A device and experimental method for passive smoking poisoning of rodent experimental animals

Abstract

The invention discloses a device and experimental method for passive smoking poisoning of rodent experimental animals. The device comprises a lower box, an upper box and a flame retardant plate. The lower box and the upper box are both bottomless boxes made of transparent materials. At least one side of the lower box is provided with a ventilation hole. The top of the lower box is provided with a smoke diffusion part, and the smoke diffusion part is provided with a plurality of diffusion holes for ventilation. The flame retardant plate is used to place the smoke. The lower box covers the flame retardant plate, and the upper box is placed on the lower box and covers the smoke diffusion part. The smoke of the smoke placed on the flame retardant plate diffuses into the upper box through the smoke diffusion part on the top of the lower box. The method uses the device to conduct experiments. The beneficial effects are: it is simple and universal, low in cost, easy to clean, can be flexibly moved, and can be poisoned without an external power supply by relying on the structure of the device. The smoke diffuses evenly, and the animals can move freely and unconstrained when they are passively smoked. At the same time, the animal status can be observed in real time.

Description

Technical Field

[0001] The present invention relates to the field of animal experiment instrument technology and method, and in particular to a device and an experimental method for passive smoking poisoning of rodent experimental animals. Background Art

[0002] Chronic obstructive pulmonary disease (COPD) is a common disease that endangers human health worldwide. It significantly increases the economic burden of medical and health care. It is also the third leading cause of death after cardiovascular disease. Reducing the incidence and mortality of COPD is of great significance to human health. However, due to the complex pathogenesis of COPD, how to effectively intervene and prevent this disease has become a difficult problem facing clinical workers and researchers. COPD disease animal models provide an important experimental carrier for the study of the occurrence and outcome mechanisms of COPD, and greatly facilitate the study of COPD disease intervention and prevention measures.

[0003] At present, the COPD disease animal model is often established by passively inhaling mainstream cigarette smoke in rodents (rats / mice). However, the existing cigarette exposure devices have the disadvantages of being large in size, complex in structure, expensive, difficult to clean, not easy to move at will, smoke needs to be induced by external power supply to diffuse, smoke diffusion is uneven, and animals need to be fixed for a long time, which greatly limits the universality of these devices. Summary of the invention

[0004] The technical problem to be solved by the present invention is that the existing cigarette exposure and poisoning devices for animal experiments have the disadvantages of large size, complex structure, high price, difficulty in cleaning, difficulty in moving the devices at will, smoke needs to be induced by external power supply to diffuse, smoke diffusion is uneven, and animals need to be fixed for a long time, which greatly limits the universality of these devices. The purpose is to provide a device and experimental method for passive smoking poisoning of rodent experimental animals, which are simple and universal, low cost, easy to clean, flexible to move, can poison animals without external power supply by relying on the structure of the device, smoke diffuses evenly, animals can move freely and unrestrained when passively smoking and poisoning, avoid physiological stress caused by restraining animals for too long and affecting the quality of experimental data, and at the same time can observe the animal status in real time.

[0005] The present invention is achieved through the following technical solutions:

[0006] A device for passive smoking exposure of rodent experimental animals, used for subjecting rodent experimental animals to passive smoking exposure, comprises a lower box, an upper box and a flame retardant plate, the lower box and the upper box are both bottomless boxes made of transparent material, at least one side of the lower box is provided with a ventilation hole, the top of the lower box is provided with a smoke diffusion part, the smoke diffusion part is provided with a plurality of diffusion holes for ventilation; at least one corner of the top surface of the upper box is provided with an upper box ventilation hole; the flame retardant plate is used for placing burning smoke; the lower box covers the flame retardant plate, the upper box is placed on the lower box and covers the smoke diffusion part; the smoke from the burning smoke placed on the flame retardant plate diffuses into the upper box through the smoke diffusion part on the top of the lower box.

[0007] Through the above technical design, when using, choose a suitable place, lay the flame retardant board, then cover the flame retardant board with the lower box, place the upper box on the lower box, and cover the smoke diffusion part of the lower box, and place the experimental animal in the upper box. At this time, the smoke generated by the cigarette on the flame retardant board rises naturally, passes through the smoke diffusion part of the lower box, and diffuses from the several diffusion holes of the smoke diffusion part to the upper box, making full use of the structural characteristics of the smoke diffusion part, turning a wisp of smoke into several wisps of smoke, and the smoke can be evenly diffused into the upper box, so that the experimental animals in the upper box are passively smoked. At the same time, the experimental animals placed in the upper box are not restrained, so that they can be infected in a free and unconstrained state. At the same time, the test state of the animal can be observed through the transparent upper box, and the burning of the internal smoke can be observed through the transparent lower box. If the smoke is extinguished, the lower box can be moved, and the cigarette can be re-lit or the smoke can be increased. After the experiment is completed, the upper box is moved away, the experimental animal is taken out, and various indicators are tested. Then the lower box, upper box and flame retardant board can be separated and cleaned. Therefore, through the above technical design, it is simple and universal, low-cost, easy to clean, flexible to move, and can be used to poison animals without external power supply, smoke diffusion is uniform, and animals can move freely and unrestrained when poisoned, avoiding physiological stress caused by restraining animals for too long and affecting the quality of experimental data, and the animal status can be observed in real time.

[0008] Furthermore, the smoke diffusion part is composed of a plurality of smoke diffusion layers; any one of the plurality of smoke diffusion layers is made of a plate having a plurality of diffusion holes, the plurality of smoke diffusion layers are arranged at intervals on the lower box body, and a cavity formed between any two adjacent smoke diffusion layers is used for smoke diffusion.

[0009] By designing a plurality of smoke diffusion layers in the smoke diffusion part, the rapid and uniform diffusion of smoke can be further improved. The diffusion principle is as follows: the burning smoke starts as a wisp, and as it rises, the smoke reaches the smoke diffusion layer of the lowest layer and is blocked by it, so that the smoke spreads along the bottom surface of the smoke diffusion layer of the lowest layer, and then passes through a plurality of diffusion holes of the smoke diffusion layer of the lowest layer, and the smoke passes through some of the diffusion holes and diffuses into a plurality of wisps of smoke; the plurality of wisps of smoke enter the cavity between the lowest layer of smoke diffusion layer and the upper layer of smoke diffusion layer for diffusion, and the diffused smoke reaches the upper layer of smoke diffusion layer and is blocked by it, so that the smoke spreads along the bottom surface of the smoke diffusion layer, and then passes through a plurality of diffusion holes of the smoke diffusion layer, so that the number of diffused wisps of smoke is further increased, and thus the smoke continues to rise to the upper layer of smoke diffusion layer, and the number of wisps increases with each rise of a smoke diffusion layer, until it passes through the upper layer of smoke diffusion layer, so that the smoke diffuses to a considerable number of wisps of smoke, and enters the upper box body, so that the experimental animals are quickly and evenly infected with poison.

[0010] Furthermore, any diffusion hole of any smoke diffusion layer among the plurality of smoke diffusion layers is staggered with any diffusion hole of an adjacent smoke diffusion layer in the vertical direction.

[0011] By staggering the diffusion holes of any two adjacent smoke diffusion layers in the vertical direction, the upper smoke diffusion layer further increases the number of smoke strands diffused by the lower smoke diffusion layer. Because the physical property of smoke is that it tends to rise vertically and linearly, the main part of a strand of smoke coming out of a diffusion hole in the lower smoke diffusion layer tends to rise vertically and linearly to the upper smoke diffusion layer. Since the diffusion holes of this layer are staggered, the strand of smoke cannot directly pass through the smoke diffusion layer of this layer. Then, the strand of smoke diffuses around the blocking point area on the bottom surface of the smoke diffusion layer, diffuses to multiple diffusion holes around the point area, and then passes through multiple diffusion holes, thus becoming multiple strands of smoke. Thus, the number of diffused smoke strands is further increased.

[0012] Furthermore, the number of diffusion holes provided in any one of the plurality of smoke diffusion layers is 18 times 20, and they are distributed in an array of 18 times 20; any one diffusion hole is a circular hole with a diameter of 2 cm, and the center distance between any one diffusion hole and an adjacent diffusion hole is 3 cm.

[0013] By designing the number of diffusion holes in each smoke diffusion layer to be distributed in an array of 18 times 20, the diffusion holes are circular holes with a diameter of 2 cm, and the center distance between any diffusion hole and the adjacent diffusion hole is 3 cm, it can meet most experimental needs, achieve rapid and uniform diffusion of multiple smoke wisps in a short time, and quickly infect experimental animals.

[0014] Preferably, any one of the plurality of smoke diffusion layers is fixedly mounted on the lower box body.

[0015] Each smoke diffusion layer is fixedly installed on the lower box body to make it a whole, which is more stable and does not need to be adjusted frequently, making it more efficient and quick to use.

[0016] Preferably, the topmost smoke diffusion layer among the plurality of smoke diffusion layers is fixedly mounted on the lower box body, and any one of the following smoke diffusion layers is detachably mounted on the lower box body.

[0017] Any of the following smoke diffusion layers can be detachably mounted on the lower box body, and can be disassembled to clean each smoke diffusion layer, making cleaning more convenient.

[0018] Furthermore, the number of the plurality of smoke diffusion layers is set to 3 layers, which are respectively a first smoke diffusion layer, a second smoke diffusion layer and a third smoke diffusion layer from bottom to top.

[0019] The smoke diffusion layer is designed to be three layers, and a wisp of smoke is quickly and evenly diffused into several wisps of smoke. The number of diffused wisps of smoke basically meets the use requirements.

[0020] Furthermore, the device for passive smoking poisoning of rodent experimental animals also includes a steel grid frame, which is placed inside the lower box body, above the flame retardant plate, and the steel grid frame is used to place the smoke; one of the sides of the lower box body is also provided with a lower box body door for placing the smoke; one of the sides of the upper box body is provided with an upper box body door for placing experimental animals.

[0021] A steel grid frame is added to place the smoke, so that the smoke burns more completely; at the same time, a lower box door is set on the lower box, through which it is more convenient to add smoke into the lower box; an upper box door is set on the upper box, through which it is more convenient to place experimental animals.

[0022] Furthermore, the four corners of the top surface of the lower box body are also provided with stoppers; the upper box body is placed between the four stoppers and is limited by the four stoppers.

[0023] The upper box is limited by setting a stopper to prevent the experimental animals inside from sliding on it.

[0024] An experimental method for passive smoking exposure of rodent experimental animals, using the above-mentioned device for passive smoking exposure of rodent experimental animals, the steps of which are as follows:

[0025] (I) Assembling the device: Select a suitable location (ground or surface), place the flame retardant board first, place the steel grid frame on the flame retardant board, and cover the flame retardant board and the steel grid frame with the lower box; place the upper box on the lower box and cover the smoke diffusion part of the lower box;

[0026] (ii) Placing animals: Select the experimental animals, open the upper box door, then place the experimental animals into the upper box through the upper box door, and then close the upper box door;

[0027] (III) Placing burning cigarettes: Light eight cigarettes, open the lower box door, and immediately place the lit cigarettes from the lower box door onto the steel grid rack inside the lower box, and close the lower box door; if the cigarettes are found to be extinguished, open the lower box door, relight the cigarettes or add new lit cigarettes;

[0028] (IV) Conducting the experiment: When eight lit cigarettes are placed in the lower box, the timing starts. When the set exposure time is reached, the exposure time is initially set to 1 hour. When 1 hour is reached, the upper box can be opened, the experimental animals can be taken out, and the experimental animals can be placed back to their original cages;

[0029] (V) Continuous experiment: Repeat steps (II) to (IV) twice a day, with an interval of more than 4 hours between the two times. After 60 days of continuous experiment, the COPD disease animal model is obtained. 24 hours after the completion of the 60th day experiment, the relevant indicators of all experimental animals are tested.

[0030] (VI) Cleaning the experimental device: After completing the passive smoking experiment on experimental animals every day, the lower box, upper box, steel grid frame, and flame retardant board can be separated and cleaned in preparation for the next experiment.

[0031] Through the above method, a simple, universal, low-cost, and easy-to-clean device is adopted. Relying on the structural characteristics of the device, animals can be poisoned without an external power supply, the smoke diffuses evenly, and the animals can move freely without restraint during the poisoning, avoiding the physiological stress caused by restraining the animals for too long and affecting the quality of experimental data. At the same time, the position of the device can be flexibly moved to conduct experiments. This experimental method is simple, fast, and efficient, and can accurately obtain experimental data of passive smoking exposure of experimental animals in a free-moving state, avoiding the physiological stress response caused by restraint and causing data distortion.

[0032] Compared with the prior art, the present invention has the following advantages and beneficial effects: a device and experimental method for passive smoking exposure of rodent experimental animals, which are simple and universal, low-cost, easy to clean, and can be flexibly moved. Relying on the structure of the device, animals can be exposed to poison without an external power supply, smoke diffuses evenly, and animals can move freely and unrestrained when passively smoking and consuming poison, avoiding physiological stress caused by binding animals for too long a time and affecting the quality of experimental data, and the animal status can be observed in real time. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The drawings described herein are used to provide a further understanding of the embodiments of the present invention, constitute a part of this application, and do not constitute a limitation of the embodiments of the present invention. In the drawings:

[0034] Figure 1 It is a perspective view of the structure of the present invention.

[0035] Figure 2 Exploded view of the assembly of the present invention.

[0036] Figure 3 It is a schematic structural diagram of the part 1 of the present invention.

[0037] Figure 4 It is a front perspective view of the part 1 of the present invention.

[0038] Figure 5 It is a top view of the part 1 of the present invention.

[0039] Figure 6 for Figure 5 AA section view.

[0040] Figure 7 for Figure 5 BB cross-sectional view.

[0041] Figure 8 This is a typical light microscopic image of the pathological morphology of the lung tissue of poison-exposed experimental mice obtained using the existing experimental equipment.

[0042] Fig. 9 This is a typical transmission electron micrograph of the ultrastructure of the lung tissue of poison-exposed experimental mice obtained using the existing experimental equipment.

[0043] Fig.10 The optical microscope image of the pathological morphology of lung tissue of typical poisoned experimental mice obtained by using the device for passive smoking poisoning of rodent experimental animals of the present invention.

[0044] Fig.11 The present invention is a transmission electron microscope image of the ultrastructure of lung tissue of a typical poisoned experimental mouse obtained by using the device for passive smoking poisoning of rodent experimental animals of the present invention.

[0045] Markings and corresponding parts names in the accompanying drawings: 1-lower box, 11-smoke diffusion part, 111-first smoke diffusion layer, 112-second smoke diffusion layer, 113-third smoke diffusion layer, 1110-diffusion hole, 12-lower box door, 13-ventilation hole, 14-block, 2-upper box, 21-upper box door, 22-upper box ventilation hole, 3-steel grid frame, 4-flame retardant board, 5-ground or object surface. DETAILED DESCRIPTION

[0046] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments and drawings. The exemplary embodiments of the present invention and their description are only used to explain the present invention and are not intended to limit the present invention.

[0047] In the following description, a large number of specific details are set forth in order to provide a thorough understanding of the present invention. However, it is apparent to one of ordinary skill in the art that these specific details are not necessarily employed to practice the present invention. In other instances, well-known structures, circuits, materials, or methods are not specifically described in order to avoid obscuring the present invention.

[0048] Throughout the specification, references to "one embodiment," "an embodiment," "an example," or "an example" mean that a particular feature, structure, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment of the present invention. Therefore, the phrases "one embodiment," "an embodiment," "an example," or "an example" appearing in various places throughout the specification do not necessarily all refer to the same embodiment or example. In addition, particular features, structures, or characteristics may be combined in one or more embodiments or examples in any suitable combination and / or sub-combination. In addition, it will be appreciated by those of ordinary skill in the art that the figures provided herein are for illustrative purposes and that the figures are not necessarily drawn to scale. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0049] In the description of the present invention, the directions or positional relationships indicated by terms such as “front”, “rear”, “left”, “right”, “up”, “down”, “vertical”, “horizontal”, “high”, “low”, “inside” and “outside” are based on the directions or positional relationships shown in the accompanying drawings and are only for the convenience of describing the present invention and simplifying the description. They do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction. Therefore, they should not be understood as limiting the scope of protection of the present invention.

[0050] Example

[0051] like Figures 1 to 7As shown, the present invention is a device for passive smoking poisoning of rodent experimental animals, which is used to make rodent experimental animals passively smoke poisoned, including a lower box body 1, an upper box body 2 and a flame retardant plate 4, the lower box body 1 and the upper box body 2 are both bottomless boxes made of transparent material, at least one side of the lower box body 1 is provided with a ventilation hole 13, the top of the lower box body 1 is provided with a smoke diffusion part 11, and the smoke diffusion part 11 is provided with a plurality of diffusion holes 1110 for ventilation; at least one corner of the top surface of the upper box body 2 is provided with an upper box ventilation hole 22; the flame retardant plate 4 is used to place burning smoke; the lower box body 1 covers the flame retardant plate 4, the upper box body 2 is placed on the lower box body 1 and covers the smoke diffusion part 11; the smoke of the burning smoke placed on the flame retardant plate 4 is diffused into the upper box body 2 through the smoke diffusion part 11 on the top of the lower box body 1.

[0052] When in use, place the flame retardant plate 4 on a suitable bottom surface or object surface 5, then place the smoke, place the upper box 2 on the lower box 1 and cover the smoke diffusion part of the lower box 1, then move the lower box 1 and the upper box 2 as a whole, move them to the flame retardant plate 4, and make the lower box 1 cover the flame retardant plate 4; then move the upper box 2 and put the experimental animal into the upper box 2, put down the upper box 2, and then the experiment can be carried out.

[0053] In some embodiments, the smoke diffusion part 11 is composed of a plurality of smoke diffusion layers; any one of the plurality of smoke diffusion layers is made of a plate, and a plurality of diffusion holes 1110 are provided on the plate. The plurality of smoke diffusion layers are arranged at intervals on the lower box 1, and the cavity formed between any two adjacent smoke diffusion layers is used for smoke diffusion. As a preferred embodiment, the smoke diffusion part 11 is designed as a plurality of smoke diffusion layers, and each smoke diffusion layer is provided with a plurality of diffusion holes, so that the number of diffusion strands of smoke is further increased. After each smoke diffusion layer, the number of smoke strands gradually increases, so that the smoke that finally enters the upper box 2 has a sufficient number of strands, so that it can be quickly and evenly diffused in the upper box 2, so that the animals can be quickly infected with the poison in a free moving state.

[0054] In some embodiments, any diffusion hole 1110 of any one of the plurality of smoke diffusion layers is staggered with any diffusion hole 1110 of the adjacent smoke diffusion layer in the vertical direction. As a preferred embodiment, the plurality of diffusion holes of two adjacent smoke diffusion layers are staggered to further increase the number of smoke diffusion strands and improve its diffusivity; if they are not staggered, a strand of smoke coming out of a diffusion hole in the lower smoke diffusion layer will have most of the strand of smoke rising straight up in a region and directly passing through the diffusion holes of the upper smoke diffusion layer, and only a small part of the strand of smoke will be diffused; if they are staggered, most of the strand of smoke will be diffused into multiple strands of smoke, so that the number of smoke diffusion strands will be further increased.

[0055] In some embodiments, the number of diffusion holes 1110 provided in any one of the plurality of smoke diffusion layers is 18 times 20, and they are distributed in an array of 18 times 20; any diffusion hole 1110 is a circular hole with a diameter of 2 cm, and the center distance between any diffusion hole 1110 and the adjacent diffusion hole 1110 is 3 cm. As a preferred embodiment, by adopting this implementation technology, a wisp of smoke can be diffused into the required number of wisps of smoke, so that it can be quickly and evenly diffused into the upper box 2.

[0056] In some embodiments, any one of the plurality of smoke diffusion layers is fixedly mounted on the lower box body 1. As a preferred embodiment, a fixed installation method is adopted to form a whole, which is more stable and more convenient to operate and carry.

[0057] In some embodiments, the topmost smoke diffusion layer among the plurality of smoke diffusion layers is fixedly mounted on the lower box 1, and any of the following smoke diffusion layers is detachably mounted on the lower box 1. As a preferred embodiment, a detachable manner is adopted to facilitate its disassembly for cleaning.

[0058] In some embodiments, the number of the plurality of smoke diffusion layers is set to 3 layers, which are respectively the first smoke diffusion layer 111, the second smoke diffusion layer 112 and the third smoke diffusion layer 113 from top to bottom. As a preferred embodiment, the number of smoke diffusion layers is set to 3 layers, so that a wisp of smoke can be diffused into a sufficient number of wisps of smoke, and diffused quickly and evenly into the upper box body 2. At the same time, using 3 layers can meet the requirements, making the structure simpler.

[0059] In some embodiments, the device for passive smoking of rodent experimental animals further includes a steel grid frame 3, which is placed inside the lower box 1, on the flame retardant board 4, and is used to place the smoke; one of the sides of the lower box 1 is also provided with a lower box door 12 for placing the smoke; one of the sides of the upper box 2 is provided with an upper box door 21 for placing experimental animals. As a preferred embodiment, the steel grid frame 3 is added, and the smoke is placed on the steel grid frame 3, so that the smoke burns more fully; the lower box door 12 is provided on the lower box 1, so that the smoke can be added to the lower box 1 through the lower box door 12, and the upper box door 21 is provided on the upper box 2, so that the experimental animals can be placed.

[0060] In some embodiments, the four corners of the top surface of the lower box 1 are further provided with stoppers 14; the upper box 2 is placed between the four stoppers 14 and is limited by the four stoppers 14. As a preferred embodiment, the stoppers 14 are added to prevent the upper box 2 from slipping and maintain its stability.

[0061] An experimental method for passive smoking poisoning of rodent experimental animals, using the device for passive smoking poisoning of rodent experimental animals as claimed in the claims, and the steps are as follows:

[0062] (I) Assembling the device: Select a suitable location (ground or surface), first place the flame retardant board 4, place the steel grid frame 3 on the flame retardant board 4, and cover the flame retardant board 4 and the steel grid frame 3 with the lower box 1; place the upper box 2 on the lower box 1 and cover the smoke diffusion part 11 of the lower box 1;

[0063] (ii) Placing animals: Selecting experimental animals, then opening the upper box door 21, and then placing the experimental animals into the upper box 2 through the upper box door 21, and then closing the upper box door 21;

[0064] (III) Placing burning cigarettes: Light eight cigarettes, open the lower box door 12, and immediately place the lit cigarettes from the lower box door 12 onto the steel grid frame 3 in the lower box 1, and close the lower box door 12; if the cigarettes are found to be extinguished, open the lower box door 12, relight the cigarettes or add new lit cigarettes;

[0065] (IV) Conducting the experiment: When eight lit cigarettes are placed in the lower box 1, the timing starts. When the set exposure time is reached, the exposure time is initially set to 1 hour. When 1 hour is reached, the upper box 2 can be opened, the experimental animals can be taken out, and the experimental animals can be placed back to their original living cages;

[0066] (V) Continuous experiment: Repeat steps (II) to (IV) twice a day, with an interval of more than 4 hours between the two times. After 60 days of continuous experiment, the COPD disease animal model is obtained. 24 hours after the completion of the 60th day experiment, the relevant indicators of all experimental animals are tested.

[0067] (VI) Cleaning the experimental device: After completing the passive smoking experiment on experimental animals every day, the lower box 1, the upper box 2, the steel grid frame 3, and the flame retardant plate 4 can be separated and cleaned to prepare for the next experiment.

[0068] After the last passive smoking exposure, the relevant indicators of the experimental animals were tested;

[0069] Fresh right lower lobe tissue was obtained, fixed with 10% neutral formaldehyde for 48 hours, dehydrated with gradient alcohol, transparentized with xylene, embedded in paraffin, sliced, stained with HE, and the pathological morphological changes of lung tissue were observed under a light microscope.

[0070] The right upper lobe was quickly removed, and the blood and other dirt around the tissue were rinsed with PBS buffer. A 1mm3 tissue block of the right hilum was retained, fixed with 2.5% glutaraldehyde, post-fixed with 1% osmium acid, dehydrated in an ethanol-acetone series, embedded in Epin812, sliced ​​conventionally with an ultrathin slicer, and double stained with lead acetate and lead citrate. Changes in the ultrastructure of the lung tissue were observed under a transmission electron microscope.

[0071] The experimental device of the present invention and the currently used experimental device were used to conduct experiments. Two groups of mice were placed in the two experimental devices for passive smoking exposure, and the morphological structure diagrams of their lung tissues were obtained, as shown in FIG. Figure 8 , shown in Figure 9, are the typical pathological morphology light microscopic images and ultrastructural transmission electron microscopic images of experimental animal lung tissue obtained by the currently available experimental equipment. Fig.10 , 11 are respectively the typical light microscopic images of pathological morphology and transmission electron microscopic images of ultrastructure of experimental animal lung tissue obtained by the experimental device of the present invention. Through comparison, it can be seen that the experimental device of the present invention has a significantly better infection effect than the existing experimental device under the same time and the same experimental method. The COPD animal model established by the experimental device of the present invention is more typical and can better replicate the clinical COPD disease characteristics. That is, the pathological morphological changes of the poisoned experimental animals obtained by the experimental device of the present invention under the same time and method are more serious, specifically manifested as: partial exfoliation of ciliated columnar epithelial cells in the bronchial pulmonary tissue, degeneration and necrosis of ciliary adhesions, goblet cell hyperplasia, infiltration of a large number of inflammatory cells in the submucosal layer and muscular layer, obvious thickening of bronchial smooth muscle, a large number of neutrophils, alveolar macrophages and chaotic fluid secretions filled in the lumen, expansion and fusion of alveoli, obvious emphysema and bullae formation, obvious thickening of some alveolar septa, in which inflammatory cell infiltration and capillary dilation and congestion can be seen; expansion and abnormal expansion of the distal air sac cavity of the terminal bronchioles, narrowing of the alveolar septum, thinning or rupture of elastic fibers; thinning of some alveolar walls, reduction of alveolar surface area, decrease in density of alveolar wall tissue, enlargement of alveolar pores, destruction of alveolar attachment, rupture or mutual fusion of alveoli to form larger cystic cavities (bullas). Due to alveolar rupture and inflammatory erosion and destruction, the capillaries inside are blocked, and the number and cross-sectional area are significantly reduced or disappeared. In addition, emphysema can also be seen with non-arteriolar inflammation associated with bronchioles, hyperplasia and thickening of the intima, hypertrophy, hyperplasia, edema, degeneration, necrosis of the medial smooth muscle, stenosis or even occlusion of the lumen, and chronic inflammation associated with small bronchi and bronchioles. A large number of macrophages (dust cells) that have engulfed smoke particles can be seen next to small bronchi and pulmonary arterioles, and some macrophages disintegrate and destroy. The distal end of the terminal bronchioles is narrowed, the respiratory bronchioles and alveolar ducts are cystic, the alveolar cavity is irregularly enlarged, and the alveolar wall is thinned, swollen, ruptured or fused to form bullae. At the same time, the ultrastructural changes are more obvious, specifically manifested in: the nuclear chromatin of type I and type II pulmonary epithelial cells is condensed, and the degree of cytoplasm cavitation is more serious.

[0072] The specific implementation methods described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific implementation method of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A device for passive smoking exposure of rodent experimental animals, used to expose rodent experimental animals to passive smoking exposure, characterized in that: The invention comprises a lower box (1), an upper box (2) and a flame retardant plate (4), wherein the lower box (1) and the upper box (2) are both bottomless boxes made of transparent materials, at least one side of the lower box (1) is provided with a ventilation hole (13), the top of the lower box (1) is provided with a smoke diffusion part (11), and the smoke diffusion part (11) is provided with a plurality of diffusion holes (1110) for ventilation; at least one corner of the top surface of the upper box (2) is provided with an upper box ventilation hole (22); the flame retardant plate (4) is used to store smoke; the lower box (1) covers the flame retardant plate (4), and the upper box (2) is placed on the lower box (1) and covers the smoke diffusion part (11); the smoke from the smoke placed on the flame retardant plate (4) diffuses into the upper box (2) through the smoke diffusion part (11) at the top of the lower box (1); The smoke diffusion part (11) is composed of a plurality of smoke diffusion layers; any one of the plurality of smoke diffusion layers is made of a plate having a plurality of diffusion holes (1110) arranged on the plate; the plurality of smoke diffusion layers are arranged on the lower box body (1) at intervals; and a cavity formed between any two adjacent smoke diffusion layers is used for smoke diffusion; Any diffusion hole (1110) of any one of the plurality of smoke diffusion layers is staggered with any diffusion hole (1110) of an adjacent smoke diffusion layer in the vertical direction; The number of diffusion holes (1110) provided in any one of the plurality of smoke diffusion layers is 18 times 20, and they are distributed in an array of 18 times 20; any one of the diffusion holes (1110) is a circular hole with a diameter of 2 cm, and the center distance between any one of the diffusion holes (1110) and the adjacent diffusion holes (1110) is 3 cm; Also included is an experimental method for passive smoking exposure of rodent experimental animals, the steps of which are as follows:

1. Assembling the device: Select a suitable location, such as the ground or the surface of an object, first place the flame retardant plate (4), place the steel grid frame (3) on the flame retardant plate (4), and cover the flame retardant plate (4) and the steel grid frame (3) with the lower box (1); place the upper box (2) on the lower box (1) and cover the smoke diffusion part (11) of the lower box (1); 2. Place the animal: select the experimental animal, open the upper box door (21), then place the experimental animal into the upper box (2) through the upper box door (21), and then close the upper box door (21); 3. Place the burning cigarettes: Light eight cigarettes, open the lower box door (12), and immediately place the lit cigarettes from the lower box door (12) onto the steel grid frame (3) in the lower box (1), and close the lower box door (12); if the cigarettes are found to be extinguished, open the lower box door (12) and relight the cigarettes or add new lit cigarettes; 4. Conducting the experiment: When eight lit cigarettes are placed in the lower box (1), the timing starts. When the set exposure time is reached, the exposure time is initially set to 1 hour. When 1 hour is reached, the upper box (2) can be opened, the experimental animals can be taken out, and the experimental animals can be placed back to their original cages; 5. Continuous experiment: Repeat steps 2 to 4 twice a day, with an interval of more than 4 hours between the two times. After 60 days of continuous experiment, obtain the COPD disease animal model. 24 hours after the completion of the 60th day experiment, test the relevant indicators of all experimental animals; 6. Cleaning the experimental device: After completing the animal passive smoking experiment every day, the lower box (1), the upper box (2), the steel grid frame (3), and the flame retardant plate (4) can be separated and cleaned to prepare for the next experiment; After the last passive smoking exposure, the relevant indicators of the experimental animals were tested; Fresh right lower lobe tissue was obtained, fixed with 10% neutral formaldehyde for 48 hours, dehydrated with gradient alcohol, transparentized with xylene, embedded in paraffin, sliced, stained with HE, and the pathological changes of lung tissue were observed under a light microscope. The right upper lobe was quickly removed, and the blood dirt around the tissue was washed away with PBS buffer, leaving 1 mm of the right hilum. 3 Large and small tissue blocks were fixed with 2.5% glutaraldehyde, post-fixed with 1% osmium acid, dehydrated in an ethanol-acetone series, embedded in Epin812, sliced ​​conventionally with an ultrathin slicer, and double-stained with lead acetate and lead citrate. Changes in the ultrastructure of lung tissue were observed under a transmission electron microscope.

2. The device for passive smoking poisoning of rodent experimental animals according to claim 1, characterized in that: Any one of the plurality of smoke diffusion layers is fixedly mounted on the lower box body (1).

3. The device for passive smoking poisoning of rodent experimental animals according to claim 1, characterized in that: The topmost smoke diffusion layer among the plurality of smoke diffusion layers is fixedly mounted on the lower box body (1), and any one of the following smoke diffusion layers is detachably mounted on the lower box body (1).

4. A device for passive smoking poisoning of rodent experimental animals according to claim 2 or 3, characterized in that: The number of the plurality of smoke diffusion layers is set to three, which are respectively a first smoke diffusion layer (111), a second smoke diffusion layer (112) and a third smoke diffusion layer (113) from top to bottom.

5. The device for passive smoking poisoning of rodent experimental animals according to claim 4, characterized in that: The invention also comprises a steel grid frame (3), wherein the steel grid frame (3) is placed inside the lower box body (1) and on the flame retardant plate (4), and the steel grid frame (3) is used to place smoke; one of the side surfaces of the lower box body (1) is also provided with a lower box body door (12) for placing smoke; and one of the side surfaces of the upper box body (2) is provided with an upper box body door (21) for placing experimental animals.

6. The device for passive smoking poisoning of rodent experimental animals according to claim 5, characterized in that: The four corners of the top surface of the lower box body (1) are also provided with stoppers (14); the upper box body (2) is placed between the four stoppers (14) and is limited in position by the four stoppers (14).

Images