CN113267389A - Lung tissue specimen fixing device and method - Google Patents

Lung tissue specimen fixing device and method Download PDF

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Publication number
CN113267389A
CN113267389A CN202110474203.4A CN202110474203A CN113267389A CN 113267389 A CN113267389 A CN 113267389A CN 202110474203 A CN202110474203 A CN 202110474203A CN 113267389 A CN113267389 A CN 113267389A
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China
Prior art keywords
lung tissue
pressure
liquid
communicating pipe
cover body
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CN202110474203.4A
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CN113267389B (en
Inventor
王祥海
郭家松
李振林
文婧
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Southern Medical University
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Southern Medical University
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q

Abstract

The invention discloses a lung tissue specimen fixing device and a method, wherein the lung tissue specimen fixing device comprises a fixing container, a sealing cover and an injector, the top of the fixing container is provided with an opening, the fixing container is also connected with a pressure regulator, and the pressure regulator is communicated with a containing cavity of the fixing container; the sealing cover comprises a cover body and a communicating pipe, wherein the first end of the communicating pipe is positioned below the cover body, and the second end of the communicating pipe penetrates through the cover body or is positioned on the cover body. The first end of the communicating pipe is inserted into the trachea of the lung tissue to be fixed, the cover body seals the opening of the fixed container, the pressure regulator is used for increasing the pressure in the fixed container, then the liquid injector is connected with the second end of the communicating pipe, then the pressure in the fixed container is reduced, and the lung tissue absorbs the stationary liquid under the action of differential pressure. The invention uses bionics to simulate the normal breathing state, changes the pressure of the pleural cavity through the movement of the thorax and the diaphragm to discharge the gas, and simultaneously can ensure that the stationary liquid is uniformly injected into the lung tissue to achieve better expansion and fixation effects.

Description

Lung tissue specimen fixing device and method
Technical Field
The invention relates to a lung tissue specimen fixing device and a method in the technical field of biological experiments.
Background
The reason that lung tissue took off the back usually because truns into malleation and self elasticity from the negative pressure that receives the thorax and presents the atrophy state, and inside still contains a large amount of gases simultaneously, leads to it to present the floating state in the stationary liquid, and the stationary liquid infiltration is slow, and fixed effect is poor, and later stage section in-process is in the atrophy state because the lung all the time, leads to section back microstructure to be clear inadequately, influences later stage clinical pathology's judgement and medical teaching effect. Early attempts to address this problem have typically applied gravity to the lung tissue to completely immerse it in the liquid, but have not addressed the situation in which the lung tissue is atrophied. Later researchers filled the lungs to normal distended state by direct pressurized perfusion of fixative and found clear tissue structure after fixation, but this method still had some drawbacks in practice. Firstly, the pressurized perfusion cannot completely exhaust the gas in the lung, and part of tissues are still isolated by the gas and cannot be contacted with the stationary liquid; in addition, the pressurized perfusion liquid flows to a larger bronchus preferentially, other tissues are further squeezed after the part of the bronchus is expanded, so that part of the tissues are not fully expanded, and meanwhile, the part of the tissues connected with the large bronchus is fractured due to over expansion.
Disclosure of Invention
The present invention is directed to solve at least one of the problems of the prior art, and provides a lung tissue specimen fixing device and method for fixing lung tissue to form a specimen.
According to an embodiment of the first aspect of the present invention, a lung tissue specimen fixing device is provided, which includes a fixing container, an opening is provided at the top of the fixing container, and a pressure regulator is further connected to the fixing container, and the pressure regulator is communicated with a cavity of the fixing container; the sealing cover comprises a cover body and a communicating pipe, the cover body is detachably connected to the opening, the first end of the communicating pipe is positioned below the cover body, the second end of the communicating pipe penetrates through the cover body or is positioned on the cover body, and the first end of the communicating pipe is used for being connected with lung tissues; and the liquid injector is used for being connected with the second end of the communicating pipe.
According to the embodiment of the first aspect of the present invention, further, the injector includes a first liquid storage barrel and a first piston rod, the first piston rod is slidably mounted in the first liquid storage barrel, a first protrusion is disposed at an end of the first liquid storage barrel, a first liquid through hole is disposed on the first protrusion, and the first protrusion can be in interference fit with the second end of the communication pipe.
According to an embodiment of the first aspect of the present invention, further, the first liquid storage cylinder is provided with scales along a length direction.
According to the embodiment of the first aspect of the present invention, further, a pressure adjusting port is provided on the fixed container, the pressure adjuster includes a second liquid storage cylinder and a second piston rod, the second piston rod is slidably mounted in the second liquid storage cylinder, and a second liquid through hole is provided at an end of the second liquid storage cylinder, and the second liquid through hole can be connected to the pressure adjusting port.
According to the embodiment of the first aspect of the present invention, a pressure adjusting pipe is disposed outside the pressure adjusting port, a second protrusion is disposed at an end of the second liquid storage cylinder, the second through hole is located on the second protrusion, and the second protrusion is in interference fit with the pressure adjusting pipe.
According to an embodiment of the first aspect of the present invention, further, an opening connecting pipe is disposed on a top surface of the opening, and the cover body is detachably connected to the opening connecting pipe.
According to a second aspect of the present invention, there is provided a lung tissue specimen fixing method, including the steps of: s10, fixing lung tissues below a sealing cover to enable a lung trachea to be communicated with a first end of a communicating pipe; s20, extending the lung tissue into a fixing container filled with fixing liquid through an opening to enable a cover body to be in sealing connection with the opening; s30, increasing the pressure in the fixed container until the lung tissue is shrunk; s40, connecting the liquid injector with the second end of the communicating pipe, and then reducing the pressure in the fixed container.
According to the embodiment of the second aspect of the present invention, further, in step S40, when the injector injects the volume of the stationary liquid into the lung tissue to be equal to the lung volume, the adjustment of the pressure in the stationary container is stopped.
The invention has the beneficial effects that: the first end of the communicating pipe is inserted into the trachea of the lung tissue to be fixed, the cover body seals the opening of the fixed container, the pressure regulator is used for increasing the pressure in the fixed container, then the liquid injector is connected with the second end of the communicating pipe, then the pressure in the fixed container is reduced, and the lung tissue absorbs the stationary liquid under the action of differential pressure. The invention uses bionics to simulate the normal breathing state, changes the pressure of the pleural cavity through the movement of the thorax and the diaphragm to discharge the gas, and simultaneously can ensure that the stationary liquid is uniformly injected into the lung tissue to achieve better expansion and fixation effects.
Drawings
In order to more clearly illustrate the technical solution in the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly described below. It is clear that the described figures are only some embodiments of the invention, not all embodiments, and that a person skilled in the art can also derive other designs and figures from them without inventive effort.
FIG. 1 is a schematic view of a stationary container in an embodiment of the invention;
FIG. 2 is a schematic view of a sealing cap in an embodiment of the invention;
FIG. 3 is a schematic view of a syringe in an embodiment of the invention;
FIG. 4 is a schematic diagram of a pressure regulator in an embodiment of the present invention.
Detailed Description
Reference will now be made in detail to the present preferred embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to like elements throughout.
In the description of the present invention, it should be understood that the orientation or positional relationship referred to in the description of the orientation, such as the upper, lower, front, rear, left, right, etc., is based on the orientation or positional relationship shown in the drawings, and is only for convenience of description and simplification of description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.
In the description of the present invention, the meaning of a plurality of means is one or more, the meaning of a plurality of means is two or more, and larger, smaller, larger, etc. are understood as excluding the number, and larger, smaller, inner, etc. are understood as including the number. If the first and second are described for the purpose of distinguishing technical features, they are not to be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of the technical features indicated.
In the description of the present invention, unless otherwise explicitly limited, terms such as arrangement, installation, connection and the like should be understood in a broad sense, and those skilled in the art can reasonably determine the specific meanings of the above terms in the present invention in combination with the specific contents of the technical solutions.
The lung tissue specimen fixing device in the first aspect of the invention comprises a fixing container 10, a sealing cover 20 and an injector 30, wherein the sealing cover 20 comprises a cover body 21 and a communicating pipe 22. The top of the fixed container 10 is provided with an opening, the fixed container 10 is also connected with a pressure regulator 40, after the lung tissue is fixed on the communicating pipe 22, the cover body 21 closes the opening, so that the lung tissue is put into the fixed container 10, the liquid injector 30 is connected with the communicating pipe 22, the pressure in the fixed container 10 is changed by controlling the pressure regulator 40, and the stationary liquid in the liquid injector 30 is filled into the lung tissue.
Referring to fig. 1, the fixing liquid is charged into or discharged from the fixing container 10 through the opening, and the lung tissue can also be charged into the fixing container 10 through the opening. Optionally, the fixing container 10 is a transparent member to facilitate observation of the fixed condition of the lung tissue and the fixed lung tissue. The pressure regulator 40 communicates with the chamber of the fixed vessel 10 to regulate the pressure within the fixed vessel 10. It can be understood that the opening connecting pipe 11 is provided on the top surface of the opening, and the cover body 21 is detachably connected to the opening connecting pipe 11. Optionally, the cover 21 is screwed to the opening connecting pipe 11, and an elastic sealing member, such as a silicone ring, is disposed between the cover 21 and the opening connecting pipe 11. The sealing of the cover 21 with the open connection tube 11 facilitates the pressure control in the fixing container 10, thereby ensuring the fixing effect of the lung tissue.
In some embodiments, the fixed container 10 is provided with a pressure adjusting port, and referring to fig. 4, the pressure regulator 40 includes a second liquid storage cylinder 41 and a second piston push rod 42, the second piston push rod 42 is slidably installed in the second liquid storage cylinder 41, and the end of the second liquid storage cylinder 41 is provided with a second liquid through hole, and the second liquid through hole can be connected with the pressure adjusting port. With the movement of the second piston rod 42 in the second fluid storage cylinder 41, the stationary fluid flows out of or into the second fluid storage cylinder 41 through the second fluid passage hole, thereby changing the pressure of the chamber of the stationary container 10. Optionally, the second liquid storage cylinder 41 is provided with scales along the length direction, so that an operator can visually observe the pressure change in the cavity of the fixed container 10, and the effect of quantitative accurate control is achieved.
Further, a pressure adjusting pipe 12 is arranged on the outer side of the pressure adjusting opening, a second protrusion 43 is arranged at the end of the second liquid storage cylinder 41, the second through hole is located on the second protrusion 43, and the second protrusion 43 is in interference fit with the pressure adjusting pipe 12. Because the second bulge 43 is in interference fit with the pressure regulating tube 12, the sealing connection between the second liquid storage cylinder 41 and the fixed container 10 can be ensured, and the pressure in the cavity of the fixed container 10 can be controlled more accurately.
Referring to fig. 2, the cover 21 is detachably attached to the opening, and the sealing cap 20 can fix lung tissue while covering the opening. Specifically, a first end of the communication pipe 22 is located below the cover 21, and a second end of the communication pipe 22 penetrates the cover 21 or is located on the cover 21, so that the communication pipe 22 can communicate the space inside and outside the cover 21. The first end of the communication tube 22 is used for connecting with the lung tissue, and the second end of the communication tube 22 is connected with the injector 30. In this embodiment, the first end of the communication tube 22 penetrates the trachea of the lung tissue and is connected to the trachea, and the fixing liquid in the injector 30 flows into the lung tissue through the communication tube 22, so that the lung tissue is sufficiently expanded and fixed.
In some embodiments, referring to fig. 3, the syringe 30 includes a first liquid storage barrel 31 and a first piston rod 32, the first piston rod 32 is slidably mounted in the first liquid storage barrel 31, a first protrusion 33 is disposed on an end portion of the first liquid storage barrel 31, a first liquid through hole is disposed on the first protrusion 33, and the first protrusion 33 can be in interference fit with the second end of the communication pipe 22. By pushing and pulling the first piston rod 32, the first piston rod 32 moves in the first reservoir 31, and the fixing liquid flows into the communicating tube 22 through the first liquid passing hole, and then enters the lung tissue. Because the first bulge 33 is in interference fit with the communicating pipe 22, the stationary liquid can not leak out from the gap between the liquid injector 30 and the communicating pipe 22, and the first liquid storage cylinder 31 can not reduce air and fill the lung tissue.
Furthermore, the first liquid storage cylinder 31 is provided with scales along the length direction, and the volume of the fixing liquid filled in the lung tissue can be judged according to the scales, so that whether the lung tissue needs to be continuously perfused or stopped perfused is judged, the lung tissue is fully expanded, and the phenomenon that the lung alveoli are broken and the manufacturing effect of the lung tissue specimen is influenced due to the fact that other tissues are extruded due to the fact that part of the bronchus is excessively expanded is avoided.
Taking an SD rat of 200g as an example, the lung tissue specimen fixing method in the second aspect of the invention comprises the following steps:
s10, fixing the lung tissue below the sealing cover 20, specifically, binding the lung tissue at a first end of the communicating pipe 22, so that the lung trachea is communicated with the first end of the communicating pipe 22.
S20, extending the lung tissue into the fixing container 10 filled with the fixing liquid through the opening, and enabling the cover body 21 to be connected with the opening in a sealing mode.
S30, increasing the pressure in the fixed container 10 through the air pressure regulator until the lung tissue is shrunk. Specifically, the second protrusion 43 is connected to the pressure-regulating tube 12 before the fixing container 10 is filled with the fixing liquid. When the pressure in the fixed container 10 is to be increased, the second piston rod 42 is pushed to make the stationary liquid in the second liquid storage cylinder 41 flow into the fixed container 10 through the second liquid through hole, so that the pressure in the fixed container 10 is increased. Due to the pressure increase in the stationary container 10, the rest of the lung tissue is discharged out of the stationary container 10 through the communicating tube 22 under the pressure, so that the lung tissue gradually shrinks until it becomes atrophic.
S40, when the lung tissue shrinks, the liquid injector 30 is connected with the second end of the communicating pipe 22, and then the pressure in the fixed container 10 is reduced. Specifically, the second piston rod 42 is pulled to flow the stationary liquid in the stationary container 10 into the second reservoir 41 through the second through hole, and the pressure in the stationary container 10 is reduced. The lung tissue can be uniformly expanded through the negative pressure effect, meanwhile, the liquid in the first liquid storage cylinder 31 flows into the lung tissue through the communicating pipe 22 under the action of pressure difference, the fixing liquid flows to each bronchus from the trachea, the circulation of gas in the lung during the respiration of animals is simulated, and various structures in the lung tissue can be kept in normal and good shapes. Because the lung tissue actively absorbs the stationary liquid in the first liquid storage cylinder 31 under the action of the pressure difference, the lung tissue is prevented from passively receiving the stationary liquid when the liquid injector 30 injects liquid into the lung tissue, and the stationary liquid can damage the structure of the lung tissue under the action of hydraulic pressure; in addition, the inside and the outside of the lung tissue are soaked in the fixing liquid, so that a better fixing effect can be achieved, and the microstructure after slicing is closer to the physiological state.
Optionally, when the injector 30 injects the lung tissue with the volume of the stationary liquid equal to the lung volume, the adjustment of the pressure in the stationary container 10 is stopped, so as to avoid the tissue damage caused by over-expansion of the lung tissue. In this embodiment, the lung volume is calculated according to the body weight of the experimental animal, and the formula is: the lung volume (ml) is 0.981+0.038 × body weight (g), and the fixed amount of liquid to be injected into the lung tissue by the injector 30 can be obtained.
In the fixing method, the mode of discharging residual gas in lung tissues by positive pressure and then sucking fixing liquid by negative pressure is adopted, bionics is applied to simulate the movement of the thorax and the diaphragm under the normal breathing state, the pressure of the pleural cavity is changed to discharge the gas, and the fixing liquid is uniformly injected into the lung tissues, so that better expansion and fixing effects are achieved. Compared with a control group which is only subjected to conventional perfusion, the lung tissue fixed by the fixing method has a normal structure, a loose alveolar structure of the lung tissue is clear, the alveolar space is thinner, various structures are kept in a normal and good shape, and no obvious distortion change of collapse, shrinkage or damage of the lung tissue structure is seen.
While the preferred embodiments of the present invention have been described in detail, it is to be understood that the invention is not limited to the precise embodiments, and that various equivalent changes and modifications may be effected therein by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (8)

1. A lung tissue specimen holding device, comprising:
the top of the fixed container is provided with an opening, the fixed container is also connected with a pressure regulator, and the pressure regulator is communicated with the accommodating cavity of the fixed container;
the sealing cover comprises a cover body and a communicating pipe, the cover body is detachably connected to the opening, the first end of the communicating pipe is positioned below the cover body, the second end of the communicating pipe penetrates through the cover body or is positioned on the cover body, and the first end of the communicating pipe is used for being connected with lung tissues;
and the liquid injector is used for being connected with the second end of the communicating pipe.
2. The lung tissue specimen fixation device according to claim 1, wherein: the liquid injector comprises a first liquid storage cylinder and a first piston push rod, the first piston push rod is slidably mounted in the first liquid storage cylinder, a first bulge is arranged at the end part of the first liquid storage cylinder, a first liquid through hole is formed in the first bulge, and the first bulge can be in interference fit with a second end of the communicating pipe.
3. The lung tissue specimen holding device according to claim 2, wherein: the first liquid storage cylinder is provided with scales along the length direction.
4. The lung tissue specimen fixation device according to claim 1, wherein: the fixed container is provided with a pressure adjusting port, the pressure adjuster comprises a second liquid storage cylinder and a second piston push rod, the second piston push rod is slidably mounted in the second liquid storage cylinder, the end part of the second liquid storage cylinder is provided with a second liquid through hole, and the second liquid through hole can be connected with the pressure adjusting port.
5. The lung tissue specimen holding device according to claim 4, wherein: the outside of pressure regulation mouth is equipped with the pressure regulation pipe, the tip of second liquid storage cylinder is equipped with the second arch, the second through-flow hole is located on the second arch, the second arch with pressure regulation pipe interference fit.
6. The lung tissue specimen fixation device according to claim 1, wherein: the top surface of the opening is provided with an opening connecting pipe, and the cover body is detachably connected with the opening connecting pipe.
7. A lung tissue specimen fixing method is characterized by comprising the following steps:
s10, fixing lung tissues below a sealing cover to enable a lung trachea to be communicated with a first end of a communicating pipe;
s20, extending the lung tissue into a fixing container filled with fixing liquid through an opening to enable a cover body to be in sealing connection with the opening;
s30, increasing the pressure in the fixed container until the lung tissue is shrunk;
s40, connecting the liquid injector with the second end of the communicating pipe, and then reducing the pressure in the fixed container.
8. The method for fixing a lung tissue specimen according to claim 7, wherein: in step S40, when the injector injects the lung tissue with the volume of the stationary liquid equal to the lung volume, the adjustment of the pressure in the stationary container is stopped.
CN202110474203.4A 2021-04-29 2021-04-29 Lung tissue specimen fixing device and method Active CN113267389B (en)

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