Method for regulating and controlling sleep and metabolic spectrum of mouse through intestinal pathway and flora transplanter

Abstract

The invention discloses a method and a flora transplanter for regulating and controlling the sleep and metabolic profile of a mouse through an intestinal pathway. The method comprises the step of applying a microbial combination extracted from a healthy donor to a sleep disorder receptor. Experiments show that when the intake of antibiotics causes the sleep disorder of mice, the microbial composition of healthy mice is applied, so that the total sleep time of the mice with the sleep disorder can be effectively prolonged, the sleep efficiency can be improved, the mice can fall asleep in a shorter time, the proportion of non-rapid eye movement sleep in the total sleep time is increased, and the sleep quality of the mice is improved. The sleep quality is effectively improved.

Description

Technical Field

[0001] The present invention relates to the technical field of regulating bacterial flora metabolic profiles, and in particular to a method for regulating mouse sleep and metabolic profiles through an enterogenic pathway and a bacterial flora transplanter. Background Art

[0002] Antibiotics play an important role in treating bacterial infections, but they can also have adverse effects on the host's intestinal microbiota, leading to dysbiosis (imbalance of microbial populations). This imbalance not only affects intestinal health, but can also trigger metabolic disorders and even affect sleep quality. Sleep disorders are closely related to health conditions, and antibiotic-induced dysbiosis often leads to changes in the host's metabolic profile, further exacerbating health problems.

[0003] Previous studies have shown that intestinal microbes play a key role in regulating sleep and maintaining metabolic homeostasis. However, the specific mechanism by which microbiota transplantation can alleviate sleep disorders and metabolic abnormalities is still unclear. This invention proposes a new method to solve these problems through microbial reconstruction.

[0004] In addition, the microbiota transplant device is generally composed of an injection device and a microbiota transplant tube. The drug is inside the syringe, and the microbiota transplant tube extends to the recipient's stomach. Then, when the injection device is working, the liquid enters the recipient's stomach through the gavage tube. However, this type of gavage method has the problem that the gas inside the gavage tube will first enter the recipient's stomach in large quantities. When a small dose is injected, the amount of liquid is difficult to reach the recipient's stomach through the long gavage tube. Summary of the invention

[0005] In view of the problem in the prior art that antibiotic-induced dysbiosis often leads to changes in the host metabolic profile, the present invention is proposed.

[0006] Therefore, the object of the present invention is to provide a method for regulating sleep and metabolic profiles in mice via a gut-derived pathway.

[0007] To solve the above technical problems, the present invention provides the following technical solution: a method for regulating the sleep and metabolic profile of mice through an enterogenic pathway, comprising administering a combination of microorganisms extracted from a healthy donor to a sleep disorder recipient.

[0008] As a preferred embodiment of the method for regulating sleep and metabolic profile of mice via an enterogenic pathway according to the present invention, the microbial combination is extracted from fecal samples of healthy donors.

[0009] As a preferred embodiment of the method for regulating sleep and metabolic profile of mice via an enterogenic pathway according to the present invention, the microbial combination is administered to a healthy donor via oral gavage.

[0010] As a preferred embodiment of the method for regulating sleep and metabolic profile of mice through an enterogenic pathway as described in the present invention, the microbial combination is extracted through fecal samples of healthy donors, including collecting fecal samples, preparing fecal homogenate through the fecal samples, filtering and centrifuging the fecal homogenate to obtain a precipitate containing microorganisms, and resuspending the precipitate to reach the desired concentration.

[0011] As a preferred embodiment of the method for regulating sleep and metabolic profile of mice via an enterogenic pathway as described in the present invention, individuals without disease and history of antibiotic treatment are selected as healthy donors.

[0012] The beneficial effects of this method of regulating the sleep and metabolic profile of mice through the enteric pathway are shown in experiments. When the intake of antibiotics causes sleep disorders in mice, the administration of a microbial combination of healthy mice can effectively increase the total sleep time of sleep-disordered mice, improve sleep efficiency, allow mice to fall asleep in a shorter time, and increase the proportion of non-rapid eye movement sleep in the total sleep time, effectively improving sleep quality.

[0013] The present invention also discloses a microbiota transplanter used in the above-mentioned method of regulating the sleep and metabolic spectrum of mice through the enterogenic pathway, comprising a syringe; an extension mechanism connected to the syringe; a medicine chamber, the medicine chamber is arranged at the end of the extension mechanism away from the syringe, and comprises a drug outlet and a compression movable part arranged inside the medicine chamber; a gas delivery pipe, arranged between the syringe and the medicine chamber; when the gas inside the syringe enters the medicine chamber through the gas delivery pipe, it can push the compression movable part to discharge the drug inside the medicine chamber from the drug outlet.

[0014] As a preferred embodiment of the microbiota transplanter of the present invention, the syringe comprises a shell having a cavity therein; a push head disposed inside the cavity of the shell; a push rod connected to the push head; a connecting port disposed on the shell and connected to the inner cavity of the shell; the push head divides the cavity of the shell, an injection chamber is formed between the push head and the connecting port, the compression movable part divides the inner space of the medicine chamber into a gas chamber and a liquid chamber, the medicine outlet is opened on the side wall of the liquid chamber, the gas delivery pipe is connected to the gas chamber, and the medicine chamber is also provided with a medicine injection port in the liquid chamber.

[0015] As a preferred embodiment of the microbiota transplanter of the present invention, the opening and closing mechanism includes a core plug, a core column connected to the core plug and passing through the injection port, a core head arranged at the end of the core column away from the core plug, and an elastic member abutting against the core head; a separating ring is arranged inside the medicine cabin, and the separating ring separates the internal space of the medicine cabin into an independent accommodating chamber; the core plug and the elastic member are both inside the accommodating chamber.

[0016] As a preferred embodiment of the bacterial colony transplanter of the present invention, the extension mechanism is a tubular structure, one end of which is provided with a pressure piece, and is connected to the syringe through the pressure piece;

[0017] An inner groove is formed on the push head, and a limiter is provided on the radial outer wall of the push rod, and the limiter is movably arranged inside the inner groove;

[0018] The shell is also provided with a through hole, the pressure piece is connected to the through hole, and the end of the push rod can pass through the through hole and enter the interior of the pressure piece.

[0019] As a preferred embodiment of the bacterial colony transplanter of the present invention, a pressing plug is arranged inside the pressure piece, and the pressure piece between the pressing plug and the core plug, the extension mechanism, and the internal area of ​​the accommodating cavity constitute an air pressure cavity.

[0020] The beneficial effects of the present microbiota transplanter are as follows: the liquid is stored in the medicine chamber, which can be directly pushed to the stomach of the recipient through an extension mechanism, and then the gas inside the syringe is injected into the interior of the gas delivery tube, and the gas inside the gas delivery tube enters the medicine chamber, pushing the compression movable part to move, thereby injecting the liquid stored in the medicine chamber into the stomach of the recipient, thus avoiding the problem of a large amount of gas being injected into the stomach of the recipient and reducing the discomfort of the recipient. In addition, because the liquid and solvent to be injected are located in the medicine chamber, they will directly enter the interior of the recipient after being discharged from the medicine outlet of the medicine chamber, thus reducing the loss of liquid and solvent inside the gavage tube. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0022] Figure 1 This is a diagram of the sleep-wake duration and structural changes in mice after antibiotic intervention in the present invention.

[0023] Figure 2 This is a diagram showing the changes in c-Fos expression in the mouse brain after antibiotic intervention in the present invention.

[0024] Figure 3 This is a diagram of the diversity and abundance of intestinal flora in mice after antibiotic intervention and flora transplantation in the present invention.

[0025] Figure 4 This is a graph showing changes in sleep-wake duration and c-Fos expression after bacterial flora transplantation in the present invention.

[0026] Figure 5This is a diagram showing changes in the serum metabolic profile of mice after antibiotic intervention and bacterial flora transplantation in the present invention.

[0027] Figure 6 It is a schematic diagram of the overall structure of the bacterial colony transplanter in the present invention.

[0028] Figure 7 It is a schematic diagram of the internal structure of the flora transplanter in the present invention.

[0029] Figure 8 It is a schematic diagram of the structure of the bacterial colony transplanter and the medicine cabin in the present invention.

[0030] Fig. 9 It is a schematic diagram of the medicine cabin structure described in the present invention.

[0031] Fig.10 It is a schematic diagram of the structure of the opening and closing mechanism described in the present invention.

[0032] Fig.11 It is a schematic diagram of the structure of the bacterial colony transplanter and the pressure piece described in the present invention. DETAILED DESCRIPTION

[0033] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the accompanying drawings.

[0034] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0035] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0036] Secondly, the present invention is described in detail with reference to the schematic diagram. When describing the embodiments of the present invention in detail, for the sake of convenience, the cross-sectional diagrams showing the device structure will not be partially enlarged according to the general scale, and the schematic diagrams are only examples, which should not limit the scope of protection of the present invention. In addition, in actual production, the three-dimensional dimensions of length, width and depth should be included.

[0037] Example 1 provides a method for regulating the sleep and metabolic profile of mice through an enterogenic pathway, comprising administering a combination of microorganisms extracted from a healthy donor to a sleep disorder recipient. Taking mice as an example, the sleep disorder recipients are individuals who have sleep disorders caused by taking antibiotics, and individuals who are disease-free and have no history of antibiotic treatment are selected as healthy donors, that is, mice that are disease-free and have no history of antibiotic treatment are selected as healthy donors.

[0038] The microbial composition was extracted from stool samples of healthy donors.

[0039] The specific microbial combination is extracted through stool samples from healthy donors, which includes collecting stool samples in a sterile environment. The stool samples need to be processed in a timely manner after collection to ensure the activity of the microorganisms.

[0040] After obtaining the stool sample, place the stool sample in a sterile mortar and add an appropriate amount of sterile saline or phosphate buffered saline (PBS). The ratio of stool sample to water or phosphate buffered saline (PBS) is usually 1:5 to 1:10 (weight / volume). Use a pestle or homogenizer to mix the stool and liquid to form a uniform stool slurry.

[0041] The fecal homogenate is filtered and centrifuged to obtain a precipitate containing microorganisms. Specifically, the fecal homogenate is filtered through a sterile filter to remove unbroken solid particles, and the filtered homogenate is centrifuged to precipitate microbial cells. Low-speed centrifugation is usually used to avoid damaging the microbial cells.

[0042] The precipitate is resuspended to a desired concentration. Specifically, the precipitate is resuspended in a new sterile liquid medium to achieve a desired concentration. A vortex mixer or a pipette may be used to gently pipette to ensure that the microbial cells are evenly dispersed.

[0043] Throughout the extraction process, aseptic operation and microbial contamination testing are performed regularly to ensure that the suspension does not contain harmful microorganisms or pathogens.

[0044] Furthermore, the microbial combination was administered to healthy donors via oral gavage.

[0045] A dysbiosis mouse model was established by intervening with antibiotics for three weeks. Figure 1 A), To determine whether gut dysbiosis is sufficient to induce sleep disturbances, we performed EEG-EMG recordings.

[0046] The data showed that antibiotic-treated mice exhibited significant disruptions in their sleep-wake rhythms. Specifically, AIMD mice (mice treated with antibiotics) showed significant differences in their sleep-wake patterns during the light phase.

[0047] Compared with the control group, AIMD mice had a 27.68% increase in wakefulness during the 12-hour light period and a 26.87% decrease in wakefulness during the 12-hour dark period ( Figure 1 B). NREM sleep decreased by 11.9% during the 12-hour light period, increased by 35.20% during the 12-hour dark period, and increased by 12.73% during the 24-hour period ( Figure 1 C). REM sleep in AIMD mice decreased by 26.30% during the 12-hour light period and increased by 17.62% during the 12-hour dark period ( Figure 1 D).

[0048] like Figure 1 As shown in E, the increase in wakefulness time in AIMD mice during the 12-hour light period compared to the control group is due to an increase in its fragmentation. In contrast, the decrease in wakefulness time during the 12-hour dark phase is due to a decrease in the duration of the episodes. The decrease in NREM sleep in AIMD mice during the 12-hour light period is due to a decrease in its occurrence, while the increase in NREM sleep during the 12-hour dark phase is due to an increase in the number of occurrences. The decrease in REM sleep in AIMD mice during the 12-hour light period is also due to a decrease in the number of occurrences ( Figure 1 EJ). Taken together, the findings suggest that disruptions in the gut microbiota can lead to alterations in sleep-wake behavior.

[0049] To investigate the neural mechanisms underlying antibiotic-induced disruptions in sleep-wake behavior, we examined the expression of c-Fos, a molecular marker of neuronal activity, in brain regions associated with sleep and wake regulation. In this study, we found that depletion of gut microbiota by antibiotics reduced c-Fos expression in the prelimbic cortex (PrL) and infralimbic cortex (IL). However, in the AIMD group, c-Fos expression was increased in the paraventricular nucleus of the thalamus (PVT), lateral habenula (LHb), and basolateral amygdala (BLA). In addition, antibiotics did not affect c-Fos expression in sleep-wake brain regions such as the stria terminalis (BNST), central amygdala (CeA), and nucleus accumbens (NAc) ( Figure 2 A, B). These findings demonstrate that depletion of the gut microbiota by antibiotics leads to significant alterations in the expression pattern of c-Fos in the brain, highlighting changes in neuronal activity within specific brain regions involved in sleep-wake regulation.

[0050] After two weeks of antibiotic intervention, fecal samples were collected from mice for 16S rDNA high-throughput sequencing. As expected, antibiotic exposure induced significant changes in the composition of the gut microbiota.

[0051] Antibiotic-treated mice exhibited lower alpha diversity (species diversity) as measured by the Chao and Shannon index ( Figure 3A, B). Principal coordinates analysis (PCoA) based on Bray-Curtis distance clearly showed different bacterial community structures between the control group and the antibiotic-treated group ( Figure 3 C).

[0052] Specifically, at the phylum level, antibiotic-treated mice showed a decrease in Lactobacillus and an increase in Salmonella ( Figure 3 D).

[0053] Therefore, we investigated whether reconstruction of normal intestinal flora could restore the intestinal microbial ecology of AIMD mice.

[0054] We transplanted the microbiota of age-matched healthy mice into antibiotic-treated mice with depleted microbiota ( Figure 3 E).

[0055] Two weeks after transplantation, we performed 16S rRNA sequencing on the recipient mice. We found that the intestinal flora had been significantly restored, with significant improvements in both the α-diversity and β-diversity of the mice, and the relative abundance of species was restored ( Figure 3 FI). These results suggest that antibiotic-induced disruption of the gut microbiota can be restored by FMT. In addition, we investigated whether fecal microbiota transplantation (FMT) could restore normal sleep patterns ( Figure 4 A) Using electroencephalogram (EEG)-electromyogram (EMG) recordings of recipient mice, we show that sleep patterns in antibiotic-treated mice were significantly improved following gut microbiota transplantation from healthy, age-matched SPF mice.

[0056] Analysis of EEG-EMG signals showed that wakefulness and REM sleep patterns were restored after transplantation ( Figure 4 B, D). Despite differences in total NREM sleep time within a 24-hour period, a significant restoration of NREM sleep was observed during both the dark and light phases after transplantation ( Figure 4 C).

[0057] These results suggest that restoring the gut microbiota can significantly improve sleep behavior.

[0058] Restoration of normal gut microbiota restored c-Fos expression in multiple brain regions and host serum metabolites in antibiotic-treated mice. Restoration of normal gut microbiota also rescued altered c-Fos expression in multiple brain regions in antibiotic-treated mice. Figure 4 EF). This further emphasizes the critical role of the gut microbiota in maintaining normal brain function. Reconstitution may restore the production of microbial metabolites, thereby normalizing neuronal activity.

[0059] This highlights the therapeutic potential of microbiota-based interventions in treating sleep disorders associated with gut microbiota dysbiosis.

[0060] Antibiotic treatment is known to alter the composition of the gut microbiota, which may in turn influence host behavior via the gut-brain axis.

[0061] To further explore the mechanisms by which gut microbiota restoration affects sleep recovery, we used a targeted metabolomics approach to analyze serum samples from control, AIMD, control, and FMT-AIMD mice.

[0062] Metabolite classification showed that organic acids, amino acids, fatty acids, short-chain fatty acids (SCFAs), and carnitine were the top five most abundant metabolites ( Figure 5 A, C). The results showed that 40 metabolites were significantly different between the AIMD group and the control group.

[0063] After microbiota transplantation, serum metabolites of recipient mice were significantly restored ( Figure 5 B, D). Venn diagram showing that 37 metabolites that changed after AIMD were restored after microbiota transplantation ( Figure 5 E).

[0064] This suggests that these metabolites may be driven by the gut microbiota and may regulate sleep.

[0065] We identified the top eight metabolites that were most significantly altered in AIMD mice.

[0066] We listed the metabolites that showed significant changes in AIMD mice and were restored after FMT ( Figure 5 F,G). By identifying metabolites that showed significant changes in AIMD mice and were restored after FMT, we highlighted potential metabolic pathways by which the gut microbiota influences sleep behavior.

[0067] These results suggest that the gut microbiota plays a key role in regulating neural activity and systemic metabolic profiles, potentially influencing sleep behavior through gut-brain communication pathways.

[0068] Experiments have shown that when antibiotic intake causes sleep disorders in mice, the administration of a microbial combination from healthy mice can effectively increase the total sleep time of sleep-disordered mice, improve sleep efficiency, allow mice to fall asleep in a shorter time, and increase the proportion of non-rapid eye movement sleep in total sleep time, effectively improving sleep quality.

[0069] Example 2, reference Figures 6 to 8This embodiment is different from the first embodiment in that: a microbiota transplanter applied to the above-mentioned method of regulating the sleep and metabolic spectrum of mice through the enterogenic pathway is also disclosed. In the prior art, the microbiota transplanter is generally composed of an injection device and a microbiota transplantation pipeline. The drug is inside the syringe 100, and the microbiota transplantation pipeline extends to the stomach of the recipient. Then, when the injection device is working, the liquid enters the stomach of the recipient through the microbiota transplantation pipeline. However, this type of microbiota transplantation method has the problem that the gas inside the microbiota transplantation pipeline will first enter the stomach of the recipient in large quantities. When a small dose is injected, it is difficult for the amount of liquid to reach the stomach of the recipient through the long microbiota transplantation pipeline.

[0070] The flora transplanter proposed in this embodiment includes a syringe 100 , whose basic structure is substantially the same as that of the existing syringe 100 . However, in this embodiment, the liquid solvent to be injected into the stomach of the recipient is not stored inside the syringe 100 .

[0071] The microbiota transplanter includes an extension mechanism 200, which is connected to the syringe 100. In this embodiment, the function of the extension mechanism 200 is the same as that of the microbiota transplantation pipeline in the prior art. The difference is that the extension mechanism 200 does not need to be responsible for delivering drugs, but only needs to increase the length so that the end away from the syringe 100 can extend to the stomach of the recipient.

[0072] The microbiota transplanter also includes a medicine chamber 300, which is arranged at the end of the extension mechanism 200 away from the syringe 100, and includes a medicine outlet 301 and a compression movable part 302 arranged inside the medicine chamber 300; a gas delivery tube 400, which is arranged between the syringe 100 and the medicine chamber 300; when the gas inside the syringe 100 enters the medicine chamber 300 through the gas delivery tube 400, it can push the compression movable part 302 to discharge the medicine inside the medicine chamber 300 from the medicine outlet 301.

[0073] Therefore, when liquid needs to be injected into the recipient's stomach, the liquid is stored in the medicine chamber 300, and the medicine chamber 300 can be directly pushed to the recipient's stomach through the extension mechanism 200. Then, the gas inside the syringe 100 is injected into the gas delivery tube 400, and the gas inside the gas delivery tube 400 enters the medicine chamber 300, pushing the compression movable part 302 to move, so that the liquid stored in the medicine chamber 300 can be poured into the recipient's stomach.

[0074] Through the above scheme, the problem of a large amount of gas being injected into the recipient's stomach can be avoided, thereby reducing the recipient's discomfort. In addition, because the liquid and solvent to be injected are located in the medicine cabin 300, they will directly enter the interior of the recipient after being discharged from the medicine outlet 301 of the medicine cabin 300, reducing the loss of liquid and solvent inside the flora transplantation pipeline.

[0075] Specifically, the syringe 100 includes a shell 101 having a cavity therein; a push head 102 disposed inside the cavity of the shell 101; a push rod 103 connected to the push head 102; a connecting port 104 disposed on the shell 101 and connected to the internal cavity of the shell 101; the push head 102 separates the cavity of the shell 101, and an injection chamber S1 is formed between the push head 102 and the connecting port 104. In the prior art, the injection chamber S1 is loaded with liquid or solvent, but in the present embodiment, only air exists inside the injection chamber S1. When the push head 102 is pushed toward the connecting port 104 by the push rod 103, the gas originally in the injection chamber S1 can be pushed into the gas delivery pipe 400 through the connecting port 104, and finally reaches the medicine cabin 300 through the gas delivery pipe 400, thereby achieving the effect of driving the compression movable part 302.

[0076] The compression movable part 302 divides the internal space of the medicine chamber 300 into a gas chamber S2 and a liquid chamber S3. The medicine outlet 301 is opened on the side wall of the liquid chamber S3. The gas delivery pipe 400 is connected to the gas chamber S2. The medicine chamber 300 is also provided with a medicine injection port 303 in the liquid chamber S3.

[0077] Therefore, when the gas inside the gas delivery tube 400 enters the medicine cabin 300, it will enter the gas chamber S2, and the medicine is stored in the liquid chamber S3. As the gas is injected, the gas pressure inside the gas chamber S2 increases, thereby pushing the compression movable part 302. The compression movable part 302 can now push the solvent or liquid inside the liquid chamber S3 out through the medicine outlet 301.

[0078] A rubber plug is provided inside the medicine injection port 303, and the liquid and solvent to be injected can enter the liquid cavity S3 of the medicine cabin 300 through the rubber plug via an injection needle.

[0079] The rest of the structure is the same as that of Example 1.

[0080] Example 3, reference Figures 9 to 11, this embodiment is different from the above embodiments in that: when injecting liquid or solvent into the recipient's stomach, in order to avoid the problem that the injected liquid or solvent has a fast flow rate and impacts the recipient's stomach and causes damage to the recipient, it is necessary to increase the overall inner diameter of the drug outlet 301 in this embodiment, thereby reducing the flow rate of the liquid or solvent discharged from the drug outlet 301. However, the increase in the overall inner diameter of the drug outlet 301 may cause the liquid or solvent in the liquid cavity S3 to leak before reaching the recipient's stomach, so the drug chamber 301 is 00 is also provided with an opening and closing mechanism 500 inside, the opening and closing mechanism 500 includes a core plug 501, a core column 502 connected to the core plug 501 and passing through the medicine injection port 303, a core head 503 arranged at the end of the core column 502 away from the core plug 501, and an elastic member 504 abutting against the core head 503; a separating ring 304 is provided inside the medicine capsule 300, and the separating ring 304 separates the internal space of the medicine capsule 300 into an independent accommodating chamber S4; the core plug 501 and the elastic member 504 are both inside the accommodating chamber S4.

[0081] The elastic member 504 is a spring, one end of which contacts the separation ring 304 , and the other end of which contacts the core plug 501 . When the core plug 501 moves toward the separation ring 304 , it presses the elastic member 504 to deform it.

[0082] The core head 503 cannot enter the inner side of the liquid cavity S3 through the medicine outlet 301. Under the push of the elastic member 504, the core plug 501 transmits the thrust to the core head 503 through the core column 502, so that the core head 503 hits the medicine outlet 301. When the core head 503 hits the medicine outlet 301, the liquid or solvent inside the liquid cavity S3 will not leak. Therefore, the problem of the liquid or solvent inside the liquid cavity S3 leaking before reaching the recipient's stomach can be avoided.

[0083] Specifically, the extension mechanism 200 is a tubular structure, one end of which is provided with a pressure piece 201, which is connected to the syringe 100 through the pressure piece 201; an inner groove 102a is provided on the pushing head 102, and a limit body 103a is provided on the radial outer wall of the push rod 103, and the limit body 103a is movably arranged inside the inner groove 102a; a through port 101a is also provided on the shell 101, and the pressure piece 201 is connected to the through port 101a, and the end of the push rod 103 can penetrate the through port 101a to enter the interior of the pressure piece 201.

[0084] Among them, the inner groove 102a has two ends, which are used to limit the limiting body 103a. One end is close to the through port 101a, which is the pushing end 102a-1, and the other end is away from the through port 101a, which is the pulling end 102a-2. When the push rod 103 is pushed to move toward the through port 101a, the push rod 103 will move toward the through port 101a relative to the pushing head 102. At this time, the limiting body 103a moves toward the pushing end 102a-1 on the inner side of the inner groove 102a. At this time, the push rod 103 enters the inner side of the pressure piece 201, and the applied pressure is transmitted to the core plug 501 through the extension mechanism 200, so that the core plug 501 presses the elastic piece 504, so that the core head 503 is separated from the medicine outlet 301, thereby opening the medicine outlet 301.

[0085] During this process, the limiting body 103a moves on the inner side of the inner groove 102a toward the pushing end 102a-1. After the medicine outlet 301 is opened, the limiting body 103a contacts the pushing end 102a-1, pushing the pushing head 102 to press the air inside the injection chamber S1, so that the air enters the gas chamber S2 through the connecting port 104 and the gas delivery tube 400, and then pushes the compression movable part 302 to move, pushing the liquid or solvent out of the liquid chamber S3.

[0086] Furthermore, a pressing plug 201 a is disposed inside the pressure member 201 , and the pressure member 201 between the pressing plug 201 a and the core plug 501 , the extending mechanism 200 , and the inner area of ​​the accommodating chamber S4 constitute the air pressure chamber S5 .

[0087] When the push rod 103 enters the pressure piece 201, it pushes the pressing plug 201a. The pressing plug 201a moves and presses the gas inside the air pressure chamber S5, causing the gas pressure inside the air pressure chamber S5 to increase, thereby transmitting the pressure to the core plug 501, so that the core plug 501 can push the elastic piece 504, causing the elastic piece 504 to deform, and the core column 502 pushes the core head 503 to separate from the medicine outlet 301.

[0088] The rest of the structure is the same as that of Example 2.

[0089] Importantly, it should be noted that the construction and arrangement of the present application shown in a plurality of different exemplary embodiments are only exemplary. Although only a few embodiments are described in detail in this disclosure, it should be readily understood by those who refer to this disclosure that many modifications are possible (e.g., the size, scale, structure, shape and proportion of various elements, as well as parameter values ​​(e.g., temperature, pressure, etc.), mounting arrangements, use of materials, color, directional changes, etc.) without substantially departing from the novel teachings and advantages of the subject matter described in the application. For example, the element shown as integrally formed can be composed of multiple parts or elements, the position of the element can be inverted or otherwise changed, and the nature or number or position of the discrete element can be changed or changed. Therefore, all such modifications are intended to be included in the scope of the present invention. The order or sequence of any process or method steps can be changed or reordered according to alternative embodiments. In the claims, any "device plus function" clause is intended to cover the structure described herein that performs the function, and is not only structurally equivalent but also equivalent structure. Without departing from the scope of the present invention, other replacements, improvements, changes and omissions can be made in the design, operating conditions and arrangement of the exemplary embodiments. Therefore, the invention is not limited to a specific embodiment, but extends to numerous modifications still falling within the scope of the appended claims.

[0090] Additionally, in order to provide a concise description of exemplary embodiments, all features of an actual embodiment (ie, those features that are not relevant to the best mode presently contemplated for carrying out the invention or those that are not relevant to implementing the invention) may not be described.

[0091] It should be understood that in the development of any actual implementation, as in any engineering or design project, numerous implementation-specific decisions may be made. Such a development effort may be complex and time-consuming, but for those of ordinary skill having the benefit of this disclosure, without undue experimentation, the development effort will be a routine task of design, fabrication, and production.

[0092] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A method for regulating sleep and metabolic profiles in mice via an enterogenic pathway, characterized in that: It involves administering a combination of microorganisms extracted from healthy donors to recipients with sleep disorders.

2. The method for regulating sleep and metabolic profile of mice via an enterogenic pathway according to claim 1, characterized in that: The microbial composition was extracted from stool samples of healthy donors.

3. The method for regulating sleep and metabolic profile of mice via an enterogenic pathway according to claim 1 or 2, characterized in that: The microbial combination is administered to a healthy donor via a flora transplant.

4. The method for regulating sleep and metabolic profile of mice via an enterogenic pathway according to claim 3, characterized in that: The extraction of microbial composition from stool samples of healthy donors includes collecting stool samples, preparing stool homogenate from the stool samples, filtering and centrifuging the stool homogenate to obtain a precipitate containing microorganisms, and resuspending the precipitate to reach a desired concentration.

5. The method for regulating sleep and metabolic profile of mice via an enterogenic pathway according to claim 1 or 4, characterized in that: Individuals with no disease and no history of antibiotic treatment were selected as healthy donors.

6. A microbiota transplanter used in the method for regulating sleep and metabolic profile of mice via an enterogenic pathway as described in any one of claims 1 to 5, characterized in that: include, Syringe (100); an extension mechanism (200) connected to the syringe (100); A medicine cabin (300), the medicine cabin (300) being arranged at the end of the extension mechanism (200) away from the syringe (100), and comprising a medicine outlet (301) and a compression movable part (302) arranged inside the medicine cabin (300); A gas delivery tube (400) is provided between the injector (100) and the medicine cabin (300); When the gas inside the syringe (100) enters the medicine chamber (300) through the gas delivery tube (400), it can push the compression movable part (302) to discharge the medicine inside the medicine chamber (300) from the medicine outlet (301).

7. The microbiota transplanter according to claim 6, characterized in that: The syringe (100) comprises: A housing (101) having a cavity therein; A push head (102) disposed inside the cavity of the housing (101); A push rod (103), which is connected to the push head (102); A communication port (104) is provided on the shell (101) and is communicated with the internal cavity of the shell (101); The push head (102) divides the cavity of the shell (101), and an injection chamber (S1) is formed between the push head (102) and the connecting port (104). The compression movable part (302) divides the internal space of the medicine cabin (300) into a gas chamber (S2) and a liquid chamber (S3). The medicine outlet (301) is opened on the side wall of the liquid chamber (S3), and the gas delivery pipe (400) is connected to the gas chamber (S2). The medicine cabin (300) is also provided with a medicine injection port (303) in the liquid chamber (S3).

8. The microbiota transplanter according to claim 7, characterized in that: An opening and closing mechanism (500) is also provided inside the medicine chamber (300), and the opening and closing mechanism (500) comprises a core plug (501), a core column (502) connected to the core plug (501) and penetrating the medicine injection port (303), a core head (503) provided at the end of the core column (502) away from the core plug (501), and an elastic member (504) abutting against the core head (503); A separation ring (304) is provided inside the medicine cabin (300), and the separation ring (304) separates the internal space of the medicine cabin (300) into an independent accommodating chamber (S4); The core plug (501) and the elastic member (504) are both located inside the accommodating cavity (S4).

9. The microbiota transplanter according to claim 8, characterized in that: The extension mechanism (200) is a tubular structure, one end of which is provided with a pressure piece (201), and is connected to the syringe (100) through the pressure piece (201); The push head (102) is provided with an inner groove (102a), and the radial outer wall of the push rod (103) is provided with a limiter (103a), and the limiter (103a) is movably arranged inside the inner groove (102a); The housing (101) is also provided with a through port (101a), the pressure piece (201) is connected to the through port (101a), and the end of the push rod (103) can penetrate the through port (101a) and enter the interior of the pressure piece (201).

10. The microbiota transplanter according to claim 9, characterized in that: A pressing plug (201a) is arranged inside the pressure piece (201), and the pressure piece (201) between the pressing plug (201a) and the core plug (501), the extension mechanism (200), and the internal area of ​​the accommodating chamber (S4) constitute an air pressure chamber (S5).

Images