CN107197828A - A kind of construction method of new osteoporosis model and the raising formula of use - Google Patents
A kind of construction method of new osteoporosis model and the raising formula of use Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
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- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/25—Animals on a special diet
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention discloses a kind of construction method of new osteoporosis model and the raising formula of use.The invention is to feed ultra-pure water to having carried out the Female Rhesus Monkey of bilateral ovaries excising operation, and feeding produces osteoporosis model in 24~28 weeks.The present invention can be successfully established rhesus macaque osteoporosis model, and model is high with clinical patient similarity, and operating process is simple, easily repeats, can be used for the evaluation and screening for the biotechnology new drug that rodent and other animal models can not be completed.
Description
Technical field
The present invention relates to a kind of construction method of new osteoporosis model and the raising formula of use.
Background technology
Have been reported that with animal coping osteoporosis models such as mouse, rat, rabbit, sheep, monkeys.The bone of wherein big mouse breathes out Freund
Reconstruct is not obvious, and skeletal diseases have larger difference after 12 months, and the blood volume of big mouse is smaller, it is impossible to continuously adopted
The analysis of blood and many indexes.And rabbit, sheep, pig, dog show without obvious sexual cycle and menopause, and to removing the histology of ovary
Reaction limits its application with the not equal factor of the mankind.Rhesus macaque keeps regular orthostatic body position, and biomethanics and the mankind are most
To be similar；Intracorporeal organ system includes immune system, internal system, and Bone m etabolism is approached with the mankind.Rhesus macaque has and mankind female
Property similar sexual cycle and menopause situation.The loose medicine of novel bone, research are evaluated using rhesus macaque osteoporosis model
Exploitation receptor target medicine simultaneously evaluated by effect of drugs, and this has important guiding meaning to future therapeutic scheme and clinical practice
Justice.Because current osteoporosis therapy medicine and method can not effectively cure osteoporosis, therefore it is long to set up non-human spirit
The loose model of osteoid is that newtype drug and targeted therapy provide evaluation and have active demand.
The content of the invention
In order to improve above mentioned problem, the present invention provides a kind of construction method of new osteoporosis model and the feeding of use
Support formula.
To achieve these goals, the technical solution adopted by the present invention is as follows：
A kind of construction method of new osteoporosis model, the Female Rhesus Monkey to having carried out bilateral ovaries excising operation
Ultra-pure water is fed, feeding produces osteoporosis model in 24~28 weeks.
Specifically include following steps：
(1) 8~10 years old Female Rhesus Monkey is taken, bilateral ovaries excising operation is carried out；
(2) according to the recovery situation of rhesus macaque, give penicillin and carry out anti-inflammation, the postoperative town of tramadol hydrochloride progress
Bitterly, etamcylate heals to help surgical wound；
(3) postoperative daily feeding ultra-pure water, feeding produces osteoporosis model in 24~28 weeks.
Further, drunk before the Female Rhesus Monkey carries out bilateral ovaries excising operation and after operation using low Ca
Food raise 24~28 weeks.
Yet further, before and after bilateral ovaries excising operation is carried out, the rearing conditions of Female Rhesus Monkey are：Raising temperature
18~26 DEG C, relative humidity 40%~70%, rate of ventilation is not less than 8 times per hour；Feeding time：The morning 08:00-09:00,
Feeding 100g, afternoon 14:00-15:00, feeding 100g.
Further, penicillin injection amount is 800,000 U/ time, 2 times/day；The injection volume of tramadol hydrochloride is 50mg/ times,
1 times/day；The injection volume of etamcylate 0.5g/ times, 2 times/day.
The raising formula used in a kind of construction method of new osteoporosis model, hand is extractd carrying out bilateral ovaries
Fed after preoperative using following raw material, by weight percentage, wheat bran 10~13%, wheat-middlings 28~32%, corn
35~40%, rice protein powder 14~18%, corn oil 3~5%, premix 1~3%.
Further, the wheat bran 11%, wheat-middlings 30%, corn 37%, rice protein powder 16%, corn oil 4%, pre-
Batch mixing 2%.
Yet further, it is not added with Ca agent in the premix.
The present invention compared with prior art, with advantages below and beneficial effect：
The present invention can be successfully established rhesus macaque osteoporosis model, and model is high with clinical patient similarity, operating process
Simply, easily repeat, can be used for the evaluation and screening for the biotechnology new drug that rodent and other animal models can not be completed.
Embodiment
With reference to embodiment, the invention will be further described, and embodiments of the present invention include but is not limited to following reality
Apply example.
Experimental study of the present invention is as follows：
Rhesus macaque feeding environment condition：
Raising temperature：18~26 DEG C；Relative humidity：40%~70%；
Ventilation：Rate of ventilation is not less than 8 times per hour, uses 100% all-fresh air (no air circulation)；
Lighting hours：Automatic illumination, per 12hr light and shades alternating, 7:30PM turns off the light, next day 7:30AM turns on light.
Animal cage：700*860*2100 double-layer stainless steel cages；Stocking density：1/cage.
Feeding time：The morning 08:00-09:00, give food 100g；Afternoon 14:00-15:00, give food 100g.
Other experimental problems are fed as stated above before experimental condition is reached, animal doctor's close observation experiment body shape
State.
When carrying out structure osteoporosis model, fed using low Ca diet, be then removed bilateral ovaries.
Specific feeding formula is as follows before being performed the operation：By weight percentage, wheat bran 11%, wheat-middlings 30%, corn 37%, big
Rice gluten powder 16%, corn oil 4%, premix (being not added with Ca agent) 2%.
What deserves to be explained is, it is configured to crude protein 19%, crude fat 6%, crude fibre 3%, Ca using above-mentioned raw materials
0.09%th, in carbohydrate 72%, and premix without any Ca agent mixed feed.The Ca amounts of adult's daily requirement
For 800mg/ days, the metabolic rate of rhesus macaque was 3 times of people, and rhesus macaque is about 2400mg/ days per Ca amounts day by day.Inventor is every
It gives food 200g, according to the formula of design, and rhesus macaque Ca daily intake is 180mg/ days in experiment, well below requirement
2400mg.
First, test material and method
1.1 test material：8-10 Sui Female Rhesus Monkey 12, no tubercle bacillus, parasite, salmonella, Shigella
Infected with B viruses；
1.2 main agents：Ultra-pure water, other conventional medications, above-mentioned feeding formula；
1.3 key instrument：Water purification machine：GN-RO-80 types；Dual-energy x-ray borne densitometers：GE Prodigy；Homeothermal animal hand
Art platform：DWV-ⅡHW；Patient monitor：GE B40i.
1.4 test method
1.4.1 rhesus macaque L1-L4 lumbar spine bmds are detected, before formally model is built, use dual-energy x-ray bone density
Instrument test experience monkey L1-L4 lumbar vertebraes, exclude the possibility of experimental monkey groups itself bone density relevant disease.
1) according to the last body weight of animal with the ketamine intramuscular injection of 10mg/kg dosage, if animal was checking
Revive or struggle in advance in journey, animal doctor according to circumstances supplements anesthesia, but supplements every time no more than injection dosage first in principle
1/3.About 5min is anaesthetized animal before inspection.
2) rhesus macaque upper and lower extremities are fixed on inspection desk, rhesus macaque vertebra position is straightened.
3) start GE Prodigy dual-energy x-rays borne densitometers and bone density scan is carried out to rhesus macaque L1-L4, and record is deposited
Shelves.
4) disposable vein blood sampling pin puncture veins of upper extremity or lower limb saphena, 5mL vacuum blood collection tubes (yellow head promotees solidifying pipe)
Take a blood sample 5mL, extracts disposable vein blood taking needle, sterilized cotton ball hemostasis, mark, as later stage BGP, β-glue connection catabolite out
The detection of (β-CTX).
5) after detection is finished, the experimental monkey groups in anesthesia are transferred in transfer cage, observation is until revival.
1.4.2 bilateral ovaries are extractd and ultra-pure water induces osteoporosis model
1) bilateral ovaries excising operation, according to the yellow Jackets intravenous injection anesthetized animal of 20mg/kg dosage, will perform the operation
The hair at position is shaved totally, is taken off using 75% alcohol is reused after iodine tincture disinfection after iodine, sanitized, wound towel is laid, with operation
Knife along median line 4~5cm of opening, will cut muscle peritonaeum, exposure abdominal cavity at 2cm below rhesus macaque navel.By the knot in abdominal cavity
After intestines are removed, you can see that ovary, uterus, bladder etc. are organized.After 1 side ovary is gently raised with tweezers, medical PGA3- is used
0 absorbability surgical sutures tubal ligation and blood vessel, are then cut with scissors, ovary are taken out.In kind take out another
Outer 1 side ovary.After 2 side ovaries all take out, wound is subjected to two layers of suture, one layer is muscle and peritonaeum, using medical PGA3-
0 absorbability surgical sutures, suture pattern is using simple continuous suture.One layer is skin, using medical silk litzendraht wire
2-0 is sutured, and suture pattern uses simple interrupted suture.In surgical procedure, the vital signs of animal, observation are carried out by animal doctor
Animal breath, pulse and narcosis, and using animal blood pressure in patient monitor monitoring art, and record.Animal doctor is according to operation
During animal state, judge whether to add anesthetic, with ensure animal in whole surgical procedure in completely fiber crops
Liquor-saturated state.
2) rhesus macaque is transferred in rearging cage, observation to monkey revival.
3) animal doctor is according to animal recovery situation, gives penicillin (800,000 U/ times, 2 times/day) and carries out anti-inflammation, salt love song
Many (50mg/ times, 1 times/day) the progress Postoperative Analgesia Afters of horse, etamcylate (0.5g/ times, 2 times/day) heal to help surgical wound.
4) state of mind, breathing and the wound healing status of postoperative daily observation animal, and recording.
5) postoperative daily feeding ultra-pure water and low Ca diet (by weight percentage, wheat bran 11%, wheat-middlings 30%, jade
Rice 37%, rice protein powder 16%, corn oil 4%, premix (being not added with Ca agent) 2%), feeding produces sclerotin for 24~28 weeks and dredged
Rapping type.
1.4.3 build after model to dual intensity X-ray Bone mineral density, determine bone density intensity of variation, assessment models whether into
Work(, the same 1.4.1 of test method.
By the research and detection to model, following examples are first provided：
Embodiment 1
A kind of construction method of new osteoporosis model, the Female Rhesus Monkey to having carried out bilateral ovaries excising operation
Ultra-pure water is fed, feeding produces osteoporosis model in 24~28 weeks.Specifically include following steps：
(1) 8~10 years old Female Rhesus Monkey is taken, bilateral ovaries excising operation is carried out；
(2) according to the recovery situation of rhesus macaque, give penicillin and carry out anti-inflammation, the postoperative town of tramadol hydrochloride progress
Bitterly, etamcylate heals to help surgical wound；
(3) postoperative daily feeding ultra-pure water, feeding produces osteoporosis model in 24~28 weeks.
Specifically, low Ca diet is used before the Female Rhesus Monkey carries out bilateral ovaries excising operation and after operation
Raise 24~28 weeks.
Specifically, before and after bilateral ovaries excising operation is carried out, the rearing conditions of Female Rhesus Monkey are：Raising temperature 18~
26 DEG C, relative humidity 40%~70%, rate of ventilation is not less than 8 times per hour；Feeding time：The morning 08:00-09:00, feeding
100g, afternoon 14:00-15:00, feeding 100g.
Specifically, penicillin injection amount is 800,000 U/ time, 2 times/day；The injection volume of tramadol hydrochloride be 50mg/ times, 1 time/
My god；The injection volume of etamcylate 0.5g/ times, 2 times/day.
Embodiment 2
The raising formula used in a kind of construction method of new osteoporosis model, hand is extractd carrying out bilateral ovaries
Fed after preoperative using following raw material, by weight percentage, the raw material of feeding is：Wheat bran 11%, wheat-middlings 30%,
Corn 37%, rice protein powder 16%, corn oil 4%, premix 2%.Specifically, it is not added with Ca agent in the premix.
Embodiment 3
The raising formula used in a kind of construction method of new osteoporosis model, hand is extractd carrying out bilateral ovaries
Fed after preoperative using following raw material, by weight percentage, the raw material of feeding is：Wheat bran 12%, wheat-middlings 29%,
Corn 38%, rice protein powder 15%, corn oil 3%, premix 3%.Specifically, it is not added with Ca agent in the premix.
Embodiment 4
The raising formula used in a kind of construction method of new osteoporosis model, hand is extractd carrying out bilateral ovaries
Fed after preoperative using following raw material, by weight percentage, the raw material of feeding is：Wheat bran 13%, wheat-middlings 28%,
Corn 36%, rice protein powder 17%, corn oil 4%, premix 2%.Specifically, it is not added with Ca agent in the premix.
According to above-described embodiment, the present invention just can be realized well.
Claims (7)
1. a kind of construction method of new osteoporosis model, it is characterised in that to having carried out bilateral ovaries excising operation
Female Rhesus Monkey feeds ultra-pure water, and feeding produces osteoporosis model in 24~28 weeks.
2. the construction method of a kind of new osteoporosis model according to claim 1, it is characterised in that including following
Step：
(1) 8~10 years old Female Rhesus Monkey is taken, bilateral ovaries excising operation is carried out；
(2) according to the recovery situation of rhesus macaque, give penicillin and carry out anti-inflammation, tramadol hydrochloride progress Postoperative Analgesia After, stop
Blood is quick to heal to help surgical wound；
(3) it is postoperative it is daily need to feed ultra-pure water, feeding produces osteoporosis model in 24~28 weeks.
3. the construction method of a kind of new osteoporosis model according to claim 1 or 2, it is characterised in that described
Carry out raising 24~28 weeks using low Ca diet before Female Rhesus Monkey carries out bilateral ovaries excising operation and after operation.
4. the construction method of a kind of new osteoporosis model according to claim 1 or 2, it is characterised in that entering
Before and after row bilateral ovaries excising operation, the rearing conditions of Female Rhesus Monkey are：18~26 DEG C of raising temperature, relative humidity 40%~
70%, rate of ventilation is not less than 8 times per hour；Feeding time：The morning 08:00-09:00, feeding 100g, afternoon 14:00-15:
00, feeding 100g.
5. the raising formula used in a kind of construction method of new osteoporosis model, it is characterised in that carrying out bilateral
Ovariectomy perioperatively is fed using following raw material, by weight percentage, wheat bran 10~13%, wheat-middlings 28
~32%, corn 35~40%, rice protein powder 14~18%, corn oil 3~5%, premix 1~3%.
6. the raising formula used in a kind of construction method of new osteoporosis model according to claim 5, its
It is characterised by, the wheat bran 11%, wheat-middlings 30%, corn 37%, rice protein powder 16%, corn oil 4%, premix 2%.
7. the raising formula used in a kind of construction method of new osteoporosis model according to claim 5, its
It is characterised by, Ca agent is not added with the premix.
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