CN117653488A - Novel EPR animal experiment platform and application method thereof - Google Patents
Novel EPR animal experiment platform and application method thereof Download PDFInfo
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Abstract
The invention discloses a novel EPR animal experiment platform, which comprises a peristaltic pump, a membrane oxygenator, a variable-temperature water tank, a constant-temperature box and a heat exchanger, wherein the constant-temperature box comprises a shell, a base and a refrigerating system, an air circulation fan is arranged at the top inside the shell, an operation window is arranged on the front surface of the shell, an observation window is arranged above the operation window, an oxygen delivery port is arranged on one side of the shell, and a heatable animal fixing table is arranged on the base. The invention also discloses a using method of the novel EPR animal experiment platform. The novel EPR animal experiment platform and the application method thereof realize high-quality cardiopulmonary support, rapid cooling and controllable rewarming, and are beneficial to improving the treatment rate and survival rate of traumatic cardiac arrest and good nervous system recovery.
Description
Technical Field
The invention relates to the technical field of animal experiments, in particular to a novel EPR animal experiment platform and a using method thereof.
Background
The novel emergency preservation and resuscitation (Emergency preservation and resuscitation, EPR) platform for traumatic cardiac arrest has the main functions of reducing the basic metabolism of organisms through rapid cooling and replacing cardiopulmonary functions through in-vitro cardiopulmonary bypass, so that rapid hemostasis can be realized before irreversible nervous system and other tissue and organ injuries occur, longer rescue and treatment windows can be obtained for doctors and patients, and controllable core temperatures can be set according to various examination indexes and vital signs of experimental objects for traumatic cardiac arrest. At present, the EPR platform mainly utilizes the measures of external circulation, such as water tank, physical cooling (ice bag and ice cap) or heating to finish rapid cooling and rewarming, and the heart and lung functions cannot be ensured in the process, so that adverse effects such as heart failure, cardiac arrest, pulmonary edema and respiratory failure cannot be avoided. High quality temperature management and preservation of cardiopulmonary function is critical for the specificity of traumatic cardiac arrest and individual variability of the subject.
Most of the low-temperature animal experiment methods at present use a body surface cooling strategy, and compared with an extracorporeal circulation perfusion autologous blood method, the strategy is difficult to stably reach a target temperature in a short time, and the induced cooling rate is difficult to control. Therefore, high-quality cardiopulmonary support, rapid cooling and controllable rewarming are realized, and the treatment rate, the survival rate and the good nervous system recovery of traumatic cardiac arrest are improved.
Disclosure of Invention
The invention aims to provide a novel EPR animal experiment platform, which realizes high-quality cardiopulmonary support, rapid cooling and controllable rewarming, and is beneficial to improving the cure rate and survival rate of traumatic cardiac arrest and good nervous system recovery. The invention further aims to provide a using method of the novel EPR animal experiment platform.
In order to achieve the above purpose, the invention provides a use method of a novel EPR animal experiment platform, which comprises the following steps:
s1, preparing before molding and starting an experimental platform, wherein the starting of the experimental platform and the preparing before molding are performed simultaneously;
s2, rapidly cooling, using with an incubator, starting a variable-temperature water tank circulating pump when cooling begins, starting a peristaltic pump, starting an air circulating fan in the incubator, starting an incubator base refrigerating system, and automatically reducing the power of the refrigerating system by the incubator after the experimental temperature is reached, so as to maintain the target experimental temperature;
s3, real-time physiological monitoring is mainly responsible for summarizing all vital sign index parameters of experimental animals, wherein the index parameters comprise electrocardio, heart rate, respiratory rate, arterial pressure, temperature and blood flow;
s4, controllable re-warming, closing a refrigerating system of the incubator, setting the incubator temperature according to the requirement, and adjusting the temperature of the variable-temperature water tank to be the target re-warming temperature;
s5, targeted resuscitation and treatment, wherein a resuscitation mode, a treatment method and maintenance core temperature are determined according to vital signs and monitoring index results of experimental animals, and the machine is continuously turned or removed.
Preferably, in the step S1, the specific operation steps of preparation before molding are as follows:
(1) Anesthesia: placing the experimental animal on the incubator on a heatable animal fixing table after injection anesthesia, fixing the limbs firmly, and connecting an animal respiratory anesthesia machine to maintain an anesthesia and sedation state;
(2) Vascular separation: performing neck unilateral vein separation and unilateral femoral artery separation on experimental animals, performing intubation operation, and separating one femoral artery/carotid artery for real-time arterial pressure monitoring, and synchronously separating esophagus for recording the core body temperature of the animals when separating neck veins;
(3) And (3) tube placement: the vessel is clamped by using an arterial clamp through a surgical tube, and the hose is respectively placed into the jugular vein and the femoral artery separated from the experimental animal;
(4) Platform equipment pipeline connection: the silica gel hose leads the blood at the neck vein cannula of the experimental animal to the peristaltic pump, the blood passes through the peristaltic pump and then the membrane oxygenator, the venous blood enters the outer cavity of the heat exchanger after being oxygenated, and enters the femoral artery after being cooled by the heat exchanger.
Preferably, in the step S1, the experimental platform is started to start the incubator, adjust to a constant temperature mode of 35 ℃, maintain the body temperature of the experimental animal to a normal temperature, and start the variable-temperature water tank to start precooling.
Preferably, in the step S3, the temperature monitoring point includes: the temperature of esophagus, anus, both sides ear, venous drainage and femoral artery delivery, and the target temperature of the experimental animal is referred to as the temperature of esophagus and the temperature of venous drainage.
Preferably, in the step S3, the target temperature of the experimental animal is referred to as esophageal temperature.
The utility model provides a novel EPR animal experiment platform, includes peristaltic pump, diaphragm type oxygenator, alternating temperature water tank, still includes thermostated container, heat exchanger, thermostated container case, base and refrigerating system, the inside top of shell is equipped with air circulation fan, and the shell openly is equipped with operating window, the operating window top is equipped with the observation window, shell one side is equipped with the oxygen therapy mouth, be equipped with heatable animal fixed station on the base.
Preferably, the heat exchanger comprises an inner cavity and an outer cavity, the inner cavity and the outer cavity are coaxially arranged, the inner cavity is a cooling liquid passage and is directly connected with the variable-temperature water tank, the outer cavity is a blood passage, and the heat transfer between the blood in the outer cavity and the cooling liquid in the inner cavity is reduced.
The novel EPR animal experiment platform and the application method thereof have the advantages that:
the invention provides a high-efficiency and controllable rapid cooling method for extracorporeal circulation perfusion autologous blood, venous blood led out from a jugular vein is input through a femoral artery through a cooling intubation position, a variable-temperature water tank is connected with heat exchanger extracorporeal blood, high-quality cardiopulmonary support, rapid cooling and controllable rewarming are realized, and the rapid cooling method is beneficial to improving the treatment rate and survival rate of traumatic cardiac arrest and good nervous system recovery.
The technical scheme of the invention is further described in detail through the drawings and the embodiments.
Drawings
FIG. 1 is a schematic view of the structure of the incubator of the present invention;
FIG. 2 is an exploded view of the incubator of the present invention;
FIG. 3 is a flow chart of the rapid cooling process according to the present invention.
Reference numerals
1. An observation window; 2. an operation window; 3. a base and a refrigeration system; 4. a heatable animal holding station; 5. an oxygen delivery port; 6. a housing; 7. an air circulation fan; 8. a constant temperature box.
Detailed Description
The technical scheme of the invention is further described below through the attached drawings and the embodiments.
Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs. The terms "first," "second," and the like, as used herein, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "upper", "lower", "left", "right", etc. are used merely to indicate relative positional relationships, which may also be changed when the absolute position of the object to be described is changed.
The utility model provides a novel EPR animal experiment platform, includes peristaltic pump, diaphragm type oxygenator, alternating temperature water tank, still includes thermostated container 8, heat exchanger, and thermostated container 8 includes shell 6, base and refrigerating system 3, and the inside top of shell 6 is equipped with air circulation fan 7, and shell 6 openly is equipped with operating window 2, and operating window 2 top is equipped with observation window 1, and shell 6 one side is equipped with oxygen therapy mouth 5, is equipped with heatable animal fixed station 4 on the base.
Example 1
The application method of the novel EPR animal experiment platform comprises the following steps:
s1, preparation before molding and starting an experimental platform, wherein the starting of the experimental platform and the preparation before molding are carried out simultaneously, vital signs of a base line level, esophageal temperature, anal temperature, mean arterial pressure, electrocardio, respiration and the like need to be monitored before molding.
The specific operation steps of preparation before molding are as follows:
(1) Anesthesia: placing the experimental animal on the incubator 8 on the heatable animal fixing table 4 after injection anesthesia, fixing the limbs firmly, and connecting with an animal respiratory anesthesia machine to maintain an anesthesia and sedation state;
(2) Vascular separation: performing neck unilateral vein separation and unilateral femoral artery separation on experimental animals, performing intubation operation, and separating one femoral artery/carotid artery for real-time arterial pressure monitoring, and synchronously separating esophagus for recording the core body temperature of the animals when separating neck veins;
(3) And (3) tube placement: the vessel is clamped by using an arterial clamp through a surgical tube, and the hose is respectively placed into the jugular vein and the femoral artery separated from the experimental animal;
(4) Platform equipment pipeline connection: the silica gel hose is used for draining blood at the neck vein cannula of the experimental animal to the peristaltic pump, the flow range is 0-80 mL/min, the forward and reverse rotation is adjustable, the blood passes through the peristaltic pump, then passes through the membrane oxygenator, the blood volume of the membrane oxygenator is 15mL, the oxygen supply of the blood required by the animal can be maintained in a low-temperature state, the venous blood enters the outer cavity of the heat exchanger after being oxygenated, enters the femoral artery after being cooled by the heat exchanger, the heat exchanger comprises an inner cavity and an outer cavity, the inner cavity and the outer cavity are coaxially arranged, the inner cavity is a cooling liquid passage and is directly connected with a variable-temperature water tank, the outer cavity is a blood passage, and the heat transfer cooling of the blood in the outer cavity and the cooling liquid in the inner cavity is realized.
The experimental platform is started to start the incubator 8, the temperature is regulated to 35 ℃ in a constant temperature mode, the body temperature of an experimental animal is maintained to be normal temperature, the animal is prevented from losing temperature before the experiment starts, the experimental result is influenced, the variable-temperature water tank is started to start precooling, and the target temperature of the cooling liquid in the variable-temperature water tank is regulated to be 0 ℃ to minus 30 ℃ according to the required cooling speed. The temperature of the water tank and the constant temperature box is not reduced to 35 ℃ in the rewarming process, but is slowly adjusted according to the real-time temperature of the experimental animal. If the body temperature of the experimental animal at the beginning of the rewarming is 16 ℃, the temperature of the water tank and the incubator is adjusted to about 18 or 20 ℃. It should be noted that the temperature of the incubator and the water tank cannot exceed 10 ℃ of the real-time temperature of the experimental animal every time, and the pump speed is adjusted in real time, generally the pump speed is adjusted to be about 10-20mL/min when the deep low temperature is maintained, and the speed is also approximately maintained in the rewarming process. There may be some differences depending on the different tanks, the different lines and the different pumps.
S2, rapidly cooling, using with the incubator 8, starting a variable-temperature water tank circulating pump when cooling starts, enabling the circulating flow of the cooling liquid to be 10-20L/min, adjusting the pump speed according to the requirement, starting a peristaltic pump, enabling the blood flow to be 40-80 mL/min, starting an air circulating fan 7 in the incubator 8, starting a base refrigerating system of the incubator 8, placing ice bags according to the requirement to accelerate cooling, setting the minimum temperature of the internal environment of the incubator 8 to be 0 ℃, the maximum temperature to be 40 ℃, enabling the fastest cooling rate to reach-20 ℃/20 min, and reducing the cooling speed according to the requirement relative to the normal body temperature of an experimental animal, wherein the incubator 8 can automatically reduce the power of the refrigerating system and maintain the target experimental temperature after the experimental temperature is reached.
S3, real-time physiological monitoring is mainly responsible for summarizing all vital sign index parameters of experimental animals, wherein the index parameters comprise electrocardiograph, heart rate, respiratory rate and arterial pressure monitored by an electrocardiograph monitoring anesthesia integrated machine, temperature is monitored by a multipath temperature monitor, blood flow is observed in real time by a pump, and temperature monitoring points comprise: the temperature of esophagus, anus, both sides ear, venous drainage and femoral artery delivery, and the target temperature of the experimental animal is referred to as the temperature of esophagus and the temperature of venous drainage. Referring primarily to the esophageal temperature, the esophageal temperature represents the core temperature. Bubbles are formed easily when the operation is improper during the blood temperature monitoring, and the bubbles are large or can cause air embolism to cause death of experimental animals. The incubator 8 can set the incubator temperature according to the requirement, and the incubator temperature display is arranged outside the incubator, so that the temperature in the incubator can be monitored in real time, and the experiment is convenient to record the environmental temperature.
S4, controllable re-warming, closing a refrigerating system of the incubator 8, setting the incubator temperature of the incubator 8 according to the requirement, and adjusting the temperature of the variable-temperature water tank to be the target re-warming temperature.
S5, targeted resuscitation and treatment, wherein a resuscitation mode, a treatment method and maintenance core temperature are determined according to vital signs and monitoring index results of experimental animals, and the machine is continuously turned or removed. The resuscitation mode comprises chest compression, electric defibrillation, administration of vasoactive drugs such as epinephrine and norepinephrine, tracheal intubation or mask for oxygen administration, and the treatment method comprises symptomatic treatment such as application of booster drugs such as dopamine and dobutamine, fluid infusion, blood transfusion, anticoagulation, heart strengthening, and diuresis.
Example 2
Experimental procedure of novel EPR animal experimental platform:
1. preparation before molding
1. Weighing, inducing anesthesia and fixing experimental animals;
2. separating jugular vein, carotid artery, trachea, esophagus and femoral artery;
3. arteriovenous intubation, trachea intubation (opening a respiratory anesthesia integrated machine, observing the respiratory condition of experimental animals and maintaining anesthesia in real time), oesophageal insertion temperature probes, oesophageal electrodes, anus insertion temperature probes and electrocardio connection;
4. the carotid artery is connected with a plurality of physiological instruments to monitor real-time arterial pressure and heart rate;
5. the electrocardio, blood pressure, temperature and blood gas at the baseline level of blood sampling monitoring are recorded without any treatment for observing for a certain time.
2. Establishing a traumatic cardiac arrest animal model
1. The bleeding of femoral artery (the bleeding speed is controlled by peristaltic pump), the average arterial pressure and the electrocardio change of the experimental animal are observed in real time by a multi-channel physiological instrument and an electrocardio monitor, the blood drawing is stopped and observed when the average arterial pressure is reduced to 20mmHg, and the arterial pressure is maintained to 20mmHg and the electrocardio shows myocardial ischemia during the observation.
2. A chamber tremor model is built through electric shock esophagus, and arterial pressure and electrocardiographic change are monitored in real time through a multichannel physiological instrument and an electrocardiographic monitor. The mean artery was maintained at 10-20mmHg and the electrocardiographic ventricular fibrillation during the observation period.
3. Fast cooling
Real-time observing arterial pressure by using normal saline (4 ℃) with rapid ice supplementing; the jugular vein catheter is connected with a pump and connected with a membrane lung, a peristaltic pump (the peristaltic pump speed is adjusted in real time according to arterial pressure and temperature of an experimental animal) is started, and blood enters the animal body through femoral artery after being oxygenated by the membrane lung; placing the experimental animal into a constant temperature box 8, and observing the temperature of the experimental animal and the temperature in the box in real time; the core temperature (esophagus temperature) of the experimental animal is quickly reduced to about 15 ℃ by means of the variable-temperature water tank and the constant-temperature box 8, and the time is 15-16min. The peristaltic pump flow rate and the temperature of the incubator 8 are controlled to be maintained at about 15-17 ℃ for a certain time.
4. Controlled resuscitation and resuscitation
The core temperature of the experimental animal is slowly increased to 36-37 ℃ by means of the variable-temperature water tank and the constant-temperature box 8, the pump speed, the variable-temperature water tank and the temperature of the constant-temperature box 8 are adjusted in real time according to the core temperature of the experimental animal, and the symptomatic treatment such as epinephrine, chest compression, defibrillation and the like is given according to the electrocardio and blood pressure change of the experimental animal. When the temperature of the experimental animal is raised to 36-37 ℃, observing for a certain time, taking blood, and killing a tissue sample. The blood gas at the baseline level, the blood gas after modeling and the blood gas after rewarming and resuscitation are measured in the whole experimental process.
Preparation before molding: the electrocardiographic blood pressure data of the experimental animal are recorded before molding, the electrocardiograph shows sinus rhythm, the heart rate is 228 times/min, and the average arterial pressure is 68mmHg.
And (3) molding: the electrocardiographic blood pressure data of the experimental animal was recorded after the femoral artery was subjected to constant speed bleeding (4 mL/min), and the electrocardiograph showed myocardial ischemia manifestation, heart rate 194 times/min and average arterial pressure of 20mmHg. After 5min observation, a ventricular fibrillation model is established through esophageal electric shock, and the electrocardio blood pressure is observed, wherein the electrocardiogram becomes ventricular fibrillation, the heart rate is 153 times/min, and the average arterial pressure is 18mmHg.
After rewarming and resuscitating: electrocardiogram blood pressure data were recorded showing sinus rhythm, heart rate 208 times/min, mean arterial pressure 58-63mmHg. Sinus rhythm is a normal heart rhythm, and the normal heart rate of rabbits is generally 180-250 times/min, if the heart rate is lower than 180 electrocardio, bradycardia is reported, and if the heart rate is about 300, the blood pressure is maintained at 50-60mmHg in traumatic cardiac arrest caused by blood loss, so that the circulatory state is stable. Blood pressure is one of the three elements of the body that maintains a steady circulatory state, and blood pressure is relatively easy to obtain, so blood pressure monitoring is most commonly used.
Rapid cooling and temperature control, and temperature change is shown in table 1.
TABLE 1 temperature variation results
| Experimental stage | Average time (min) | Esophageal temperature (DEG C) |
| Baseline (Rapid cooling front) | 77 | 37.6 |
| Fast cooling | 16 | 16.8 |
| Maintaining deep cryogenic temperatures | 30 | 15～17 |
| Rewarming | 245.5 | 37.1 |
Therefore, the novel EPR animal experiment platform and the application method thereof realize high-quality cardiopulmonary support, rapid cooling and controllable rewarming, and are beneficial to improving the cure rate and survival rate of traumatic cardiac arrest and good nervous system recovery.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting it, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that: the technical scheme of the invention can be modified or replaced by the same, and the modified technical scheme cannot deviate from the spirit and scope of the technical scheme of the invention.
Claims (7)
1. The application method of the novel EPR animal experiment platform is characterized by comprising the following steps of:
s1, preparation before molding and starting an experimental platform, wherein the starting of the experimental platform and the preparation before molding are performed simultaneously, and vital signs of a baseline level need to be monitored before molding;
s2, rapidly cooling, using with an incubator, starting a variable-temperature water tank circulating pump when cooling begins, starting a peristaltic pump, starting an air circulating fan in the incubator, starting an incubator base refrigerating system, and automatically reducing the power of the refrigerating system by the incubator after the experimental temperature is reached, so as to maintain the target experimental temperature;
s3, real-time physiological monitoring is mainly responsible for summarizing all vital sign index parameters of experimental animals, wherein the index parameters comprise electrocardio, heart rate, respiratory rate, mean arterial pressure, temperature and blood flow;
s4, controllable re-warming, closing a refrigerating system of the incubator, setting the incubator temperature according to the requirement, and adjusting the temperature of the variable-temperature water tank to be the target re-warming temperature;
s5, targeted resuscitation and treatment, wherein a resuscitation mode, a treatment method and maintenance core temperature are determined according to vital signs and monitoring index results of experimental animals, and the machine is continuously turned or removed.
2. The method for using the novel EPR animal experiment platform according to claim 1, which is characterized in that: in the step S1, the specific operation steps of preparation before molding are as follows:
(1) Anesthesia: placing the experimental animal on the incubator on a heatable animal fixing table after injection anesthesia, fixing the limbs firmly, and connecting an animal respiratory anesthesia machine to maintain an anesthesia and sedation state;
(2) Vascular separation: performing neck unilateral vein separation and unilateral femoral artery separation on experimental animals, performing intubation operation, and separating one femoral artery/carotid artery for real-time arterial pressure monitoring, and synchronously separating esophagus for recording the core body temperature of the animals when separating neck veins;
(3) And (3) tube placement: the vessel is clamped by using an arterial clamp through a surgical tube, and the hose is respectively placed into the jugular vein and the femoral artery separated from the experimental animal;
(4) Platform equipment pipeline connection: the silica gel hose leads the blood at the neck vein cannula of the experimental animal to the peristaltic pump, the blood passes through the peristaltic pump and then the membrane oxygenator, the venous blood enters the outer cavity of the heat exchanger after being oxygenated, and enters the femoral artery after being cooled by the heat exchanger.
3. The method for using the novel EPR animal experiment platform according to claim 1, which is characterized in that: in the step S1, the experimental platform is started to start the incubator, the temperature of the experimental animal is maintained to be normal by adjusting the temperature to a constant temperature mode of 35 ℃, and the temperature-changing water tank is started to start precooling.
4. The method for using the novel EPR animal experiment platform according to claim 1, which is characterized in that: in the step S3, the temperature monitoring point includes: the temperature of esophagus, anus, both sides ear, venous drainage and femoral artery delivery, and the target temperature of the experimental animal is referred to as the temperature of esophagus and the temperature of venous drainage.
5. The method for using the novel EPR animal experiment platform according to claim 4, wherein the method comprises the following steps: the target temperature of the experimental animal is referred to as esophageal temperature.
6. The utility model provides a novel EPR animal experiment platform, includes peristaltic pump, diaphragm type oxygenator, alternating temperature water tank, its characterized in that: still include thermostated container, heat exchanger, thermostated container case, base and refrigerating system, the inside top of shell is equipped with air circulation fan, and the shell openly is equipped with operating window, the operating window top is equipped with the observation window, shell one side is equipped with the oxygen therapy mouth, be equipped with heatable animal mount table on the base.
7. The novel EPR animal experiment platform according to claim 6, wherein: the heat exchanger comprises an inner cavity and an outer cavity, the inner cavity and the outer cavity are coaxially arranged, the inner cavity is a cooling liquid passage and is directly connected with a variable-temperature water tank, the outer cavity is a blood passage, and the heat transfer between the blood in the outer cavity and the cooling liquid in the inner cavity is reduced.
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