A system and device for constructing an animal model of high altitude cerebral edema

Abstract

This invention discloses a system and apparatus for constructing an animal model of high-altitude cerebral edema (HACE). The resulting animal model successfully simulates the cognitive impairment and neuroinflammatory response associated with HACE, effectively reproducing key pathological features of HACE, such as oxidative stress, neuroinflammation, and neurological impairment. This model can be used to study the pathogenesis, early diagnosis, and potential treatment strategies for HACE. This invention provides a reliable animal model for experimental research and drug screening of HACE-related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of intelligent medical technology, and specifically relates to a system and device for constructing an animal model of high altitude cerebral edema. Background Art

[0002] High Altitude Cerebral Edema (HACE) is a brain disorder that occurs when individuals rapidly ascend from low altitude to high altitude and fail to acclimate to the low oxygen environment. Its main symptoms include headache, vomiting, and gait disturbances. In severe cases, HACE can also lead to cognitive impairment, such as difficulty concentrating and memory loss. Given the acute onset of HACE and its potentially severe long-term consequences, understanding its mechanisms is crucial for early intervention and improving patient outcomes.

[0003] The animal model of high-altitude cerebral edema (HACE) is the basis for animal experimental research on HACE and is of great significance for studying the pathophysiological changes and clinical treatment of HACE. The existing method for constructing an animal model of high-altitude cerebral edema is to place the modeling animal in a low-pressure, low-oxygen chamber for several days, during which time the animal is maintained on a normal diet and can be driven away. However, experimental animals have different susceptibilities to low pressure and / or low oxygen. For example, rats are less likely to develop high-altitude cerebral edema (HACE) or have less obvious symptoms of HACE under low pressure and / or low oxygen conditions. Therefore, there is an urgent need for a new method for constructing an animal model of high-altitude cerebral edema to overcome the problem of different susceptibilities of animals to low pressure and / or low oxygen. Summary of the Invention

[0004] In order to make up for the deficiencies of the prior art, the present invention aims to provide a novel system and device for constructing an animal model of high altitude cerebral edema.

[0005] A first aspect of the present invention provides a system for constructing an animal model of high altitude cerebral edema, the system comprising:

[0006] a fixing unit for fixing the animal's head on the stereotaxic apparatus;

[0007] The first processing unit is used for shaving and disinfecting, cutting the scalp along the midline, peeling off the subcutaneous tissue, and exposing the skull surface;

[0008] The second processing unit is used to determine the intracranial drug delivery cannula implantation coordinates according to the brain atlas, drill holes at the intracranial drug delivery cannula implantation coordinates using a micro drill, and vertically insert the intracranial drug delivery cannula into the target area. The intracranial drug delivery cannula implantation coordinates are the bilateral DG areas.

[0009] The third treatment unit is used for scalp suture and postoperative care and recovery;

[0010] The fourth processing unit is used to inject exosomes derived from patients with high altitude cerebral edema into the animal's brain through an intracranial drug delivery cannula after the animal recovers from surgery, thereby obtaining an animal model of high altitude cerebral edema.

[0011] The term "unit" in the present invention refers to a software or hardware component that plays a certain role, such as a field programmable gate array (FPGA), an application specific integrated circuit (ASIC), an operable medical component, a visualization component, etc. However, the term "unit" is not limited to software or hardware. The term "unit" can be configured in an addressable storage medium, or can be configured to reproduce one or more processors. Therefore, for example, the term "unit" can refer to components such as software components, object-oriented software components, class components, and task components, and can include processes, functions, properties, procedures, subroutines, program code segments, drivers, firmware, microcode, circuits, data, databases, data structures, tables, arrays, and variables. The functions provided in components and "units" can be combined into fewer components and "units", or can be further divided into additional components and "units". In addition, components and "units" can be implemented to operate one or more central processing units (CPUs) in a device or a secure multimedia card.

[0012] Furthermore, the coordinates of the bilateral DG regions are defined as the origin at the anterior bregma point, and the coordinates relative to Bregma are: AP = -2.1 mm, ML = ±1.7 mm, DV = -2.1 mm.

[0013] Furthermore, an intracranial drug administration cannula fixing unit is included before the third processing unit, for fixing the base of the intracranial drug administration cannula to the skull surface.

[0014] Furthermore, the fixing method includes using medical biological glue or dental cement for fixation.

[0015] Furthermore, the diameter of the micro drill is 0.5 mm to 1 mm.

[0016] Furthermore, the depth of the drilling is 0.1-0.2 mm.

[0017] Furthermore, the postoperative care and recovery includes applying antibiotic ointment to the animal's suture area and / or the area surrounding the suture area.

[0018] Furthermore, the postoperative care and recovery also includes administering analgesics to relieve pain after surgery.

[0019] Furthermore, the analgesic is ibuprofen.

[0020] Furthermore, the dosage of ibuprofen is 0.1 mg / kg.

[0021] Furthermore, the postoperative care and recovery also includes monitoring the mouse's activity, body weight and / or incision healing.

[0022] Furthermore, the injecting of exosomes derived from patients with high altitude cerebral edema into the animal brain through the intracranial administration cannula includes connecting the intracranial administration cannula and a microsyringe through a catheter 3 to 7 days after the animal recovers from surgery, and injecting the exosomes derived from patients with high altitude cerebral edema with the microsyringe.

[0023] Furthermore, the injection cycle of the exosomes derived from patients with high altitude cerebral edema is once every 4 days, for a total of 4 injections.

[0024] Furthermore, the number of particles of the exosomes derived from patients with high altitude cerebral edema per injection was 2.8×10 9 indivual.

[0025] Furthermore, the construction system further comprises an anesthesia unit for anesthetizing the animal.

[0026] Furthermore, the anesthesia is inhalation anesthesia.

[0027] Furthermore, the anesthetic drug is isoflurane.

[0028] Furthermore, the concentration of isoflurane is 2% to 3%.

[0029] Furthermore, the animal model is a mouse model.

[0030] Furthermore, the mice are C57BL / 6 mice.

[0031] The term "animal" in the present invention refers to any organism, including mammals, and the steps mentioned in the present invention can be adaptively adjusted according to different animals.

[0032] The second aspect of the present invention provides any of the following applications:

[0033] 1) Application of the animal model constructed by the construction system described in the first aspect of the present invention in screening drugs for preventing or treating high altitude cerebral edema; and / or in studying the mechanism of high altitude cerebral edema;

[0034] 2) Application of exosomes derived from patients with high altitude cerebral edema in constructing animal models of high altitude cerebral edema.

[0035] A third aspect of the present invention provides a device for constructing an animal model of high altitude cerebral edema, the device comprising: a single or multiple processors, and a memory, the memory being configured to store a single or multiple computer programs, which, when executed by the single or multiple processors, implement:

[0036] Operation 1 is used to fix the animal's head on the stereotaxic apparatus;

[0037] Operation 2 is used to shave and disinfect, incise the scalp along the midline, peel off the subcutaneous tissue, and expose the skull surface;

[0038] Operation 3 is used to determine the intracranial drug delivery cannula implantation coordinates based on the brain atlas, drill a hole at the intracranial drug delivery cannula implantation coordinates with a microdrill, and vertically insert the intracranial drug delivery cannula into the target area. The intracranial drug delivery cannula implantation coordinates are the bilateral DG regions; the coordinates of the bilateral DG regions are defined as the origin at the anterior bregma point, and the coordinates relative to Bregma are: AP = -2.1 mm, ML = ±1.7 mm, DV = -2.1 mm;

[0039] Operation 4 is used to suture the scalp and provide postoperative care and recovery;

[0040] Operation 5 is used to inject exosomes derived from patients with high altitude cerebral edema into the animal's brain through an intracranial drug delivery cannula after the animal recovers from surgery, thereby obtaining an animal model of high altitude cerebral edema.

[0041] Furthermore, the injection cycle of the exosomes derived from patients with high altitude cerebral edema is once every 4 days, for a total of 4 injections.

[0042] Furthermore, the number of particles of the exosomes derived from patients with high altitude cerebral edema per injection was 2.8×10 9 indivual.

[0043] Furthermore, the construction device further comprises: a depilatory component, a disinfecting component, a cutting component, a medication component, and a suturing component;

[0044] The hair removal parts include: a shaver and surgical scissors;

[0045] The cutting member includes: scissors;

[0046] The drug delivery device includes: a micro syringe;

[0047] The suture piece includes a suture thread.

[0048] The term "device" in the present invention is not limited to one or a specific number of physical objects. As used herein, a device can be any medical or electronic device having multiple components that can implement at least some aspects of the present disclosure. Although the following description and examples use the term "device" to describe certain aspects of the present disclosure, the term "device" is not limited to a specific configuration, type, or number of objects.

[0049] The term "processor" or "memory" in the present invention includes a computing device having one processor or one memory, as well as a device having multiple processors or multiple memories, which can be used to perform some or all of the steps described. A "processor" may include more than one processor, for example, a multi-core design or multiple processors each having a multi-core design.

[0050] Beneficial effects of this application:

[0051] This application discloses a novel system for constructing an animal model of high-altitude cerebral edema (HACE). The resulting animal model successfully mimics the cognitive impairment and neuroinflammatory responses associated with HACE, effectively reproducing key pathological features of HACE, such as oxidative stress, neuroinflammation, and neurological impairment. This model can be used to study the pathogenesis, early diagnosis, and potential treatment strategies for HACE. This invention provides a reliable animal model for experimental research and drug screening of HACE-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 A schematic diagram of the system flow for constructing an animal model of high altitude cerebral edema;

[0053] Figure 2 A schematic diagram of a device for constructing an animal model of high altitude cerebral edema;

[0054] Figure 3 Figure 1 shows the experimental results of cognitive dysfunction in mice induced by injection of exosomes from HACE patients; A: Schematic diagram of exosome injection; B: Flowchart of animal experiment; C: Results of open field test; D: Results of novel object recognition test; E: Results of Y-maze test; F: Results of novel environment feeding inhibition test; G: Results of fear conditioning reflex;

[0055] Figure 4 Experimental results on oxidative stress response in mice induced by injection of exosomes from HACE patients: A: expression level of NO in the serum of mice in each group; B: expression level of SOD in the serum of mice in each group; C: expression level of GSH in the serum of mice in each group; D: expression level of MDA in the serum of mice in each group; E: expression level of CAT in the serum of mice in each group; F: expression level of T-AOC in the serum of mice in each group; G: expression level of NO in the hippocampus and prefrontal cortex of mice in each group; H: expression level of SOD in the hippocampus and prefrontal cortex of mice in each group; I: expression level of GSH in the hippocampus and prefrontal cortex of mice in each group; J: expression level of MDA in the hippocampus and prefrontal cortex of mice in each group; K: expression level of CAT in the hippocampus and prefrontal cortex of mice in each group; L: expression level of T-AOC in the hippocampus and prefrontal cortex of mice in each group;

[0056] Figure 5Figures show the experimental results of abnormal hippocampal neuronal death in mice induced by injection of exosomes derived from HACE patients; A: Schematic diagram of Golgi staining in the DG region; B: The number of dendritic spines in the DG region of mice in the HACE group was significantly reduced; C: The number of MAP2-positive cells in the DG region of mice in the HACE group was significantly reduced; D: Schematic diagram of MAP2 immunofluorescence in the DG region;

[0057] Figure 6 Figure 3: Experimental results showing the proliferation and activation of microglia in the hippocampus and prefrontal cortex of mice induced by injection of exosomes derived from HACE patients. A: Schematic diagram of IBA1 immunofluorescence in the DG region. B: Increased number of microglia in the hippocampus and prefrontal cortex of mice in the HACE group. CF: Schematic diagram of IBA1 immunofluorescence in the CA1, CA2, CA3 and mPFC regions. DETAILED DESCRIPTION

[0058] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention and the accompanying drawings. It should be understood that the embodiments described in the present invention are only a part of the embodiments that can be implemented in the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention.

[0059] The present invention has been described in terms of operational procedures in the specification, the drawings and even the claims, which include a plurality of operational procedures. It should be clearly understood that these operational procedures may not be executed in the order in which they appear in this document or in the drawings, or may be executed in parallel. The numbers of the operational steps, such as 101, 102, 103, etc., are only used to distinguish different operations, and the numbers themselves do not represent any execution order. In addition, operations other than the numbers may also be added to the operational procedures of the present invention, and may be executed in sequence or in parallel. In addition, all operations appearing in this document may be partially omitted when necessary. It should be noted that the descriptions such as "first" and "second" mentioned in the present invention are only used to distinguish different operations, devices, modules, messages, etc., and do not limit the order of precedence or the specific type.

[0060] Figure 1 1 is a flow chart of a system for constructing an animal model of high altitude cerebral edema provided by an embodiment of the present invention. Specifically, the system includes:

[0061] A fixing unit 101 is used to fix the animal's head on the stereotaxic apparatus;

[0062] The first processing unit 102 is used for shaving and disinfecting, cutting the scalp along the midline, peeling off the subcutaneous tissue, and exposing the skull surface;

[0063] The second processing unit 103 is configured to determine the intracranial drug delivery cannula implantation coordinates based on the brain atlas, drill holes at the intracranial drug delivery cannula implantation coordinates using a micro drill, and vertically insert the intracranial drug delivery cannula into the target area. The intracranial drug delivery cannula implantation coordinates are the bilateral DG areas.

[0064] The third treatment unit 104 is used for scalp suturing and postoperative care and recovery;

[0065] The fourth processing unit 105 is used to inject exosomes derived from patients with high altitude cerebral edema into the animal's brain through a catheter after the animal recovers from surgery, so as to obtain an animal model of high altitude cerebral edema.

[0066] Furthermore, the coordinates of the bilateral DG regions are defined as the origin at the anterior bregma point, and the coordinates relative to Bregma are: AP = -2.1 mm, ML = ±1.7 mm, DV = -2.1 mm.

[0067] In some embodiments, each processing unit, fixing unit, and disinfection unit in the system for constructing an animal model of high-altitude cerebral edema may have an independent module for positioning or judgment. The independent module for positioning or judgment can independently or assist the processing unit, fixing unit, and disinfection unit in completing the procedures to be executed by the construction system.

[0068] In one embodiment, the construction system further comprises an anesthesia unit for anesthetizing the animal.

[0069] In one embodiment, the animal is a mouse. Male C57BL / 6 mice, 11 weeks old and weighing 22±1.5 g, were obtained from the Beijing Fangyuan Breeding Center and housed in an environment with a temperature of 23±1°C and a relative humidity of 50±1%. They were housed in a 12-hour light-dark cycle (lights on from 8:00 AM to 8:00 PM daily) and had free access to food and water. All animal experiments followed the Guide for the Care and Use of Animals of the National Institutes of Health (NIH Publication No. 80-23) and were approved by the Animal Care and Use Committee of Minzu University of China.

[0070] In a specific embodiment, the present invention adopts the following construction method to construct an animal model of high-altitude cerebral edema: 11-week-old male C57BL / 6 mice (22±1.5g) are selected, and after 1 week of adaptive feeding, catheter implantation is prepared to reduce the pain caused by multiple injections. The specific operation of catheter implantation is as follows: fasting for 12 hours before surgery to ensure safety during surgery, anesthesia is performed with isovolumetric isoflurane (2%~3%), and respiration and heart rate are monitored; then the mouse is fixed on a stereotaxic instrument (RWD 68025), the scalp is shaved, and the surgical area is disinfected with iodine and 75% ethanol; the scalp is incised along the midline (about 1 cm long), and the subcutaneous tissue is gently peeled off with a sterile cotton swab to expose the skull surface; with reference to the brain atlas, the coordinates of the bilateral DG areas are determined (AP -2.1 mm, ML ±1.7 mm, DV -2.1 The patient was followed up with a micro-drill (approximately 0.5 mm in diameter) at the identified location to avoid damaging the dura mater. The intracranial drug delivery cannula was then slowly inserted into the target brain area, and the base of the cannula was fixed to the skull surface using medical glue or dental cement to prevent displacement. The drug delivery cannula interface was sealed with a sealing cap to prevent postoperative infection. The scalp was sutured and antibiotic ointment was applied. After surgery, sustained-release ibuprofen (0.1 mg / kg) was administered to relieve pain, and the patient was monitored continuously for 3 days. After a five-day recovery period, the drug delivery cannula was connected to a micro-syringe via a catheter, and exosomes from patients with high-altitude cerebral edema were injected every four days using a micro-syringe, with 2.8 × 10 8 The exosomes from the granules were injected four times to obtain an animal model of high altitude cerebral edema. In one embodiment, the exosomes from patients with high altitude cerebral edema can be injected through a catheter connected to a microinjection pump, preferably at a flow rate of 100 nL per minute.

[0071] In one example, exosomes derived from HACE patients were obtained from the blood of 11 patients diagnosed with HACE by physicians at the Tibet Military Region General Hospital. Eleven healthy volunteers were also recruited as a control group (Ctr). All patients provided written informed consent. The study protocol was approved by the Ethics Committee of Minzu University of China and adhered to the guidelines of the Declaration of Helsinki. Figure 3 Figure A shows a schematic diagram of exosome extraction and injection. Figure 3 Figure B outlines the timeline for model development.

[0072] In one example, blood samples from HACE patients and healthy individuals were collected using BD Vacutainer™ serum tubes (Thermo Fisher Scientific, BD367895). These tubes do not contain anticoagulants, allowing the blood to clot naturally. After collection, the samples were left at room temperature for 60 minutes to complete the clotting process, and then centrifuged at 1300 × g for 10 minutes to separate the serum. The separated serum was transferred to a new EP tube and immediately stored at −80°C. Exosomes were extracted using the qEVoriginal 70 nm Gen 2 system (IZON, ICO70-13030) according to the manufacturer's instructions and purified with Vivaspin. ® Exosomes were concentrated using 20 ultrafiltration units (Sartorius, 30,000 MWCO PES), and the size distribution of exosomes was determined by nanoparticle tracking analysis using the NanoSight system (NanoSight, London, UK).

[0073] In one embodiment, sera from 2 to 4 patients are combined to extract exosomes, and the extracted exosomes are evenly distributed to the corresponding mouse groups to ensure that the observed effects more accurately reflect the overall group characteristics rather than the variability of individual human samples.

[0074] In one embodiment, the construction system further includes a verification unit for verifying whether the animal model is successfully constructed.

[0075] In one embodiment, the success of the model construction is verified by evaluating the cognitive function of the mice. After 4 injections, the cognitive function of the mice is evaluated by a series of behavioral tests.

[0076] In one embodiment, the cognitive function of mice is assessed by an open field test. The open field test (OFT) is a commonly used method to assess the motor activity and exploratory behavior of mice. The experimental apparatus is divided into 16 equal areas, and the mice are initially placed in the central position and adapted to the environment for 2 minutes. This process allows the mice to adapt to the new environment, thereby ensuring their natural reactions. After the adaptation period, the following three indicators are recorded: the total distance walked, the time spent in the central area, and the number of times the central area is entered, and the recording time is 3 minutes. In order to maintain the objectivity of the analysis, the experiment was performed by an experimenter who was unaware of the grouping of the mice. The results showed that the mice injected with exosomes from HACE patients had significantly reduced exploration time in the central area in the open field test (OFT) ( Figure 3 (C) shows impaired exploratory behavior. In contrast, mice injected with exosomes from healthy individuals or saline showed higher activity levels.

[0077] In one embodiment, the cognitive function of mice is assessed by a novel object recognition test (NOR). The novel object recognition test assesses cognitive function through the natural tendency of mice to explore unfamiliar objects. In the experiment, mice were allowed to explore two identical objects for 10 minutes. After a one-hour rest, one of the objects was replaced with a new object, and the mice re-entered the experimental area and explored the new object for 10 minutes. The exploration time and interaction frequency of the mice with the new objects and familiar objects were recorded to assess the recognition memory of the mice. The results showed that the time required for mice in the HACE group to recognize new objects was significantly prolonged, indicating that exosomes from HACE patients had a negative impact on learning, memory and cognitive function ( Figure 3 D in the figure).

[0078] In one embodiment, the cognitive function of mice was assessed by a Y-maze test. The Y-maze test measures spatial learning and memory abilities by tracking the animals' exploratory behavior in a maze. Mice were placed in the starting arm of the maze and allowed to explore freely. The time spent in each arm and the order in which the arms were entered were recorded. As an indicator of working memory, the spontaneous alternation rate was calculated as: [(number of alternations) / (total number of arm entries-2)]×100. The results showed that the number of alternations in the HACE group of mice was lower, further reflecting the defects in cognition and memory ( Figure 3 E in).

[0079] In one embodiment, the cognitive function of mice was assessed by the novelty-suppressed feeding test (NSF). The novelty-suppressed feeding test assesses the motivation and anxiety levels of mice when they are exposed to a new environment after a 24-hour fast. In this test, food was placed on a piece of white filter paper and placed in the center of a 50×50×45 cm apparatus. Each mouse was placed individually in a corner of the experimental arena and allowed to explore. The latency to start eating was recorded to reveal the anxiety and motivation of the animals in the new environment. The results showed that the mice in the HACE group spent a longer time approaching the food, further indicating that their exploratory ability was significantly reduced ( Figure 3 F in ).

[0080] In one embodiment, the cognitive function of mice was assessed by fear conditioning. The fear conditioning experiment consisted of two apparatuses: a refrigerated monitoring box (23 × 23 × 30 cm) located inside a larger soundproof box (30 × 30 × 37 cm). The refrigerated monitoring box was equipped with a metal grid for electric shock and recording the vertical and horizontal movements of the animals. Conditioning training was performed on the first day, following a specific protocol: the animals remained still for 60 seconds and then received 12 stimuli. Each stimulus consisted of a 30-second conditioned stimulus at 75 dB, followed by a 30-second interstimulus interval, and finally a 2-second foot shock (30 mA) and 15 seconds of stillness. Six hours later, a short-term memory test was performed, reducing the number of stimuli to six. Each trial consisted of a conditioned stimulus and a 30-second follow-up interval, and ended with no foot shock. Long-term memory testing was performed on the second day, the day after the short-term test, and the same method was used to assess long-term memory quality. The results showed that mice in the HACE group showed less freezing in response to auditory stimuli, while no abnormalities were observed in the other groups ( Figure 3 G in ).

[0081] In summary, multiple behavioral tests revealed significant cognitive impairment in the HACE group, while mice injected with healthy exosomes showed no such deficits. In fact, some cognitive indicators in the healthy exosome group were even better than those in the saline group. This suggests that exosomes from HACE patients induce the same cognitive impairment seen in patients with high-altitude cerebral edema (HACE) in mice, demonstrating the successful establishment of a HACE mouse model.

[0082] In one embodiment, physiological assessments of mouse brain tissue and blood were further performed to verify the successful establishment of the model. Studies have shown that an imbalance in oxidative stress homeostasis is involved in the pathogenesis of HACE. This imbalance primarily stems from excessive production of reactive oxygen species (ROS) or insufficient antioxidant defenses. These factors promote abnormal microglial activation and disrupt endothelial tight junctions, ultimately affecting the integrity of the neurovascular unit. This disruption leads to irreversible neuronal death and blood-brain barrier damage, which in turn causes cerebral edema. To assess whether HACE exosomes induce abnormalities in oxidative capacity and ROS levels, we measured the expression levels of NO, SOD, GSH-Px, MDA, CAT, and T-AOC in mouse serum and in the hippocampus (Hip) and medial prefrontal cortex (mPFC).

[0083] NO is a redox-active molecule that plays a key role in regulating endothelial tension, improving blood flow and oxygen supply, and influencing oxidative stress and inflammatory responses. These functions significantly affect the occurrence and progression of HACE. We measured NO levels in the serum, hippocampus, and prefrontal cortex of mice and found that NO levels in the serum and hippocampus of mice injected with HACE exosomes were significantly increased ( Figure 4 A in Figure 4G in ).

[0084] SOD is an antioxidant enzyme that catalyzes the conversion of superoxide to hydrogen peroxide, thereby affecting oxidative stress levels. Studies have shown that SOD expression is significantly elevated in the brain tissue of experimental mice exposed to high-altitude cerebral edema. We conducted a detailed examination of the SOD levels in mice injected with HACE exosomes and found that SOD levels in the hippocampus, prefrontal cortex, and serum of mice in the HACE group were abnormally elevated ( Figure 4 B in Figure 4 H in ).

[0085] GSH-Px is an antioxidant that can neutralize excessive ROS and prevent oxidative damage. It was significantly decreased in the hippocampus of HACE group mice ( Figure 4 C in Figure 4 I in the ).

[0086] MDA is a byproduct of lipid peroxidation and an indirect marker of oxidative stress. Studies have shown that MDA levels are significantly increased in the hippocampus and cortex of HACE rats. Similarly, we observed a significant increase in MDA levels in the blood, prefrontal cortex, and hippocampus of mice in the HACE group ( Figure 4 D in Figure 4 J in ).

[0087] CAT primarily decomposes hydrogen peroxide into water and oxygen, protecting cells from oxidative stress. A decrease in CAT levels can lead to increased oxidative stress. In the HACE group of mice, we observed a slight increase in CAT expression ( Figure 4 The E in Figure 4 K in ).

[0088] AOC reflects the overall antioxidant status of various substances and enzymes and is an important indicator of oxidative stress. T-AOC levels in the hippocampus of mice in the HACE group were significantly altered. Overall, the HACE group exhibited active lipid peroxidation and redox imbalance in serum and brain tissue.

[0089] In one example, to evaluate whether exosomes from HACE patients cause neuronal damage in mice, we performed Golgi staining on the DG region of the mouse hippocampus. The results showed that the dendritic spine density of neurons in the HACE group of mice was significantly reduced ( Figure 5 A in Figure 5 In addition, immunofluorescence staining of MAP2, which marks neurons in the DG region of the hippocampus, showed a significant decrease in the number of neurons in the HACE group. These findings suggest that exosomes derived from HACE patients severely damage hippocampal neurons in mice.

[0090] In one example, to further investigate the number and activation status of microglia in the hippocampus, we labeled microglia with IBA1 and CD68. The results showed that the HACE group mice had a significantly decreased number of microglia in the DG ( Figure 6 A in), CA1 ( Figure 6 C in), CA2 ( Figure 6 D in), CA3 ( Figure 6 E) and mPFC ( Figure 6 F) The number of microglia increased in the region, and the number of activated microglia increased significantly ( Figure 6 In addition, these microglia displayed abnormal features such as enlarged cell bodies and shortened processes.

[0091] In summary, the above results show that mice injected with HACE patient-derived exosomes showed cognitive dysfunction and redox imbalance in the mouse brain tissue, indicating that injecting HACE patient-derived exosomes into the mouse brain can cause mice to develop symptoms related to high-altitude cerebral edema, indicating that the HACE mouse model was successfully constructed.

[0092] Figure 2 1 is a schematic diagram of a device for constructing an animal model of high altitude cerebral edema provided by an embodiment of the present invention. Specifically, the device comprises: a single or multiple processors, and a memory, wherein the memory is used to store a single or multiple computer programs. When the single or multiple computer programs are executed by the single or multiple processors, the following are implemented:

[0093] Operation 1 is used to fix the animal's head on the stereotaxic apparatus;

[0094] Operation 2 is used to shave and disinfect, incise the scalp along the midline, peel off the subcutaneous tissue, and expose the skull surface;

[0095] Operation 3 is used to determine the intracranial drug delivery cannula implantation coordinates based on the brain atlas, drill a hole at the intracranial drug delivery cannula implantation coordinates with a microdrill, and vertically insert the intracranial drug delivery cannula into the target area. The intracranial drug delivery cannula implantation coordinates are the bilateral DG regions; the coordinates of the bilateral DG regions are defined as the origin at the anterior bregma point, and the coordinates relative to Bregma are: AP = -2.1 mm, ML = ±1.7 mm, DV = -2.1 mm;

[0096] Operation 4 is used to suture the scalp and provide postoperative care and recovery;

[0097] Operation 5 is used to inject exosomes derived from patients with high altitude cerebral edema into the animal's brain through an intracranial drug delivery cannula after the animal recovers from surgery, thereby obtaining an animal model of high altitude cerebral edema.

[0098] In one embodiment, the injection cycle of the exosomes derived from patients with high altitude cerebral edema is once every 4 days, for a total of 4 injections.

[0099] In one embodiment, the number of particles of the exosomes derived from patients with high altitude cerebral edema per injection is 2.8×10 9 indivual.

[0100] Those skilled in the art will clearly understand that, for the convenience and brevity of description, the specific working processes of the systems, devices and units described above can refer to the corresponding processes in the aforementioned method embodiments and will not be repeated here.

[0101] In the several embodiments provided in this application, it should be understood that the disclosed systems, devices and methods can be implemented in other ways. For example, the device embodiments described above are merely schematic. For example, the division of the units is merely a logical function division. In actual implementation, there may be other division methods, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be an indirect coupling or communication connection through some interfaces, devices or units, which can be electrical, mechanical or other forms.

[0102] The units described as separate components may or may not be physically separate, and the components shown as units may or may not be physical units, that is, they may be located in one place or distributed across multiple network units. Some or all of these units may be selected to achieve the purpose of this embodiment according to actual needs.

[0103] In addition, the functional units in the various embodiments of the present invention may be integrated into a single processing unit, each unit may exist physically separately, or two or more units may be integrated into a single unit. The aforementioned integrated units may be implemented in the form of hardware or software functional units.

[0104] Those skilled in the art will understand that all or part of the steps in the various methods of the above embodiments can be completed by instructing related hardware through a program, and the program can be stored in a computer-readable storage medium, which may include: read-only memory (ROM), random access memory (RAM), disk or optical disk, etc.

[0105] Those skilled in the art will understand that all or part of the steps in the above-mentioned embodiment method can be implemented by instructing the relevant hardware through a program, and the program can be stored in a computer-readable storage medium. The above-mentioned storage medium can be a read-only memory, a disk or an optical disk, etc.

[0106] The above is a detailed introduction to a computer device provided by the present invention. For those skilled in the art, according to the concept of the embodiments of the present invention, there may be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A system for constructing a high altitude cerebral edema mouse model, characterized in that: The build system includes: A fixing unit, used to fix the mouse head on the stereotaxic apparatus; The first processing unit is used for shaving and disinfecting, cutting the scalp along the midline, peeling off the subcutaneous tissue, and exposing the skull surface; The second processing unit is used to determine the intracranial drug delivery cannula implantation coordinates according to the brain atlas, drill holes at the intracranial drug delivery cannula implantation coordinates using a micro drill, and vertically insert the intracranial drug delivery cannula into the target area. The intracranial drug delivery cannula implantation coordinates are the bilateral DG areas. The third treatment unit is used for scalp suture and postoperative care and recovery; The fourth processing unit is used to connect the intracranial drug delivery cannula and the microinjector through the catheter after the mouse recovers from surgery, and inject the exosomes derived from the high altitude cerebral edema patient with the microinjector to obtain a high altitude cerebral edema mouse model; the injection cycle of the exosomes derived from the high altitude cerebral edema patient is once every 4 days, for a total of 4 injections; the number of particles of the exosomes derived from the high altitude cerebral edema patient injected each time is 2.8×10 9 indivual.

2. The construction system according to claim 1, wherein: The coordinates of the bilateral DG regions were defined as the origin at the anterior bregma point, and the coordinates relative to Bregma were: AP = -2.1 mm, ML = ±1.7 mm, and DV = -2.1 mm.

3. The construction system according to claim 1, wherein: The third processing unit further includes an intracranial drug administration cannula fixing unit for fixing the base of the intracranial drug administration cannula to the surface of the skull.

4. The construction system according to claim 3, characterized in that The method of fixing the base of the intracranial drug delivery cannula to the skull surface comprises fixing with medical biological glue or dental cement.

5. The construction system according to claim 1, wherein: The diameter of the micro drill is 0.5 mm to 1 mm.

6. The construction system according to claim 1, wherein: The depth of the drilling is 0.1-0.2 mm.

7. The construction system according to claim 1, wherein: The postoperative care and recovery included applying antibiotic ointment to the suture area and / or the area surrounding the suture area of ​​the mouse.

8. The construction system according to claim 1, wherein: The postoperative care and recovery also includes administering analgesics to relieve pain after surgery.

9. The construction system according to claim 8, characterized in that The analgesic is ibuprofen.

10. The construction system according to claim 9, characterized in that The dosage of ibuprofen was 0.1 mg / kg.

11. The construction system according to claim 1, wherein: The postoperative care and recovery also includes monitoring the mouse's activity, body weight and / or wound healing.

12. The construction system according to claim 1, wherein: An anesthesia unit is also included for anesthetizing mice.

13. The construction system according to claim 12, wherein: The anesthesia is inhalation anesthesia.

14. The construction system according to claim 12 or 13, characterized in that: The anesthetic drug is isoflurane.

15. The construction system according to claim 14, wherein: The concentration of isoflurane is 2% to 3%.

16. The construction system according to claim 1, wherein: The mice are C57BL / 6 mice.

17. A device for constructing a high altitude cerebral edema mouse model, characterized in that: The construction device includes: a single or multiple processors, and a memory, wherein the memory is used to store a single or multiple computer programs, and when the single or multiple computer programs are executed by the single or multiple processors, the following are implemented: Operation 1 is used to fix the mouse head on the stereotaxic instrument; Operation 2 is used to shave and disinfect, incise the scalp along the midline, peel off the subcutaneous tissue, and expose the skull surface; Operation 3 is used to determine the intracranial drug delivery cannula implantation coordinates based on the brain atlas, drill a hole at the intracranial drug delivery cannula implantation coordinates with a microdrill, and vertically insert the intracranial drug delivery cannula into the target area. The intracranial drug delivery cannula implantation coordinates are the bilateral DG regions; the coordinates of the bilateral DG regions are defined as the origin at the anterior bregma point, and the coordinates relative to Bregma are: AP = -2.1 mm, ML = ±1.7 mm, DV = -2.1 mm; Operation 4 is used to suture the scalp and provide postoperative care and recovery; Operation 5 is used to connect the intracranial drug delivery cannula and the microinjector through the catheter after the mouse recovers from surgery, and inject the exosomes derived from the high altitude cerebral edema patient with the microinjector to obtain a high altitude cerebral edema mouse model; the injection cycle of the exosomes derived from the high altitude cerebral edema patient is once every 4 days, for a total of 4 injections; the number of particles of the exosomes derived from the high altitude cerebral edema patient injected each time is 2.8×10 9 indivual.

18. The construction device according to claim 17, characterized in that The construction device also includes: a hair removal unit, a disinfection unit, a cutting unit, a drug delivery unit, and a suturing unit; The hair removal parts include: a shaver and surgical scissors; The cutting member includes: scissors; The drug delivery device includes: a micro syringe; The suture piece includes a suture thread.

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