by the invention has high antler growth rate, does not influence the reproductive performance of the stag, and greatly improves the economic added value of the stag.
Description
Technical Field
The invention belongs to the technical field of agricultural production, and particularly relates to a product for inducing female deer to grow antler, a preparation method thereof, a method for inducing female deer to grow antler and application thereof.
Background
The main economic source of the breeding industry of the spotted deer is to sell the antler, but only male spotted deer generates antler, and female does not generate antler. Research shows that exogenous male hormone can induce female sika deer to produce antler, but androgen can affect reproductive performance of female sika deer. Therefore, it cannot be applied to actual production. In order to further improve the economic benefit, a method which can make the female deer antler but does not influence the reproductive capacity is urgently needed.
Disclosure of Invention
In order to solve the problems, the invention provides a product for inducing the antler growth of a female deer, a preparation method thereof, a method for inducing the antler growth of the female deer and application thereof. The product for inducing the stag to grow the antler provided by the invention has high antler growth rate, does not influence the reproductive performance of the stag, and greatly improves the economic added value of the stag.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a product for inducing antler growth of a stag, which comprises the components of cytokines, wherein the cytokines comprise a chemotactic factor CCL2 and/or a chemotactic factor CCL3, and the mass concentration of the cytokines in the product is 1 ng/ml-1 mg/ml.
Preferably, the mass concentration of the cell factor in the product is 10 ng/ml-10 ug/ml.
Preferably, the mass concentration ratio of the chemokine CCL2 to the chemokine CCL3 is (1-3): 1.
preferably, the ingredients of the product further comprise an emulsifier.
Preferably, the emulsifier comprises liquid paraffin and lanolin, and the mass ratio of the liquid paraffin to the lanolin is (1-5): 1.
The invention also provides a preparation method of the product, which comprises the following steps: and (3) preparing the cytokine to ensure that the mass concentration of the cytokine in the product is 1 ng/ml-1 mg/ml.
Preferably, when the ingredients of the product include an emulsifier, the preparation method further comprises: mixing the emulsifier and the cell factor and emulsifying.
Preferably, the emulsification method comprises bidirectional injector emulsification or oscillator oscillation emulsification.
The invention also provides a method for inducing the antler growth of the stag, the product or the product prepared by the method is injected into the antler growth area of the stag, and the injection dosage of each antler growth area is 0.05-0.5 ml.
The invention also provides application of the chemotactic factor CCL2 and/or the chemotactic factor CCL3 or the product prepared by the preparation method in inducing the antler growth of the stag.
The invention provides a product for inducing antler growth of a stag, which comprises the components of cytokines, wherein the cytokines comprise a chemotactic factor CCL2 and/or a chemotactic factor CCL3, and the mass concentration of the cytokines is 1 ng/ml-1 mg/ml. The product for inducing the stag to grow the antler provided by the invention has high antler growth rate, does not influence the reproductive performance of the stag, and greatly improves the economic added value of the stag.
Drawings
FIG. 1 is a graph comparing induction and non-induction of cornu Cervi Pantotrichum in application example 2, wherein A in FIG. 1 is the result of induction treatment; in FIG. 1, B is the result of no induction treatment.
Detailed Description
The invention provides a product for inducing antler growth of a stag, which comprises the components of cytokines, wherein the cytokines comprise a chemotactic factor CCL2 and/or a chemotactic factor CCL3, and the mass concentration of the cytokines in the product is 1 ng/ml-1 mg/ml.
In the invention, the cell factor can attract immune cells, stimulate the immune cells to release cell growth stimulating factors to stimulate stem cells in the periosteum of the antler to be activated, thereby inducing the antler growth of the stag. In the invention, the mass concentration of the cell factor in the product is preferably 10 ng/ml-10 ug/ml; more preferably 50ng/ml to 1 ug/ml; more preferably 100ng/ml to 200 ng/ml; the cytokine is preferably formulated by dilution with physiological saline.
In the present invention, the cytokine includes one or both of chemokine CCL2 and chemokine CCL 3; preferably the chemokine comprises CCL2 and the chemokine CCL 3; preferably, the mass concentration ratio of the chemokine CCL2 to the chemokine CCL3 is (1-3): 1; more preferably 1: 1. the sources of the chemokines CCL2 and CCL3 are not particularly limited in the present invention, and conventional commercially available products of the chemokines CCL2 and CCL3 known to those skilled in the art may be used.
In the present invention, the ingredients of the product also preferably include an emulsifier. The emulsifier mainly plays a role of slow release, can slowly release the cell factors, and can achieve the aim of inducing antler generation by one-time injection.
In the invention, the emulsifier preferably comprises liquid paraffin and lanolin, and the mass ratio of the liquid paraffin to the lanolin is preferably (1-5): 1; more preferably 2: 1.
In the invention, the volume ratio of the cytokine to the emulsifier is preferably (0.5-2): 1; more preferably 1: 1.
the invention also provides a preparation method of the product, which comprises the following steps: and (3) preparing the cytokine to ensure that the mass concentration of the cytokine in the product is 1 ng/ml-1 mg/ml. In the present invention, when the ingredients of the product include an emulsifier, the preparation method further includes: mixing the emulsifier and the cell factor and emulsifying. In the present invention, the emulsification method preferably includes bidirectional syringe emulsification or oscillator oscillation emulsification. The bidirectional injector is preferably emulsified for more than 200 times, and more preferably emulsified for 300 times; the shaking emulsification of the oscillator is preferably carried out for more than 30min under the condition of 10000rpm, and more preferably for 30 min.
The method for carrying out quality inspection on the emulsion comprises the following steps: centrifuging the emulsion at 5000g/min for 5min to avoid emulsion layering, and inspecting; or dropping the emulsified product onto the water surface, wherein the emulsion drop can be kept on the water surface for more than 1 hour.
The invention also provides a method for inducing the antler growth of the stag, the product or the product prepared by the method is injected into the antler growth area of the stag, and the injection dosage of each antler growth area is 0.05-0.5 ml, and more preferably 0.1 ml. In the present invention, when the ingredients of the product do not have an emulsifier, the injection dose is preferably 100ng, and the number of injections is preferably 4.
The invention also provides application of the chemotactic factor CCL2 and/or the chemotactic factor CCL3 or the product prepared by the preparation method in inducing the antler growth of the stag.
In order to further illustrate the present invention, the following examples are provided to describe in detail the production of antler-inducing deer, the method for producing the same, and the method and use of antler-inducing deer according to the present invention, but they should not be construed as antecedent to the scope of the present invention.
Example 1:
the product comprises the following components: the cell factors are CCL2 and CCL3, and the concentration is 100 ng/ml.
The preparation method comprises the following steps:
the cell factor solution is prepared according to the concentration of the cell factor solution, and the diluent is normal saline.
Example 2:
the product comprises the following components: the cell factors are CCL2 and CCL3, and the concentration is 100 ng/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 3:
the product comprises the following components: the cell factors are CCL2 and CCL3, and the concentration is 20 ng/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 4:
the product comprises the following components: the cell factor is CCL2, and the concentration is 10 ug/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 5:
the product comprises the following components: the cell factor is CCL2, and the concentration is 100 ng/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 6:
the product comprises the following components: the cell factor is CCL2, and the concentration is 10 ng/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 7:
the product comprises the following components: the cell factor is CCL3, and the concentration is 10 ug/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 8:
the product comprises the following components: the cell factor is CCL3, and the concentration is 100 ng/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Example 9:
the product comprises the following components: the cell factor is CCL3, and the concentration is 10 ng/ml; the emulsifier is liquid paraffin and lanolin with the mass ratio of 2: 1; the volume ratio of the cell factor to the emulsifier is 1: 1.
The preparation method comprises the following steps:
mixing the emulsifier and the cytokine solution according to the proportion, and emulsifying for 200 times by using a bidirectional syringe. And centrifuging the emulsion at 5000g/min for 5min, wherein the emulsion is qualified without emulsion layering.
Application example 1
Selecting 5 stags of 3-year-old stags in the Jilin Zhenyuan deer industry in 2019 in 5 months; anaesthetizing selected doe, injecting the product of example 1 into left antler-forming area with injection dose of 0.1ml, injecting 0.9% normal saline into right control group, injecting once per week, and continuously injecting for 4 times; after 2 months, the left side of the treated 4 stags generates pilose antler, and the right side does not generate pilose antler. All the deer in autumn normally estrus and breed, and all the deer produce children normally.
Application example 2
In 2019, in 4 months, 6 does of 3-year-old are selected in a left test station; anaesthetizing selected doe, injecting the product of example 2 into the left antler-forming area with injection dose of 0.1ml, and injecting the emulsifying agent without cell factor into the right control group; after 2 months, the processed deer antlers grow out on the left side of the processed deer, and no antlers grow out on the right side, as shown in figure 1. All the deer in autumn normally estrus and breed, and all the deer produce children normally.
Application example 3
Selecting 10 female deer born by 4-6 years old in Shuangyang deer village in 6 months in 2019; the product of example 2 was injected bilaterally, each side at a dose of 0.1 ml. After 2 months, 8 deer grow antler on both sides, and on average, 0.6kg antler is produced by each deer, 0.4kg antler is grown on one side of 1 deer, and 1 deer does not grow antler.
Application example 4
In 4 months of 2020, 12 barren doe deer in Jilin Zhenyuan deer industry are injected on both sides with the product of example 3, and the injection dosage on each side is 0.1 ml. After 1 month, 8 hairs grow on both sides, 3 hairs grow on one side, and 1 hair does not grow.
Application example 5
In 2 months of 2020, 6 and 3-year-old female deer are selected in the left experimental station, 0.1ml of the product of the embodiment 4 is injected on each side of the antler growing area, 4 deer antlers begin to develop after 3 weeks, the length of the deer antlers is 10-20cm after 3 months, the weight of each deer antlers is 200 plus 300g, and the success rate is 66.67%.
Application example 6
In 2019, in 4 months, 6 3-year-old female deer are selected in the double-yang deer farm, 0.1ml of the product in the embodiment 5 is injected on each side of the antler growing area, 3 deer antlers begin to develop after 3 weeks, 3 deer antlers grow out from 3 deer antlers after 3 months, 1 deer antlers grow out from 1 deer antlers, and 2 deer antlers do not grow out. The length of the pilose antler is between 10 and 20cm, and the weight of the pilose antler is 200-300 g.
Application example 7
In 5 months in 2019, 5 3-year-old female deer are selected in the double-Yang deer farm, 0.1ml of the product in the embodiment 6 is injected on each side of an antler growing area, 2 deer antlers begin to develop after 3 weeks, the length of the deer antlers is 10-15cm after 3 months, the weight of each deer antlers is 150 plus 250g, and the success rate is 40%.
Application example 8
In 3 months of 2020, 10 female deer of 3 years old are selected in a left test station, 0.1ml of the product of the embodiment 7 is injected into each side of an antler growing area, 5 deer antlers can be seen to start to develop after 3 weeks, the length of the deer antlers is 5-15 cm after 2 months, and the success rate is 50%.
Application example 9
In the year 2020 and 4 months, 6 female deer of 3 years old are selected in the double-yang deer farm, 0.1ml of the product of the embodiment 8 is injected into each side of the antler growing area, 3 deer antlers can be seen to start to develop after 3 weeks, the length of the deer antlers ranges from 5cm to 15cm after 2 months, and the success rate is 50%.
Application example 10
In the 4 th month of 2020, the double-yang deer farm selects 3-year-old female deer antler growing areas and injects 0.1ml of the product of the embodiment 9 to each side, 1 deer antler starts to develop after 3 weeks, the length of the deer antler is 5-15 cm after 2 months, and the success rate is 33%.
According to the embodiments, the antler growth rate of the products for inducing the antler growth of the stags provided by the invention can reach 66% when the CCL2 is used alone, the antler growth rate of the CCL3 is used alone and can reach 50%, and when the CCL2 and the CCL3 are used in combination, the antler growth rate can reach more than 85%, so that the antler growth rate is high, the reproductive performance of the stags is not influenced, and the economic additional value of the stags is greatly improved.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. The product for inducing the antler growth of the stag is characterized in that the components of the product comprise cytokines, the cytokines comprise a chemokine CCL2 and/or a chemokine CCL3, and the mass concentration of the cytokines in the product is 1 ng/ml-1 mg/ml.
2. The product of claim 1, wherein the cytokine is present in the product at a mass concentration of 10ng/ml to 10 ug/ml.
3. The product according to claim 1, wherein the mass concentration ratio of the chemokine CCL2 to the chemokine CCL3 is (1-3): 1.
4. the product of claim 1, wherein the product ingredients further comprise an emulsifier.
5. The product of claim 1, wherein the emulsifier comprises liquid paraffin and lanolin, and the mass ratio of the liquid paraffin to the lanolin is (1-5): 1.
6. A process for the preparation of a product according to any one of claims 1 to 5, comprising the steps of: and (3) preparing the cytokine to ensure that the mass concentration of the cytokine in the product is 1 ng/ml-1 mg/ml.
7. The method of claim 6, wherein when the product ingredient comprises an emulsifier, the method further comprises: mixing the emulsifier and the cell factor and emulsifying.
8. The method of claim 7, wherein the emulsifying comprises: emulsifying by a bidirectional injector or oscillating and emulsifying by an oscillator.
9. A method for inducing antler growth of a female deer, comprising the step of injecting the product of any one of claims 1 to 5 or the product prepared by the method of any one of claims 6 to 8 into an area of antler growth of the female deer, wherein the injection dose per area of antler growth is 0.05 to 0.5 ml.
10. Use of chemokine CCL2 and/or chemokine CCL3, or a product according to any one of claims 1 to 5, or a product obtained by the preparation method according to any one of claims 6 to 8, for inducing antler growth in a doe.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010640880.4A CN113893331A (en) | 2020-07-06 | 2020-07-06 | Product for inducing female deer to grow antler, preparation method thereof, method for inducing female deer to grow antler and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010640880.4A CN113893331A (en) | 2020-07-06 | 2020-07-06 | Product for inducing female deer to grow antler, preparation method thereof, method for inducing female deer to grow antler and application thereof |
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| CN113893331A true CN113893331A (en) | 2022-01-07 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0121579D0 (en) * | 2001-09-06 | 2001-10-24 | Medical Res Council | Chemokines |
| WO2010051820A1 (en) * | 2008-11-10 | 2010-05-14 | Aarhus Universitet | Multiplexed cytokine vaccination |
| CN102782148A (en) * | 2009-08-28 | 2012-11-14 | Vlst公司 | Anti-factor antibodies that bind to multiple CC chemokines |
| CN109517785A (en) * | 2018-11-30 | 2019-03-26 | 中国农业科学院特产研究所 | A kind of raw fine and soft method of doe Sheng Rong area's periosteal tissue stimulation |
-
2020
- 2020-07-06 CN CN202010640880.4A patent/CN113893331A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0121579D0 (en) * | 2001-09-06 | 2001-10-24 | Medical Res Council | Chemokines |
| WO2010051820A1 (en) * | 2008-11-10 | 2010-05-14 | Aarhus Universitet | Multiplexed cytokine vaccination |
| CN102782148A (en) * | 2009-08-28 | 2012-11-14 | Vlst公司 | Anti-factor antibodies that bind to multiple CC chemokines |
| CN109517785A (en) * | 2018-11-30 | 2019-03-26 | 中国农业科学院特产研究所 | A kind of raw fine and soft method of doe Sheng Rong area's periosteal tissue stimulation |
Non-Patent Citations (3)
| Title |
|---|
| CHRISTOPH ANDREAS REICHEL ET AL: "Ccl2 and Ccl3 mediate neutrophil recruitment through induction of protein synthesis and secondary generation of lipid mediators", THE FASEB JOURNAL EXPERIMENTAL BIOLOGY 2009 MEETING ABSTRACTS, pages 762 * |
| CHRISTOPH ANDREAS REICHEL ET AL: "Ccl2 and Ccl3 mediate Neutrophil Recruitment via Induction of Protein Synthesis and Generation of Lipid Mediators", ARTERIOSCLER THROMB VASC BIOL, pages 1787 - 1801 * |
| 李大金;: "趋化因子在人母-胎界面的整合性调节作用", 现代免疫学, no. 01 * |
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