CN102274084A - Fabrication method of burnt animal model - Google Patents
Fabrication method of burnt animal model Download PDFInfo
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- CN102274084A CN102274084A CN2011101232267A CN201110123226A CN102274084A CN 102274084 A CN102274084 A CN 102274084A CN 2011101232267 A CN2011101232267 A CN 2011101232267A CN 201110123226 A CN201110123226 A CN 201110123226A CN 102274084 A CN102274084 A CN 102274084A
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Abstract
The invention relates to a fabrication method of a burnt animal model with blended fuel, and the fabrication method is characterized in that the blended fuel consists of petroleum oil refining residues 20-60% and gasoline 40-80%. The fabrication method comprises the following steps: dissolving petroleum oil refining residues in gasoline, blending, freezing at -20 DEG C, taking out, stirring, and repeating freezing and stirring for 3-7 times to obtain uniform gel; uniformly coating the blended fuel on the surface of depilated pre-burnt skin of rats, mice, guinea pigs, rabbits, pigs, dogs and other animals, firing, and continuously combusting for 15-30 s; immediately supplementing fluid for recovery, breeding with separate cages, and allowing free food taking; and observing wound surface tissue pathological sections with an optical microscope, and determining a burn scale. The model provided by the invention has the advantages of simple operation, good repeatability, high success ratio, easy control of burn severity, and so on, and can be used for researches on pathological mechanism of human burnt, screening and evaluation of therapeutic drugs, and the like.
Description
Technical field
The present invention relates to a kind of method of utilizing oil refinery's defective material to make animal model of burn, be used for pathomechanism, the screening of medicine and the research of therapeutical effect of human burn.The invention belongs to medical science and life sciences experimental study field.
Background technology
Burn is all multifactor a kind of extremely complicated traumatic diseases that acts on body and cause such as physics, chemistry, lonizing radiation, laser, although human generation of making great efforts the prevention burn, the annual burn patient quantity that receives of emergency room is still very big.According to statistics, aing burn case of U.S. every year can reach 500,000 more than.Because the complexity that clinical limitation of accumulating experience and burn take place to develop makes the pathomechanism of burn, the research of medicine remain one of emphasis direction of medicine and pharmacology research.Be subjected to morally with methodology on all restrictions, many experiments can't be with patient as subjects, causes greatly difficulty such as therapeutic effect observations such as the pathogeny of deep discussion disease and medicine.Animal model can overcome above-mentioned deficiency, is bringing into play unique effect in biomedical research.So set up animal model of burn and be very important experimental technique and means in modern burn research field basis and the applied research.Ideal burn model will help science more, the rule of development of understanding burn back each system of body and local woanded surface more accurately, explore prophylactico-therapeutic measures targetedly.
Temperature action more than 55 ℃ can cause experimental burn behind animal surface texture certain hour.Therefore, the production method kind of relevant animal model of burn is more, mainly is flame burn, electric light source method, burn wound etc.Experimental burn is hindered the warm scald method of branch point-like, direct heat scald method and vapours and is scalded method three classes.The warm scald method of point-like is created in 1949 by Sevitt.Animal pattern is not anaesthetized, and back fixation is scalded the device bottom with point-like and conducted oneself with dignity by the scald device in scalding under the plate, is close to laboratory animal and intends scalding district's enforcement scald.This method has scalding area and the degree of depth is more easy to control, and repeatability is high, and big toy such as all is suitable at advantage, is considered to the scalding model manufacture method of standard; Direct heat scald method is intended scalding the district with laboratory animal and is directly placed constant-temperature hot water to cause injury, and the degree of depth of causing injury is easy to control, good reproducibility, but the difficult control of scalding area; Vapours is scalded fado and is used to set up animal inhalation injury model.In general, the easy control of temperature of scald, even action, area is more easily grasped, generally than other burn model stability, accurate, good reproducibility.In the actual life or the flame burn that causes by chemical substances such as fuel in the military war hinder than burn more common, the burn that these reasons are caused carry out correlational study real belong to necessary.Flame burn comprises that mainly solidified gasoline duplicates the burn model and fuel combination duplicates the burn model.But burn model fuel commonly used at present all is difficult to buy, and does not have definite consumption and burn time mostly, and the burn degree is wayward.Therefore stable, accurately, the foundation of the flame burn animal model of good reproducibility is very important.
The extent of damage of experimental burn depends on height, the length of the time of causing injury, the preceding situation of wound of tissue of burn and the reactivity of organism of laboratory animal of the temperature of causing injury.For keeping causing injury the constant unanimity of condition, copy the model of extent of damage unanimity, so that carry out repetition and contrast, should be at the time of causing injury, the temperature of causing injury and the various possible influence factors in back that cause injury, take appropriate measures, reduce the influence of various factors to greatest extent.
Can evenly be coated with fuel combination in animal skin surfaces with soft brush when using this law modeling, the strict control burn time, reach once fire extinguishing simultaneously, can copy the required burn surface area and the degree of depth, the sings and symptoms of simulating human burn, and avoid determination of burn depth inconsistent, can satisfy the requirement of further experiment.It is simple that this model has modeling, draw materials conveniently, and model stability, the success rate height, favorable reproducibility, the burn degree is easy to characteristics such as control.
Summary of the invention
The object of the present invention is to provide a kind of is the method that fuel combination is made animal burn model with oil refinery's defective material and gasoline, this method has that modeling is simple, convenience of drawing materials, model stability, success rate height, favorable reproducibility, burn degree are easy to characteristics such as control, can be used for the animal model that the screening of human pathomechanism of burning, medicine and evaluation etc. are studied.
Mixed fuel formula: be that 20～60% oil refinery's defective materials are reduced crude by weight percentage, 40～80% gasoline.
Model production method: oil refinery's defective material is dissolved in the gasoline, mixing ,-20 ℃ are freezing, take out, and stir 3～7 extremely even glues of freezing repeatedly stirring; This fuel combination is applied to animal skin surfaces, igniting, sustained combustion 15～30 seconds causes burn in various degree.The recovery of fluid infusion immediately after the wound, sub-cage rearing, free diet; Adopt the wound tissue pathological section, pathological diagnosis and definite burn grade under the light microscopic.
The used animal of this model comprises rat, mice, Cavia porcellus, rabbit, pig and Canis familiaris L. etc., can simulate the sings and symptoms of mankind's burn that a variety of causes such as flame, electric light, lonizing radiation, hot water etc. cause, for scientific research and clinical treatment provide necessary animal burn disease model.
The specific embodiment
Enumerate exemplary embodiments below, the present invention is further described, but be not construed as limiting the invention in any form.
Embodiment 1: fuel combination causes the shallow II degree burn of rat 20% body surface area
The preparation of fuel combination: accurately weighing 35g oil refinery defective material is a reduced crude, adds 93
#Gasoline is put in the port grinding bottle to 100g, fully stirs.To refrigerator, the 10min that fully vibrates every day totally 7 days, is even glue.
Laboratory animal: the SD rat, male and female are regardless of, and body weight 200～250g is provided by The Fourth Military Medical University Animal Experimental Study center.
Skin treatment: (1) hair of dispelling: 3% pentobarbital sodium 30mg/kg ip anesthesia, remove by hair with electric clippers onboard rat is fixing, be slightly larger than the plan burn surface area.(2) depilation:, stand-by with the depilation of sodium sulfide paste.
Body surface area calculates: accurately weigh in fasting 16h before weighing.Calculate body surface area as follows:
S=body surface area (cm
2), W=daystart body weight (g), K=constant (12.54).
Burn surface area calculates: burn surface area (cm
2)=body surface area (cm
2) * intend burnt area %.
Modelling: animal 3% pentobarbital sodium 30mg/kg ip anesthesia will hide with wet cloth around the pre-burning skin.With hairbrush fuel combination is applied to pre-burn district uniformly, the coating area is consistent with pre-burn area, on every side with refractory material or wet gauze protection, lights immediately and with manual time-keeping, the burn time set is 15 seconds, then puts out a fire with wet gauze; Burning needs to carry out at the environment of steady air current.Lumbar injection Ringer lactate solution 5ml immediately after the wound, sub-cage rearing, free diet.
Determine depth of burn: burn wound is respectively at hindering back 1h with interior and 24h, the burned skin of big or small 1cm * 1cm taken off make histopathological examination to determine depth of burn, cut matched group holostrome skin, specimen is fixed in 10% the formalin, through dehydration, routine paraffin wax embedding, section, HE dyeing.
Diagnostic result: the wound surface redness vesicle or epidermis occur and separates with corium.Histological examination finds that epidermis destroys, and the high dermis damage is diagnosed as shallow II degree burn.
Embodiment 2: fuel combination causes rat 20% body surface area deep ii degree burn
Fuel combination preparation, laboratory animal, skin treatment etc. are all with embodiment 1.
Modelling: animal 3% pentobarbital sodium 30mg/kg ip anesthesia will hide with wet cloth around the pre-burning skin.With hairbrush fuel combination is applied to pre-burn district uniformly, the coating area is consistent with pre-burn area, lights immediately and with manual time-keeping, the burn time set is 25 seconds, then puts out a fire with wet gauze; Burning needs to carry out at the environment of steady air current.Lumbar injection Ringer lactate solution 5ml immediately after the wound, sub-cage rearing, free diet.
Determine that depth of burn is with embodiment 1.
Diagnostic result: scald the back wound surface and bleach immediately, mild swelling, gloss difference, naked eyes can obviously be distinguished burn position and normal dermatotome; The wound surface color and luster is deepened behind the 24h, sharpness of border.Histological examination: the epidermis total necrosis comes off, and skin corium collagen fiber structure is unclear, and above-mentioned change is also arranged at deep layer top, but as seen some remaining appendages of skin exist, neutral the more infiltration of carefully roaring arranged down again, and vasodilation hyperemia is diagnosed as deep ii degree burn.
Embodiment 3: fuel combination causes rat 20% body surface area III degree burn
Fuel combination preparation, laboratory animal, skin treatment etc. are all with embodiment 1.
Modelling: animal 3% pentobarbital sodium 30mg/kg ip anesthesia will hide with wet cloth around the pre-burning skin.With hairbrush fuel combination is applied to pre-burn district uniformly, the coating area is consistent with pre-burn area, lights immediately and with manual time-keeping, the burn time set is 30 seconds, then puts out a fire with wet gauze.Burning needs to carry out at the environment of steady air current.Lumbar injection Ringer lactate solution 5ml immediately after the wound, sub-cage rearing, free diet.
Determine that depth of burn is with embodiment 1.
Diagnostic result: hinder the back skin area and significantly dwindle, color is sallow and empyreumatic taste arranged.Naked eyes see that burned skin solidifies attenuation.Pathological section shows: lamina reticularis the latter half is ruined down to whole or most of, and hair follicle and sebaceous gland total necrosis are not seen remaining hair follicle, and pathological changes meets III degree burn wound feature.
Claims (4)
1. make animal burn model with fuel combination for one kind, its mixed fuel formula is characterised in that 20%～60% oil refinery's defective material that contains percentage by weight is reduced crude, 40～80% gasoline.
2. according to the fuel combination of the described manufacturing animal model of burn use of claim 1, its compound method is characterised in that oil refinery's defective material is dissolved in the gasoline, mixing, and-20 ℃ are freezing, take out, stir, 3～7 extremely even glues of freezing repeatedly stirring, standby.
3. according to described mixed fuel formula and the compound method thereof of making the employing of animal burn model of claim 1-2, its method feature of making animal burn model is to select laboratory animals such as mice, rat, Cavia porcellus, rabbit, pig and Canis familiaris L. for use, fuel combination evenly is applied to animal depilation pre-burning skin surface with soft brush, hide with wet cloth on every side, igniting, sustained combustion 15～30 seconds causes burn in various degree; The recovery of fluid infusion immediately after the wound, sub-cage rearing, free diet; Adopt morphologic observation under the wound tissue pathological section light microscopic, diagnosis is also determined the burn grade.
4. according to the described animal model of burn made from fuel combination of claim 1～3, its pathological change and clinical sign are characterised in that human body burn pathological change that this model and a variety of causes such as flame, electric light, lonizing radiation, hot water etc. cause, sings and symptoms etc. are closely similar, for the pathogenesis of research burn and the medicine etc. of estimating burn provide animal burn model; This model has easy and simple to handle, favorable reproducibility, and the success rate height, the burn degree is easy to remarkable advantages such as control.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103565536A (en) * | 2013-10-29 | 2014-02-12 | 中国人民解放军第四军医大学 | Simulation steam nozzle of uniform steam burn model, applicable to different body surface areas |
| CN111174229A (en) * | 2020-02-21 | 2020-05-19 | 中国人民解放军陆军军医大学 | Burn fire source generating device |
-
2011
- 2011-05-12 CN CN2011101232267A patent/CN102274084A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103565536A (en) * | 2013-10-29 | 2014-02-12 | 中国人民解放军第四军医大学 | Simulation steam nozzle of uniform steam burn model, applicable to different body surface areas |
| CN103565536B (en) * | 2013-10-29 | 2015-10-14 | 中国人民解放军第四军医大学 | A kind of simulation vapour lock being applicable to different body surface area uniform steam bum model |
| CN111174229A (en) * | 2020-02-21 | 2020-05-19 | 中国人民解放军陆军军医大学 | Burn fire source generating device |
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Application publication date: 20111214 |