Plateau environment cardiac contraction function simulation and detection system based on simulated jellyfish

Abstract

The invention provides a simulated jellyfish-based plateau environment systolic function model, which comprises a jellyfish-shaped framework and a cardiac muscle tissue layer cultured on the jellyfish-shaped framework, and is used for simulating the systolic function of the heart through spontaneous or rhythmic systole; the pacing point and conduction system consists of a patterned myocardial cell layer and is used for generating and / or conducting an electric signal to stimulate rhythmic contraction / relaxation of a myocardial tissue layer; the balance weight is arranged at the bottom of the center of the jellyfish-shaped frame and used for enabling the gravity borne by the simulated jellyfish in the culture solution to be larger than the buoyancy borne by the simulated jellyfish. The invention also provides a simulated jellyfish-based plateau environment systolic function simulation and detection system, and the simulated jellyfish-based plateau environment systolic function model is used as a bionic simulator and is used for analyzing the motion state, tissue pathology and metabolism conditions of the bionic simulator in a body blood component culture solution in a simulated plateau environment. And evaluating the influence of the culture solution on the heart function.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical engineering, and particularly relates to a simulation jellyfish-based highland environment heart systolic function simulation and detection system. BACKGROUND

[0002] The highland environment is mainly characterized by low pressure and low oxygen, and the human body will appear the decrease of blood oxygen saturation, the disorder of acid-base balance and metabolism, and the change of blood pressure, which can cause the increase of heart load, the impairment of systolic function, and a series of highland heart-related diseases. The research and development of the heart model in the highland environment have significant meaning for solving the highland heart disease and related complications of the population in the highland area of China.

[0003] In the prior art, the heart model for the research of highland heart disease is mainly a virtual diagnosis model constructed based on big data and artificial intelligence, and an animal model of highland heart disease constructed by using extreme highland environmental factors. However, the above models have the following problems in the research and use: the virtual model cannot meet the demand of exploring the physiological mechanism of highland heart disease. Although the animal model can judge the heart function through blood routine, blood pressure, heart rate and other overall vital signs, it cannot directly observe the morphological and histological changes of the heart, and the animal model needs to dissect the heart when detecting the pathological and physiological changes of the heart, so that the observation of the heart in each pathological development stage will be limited, and the continuity of the pathological changes of the animal model is not good enough and the culture cost is high. SUMMARY

[0004] In view of the problems in the prior art, the present application provides a simulation jellyfish-based highland environment heart systolic function simulation and detection system to solve the technical problem that the heart model for the research of highland heart disease in the prior art cannot directly observe the morphological and histological changes of the heart.

[0005] The technical scheme adopted by the present application is as follows: In a first aspect, a simulation jellyfish-based highland environment heart systolic function model is provided, comprising: A jellyfish-like frame, which is an umbrella-shaped structure of a natural jellyfish; A myocardial tissue layer cultured on the jellyfish-like frame, used to simulate the heart systolic function through spontaneous or rhythmic contraction; A pacemaker and conduction system composed of a patterned myocardial cell layer, used to generate and / or conduct electrical signal to stimulate the rhythmic contraction / diastole of the myocardial tissue layer; A counterweight arranged at the center bottom of the jellyfish-like frame, used to make the gravity of the simulation jellyfish in the culture solution greater than the buoyancy.

[0006] Further, the jellyfish-like frame is made of biocompatible light-cured resin or elastomer by 3D printing. The myocardial tissue is differentiated from primate myocardial stem cells or from human induced pluripotent stem cells.

[0007] Further, the patterned myocardial cell layer is prepared by micro-contact printing technology, including a central pacing functional zone and peripheral conduction pathways, and the conduction pathways extend from the base of each petal-like structure in the central pacing functional zone.

[0008] Further, the generated and / or conducted electrical signals are generated by a programmable multi-channel electrical stimulation subsystem, which includes: a multi-channel electrical stimulator for generating at least two independently controllable electrical pulse sequences; a master electrode located in the central pacing functional zone for regulating dominant rhythm; a plurality of auxiliary electrodes located on the conduction pathways for introducing ectopic rhythm or testing conduction performance.

[0009] In a second aspect, a simulation and detection system for high-altitude environment heart contraction function based on simulated jellyfish is provided, which uses the high-altitude environment heart contraction function model based on simulated jellyfish as described in the first aspect as a bionic simulator for heart contraction, to analyze the motion state, histopathology and metabolism of the bionic simulator in the simulated high-altitude environment body blood component culture solution, and to evaluate the influence of the culture solution on heart function.

[0010] Further, the simulation and detection system for high-altitude environment heart contraction function based on simulated jellyfish further includes: an environment simulation unit for accommodating and controlling the living environment of the simulated jellyfish; a motion monitoring and data analysis unit for real-time capturing and quantifying the motion state of the simulated jellyfish, and evaluating the contraction function according to the motion state analysis; a histopathology and metabolism analysis unit for cytological and metabolic analysis of the myocardial tissue in the simulated jellyfish.

[0011] Further, the environment simulation unit includes a culture cylinder in the shape of a cylinder, which contains a simulated high-altitude environment tissue blood or plasma component culture solution.

[0012] Further, the culture cylinder is placed in a bioreactor with adjustable oxygen partial pressure.

[0013] Further, the motion monitoring and data analysis unit includes: The camera equipment is used for recording video data of a motion state of the simulation jellyfish in the culture solution; the motion state includes frequency, speed and amplitude of upstream and downstream motion in a certain time; The image processing analysis module is electrically connected with the camera equipment, and is used for extracting one or more parameters of a contraction frequency, a contraction amplitude, a swimming speed and a bending angle of the lobe structure from the video data, and analyzing and evaluating the contraction function.

[0014] Further, the tissue pathology and metabolism analysis unit comprises: The tissue morphology observation module is used for observing and analyzing key pathological morphological changes of myocardial cell arrangement, cell interconnection, cytoplasm vacuolar degeneration, cell nucleus pyknosis and fibrosis by using tissue chemical staining and a microscope imaging device, or is used for labeling cardiac troponin I and desmin by using specific antibodies through immunofluorescence staining. The cell metabolism detection module is used for detecting concentration changes of lactic acid, pyruvic acid, ATP / ADP / AMP and active oxygen metabolites in the culture solution by using a biochemical kit or a liquid chromatography-mass spectrometry analysis platform, or is used for performing metabolic flux analysis of glycolysis and oxidative phosphorylation on a tissue sample of the myocardial cell layer, or is used for performing real-time and dynamic oxygen consumption rate and acid production rate determination on the living tissue of the myocardial cell layer by using a hippocampus biological energy analyzer.

[0015] From the above technical solutions, the beneficial technical effects of the present application are as follows: 1. The simulation jellyfish is used as a biological sensor and a pathophysiological model in the present application, and is used for simulating and detecting changes of heart function under high altitude blood, and the simulation is the contraction-diastolic blood pumping function of the heart, and the motion of the jellyfish is used as an observable alternative index of the function.

[0016] 2. The high altitude environment heart contraction function simulation and detection system based on the simulation jellyfish integrates specific body fluid environment simulation and function quantitative analysis, constitutes a complete detection system, provides a highly controllable and replicable pathological environment model, and can stably simulate biochemical environments of different altitudes and different disease stages by accurately compounding a culture solution simulating high altitude blood components. Real-time, intuitive and quantitative evaluation of the heart contraction function under the high altitude environment is realized, and research cost and ethical restrictions are significantly reduced. The motion state of the simulation jellyfish directly reflects the heart contraction function, replaces traditional live animal experiments, shortens the test cycle and reduces the test cost. A complete evaluation chain from macro function to micro mechanism is established, and through the motion monitoring and data analysis unit and the tissue pathology and metabolism analysis unit, multi-dimensional synchronous correlation observation of the same sample from the contraction function, cell morphology to energy metabolism is realized.

[0017] 3. Through the standardized preparation process and the automated analysis software, the comparative study of multiple experimental groups can be simultaneously conducted, the research efficiency is greatly improved, the technical bottleneck of low throughput and long time consumption in traditional pathological research is solved, and the high-throughput and multi-parameter parallel detection capability is realized. A reliable preclinical evaluation platform is provided for high altitude heart disease drug research and development. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual scale.

[0019] Figure 1 is a schematic diagram of the bionic jellyfish structure in the embodiment of the present application; Figure 2 is a longitudinal cutting schematic diagram of the bionic jellyfish structure in the embodiment of the present application; Figure 3 is a schematic diagram of the bionic jellyfish movement in the embodiment of the present application; Figure 4 is a working flow schematic diagram of the high altitude environment heart contraction function simulation and detection system based on the simulation jellyfish in the embodiment of the present application. DETAILED DESCRIPTION

[0020] The embodiments of the technical solutions of the present application will be described in detail below with reference to the drawings. The following embodiments are only used to more clearly illustrate the technical solutions of the present application, and therefore only serve as examples, but cannot limit the protection scope of the present application.

[0021] It should be noted that, unless otherwise specified, the technical terms or scientific terms used in the present application should be understood as the usual meanings understood by the skilled person in the field to which the present application belongs.

[0022] EMBODIMENT The embodiment provides a high altitude environment heart contraction function simulation and detection system based on a simulation jellyfish. The system uses a simulation jellyfish with rhythmic contraction function as a bionic simulator of heart contraction function. By analyzing the motion state of the simulation jellyfish, including the frequency, speed and amplitude of upstream and downstream movement over time, the influence of the body blood component culture solution simulating the high altitude environment on the heart function is evaluated. The system can directly and quantitatively evaluate the influence of the special body internal environment caused by the high altitude on the heart contraction function.

[0023] The high altitude environment heart contraction function simulation and detection system based on the simulation jellyfish specifically comprises: 1. Simulation jellyfish unit: Core functional unit, equivalent to a high-altitude environment heart systolic function model based on simulated jellyfish, used to simulate the systolic and diastolic function of the heart. As shown in Figure 1 and Figure 2 , the unit is mainly composed of a tissue-compatible jellyfish-like framework and myocardial tissue cultured on the framework, and also includes a pacemaker and a conduction system, a counterweight. The jellyfish-like framework simulates the umbrella structure of natural jellyfish, containing a central disc and multiple radially extending petal-like structures, providing attachment points for myocardial tissue and guiding its systolic-diastolic direction. In some embodiments: The jellyfish-like framework is made of biocompatible light-cured resin (such as polyethylene glycol diacrylate photosensitive hydrogel) or elastomer (such as medical-grade polyurethane, polydimethylsiloxane, or gelatin-methacryl hydrogel) by high-precision 3D printer (EnvisionTEC 3D-Bioplotter series or CELLINK BIO X), forming an elastic polymer base layer, i.e. a hydrogel layer. The jellyfish-like framework is designed to be about 1-2 centimeters in diameter, with 8 wedge-shaped petal-like structures, about 20-50 microns thick, to allow sufficient flexibility.

[0024] The myocardial tissue is differentiated from primate, such as experimental monkey-derived myocardial stem cells, or from human induced pluripotent stem cells; using primate myocardial stem cells, as they are more physiologically similar to humans, can provide a more clinically predictive model; using myocardial cells differentiated from human induced pluripotent stem cells to achieve personalized medicine and drug testing. Cell culture and seeding of myocardial tissue: a. Isolate, extract and expand myocardial stem cells or embryonic stem cells from the bone marrow or embryos of experimental monkeys (such as cynomolgus monkeys). b. By micro-contact printing or direct seeding, seed the stem cells at a high density (e.g., 1-5 x 10^6 cells / mL) onto a 3D-printed framework coated with fibronectin. c. Induce culture in a suitable myocardial cell differentiation medium (containing necessary growth factors such as VEGF, FGF, etc.) for about 1-2 weeks to form a myocardial tissue layer that can spontaneously or rhythmically contract with ordered muscle fiber arrangement. In specific embodiments, cell culture can be performed in a Thermo Scientific Heracell series CO2 incubator.

[0025] Pace-making and conduction system: The system is composed of a layer of patterned cardiomyocytes, which is fabricated by micro-contact printing technique to exhibit a central pace-making functional zone and peripheral fast conduction pathways on a flexible substrate. The central pace-making functional zone is located in the central disc region, which is enriched with a subpopulation of high-autonomic cardiomyocytes or genetically engineered cells to dominate the pace-making function. From the base of each petaloid structure radially extending out of the central pace-making functional zone are a plurality of fast conduction pathways, which are precisely printed with a pattern of cell adhesion proteins on a flexible polymer substrate by micro-contact printing technique to guide the growth of cardiomyocytes in a predetermined direction. The conduction pathways together with the cardiomyocytes growing thereon form the conduction system, which are genetically engineered to highly express Connexin 40 junction proteins to form low-resistance channels such as gap junctions to have the function of fast conduction. The conduction bundle pathways are printed with parallel and dense (such as 20 μm wide and 20 μm apart) linear stripes to achieve the growth of cardiomyocytes in high alignment to form tight junctions and form fast conduction pathways. The speed of electrical signals propagating along the long axis of the cells is more than twice faster than the transverse direction. The linear stripes are parallel to the long axis of the leaflets to ensure that the contractile force can be efficiently used to bend the leaflets.

[0026] In the present application, the simulated jellyfish includes two contraction modes, the first one is an autonomous contraction mode, which relies on the autonomic nature of the cardiomyocytes in the central pace-making functional zone to simulate the intrinsic rhythm of the heart; the second one is an external field pacing mode: through the application of an electric field by external electrodes to achieve precise control of the contraction frequency and rhythm. In the test, the rhythm of the pace-making point can be started, stopped and adjusted by external electric field stimulation, and then the contraction frequency of the simulated jellyfish can be started, stopped or adjusted. In some embodiments, the way of applying an electric field by external electrodes is realized by a programmable multi-channel electric stimulation subsystem, which specifically includes: (1) a multi-channel electric stimulator (preferably IonOptix C-Pace EP or equivalent functional device), which can generate at least two independently controllable electric pulse sequences; (2) a main control electrode located in the central pace-making functional zone for regulating the dominant rhythm; a plurality of auxiliary electrodes located on the leaflets or fast conduction pathways for introducing ectopic rhythm or testing conduction performance; (3) control software for programming and executing various different electric signal stimulations; the implementation of the control software is not limited, and any one of the existing technologies can be implemented, such as the control software of a cardiac pacemaker. The programmable multi-channel electric stimulation subsystem can realize the following functions: (1) Contraction initiation and termination: Initiate contraction by applying a sequence of electrical pulses through the master electrode, identical to the pacing rhythm; Terminate the pacing rhythm, and thus the contraction, by applying a chaotic (mimicking the development of fibrillation) or gradually slowing sequence of electrical pulses. (2) Pacing capture and suppression: Suppress the spontaneous rhythm of the pacing site by applying overdrive pacing through the master electrode, and assess the ability of the spontaneous rhythm to recover by suddenly stopping the stimulation. (3) Arrhythmia modeling: Induce arrhythmic events such as premature contractions or fibrillation by applying additional stimulation through the auxiliary electrodes on top of the basal rhythm.

[0027] For the non-conductive regions: Print an irregular lattice or grid pattern, or leave the substrate uncoated with protein. This will guide the cells to grow anisotropically, forming irregular, inefficient connections that suppress abnormal electrical conduction. Through this biomimetic design, spontaneous pacing and directional fast conduction of electrical impulses are achieved, driving overall coordinated contraction-relaxation motion, mimicking the normal electrophysiological function of the heart.

[0028] Counterweight: To stabilize the motion posture of the simulated jellyfish in the culture solution, the simulated jellyfish unit of the embodiment further comprises a balance and positioning design. Specifically, a counterweight is integrated at the center bottom of the jellyfish-shaped frame. In a preferred embodiment, the counterweight is a high-density micro metal ball (such as tungsten or stainless steel) which is completely wrapped inside the elastic polymer (such as PDMS) substrate, forming an embedded structure. This design ensures the sterility and stability of the counterweight and avoids interference with the local flow field. The weight of the counterweight is optimally selected, with the principle being to make the gravity experienced by the simulated jellyfish in the culture solution slightly greater than the buoyancy it experiences. This design ensures that the structure can maintain a vertical posture with the top pointing upwards when at rest, and can produce clear and stable in-situ up-and-down motion in the vertical direction when rhythmically contracting, thus more accurately simulating the axial blood pumping function of the heart. The counterweight can be suspended in the center of the jellyfish-shaped frame by a biocompatible fine wire. In this way, the counterweight can be replaced according to changes in the culture solution to ensure that the simulated jellyfish moves up and down in situ.

[0029] 2. Environment simulation unit: For containing and controlling the living environment of the simulated jellyfish. This unit includes a culture tank, which contains a culture solution simulating tissue blood or plasma components. The tank body is a cylinder, ensuring that the simulated jellyfish always maintains a "top-up" orientation, simulating the posture of the jellyfish in water.

[0030] For the culture solution simulating tissue blood or plasma components, the following methods are used to obtain: Firstly, the blood samples of the plateau are processed to establish the animal model of the plateau (e.g. rats or mice are placed in a low-pressure oxygen cabin to simulate an altitude of 5000 meters for 4 weeks), so as to more accurately simulate the real microenvironment of the myocardial cells when the right heart fails (common in plateau heart disease), and the blood culture medium of the simulated plateau is compounded according to the blood samples taken from the central vein or the pulmonary artery. The samples are analyzed by using the Roche cobas c 111 blood biochemical analyzer to obtain key parameters (such as pH, lactic acid, catecholamine, electrolyte, inflammatory factor, etc.). Then, based on the standard cell culture medium (such as M199 culture medium), the culture medium is compounded according to the key parameters, including: reducing the oxygen partial pressure to the plateau level (the data are monitored by using the PreSens oxygen partial pressure sensor), adding specific concentrations of lactic acid, adrenaline, etc., and monitoring the culture medium at any time, so as to add or remove components and metabolites in time to maintain the stability of the culture medium composition, and measuring the central venous pressure of the animal model of the plateau, so as to simulate the internal environment under the plateau disease state. The Sartorius precision electronic balance can be used when the culture medium is prepared.

[0031] In some embodiments, the entire culture cavity can also be placed in a bioreactor with adjustable oxygen partial pressure to directly simulate the low-oxygen partial pressure environment of the circulatory system, which is complementary to the biochemical environment simulation. In specific embodiments, the bioreactor with adjustable oxygen partial pressure is not limited in its implementation, and can be implemented in any way in the prior art, such as the PreSens test system.

[0032] In some embodiments, the environment simulation unit further comprises a pressurization system for the culture tank, the internal hydrostatic pressure of which can be obtained by measuring the central venous pressure or the pulmonary arterial pressure of the animal model of the plateau, so as to simulate the increase of the preload and the afterload of the right ventricle (mainly the afterload) at the mechanical level.

[0033] 3. Motion monitoring and data analysis unit: The unit is used for real-time capturing and quantifying the motion function of the simulated jellyfish. The unit comprises a high-speed camera device and an image processing analysis module connected thereto. The high-speed camera device is directed to the culture cavity and is used for recording the upstream and downstream motion process of the simulated jellyfish in the culture medium; the image processing analysis module extracts one or more parameters of the contraction frequency, the contraction amplitude, the swimming speed and the bending angle of the lappet structure as the indexes for evaluating the contraction function by analyzing the video data. The high-speed camera device and the image processing analysis module are not limited in their implementation, and can be implemented in any way in the prior art.

[0034] In some embodiments, in addition to the video analysis described above, a microelectrode array can be integrated to monitor the field potential signal to reflect the electrophysiological activity, or a laser Doppler vibrometer can be used to non-contact measure the vibration speed of the hydrogel layer of the simulated jellyfish to more accurately reflect the contraction force.

[0035] 4. Histopathology and metabolism analysis unit: For in-depth cytological and metabolic analysis of myocardial tissue in simulated jellyfish during and at the end of the experiment. This unit complements the motion monitoring and data analysis unit, together forming an evaluation system of "function-structure-metabolism". It includes histomorphological observation module and cell metabolism detection module: Histomorphological observation module: Through histochemical staining and microscopic imaging equipment, the jellyfish tissue is regularly sampled and observed to analyze key pathological morphological changes such as myocardial cell arrangement, intercellular connection, cytoplasmic vacuolar degeneration, karyopyknosis, and fibrosis. In some embodiments, specific antibodies can be used to label cardiac troponin I, desmin, etc. through immunofluorescence staining to more accurately display the unique structure of myocardial cells.

[0036] Cell metabolism detection module: Through biochemical kits or liquid chromatography-mass spectrometry analysis platform, the concentration changes of metabolites such as lactic acid, pyruvic acid, ATP / ADP / AMP, and active oxygen in the culture medium are detected, or the glycolysis and oxidative phosphorylation metabolic flux analysis of the myocardial cell layer tissue sample is directly performed to evaluate the energy metabolism status of myocardial cells under simulated high-altitude blood environment. In some embodiments, the oxygen consumption rate and acid production rate of the living tissue cells in the myocardial cell layer can be measured in real time and dynamically using the hippocampus bioenergy analyzer, directly reflecting the mitochondrial function and glycolytic capacity of the cells.

[0037] The above-mentioned simulated high-altitude environment heart contraction function simulation and detection system based on simulated jellyfish has the following working principle: Myocardial stem cells of experimental monkeys differentiate and self-organize on the 3D printed jellyfish-like framework to form a myocardial tissue layer with synchronous rhythmic contraction ability. When electric stimulation is applied or relying on its spontaneous beating, the myocardial contraction drives the elastic framework to bend, completing a "power stroke" that pushes the culture solution downward, generating a pushing force that moves the entire structure upward; then, the elastic restoring force of the framework makes it relax, completing a "recovery stroke", and the structure sinks, as shown in Figure 3 The process of "up and down movement in place" is a direct biomimetic simulation of the "contraction-diastole" pumping cycle of the heart.

[0038] By analyzing the blood from different animal models of high altitude diseases (such as high altitude heart disease model mice, monkeys, etc.) or high altitude heart disease patients, and reconstituting the key components (such as pH, oxygen partial pressure, specific metabolites, hormones, ion concentration, etc.) in the culture medium, and then placing the simulated jellyfish in this specific culture medium, the changes in its movement state are observed through the movement monitoring and data analysis unit. If the culture medium environment inhibits or damages the "heart function", it will directly show as a decrease in the contraction frequency, amplitude and swimming speed of the jellyfish, which can be quantified. When the contraction function of the jellyfish is irreversibly damaged (showing a decrease in contraction function that cannot be recovered in normal culture medium), the jellyfish biological component tissue cells are identified for pathological changes, and the histological changes are analyzed to deduce the pathological changes of the heart in this environment.

[0039] The core pathophysiological changes of high altitude hypoxic environment are energy metabolism disorder and oxidative stress. The invention simulates this environment by reconstituting the culture medium and acting on the simulated jellyfish. In terms of function, it shows a decrease in contraction force, while at the cellular level it will trigger a series of adaptive or damaging changes, including: Metabolic reprogramming: cells shift from efficient aerobic oxidation (oxidative phosphorylation) to inefficient anaerobic glycolysis (glycolysis), leading to increased lactic acid production and insufficient ATP production.

[0040] Oxidative stress damage: accumulation of reactive oxygen species, attacking cell membranes, proteins and DNA, leading to cell structure damage.

[0041] Morphological outcome: long-term metabolic disorder and oxidative stress will lead to edema and vacuolar degeneration of cardiomyocytes, and then progress to apoptosis or necrosis, and eventually may trigger replacement fibrosis.

[0042] The following is a specific test process to briefly describe the working process of the high altitude environment heart contraction function simulation and detection system based on the simulated jellyfish: Mature simulated jellyfish units are transferred to a culture tank containing simulated culture medium, ensuring that they are suspended "top up". The culture medium includes "normal blood culture medium" and "high altitude disease blood culture medium", and the simulated jellyfish units in the two are compared, for example, in the high altitude disease blood culture medium, if a significant decrease in contraction frequency and a decrease in contraction amplitude are observed, it indicates that the blood environment has inhibited the contraction function of the heart; the pathological development and outcome of the heart in the high altitude environment can be inferred from the histological and metabolomic cellomic test results, as follows: Record spontaneous or electrically-induced (e.g. 1 Hz, 2.5 V / cm) contraction of jellyfish in a culture tank using a high-speed camera system (e.g. Photron FASTCAM Mini AX series) at a rate of 100-200 frames / s for 5-10 minutes; electric stimulation can be applied using an IonOptix MyoPacer stimulator. Use image processing analysis module (image processing analysis software can use open source tools such as ImageJ combined with MATLAB or Python scripts, or Simi Motion) to perform the following operations: 1) Track the vertical displacement of the central disc or the apex of the lobe structure of the jellyfish, and generate a displacement-time curve; 2) Calculate the contraction frequency (times / minute); 3) Calculate the peak displacement of a single contraction (contraction amplitude); 4) Calculate the average swimming speed.

[0043] Perform histopathological and metabolic analysis: a. Set up multiple experimental groups to simulate different altitudes (e.g. 3000 meters, 5000 meters) or different disease stages (e.g. acute high altitude reaction, chronic high altitude heart disease) of blood environment.b. In each experimental group, set multiple observation time points (e.g. 1st day, 3rd day, 7th day, 14th day after exposure).c. Regular sampling: At each preset time point, randomly take out part of the simulated jellyfish units from each group.

[0044] Perform tissue morphological analysis:a. Fix and section: Fix the removed jellyfish samples with 4% paraformaldehyde, then paraffin-embedded and sectioned using Leica EG series embedding machine and RM series sectioning machine.b. Staining and observation: Hematoxylin-eosin (H&E) staining is performed using Leica Aperio AT2 digital pathology scanning system to observe the general morphological structure of cells, nuclear shape, and whether there is inflammatory cell infiltration. Masson's Trichrome staining is performed to specifically show collagen fibers (blue) for quantifying the degree of tissue fibrosis. TUNEL staining is performed to label apoptotic cells in situ.c. Image analysis: Use high-resolution pathological section scanner (e.g. Leica Aperio) to obtain digital images, and use image analysis software (e.g. Image-Pro Plus) to quantitatively analyze the percentage of fibrosis area, number of apoptotic cells, etc.

[0045] Cell metabolism analysis: a. Culture supernatant metabolite detection: collect culture supernatant at different time points, use commercial lactic acid detection kit and ATP detection kit, and quantitatively detect by BioTek Synergy microplate reader or Tecan Infinite microplate reader. b. Oxidative stress level detection: use reactive oxygen species (ROS) detection fluorescent probe (such as DCFH-DA), stain and quantify live cells by flow cytometry or fluorescence microscopy, and evaluate the oxidative stress level of cells. c. Metabolomics analysis: send tissue samples or culture supernatant to a professional platform for non-targeted or targeted metabolomics analysis to systematically reveal the changes in global metabolic network under high altitude environment disturbance.

[0046] Integrate and analyze the functional data (contraction amplitude, frequency), morphological data (fibrosis area, apoptosis index), and metabolic data (lactic acid / ATP / ROS level) obtained at different time points. For example, it can be observed that in the early exposure stage, lactic acid may first increase and ATP may decrease (metabolic compensation), followed by a decrease in contraction force (functional decompensation), and finally, significant cell apoptosis and tissue fibrosis (structural damage) may occur in the late stage. This dynamic, multi-dimensional observation can accurately locate the key nodes and potential intervention targets of disease development, providing unprecedented insights for drug development.

[0047] By regularly and dynamically observing the above indicators, the present application can directly reveal the structural and metabolic basis of impaired myocardial function in high altitude environment, and completely present the dynamic outcome process from early compensation, decompensation to irreversible damage.

[0048] The present application uses simulated jellyfish as a biosensor and pathophysiological model to simulate and detect changes in heart function under high altitude blood. The simulation is the contraction-diastolic pumping function of the heart, and the movement of the jellyfish is used as an observable substitute indicator of this function.

[0049] The simulated jellyfish-based high altitude environment heart contraction function simulation and detection system integrates specific body fluid environment simulation and functional quantitative analysis, forming a complete detection system, providing a highly controllable and reproducible pathological environment model. By accurately compounding the culture medium to simulate high altitude blood components, the biochemical environment at different altitudes and different disease stages can be stably simulated. Real-time, direct, and quantitative evaluation of heart contraction function under high altitude environment is achieved, significantly reducing research costs and ethical restrictions. The movement state of the simulated jellyfish directly reflects the heart contraction function, replacing traditional live animal experiments, shortening the test period and reducing the test cost. A complete evaluation chain from macro function to micro mechanism is established, and through the movement monitoring and data analysis unit and the histopathology and metabolism analysis unit, multi-dimensional synchronous correlation observation of the same sample from contraction function, cell morphology to energy metabolism is achieved.

[0050] At the same time, by standardizing the preparation process and automating the analysis software, comparative studies of multiple experimental groups can be carried out simultaneously, greatly improving the research efficiency, solving the technical bottleneck of low throughput and long time consumption of traditional pathological research, and realizing the high-throughput, multi-parameter parallel detection capability. A reliable preclinical evaluation platform is provided for the drug research and development of high altitude heart disease.

[0051] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application, and they should be covered in the scope of the claims and the description of the present application.

Claims

1. A cardiac contractile function model based on simulated jellyfish in a plateau environment, characterized in that, include: A jellyfish-like framework, wherein the jellyfish-like framework is the umbrella-shaped structure of a natural jellyfish; A layer of myocardial tissue, cultured on the medullary framework, is used to simulate cardiac contractile function through spontaneous or rhythmic contractions; The pacemaker and conduction system, composed of patterned layers of myocardial cells, are used to generate and / or conduct electrical signals to stimulate rhythmic contraction / relaxation of the myocardial tissue layer. The counterweight, located at the bottom center of the jellyfish-shaped frame, is used to ensure that the weight of the simulated jellyfish in the culture medium is greater than the buoyancy.

2. The plateau environment cardiac contractile function model based on simulated jellyfish as described in claim 1, characterized in that, The jellyfish-like framework is prepared by 3D printing using a biocompatible photocurable resin or elastomer. Cardiac tissue is derived from cardiac stem cells of primates or from cardiac cells derived from human induced pluripotent stem cells.

3. The plateau environment cardiac contractile function model based on simulated jellyfish as described in claim 1, characterized in that, The patterned cardiomyocyte layer is prepared using microcontact printing technology and includes a central pacing functional area and peripheral conduction pathways, with multiple conduction pathways extending radially from the base of each valve-like structure in the central pacing functional area.

4. The plateau environment cardiac contractile function model based on simulated jellyfish as described in claim 3, characterized in that, The generated and / or conducted electrical signals are generated by a programmable multichannel electrical stimulation subsystem, which includes: A multichannel electrical stimulator for generating at least two independent and controllable electrical pulse sequences; The master control electrode, located in the central pacing functional area, is used to regulate the dominant rhythm; Multiple auxiliary electrodes are placed along the conduction path to introduce ectopic rhythms or test conduction performance.

5. A system for simulating and detecting cardiac contractile function in a high-altitude environment based on simulated jellyfish, characterized in that, The plateau environment cardiac contraction function model based on simulated jellyfish as described in any one of claims 1-4 is used as a biomimetic simulator for cardiac contraction to analyze the motion state, histopathology, and metabolic status of the biomimetic simulator in the culture medium of body blood components in a simulated plateau environment, and to evaluate the effect of the culture medium on cardiac function.

6. The system for simulating and detecting cardiac contractile function in a high-altitude environment based on simulated jellyfish, as described in claim 5, is characterized in that... Also includes: An environmental simulation unit is used to contain and control the living environment of simulated jellyfish; The motion monitoring and data analysis unit is used to capture and quantify the motion state of the simulated jellyfish in real time, and to evaluate the contraction function based on the motion state analysis. The histopathology and metabolic analysis unit is used for cellular and metabolic analysis of myocardial tissue in simulated jellyfish.

7. The system for simulating and detecting cardiac contractile function in a high-altitude environment based on simulated jellyfish as described in claim 6, characterized in that, The environmental simulation unit includes a cylindrical culture tank containing a culture medium simulating tissue, blood, or plasma components in a high-altitude environment.

8. The system for simulating and detecting cardiac contractile function in a high-altitude environment based on simulated jellyfish, as described in claim 7, is characterized in that... The culture tank is placed in a bioreactor with adjustable oxygen partial pressure.

9. The system for simulating and detecting cardiac contractile function in a high-altitude environment based on simulated jellyfish as described in claim 6, characterized in that, The motion monitoring and data analysis unit includes: The camera equipment is used to record video data of the simulated jellyfish's movement in the culture medium; the movement includes the frequency, speed, and amplitude of its swimming up and down over a certain period of time. The image processing and analysis module, electrically connected to the camera device, is used to extract one or more parameters from the video data, including contraction frequency, contraction amplitude, swimming speed, and the bending angle of the petal structure, to analyze and evaluate the contraction function.

10. The system for simulating and detecting cardiac contractile function in a high-altitude environment based on simulated jellyfish according to claim 6, characterized in that, The histopathology and metabolic analysis unit includes: The tissue morphology observation module is used to sample and observe jellyfish tissue using histochemical staining and microscopic imaging equipment, and to analyze key pathological morphological changes such as cardiomyocyte arrangement, intercellular connections, cytoplasmic vacuolar degeneration, nuclear condensation, and fibrosis; or to use immunofluorescence staining to label cardiac troponin I and desmin with specific antibodies. The cell metabolism detection module is used to detect changes in the concentrations of lactate, pyruvate, ATP / ADP / AMP, and reactive oxygen metabolites in the culture medium using biochemical reagent kits or liquid chromatography-mass spectrometry analysis platforms; or to perform metabolic flux analysis of glycolysis and oxidative phosphorylation in tissue samples of cardiomyocyte layers; or to perform real-time, dynamic determination of oxygen consumption and acid production rates in living tissues of cardiomyocyte layers using a hippocampal bioenergy analyzer.

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