Traditional Chinese medicine composition with effects of improving sleep and resisting depression as well as application, preparation and preparation method of traditional Chinese medicine composition

Abstract

The invention discloses a traditional Chinese medicine composition with effects of improving sleep and resisting depression, and application, a preparation and a preparation method thereof, and relates to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition with the sleep improving and depression resisting effects is prepared from the following raw materials in parts by weight: 5 to 100 parts of semen ziziphi spinosae, 5 to 100 parts of semen persicae, 5 to 100 parts of fructus gardeniae, 5 to 100 parts of semen sojae praeparatum, 10 to 200 parts of concha ostreae, 1 to 20 parts of arillus longan and 1 to 10 parts of gamma-aminobutyric acid. The traditional Chinese medicine composition with the effects of improving sleep and resisting depression, which is provided by the technical scheme of the invention, can be used for remarkably shortening the sleep time of rats with insomnia and improving the content of 5-hydroxytryptamine, dopamine and melatonin in brain tissues, and is used for improving the symptoms such as difficulty in falling asleep, insomnia, easy waking, dreaminess and anxiety; the traditional Chinese medicine composition helps patients to improve sleep quality and accompanying depressive symptoms, and is small in side effect and stable in curative effect.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition having sleep-improving and anti-depression effects, and an application thereof, a preparation thereof, and a preparation method thereof. Background Art

[0002] With the accelerated pace of life and the increasing pressure of work and study, more and more people are troubled by sleep problems, including difficulty falling asleep, easy waking, and insufficient sleep time. Sleep disorders are strongly correlated with negative emotions and can be the cause of a series of mental health problems, such as depression and anxiety. Sleep disorders and depression overlap in the neural pathways and neurotransmitter systems (such as serotonin, dopamine, and melatonin) in brain tissue. The ability of people with depression to release serotonin in the brain is significantly lower than that of normal people. The imbalance of serotonin and dopamine in brain tissue can induce the occurrence of depression. In addition, studies have shown that the plasma melatonin level in the first-episode depression group is lower than that in the healthy control group. In addition, sleep disorders may also increase the risk of cardiovascular disease, endocrine disease, diabetes, and senile dementia.

[0003] At present, the clinical treatment of sleep disorders and depression mainly relies on chemical drug treatment. Chemical drugs are effective in the short-term treatment of insomnia, but long-term use is prone to tolerance, addiction and a series of adverse reactions, such as drowsiness, dizziness, ataxia, etc., which bring potential health risks to patients. Although there are some Chinese medicine compositions for treating insomnia or depression, for example, Chinese patent CN116139236A discloses a Chinese medicine composition for treating liver depression type insomnia, including a day prescription and a night prescription. The day prescription is made of the following raw materials in parts by weight: 3-15 parts of bupleurum, 12-24 parts of stir-fried white peony root, 20-40 parts of calcined dragon bone, 1-5 parts of amomum, 3-15 parts of processed cyperus, 3-15 parts of calamus, 2-10 parts of cicada shell, 2-10 parts of angelica, 3-15 parts of sophora flavescens, 3-15 parts of bombyx batryticatus, 1-20 parts of radix scutellariae ... The night prescription is prepared from the following raw materials in parts by weight: 3-15 parts of scutellaria, 1-5 parts of cinnamon, 3-15 parts of curcuma, 20-40 parts of salvia miltiorrhiza, 3-15 parts of processed cyperus, 3-15 parts of charred gardenia, 5-25 parts of Polygonum multiflorum, 12-24 parts of julibrissin bark, 3-15 parts of polygala, 5-25 parts of ganoderma, and 2-10 parts of licorice. The traditional Chinese medicine composition can relieve insomnia of liver depression type, but the traditional Chinese medicine composition has too many medicinal flavors, and it is necessary to distinguish between the day prescription and the night prescription, which brings inconvenience to patients. The Chinese patent CN118787701A discloses a Chinese medicine composition for improving insomnia, comprising the following raw material components by weight: 1-5 parts of Allium orchid, 1-20 parts of black red rose, 1-15 parts of Sappan wood, 1-15 parts of Magnolia grandiflora, 1-15 parts of Prunus mume and 1-15 parts of Eclipta prostrata. The Chinese medicine composition can relieve sleep disorders, but the effect of relieving sleep disorders is not obvious, and the efficacy on insomnia symptoms is not significant. In addition, some medicinal materials in the Chinese medicine composition are not easy to obtain, and it is difficult to scale up. Serving a large number of patients; Chinese patent CN114748570A discloses a traditional Chinese medicine composition for treating depression, which is mainly prepared from astragalus, codonopsis, ophiopogon, polygonum multiflorum, epimedium, kudzu root, angelica, salvia miltiorrhiza, gleditsia thorn, seaweed, kelp, oyster, and immature bitter orange. The traditional Chinese medicine composition can significantly increase the levels of 5-hydroxytryptamine and dopamine in the cortex and hippocampus of rats, and effectively treat depression. However, the traditional Chinese medicine composition requires the use of several precious medicinal materials, and the cost is high, which increases the cost of use for patients.

[0004] The causes of sleep disorders and depression are complex and diverse, and the treatment principles are also different. The use of medicines requires the combination of monarch, minister, assistant and envoy, as well as the dosage and proportion of the medicines. Traditional Chinese medicine treatment should consider the synergistic effect and integrity of different medicinal materials. For example, the Chinese medicine composition disclosed in Chinese patent CN116139236A is a prescription for solving sleep disorders for patients with insomnia caused by liver depression. It can achieve the effect of relieving depression and calming the mind by invigorating the spirit and regulating qi; for example, Chinese patent CN114748570A can achieve the effect of treating depression by invigorating qi and activating blood circulation, resolving phlegm and unblocking collaterals. Traditional Chinese medicine believes that depression is a symptom manifested by qi stagnation, liver failure to release, spleen failure to function properly, and heart failure to nourish. Both of them have the cause of "liver depression", and the treatment also has the effects of regulating qi and invigorating qi, soothing the liver and relieving depression, and calming the mind, but their target patient groups are different, the focus of treatment is different, and the prescriptions are also different.

[0005] In the context of high incidence of sleep disorders in the population and their frequent accompaniment by depression, it is of great significance to explore and develop Chinese herbal prescriptions that can be used to improve sleep disorders and accompanying depressive symptoms based on the theory of traditional Chinese medicine. Summary of the invention

[0006] The main purpose of the present invention is to provide a Chinese medicine composition with sleep-improving and anti-depressant effects, and its application, preparation and preparation method. The Chinese medicine composition with sleep-improving and anti-depressant effects can be used to treat sleep disorders and accompanying depressive symptoms, and has small toxic and side effects, can reduce the risk of addiction, has low production cost and definite efficacy.

[0007] To achieve the above object, the present invention provides a Chinese medicine composition having the effects of improving sleep and resisting depression, comprising the following raw materials in parts by weight: 5-100 parts of sour jujube seeds, 5-100 parts of peach kernels, 5-100 parts of gardenias, 5-100 parts of light black beans, 10-200 parts of oysters, 1-20 parts of longan meat, and 1-10 parts of gamma-aminobutyric acid.

[0008] In one embodiment, the Chinese medicine composition with sleep-improving and anti-depressant effects further comprises the following raw materials in parts by weight: 5-100 parts of bupleurum, 5-100 parts of julibrissin bark, and 5-100 parts of curcuma.

[0009] In one embodiment, the Chinese medicine composition with sleep-improving and anti-depressant effects further comprises the following raw materials in parts by weight: 5-100 parts of Poria cocos and 5-100 parts of Anemarrhena asphodeloides.

[0010] In one embodiment, the Chinese medicine composition having the effects of improving sleep and resisting depression comprises the following raw materials in parts by weight: 50-100 parts of spiny jujube seeds, 30-60 parts of peach kernels, 40-100 parts of gardenias, 40-100 parts of light fermented black beans, 100-200 parts of oysters, 10-20 parts of longan pulp, 50-100 parts of bupleurum, 50-100 parts of julibrissin barks, 50-100 parts of turmeric, 50-100 parts of poria, 50-100 parts of anemarrhena, and 2-4 parts of gamma-aminobutyric acid.

[0011] The present invention provides an application of the Chinese medicine composition with the effects of improving sleep and resisting depression in the preparation of a preparation for improving sleep disorders and resisting depression.

[0012] The present invention provides a method for preparing a preparation, comprising the following steps: S1. Obtaining a Chinese medicine composition according to the raw material ratio described in the above technical solution; S2, crushing the Chinese jujube seeds, peach kernels and gardenia in the traditional Chinese medicine composition respectively to obtain crushed Chinese jujube seeds, crushed peach kernels and crushed gardenia; S3, mixing the crushed material of spiny jujube seeds, crushed material of peach kernels, crushed material of gardenia, and each raw material in the Chinese medicine composition except γ-aminobutyric acid, spiny jujube seeds, peach kernels, and gardenia to obtain a mixture, soaking the mixture in water, heating and boiling, filtering the mixture to obtain a filtrate, and concentrating the filtrate under reduced pressure for the first time to obtain a first concentrated solution; S4, mixing the first concentrated solution with an ethanol solution to obtain a mixed solution, and concentrating the mixed solution under reduced pressure for a second time to obtain a second concentrated solution; S5. Mixing the second concentrated solution and γ-aminobutyric acid to obtain the preparation.

[0013] In one embodiment, in step S3: The mass of the water is 5 to 15 times that of the mixture; and / or, The soaking time is 20 to 90 minutes; and / or, The heating and boiling temperature is 80-120°C; and / or, The heating and boiling time is 40 to 120 min; and / or, The temperature of the first reduced pressure concentration is 60-90° C.; and / or, The pressure of the first reduced pressure concentration is -0.05~-0.1 MPa.

[0014] In one embodiment, in step S4: The ethanol solution is an ethanol aqueous solution with an ethanol volume fraction of 95%; and / or, The volume fraction of ethanol in the mixed solution is 20-60%; and / or, The temperature of the second reduced pressure concentration is 40-70° C.; and / or, The pressure of the second reduced pressure concentration is -0.05~-0.1 MPa.

[0015] In one embodiment, step S4 includes: The first concentrated solution is mixed with the ethanol solution to obtain a mixed solution, the mixed solution is allowed to stand for 12 to 24 hours and then filtered, the filtrate is collected, and the filtrate is concentrated under reduced pressure for a second time to obtain a second concentrated solution.

[0016] In one embodiment, step S5 comprises: The second concentrated solution, γ-aminobutyric acid, auxiliary materials and water are mixed to obtain the preparation.

[0017] In one embodiment, the auxiliary material comprises a sweetener and / or a preservative, wherein: The sweetener includes at least one of sucralose, xylitol, erythritol and aspartame; The preservative includes at least one of potassium sorbate, sorbic acid, sodium benzoate and sodium ethyl paraben.

[0018] The invention provides a preparation, which is prepared according to the preparation method.

[0019] The technical solution of the present invention has the following beneficial effects: The Chinese medicine composition with sleep-improving and anti-depressant effects provided by the technical solution of the present invention is based on the theory of traditional Chinese medicine, and is reasonably formulated. The raw materials are easy to obtain and the toxic and side effects are small. The Chinese medicine composition with sleep-improving and anti-depressant effects comprises raw materials including Chinese date seeds, peach seeds, gardenia, light black beans, oysters, longan meat and γ-aminobutyric acid, wherein Chinese date seeds belong to the liver, gallbladder, and heart meridians, nourish the heart and liver, calm the mind and tranquilize the nerves, astringe sweat, and promote body fluid; gardenia belongs to the heart, lung, and triple burner meridians, purges fire and eliminates restlessness, clears heat and removes dampness, cools blood and detoxifies; Chinese date seeds and gardenia are compatible, and play the effects of nourishing blood and astringing yin, clearing the heart and purging heat, eliminating restlessness and tranquilizing the nerves. Peach seeds belong to the heart, liver, and large intestine meridians, promote blood circulation and remove blood stasis, and moisten the intestines and promote bowel movements. Light black beans belong to the lungs and stomach meridians, relieve the exterior, eliminate restlessness, and promote the release of stagnant heat. Oysters belong to the liver, gallbladder, and kidney meridians, calm the nerves, subdue yang and replenish yin, and soften and disperse nodules. Longan meat is in the heart and spleen meridians, nourishes the heart and spleen, nourishes blood and calms the mind. γ-aminobutyric acid is an important inhibitory neurotransmitter of the central nervous system, which has physiological effects such as improving the body's sleep quality and lowering blood pressure. The Chinese medicine composition with the effect of improving sleep and anti-depression provided by the technical solution of the present invention is based on the Chinese jujube seeds as the main ingredient, nourishes the heart and liver, calms the mind and calms the mind, oysters, gardenias and light fermented black beans as the ministers, clears away heat and relieves restlessness, calms the mind, peach kernels, longan meat and γ-aminobutyric acid as the adjuvants, promotes blood circulation and nourishes blood, calms the mind and helps sleep, and all the medicines are used together to calm the mind and calm the mind, clear away heat and relieve restlessness, and can be used to solve the problem of difficulty in sleeping accompanied by depression. The Chinese medicine composition with the effect of improving sleep and anti-depression provided by the technical solution of the present invention can shorten the time for insomnia rats to fall asleep, increase the content of 5-hydroxytryptamine and dopamine in rat brain tissue, and increase the content of melatonin in rat serum, which is used to improve the insomnia symptoms and depression of insomnia rats. In addition, the present invention provides a method for preparing a preparation using the traditional Chinese medicine composition, which has a simple preparation process, strong operability, low cost, and is easy to industrialize. The content of active ingredients in the obtained preparation is high, and it is suitable for large-scale promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. 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 the structures shown in these drawings without paying creative work.

[0021] Figure 1 The result diagram of the sleep onset time of rats in the normal group, model group, Example 1 group, Example 2 group, Example 3 group and Example 4 group (* indicates the difference compared with the model group, * indicates P < 0.05, ** indicates P < 0.01); Figure 2The figure is a graph of the content of 5-hydroxytryptamine in the brain tissue of rats in the normal group, the model group, the example 1 group, the example 2 group, the example 3 group and the example 4 group (* is the difference compared with the model group, * is P < 0.05, ** is P < 0.01); Figure 3 The dopamine content in the brain tissue of rats in the normal group, model group, Example 1 group, Example 2 group, Example 3 group and Example 4 group (* indicates the difference compared with the model group, * indicates P < 0.05, ** indicates P < 0.01); Figure 4 The figure is a graph of melatonin content in the serum of rats in the normal group, model group, Example 1 group, Example 2 group, Example 3 group and Example 4 group (* is the difference compared with the model group, * is P < 0.05, ** is P < 0.01).

[0022] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0023] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] It should be noted that if the embodiments of the present invention involve directional indications (such as up, down, left, right, front, back, etc.), the directional indications are only used to explain the relative position relationship, movement status, etc. between the components in a certain specific posture. If the specific posture changes, the directional indication will also change accordingly.

[0025] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present invention, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of technical features indicated. Therefore, the features limited to "first" and "second" may explicitly or implicitly include at least one of the features. In addition, if "and / or" or "and / or" appears in the full text, its meaning includes three parallel solutions. Taking "A and / or B" as an example, it includes solution A, solution B, or solutions that satisfy both A and B. In addition, the technical solutions between the various embodiments can be combined with each other, but it must be based on the ability of ordinary technicians in this field to implement. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0026] With the accelerated pace of life and the increasing pressure of work and study, more and more people are troubled by sleep problems, including difficulty falling asleep, easy waking, and insufficient sleep time. Sleep disorders are strongly correlated with negative emotions and can be the cause of a series of mental health problems, such as depression and anxiety. Sleep disorders and depression overlap in the neural pathways and neurotransmitter systems (such as serotonin, dopamine, and melatonin) in the brain. The ability of people with depression to release serotonin in the brain is significantly lower than that of normal people. The imbalance of serotonin and dopamine in brain tissue can induce the occurrence of depression. In addition, studies have shown that the plasma melatonin level in the first-episode depression group is lower than that in the healthy control group. In addition, sleep disorders may also increase the risk of cardiovascular disease, endocrine disease, diabetes, and senile dementia.

[0027] At present, the clinical treatment of sleep disorders mainly relies on drug therapy, including benzodiazepines such as estazolam, midazolam, temazepam, lorazepam, alprazolam, diazepam, and non-benzodiazepines such as zolpidem, zopiclone, dexpiclone, and zaleplon. At present, the drugs used in the clinical treatment of depression mainly include selective serotonin reuptake inhibitors (SSRIs), tricyclic antidepressants (TCAs), monoamine oxidase inhibitors (MAOIs), serotonin and norepinephrine reuptake inhibitors (SNRIs), noradrenergic and specific serotonergic antidepressants (NaSSAs), etc. Although chemical drugs are effective in the short term in the treatment of insomnia and depression, long-term use is prone to tolerance, addiction, and a series of adverse reactions, such as drowsiness, dizziness, ataxia, etc., which bring potential health risks to patients. Although there are many traditional Chinese medicine compositions for treating insomnia, the existing traditional Chinese medicines and their compound preparations have no obvious sedative effect, and the efficacy on insomnia and depression symptoms is not significant. In addition, a variety of auxiliary materials need to be added during the production process of the preparations, which increases the patient's medication dosage and usage cost. Therefore, in the context of the high incidence of sleep disorders in the Chinese population and the frequent occurrence of depression, it is of great significance to explore and develop traditional Chinese medicine prescriptions that can be used to improve sleep disorders and the accompanying depression symptoms based on the theory of traditional Chinese medicine.

[0028] In view of this, the present invention proposes a Chinese medicine composition with the effects of improving sleep and resisting depression, wherein the Chinese medicine composition with the effects of improving sleep and resisting depression comprises the following raw materials in parts by weight: 5-100 parts of sour jujube seeds, 5-100 parts of peach kernels, 5-100 parts of gardenias, 5-100 parts of light black beans, 10-200 parts of oysters, 1-20 parts of longan meat, and 1-10 parts of gamma-aminobutyric acid.

[0029] The Chinese medicine composition with the effects of improving sleep and anti-depression provided by the technical solution of the present invention is based on the theory of traditional Chinese medicine, and is reasonably formulated. The raw materials are easy to obtain and the toxic and side effects are small. The Chinese medicine composition with the effects of improving sleep and anti-depression comprises raw materials including Chinese date seeds, peach seeds, gardenia, light black beans, oysters, longan meat and γ-aminobutyric acid. Among them, Chinese date seeds belong to the liver, gallbladder, and heart meridians, nourish the heart and liver, calm the mind and tranquilize the nerves, astringe sweat, and promote body fluid; gardenia belongs to the heart, lung, and triple burner meridians, purges fire and eliminates restlessness, clears heat and removes dampness, cools blood and detoxifies. Chinese date seeds and gardenia are compatible, and they play the effects of nourishing blood and astringing yin, clearing the heart and purging heat, eliminating restlessness and tranquilizing the nerves. Peach seeds belong to the heart, liver, and large intestine meridians, promote blood circulation and remove blood stasis, and moisten the intestines and promote bowel movements. Light black beans belong to the lungs and stomach meridians, relieve the exterior, eliminate restlessness, and promote the release of stagnant heat. Oysters belong to the liver, gallbladder, and kidney meridians, calm the nerves, subdue yang and replenish yin, and soften and disperse nodules. Longan meat belongs to the heart and spleen meridians, nourishes the heart and spleen, nourishes blood and calms the mind. γ-aminobutyric acid is an important inhibitory neurotransmitter of the central nervous system, which can be used to improve the body's sleep quality and lower blood pressure. The Chinese medicine composition with the effects of improving sleep and anti-depression, with Chinese jujube seeds as the main, nourishing the heart and liver, calming the mind and calming the mind, oysters, gardenias, light black beans, etc. as the ministers, clearing away heat and relieving restlessness, calming the mind, peach kernels, longan meat and γ-aminobutyric acid as the adjuvants, promoting blood circulation and nourishing blood, calming the mind and helping sleep, all the medicines are used together, calming the mind and calming the mind, clearing away heat and relieving restlessness, and can be used to solve the problem of difficulty in sleeping accompanied by depression. The Chinese medicine composition provided by the technical solution of the present invention can shorten the time for insomnia rats to fall asleep, increase the content of 5-hydroxytryptamine and dopamine in rat brain tissue, and increase the content of melatonin in rat serum, which is used to improve the insomnia symptoms and depression of insomnia rats.

[0030] In an embodiment of the present invention, the Chinese medicine composition with sleep improving and anti-depressant effects comprises the following raw materials in parts by weight: 5-100 parts of Chinese jujube seeds, 5-100 parts of peach kernels, 5-100 parts of gardenias, 5-100 parts of light black beans, 10-200 parts of oysters, 1-20 parts of longan pulp, 5-100 parts of bupleurum, 5-100 parts of julibrissin bark, 5-100 parts of curcuma, and 1-10 parts of γ-aminobutyric acid.

[0031] In order to further improve the effect of improving insomnia symptoms and depression in insomnia rats, the Chinese medicine composition with sleep improvement and anti-depression effects also includes the following raw materials in parts by weight: 5-100 parts of bupleurum, 5-100 parts of jujube bark, and 5-100 parts of turmeric, that is, the raw materials included in the Chinese medicine composition are spiny jujube seeds, peach kernels, gardenia, light black beans, oysters, longan meat, bupleurum, jujube bark, turmeric and gamma-aminobutyric acid. Spiny jujube seeds belong to the liver, gallbladder, and heart meridians, nourish the heart and liver, calm the mind and tranquilize the nerves, astringe sweat, and promote body fluid; gardenia belongs to the heart, lung, and triple burner meridians, purge fire and eliminate restlessness, clear heat and remove dampness, cool blood and detoxify. Spiny jujube seeds and gardenia are compatible, and they play the effects of nourishing blood and astringing yin, clearing the heart and purging heat, and eliminating restlessness and tranquilizing the nerves. Peach kernel belongs to the heart, liver, and large intestine meridians, promotes blood circulation and removes blood stasis, and moistens the intestines and promotes bowel movements. Light black beans belong to the lung and stomach meridians, relieve the exterior, eliminate restlessness, and promote the release of stagnant heat. Oysters belong to the liver, gallbladder, and kidney meridians, calming the nerves, suppressing yang and replenishing yin, softening and dispersing nodules. Longan meat belongs to the heart and spleen meridians, nourishing the heart and spleen, nourishing blood and calming the nerves. Bupleurum belongs to the liver, gallbladder, and lung meridians, reconciling the exterior and interior, soothing the liver and relieving depression; Albizzia bark belongs to the heart, liver, and lung meridians, relieving depression and calming the nerves, promoting blood circulation and reducing swelling; Bupleurum and Albizzia bark are used together to soothe the liver and relieve depression, and relieve depression and calm the nerves. Curcuma belongs to the liver, heart, and lung meridians, promoting qi and relieving depression, clearing the heart and cooling blood. γ-Aminobutyric acid is an important inhibitory neurotransmitter in the central nervous system, which can be used to improve the body's sleep quality and lower blood pressure. The Chinese medicine composition uses Semen Ziziphi Spinosae as the main ingredient, nourishing the heart and liver, calming the mind and tranquilizing the nerves; Oyster, Gardenia and Fructus Sojae are the subordinate ingredients, clearing away heat and relieving restlessness, calming the nerves and tranquilizing the nerves; Peach kernel, Longan pulp and γ-aminobutyric acid are the adjuvants, promoting blood circulation and nourishing blood, tranquilizing the nerves and promoting sleep; Bupleurum, Albizzia Julibrissin and Curcuma Radix are used to enhance the effect of soothing the liver and relieving depression. All the medicines are used together to calm the mind and tranquilize the nerves, clear away heat and relieve restlessness, and can be used to solve the problem of difficulty in sleeping accompanied by depression. The raw materials of the prescription are easy to obtain, the toxic and side effects are small, and the sleeping time of insomnia rats can be shortened, the content of serotonin and dopamine in the rat brain tissue is increased, and the content of melatonin in the rat serum is increased, and it is used to improve the insomnia symptoms and depression of insomnia rats.

[0032] In an embodiment of the present invention, the Chinese medicine composition having the effects of improving sleep and resisting depression comprises the following raw materials in parts by weight: 5-100 parts of spiny jujube seeds, 5-100 parts of peach kernels, 5-100 parts of gardenias, 5-100 parts of light fermented black beans, 10-200 parts of oysters, 1-20 parts of longan pulp, 5-100 parts of bupleurum, 5-100 parts of julibrissin barks, 5-100 parts of turmeric, 5-100 parts of poria, 5-100 parts of anemarrhena, and 1-10 parts of gamma-aminobutyric acid.

[0033] In order to further improve the effect of improving insomnia symptoms and depression in insomnia rats, the Chinese medicine composition with sleep improvement and anti-depression effects also includes the following raw materials in parts by weight: 5-100 parts of Poria cocos and 5-100 parts of Rhizoma Anemarrhenae, that is, the raw materials included in the Chinese medicine composition formula are Semen Ziziphi Spinosae, Semen Pruni, Gardenia, Fructus Sojae, Oyster, Longan Meat, Bupleurum, Cortex Albizziae, Curcuma, Poria cocos, Rhizoma Anemarrhenae and γ-aminobutyric acid. Semen Ziziphi Spinosae belongs to the liver, gallbladder, and heart meridians, nourishes the heart and liver, calms the mind and tranquilizes the nerves, astringes sweat, and promotes fluid production; Gardenia jasminoides belongs to the heart, lung, and triple burner meridians, purges fire and eliminates restlessness, clears heat and removes dampness, cools blood and detoxifies. Semen Ziziphi Spinosae and Gardenia jasminoides are compatible, and they play the effects of nourishing blood and astringing yin, clearing the heart and purging heat, and eliminating restlessness and tranquilizing the nerves. Semen Pruni belongs to the heart, liver, and large intestine meridians, promotes blood circulation and removes blood stasis, and moistens the intestines and promotes bowel movements. Fructus Sojae Sojae belongs to the lung and stomach meridians, relieves exterior symptoms, eliminates restlessness, and promotes the release of stagnant heat. Oysters belong to the liver, gallbladder, and kidney meridians, calming the mind, suppressing yang and replenishing yin, softening and dispersing nodules. Longan meat belongs to the heart and spleen meridians, nourishing the heart and spleen, nourishing blood and calming the mind. Bupleurum belongs to the liver, gallbladder, and lung meridians, reconciling the exterior and interior, soothing the liver and relieving depression; Albizzia bark belongs to the heart, liver, and lung meridians, relieving depression and calming the mind, promoting blood circulation and reducing swelling; Bupleurum and Albizzia bark are used together to soothe the liver and relieve depression, and relieve depression and calm the mind. Curcuma belongs to the liver, heart, and lung meridians, promoting qi and relieving depression, clearing the heart and cooling blood. Poria belongs to the lung, stomach, and kidney meridians, clearing heat and purging fire, nourishing yin and moistening dryness; Oysters belong to the liver, gallbladder, and kidney meridians, calming the mind, suppressing yang and replenishing yin, softening and dispersing nodules; Poria and oysters are used together to strengthen the spleen and calm the mind, nourish yin, and calm the mind. Anemarrhena belongs to the lung, stomach, and kidney meridians, clearing heat and purging fire, nourishing yin and moistening dryness. γ-aminobutyric acid is an important inhibitory neurotransmitter of the central nervous system, which has physiological effects such as improving the quality of sleep and lowering blood pressure. The Chinese medicine composition uses Semen Ziziphi Spinosae as the main ingredient, nourishing the heart and liver, calming the mind and tranquilizing the nerves, oyster, gardenia and light fermented black beans as the subordinate ingredients, clearing away heat and relieving restlessness, calming the nerves, peach kernel, longan meat and γ-aminobutyric acid as the assistant ingredients, promoting blood circulation and nourishing blood, tranquilizing the nerves and helping sleep, Bupleurum, Albizzia Julibrissin and Curcuma Radix are used to enhance the effect of soothing the liver and relieving depression, Poria and Rhizoma Anemarrhenae are used to enhance the effect of removing dampness and removing heat, tranquilizing the nerves, and taking into account the coordination of internal organs such as spleen, lung and kidney. All the medicines are used together to calm the mind and tranquilize the nerves, clear away heat and relieve restlessness, and can be used to solve the problem of difficulty in sleeping accompanied by depression. The raw materials of the prescription are easy to obtain, the toxic and side effects are small, and the time of falling asleep of insomnia rats can be shortened, the content of 5-hydroxytryptamine and dopamine in rat brain tissue can be increased, and the content of melatonin in rat serum can be increased, and the insomnia symptoms and depression of insomnia rats can be improved.

[0034] In an embodiment of the present invention, the Chinese medicine composition having the effects of improving sleep and resisting depression comprises the following raw materials in parts by weight: 50-100 parts of spiny jujube seeds, 30-60 parts of peach kernels, 40-100 parts of gardenia, 40-100 parts of light fermented black beans, 100-200 parts of oysters, 10-20 parts of longan pulp, 50-100 parts of bupleurum, 50-100 parts of albizzia bark, 50-100 parts of turmeric, 50-100 parts of tuckahoe, 50-100 parts of rhizome of anemarrhena, and 2-4 parts of gamma-aminobutyric acid. Under this raw material ratio, the Chinese medicine composition with sleep-improving and anti-depressant effects has a better effect in treating insomnia or depression.

[0035] The present invention provides an application of the Chinese medicine composition with the effects of improving sleep and resisting depression in the preparation of a preparation for improving sleep disorders and resisting depression.

[0036] The traditional Chinese medicine composition provided by the technical solution of the present invention can shorten the sleep time of insomnia rats, increase the content of serotonin and dopamine in the rat brain tissue, increase the content of melatonin in the rat serum, and finally improve the insomnia symptoms and depression of the insomnia rats, and can be used to prepare a preparation for treating insomnia or depression.

[0037] The present invention provides a method for preparing a preparation, comprising the following steps: S1. Obtaining a Chinese medicine composition according to the raw material ratio; S2, crushing the Chinese jujube seeds, peach kernels and gardenia in the traditional Chinese medicine composition respectively to obtain crushed Chinese jujube seeds, crushed peach kernels and crushed gardenia; S3, mixing the crushed material of spiny jujube seeds, crushed material of peach kernels, crushed material of gardenia, and each raw material in the Chinese medicine composition except γ-aminobutyric acid, spiny jujube seeds, peach kernels, and gardenia to obtain a mixture, soaking the mixture in water, heating and boiling, filtering the mixture to obtain a filtrate, and concentrating the filtrate under reduced pressure for the first time to obtain a first concentrated solution; S4, mixing the first concentrated solution with an ethanol solution to obtain a mixed solution, and concentrating the mixed solution under reduced pressure for a second time to obtain a second concentrated solution; S5. Mixing the second concentrated solution and γ-aminobutyric acid to obtain the preparation.

[0038] In the technical solution of the present invention, the preparation method provided first extracts and concentrates the Chinese medicine composition under reduced pressure for the first time, and then uses an ethanol solution to perform a second reduced pressure concentration to achieve secondary extraction, thereby improving the extraction rate of the Chinese medicine composition, improving the clarity of the final prepared preparation, and reducing precipitation during subsequent storage. The preparation method of the present invention does not require the use of special extraction reagents, has a simple preparation process, strong operability, low cost, and is easy to industrialize. The content of active ingredients in the obtained preparation is high, and it is suitable for large-scale promotion.

[0039] In an embodiment of the present invention, in step S3, the mass of the water is 5 to 15 times that of the mixture.

[0040] In an embodiment of the present invention, in step S3, the soaking treatment time is 20 to 90 minutes.

[0041] In an embodiment of the present invention, in step S3, the temperature of the heating and boiling is 80-120°C.

[0042] In an embodiment of the present invention, in step S3, the heating and boiling time is 40 to 120 minutes.

[0043] In an embodiment of the present invention, in step S3, the temperature of the first vacuum concentration is 60-90°C; the pressure of the first vacuum concentration is -0.05--0.1 MPa. After the first vacuum concentration, the mass ratio of the first concentrated liquid to the mixture is 1:1-3:1.

[0044] In an embodiment of the present invention, in step S4, the ethanol solution is an ethanol aqueous solution with an ethanol volume fraction of 95%.

[0045] In an embodiment of the present invention, in step S4, the volume fraction of ethanol in the mixed solution is 20-60%. By controlling the amount of ethanol solution added so that the volume fraction of ethanol in the mixed solution is 20-60%, the volume fraction of ethanol in the mixed solution is controlled within the above range, and the extraction efficiency of the second reduced pressure concentration is higher.

[0046] In an embodiment of the present invention, in step S4, the temperature of the second vacuum concentration is 40-70° C., and the pressure of the second vacuum concentration is -0.05--0.1 MPa. After the second vacuum concentration, the second concentrated liquid does not contain ethanol.

[0047] In an embodiment of the present invention, step S4 includes: The first concentrated solution is mixed with the ethanol solution to obtain a mixed solution, the mixed solution is allowed to stand for 12 to 24 hours and then filtered, the filtrate is collected, and the filtrate is concentrated under reduced pressure for a second time to obtain a second concentrated solution. Precipitating the mixed solution can improve the extraction rate of the Chinese medicine composition, improve the clarity of the final prepared preparation, and reduce precipitation during subsequent storage.

[0048] In an embodiment of the present invention, step S5 includes: The second concentrated solution, γ-aminobutyric acid, auxiliary materials and water are mixed to obtain the preparation.

[0049] In an embodiment of the present invention, the auxiliary material comprises a sweetener and / or a preservative, wherein: The sweetener includes at least one of sucralose, xylitol, erythritol and aspartame; The preservative includes at least one of potassium sorbate, sorbic acid, sodium benzoate and sodium ethyl paraben.

[0050] In an embodiment of the present invention, the sweetener is composed of sucralose and xylitol, and the preservative is potassium sorbate, wherein: The weight proportion of the sucralose is 0.10-0.20 parts, the weight proportion of the xylitol is 20-60 parts, and the weight proportion of the preservative is 0.10-0.60 parts.

[0051] The invention provides a preparation, which is prepared according to the preparation method.

[0052] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments and drawings. It should be understood that the following embodiments are only used to explain the present invention and are not used to limit the present invention.

[0053] Example 1 A Chinese medicine composition having the effects of improving sleep and resisting depression, the Chinese medicine composition comprising the following raw materials in parts by weight: 50 parts of spiny jujube seeds, 30 parts of peach kernels, 50 parts of gardenias, 50 parts of light fermented black beans, 100 parts of oysters, 10 parts of longan pulps, and 2 parts of gamma-aminobutyric acid.

[0054] Example 2 A Chinese medicine composition having the effects of improving sleep and resisting depression, the Chinese medicine composition comprising the following raw materials in parts by weight: 100 parts of spiny jujube seeds, 60 parts of peach kernels, 100 parts of gardenias, 100 parts of light fermented black beans, 200 parts of oysters, 20 parts of longan pulps, and 4 parts of gamma-aminobutyric acid.

[0055] Example 3 A Chinese medicine composition having the effects of improving sleep and resisting depression, the Chinese medicine composition comprising the following raw materials in parts by weight: 50 parts of spiny jujube seeds, 30 parts of peach kernels, 50 parts of gardenias, 50 parts of light fermented black beans, 100 parts of oysters, 10 parts of longan pulps, 50 parts of bupleurum, 50 parts of julibrissin barks, 50 parts of turmeric, and 2 parts of gamma-aminobutyric acid.

[0056] Example 4 A Chinese medicine composition having the effects of improving sleep and resisting depression, the Chinese medicine composition comprising the following raw materials in parts by weight: 50 parts of spiny jujube seeds, 30 parts of peach kernels, 50 parts of gardenias, 50 parts of light fermented black beans, 100 parts of oysters, 10 parts of longan pulps, 50 parts of bupleurum, 50 parts of julibrissin barks, 50 parts of turmeric, 50 parts of poria, 50 parts of anemarrhena, and 2 parts of gamma-aminobutyric acid.

[0057] Example 5 A method for preparing a preparation comprises the following steps: (1) preparing the Chinese medicine composition of Example 1; (2) crushing the Chinese jujube seeds, peach kernels and gardenia jasminoides in the traditional Chinese medicine composition to obtain crushed Chinese jujube seeds, crushed peach kernels and crushed gardenia jasminoides; (3) adding the above-mentioned crushed material and the remaining raw materials except γ-aminobutyric acid into water with the mass of water being 10 times the total weight of the raw materials, boiling once for 1 hour, filtering, and concentrating the filtrate under reduced pressure for the first time to obtain a first concentrated solution; (4) mixing the first concentrated solution with a 95% by volume ethanol solution so that the volume fraction of ethanol in the mixed solution is 30%, leaving the solution at room temperature for 24 hours, centrifuging, filtering, collecting the filtrate, and concentrating the filtrate under reduced pressure for a second time to obtain a second concentrated solution; (5) The second concentrated solution, γ-aminobutyric acid, 0.10 parts of sucralose, 40 parts of xylitol and 0.30 parts of potassium sorbate were mixed, purified water was added to make up to 1000 mL, and sterilized to obtain the preparation.

[0058] Animal experiments 1. Materials 1.1 Drugs and reagents Test drug 1: a preparation prepared from the Chinese medicine composition of Example 1 according to the method of implementation 5, test drug 2: a preparation prepared from the Chinese medicine composition of Example 2 according to the method of Example 5, test drug 3: a preparation prepared from the Chinese medicine composition of Example 3 according to the method of Example 5, test drug 4: a preparation prepared from the Chinese medicine composition of Example 4 according to the method of Example 5, normal saline, PBS (0.01 M. pH = 7.4), p-chlorophenylalanine PCPA (Aladdin), Tween 80 (Aladdin), chloral hydrate (McLean), dopamine and serotonin kits, melatonin enzyme-linked kit.

[0059] 1.2 Animals The experimental mice were 12 SPF-grade 6-week-old male rats weighing 180±15 g and 12 6-week-old female rats weighing 150±12 g. They were adaptively fed for 1 week and entered the experimental period after adapting to the environment. All animal experiments were in accordance with and strictly abided by the relevant guidelines of the China Animal Protection Association.

[0060] 2. Experimental Methods 2.1 Grouping, modeling and drug administration Rats were randomly divided into a normal group, a model group, an example 1 group, an example 2 group, an example 3 group, and an example 4 group, with 4 rats in each group. The animals in the model group, the example 1 group, and the example 2 group were intraperitoneally injected with a suspension of PCPA (60 mg / mL) at 8:00 every morning for 3 consecutive days, with an injection dose of 0.5 mL / 100g, and the normal group was intraperitoneally injected with the same volume of saline as PCPA. When the model group rats showed disappearance of circadian rhythm and continuous activity during the day and night, it indicated that the insomnia model was successfully established. Chloral hydrate was used for auxiliary verification, and the time of disappearance of the righting reflex in the model group and the normal group was recorded.

[0061] After successful modeling, medication was started. The blank group and the model group were gavaged with normal saline for 7 consecutive days; the Example 1 group was gavaged with test drug 1 for 7 consecutive days at 0.5 mL / 100 g / d; the Example 2 group was gavaged with test drug 2 for 7 consecutive days at 0.5 mL / 100 g / d; the Example 3 group was gavaged with test drug 3 for 7 consecutive days at 0.5 mL / 100 g / d; the Example 4 group was gavaged with test drug 4 for 7 consecutive days at 0.5 mL / 100 g / d. The dosage was obtained by converting the dosage for animals to that for humans.

[0062] 2.2 Sleep latency measurement 30 minutes after the last oral administration of the test sample, the animals in each group were intraperitoneally injected with 0.45 mL / 100 g of 10% chloral hydrate. The sleep latency of the rats in each group was observed and recorded based on the disappearance of the righting reflex.

[0063] 2.3 Determination of 5-HT and dopamine content in brain tissue The brain tissue was removed and the cerebral cortex, hypothalamus, hippocampus and brain stem were separated. After being washed with normal saline / PBS, the weights were weighed and recorded respectively. The contents of 5-hydroxytryptamine and dopamine in the brain tissue were determined by Elisa method.

[0064] 2.4 Serum melatonin content determination After anesthesia, the abdominal cavity was opened, blood was collected from the abdominal aorta, and centrifuged at 3000 r / min for 10 min at 4°C. The supernatant was taken and the melatonin content in serum was determined by Elisa method.

[0065] 3. Results 3.1 Sleep latency Figure 1 Result diagram of the sleep onset time of rats in the normal group, model group, Example 1 group, Example 2 group, Example 3 group and Example 4 group; Figure 1It can be seen that compared with the model group, the sleep latency of rats in Example 1, Example 2, Example 3 and Example 4 groups was reduced, among which the Example 2 group was significantly reduced (P < 0.01). This result shows that the test drug 1, the test drug 2, the test drug 3 and the test drug 4 can shorten the sleep time of insomnia rats. Figure 1 It can also be seen that the sleep time of Example 3 group and Example 4 group is shorter than that of Example 1 group. This result shows that the effect of test drug 3 and test drug 4 in shortening the sleep time is better than that of test drug 1.

[0066] 3.2 Contents of 5-HT and dopamine in brain tissue Figure 2 is a graph showing the content of 5-hydroxytryptamine in the brain tissue of rats in the normal group, the model group, the Example 1 group, the Example 2 group, the Example 3 group, and the Example 4 group. Figure 3 The figure is a graph showing the dopamine content in the rat brain tissue in the normal group, the model group, the Example 1 group, the Example 2 group, the Example 3 group, and the Example 4 group. Figure 2 and Figure 3 It can be seen that compared with the normal group, the 5-hydroxytryptamine content and dopamine content in the brain tissue (hypothalamus, hippocampus and brainstem) of the model group were significantly reduced (P < 0.05); compared with the model group, the 5-hydroxytryptamine and dopamine content in the brain tissue of rats in Example 1, Example 2, Example 3 and Example 4 groups were increased, among which the content in Example 2, Example 3 and Example 4 groups was significantly increased (P < 0.05). This result shows that test drug 1, test drug 2, test drug 3 and test drug 4 can effectively improve the levels of 5-hydroxytryptamine and dopamine in the brain tissue of rats in the model group, thereby having a certain improvement effect on the depression of insomnia rats. Figure 2 and Figure 3 It can also be seen that the 5-hydroxytryptamine content of Example 3 and Example 4 groups is higher than that of Example 1 group, and the dopamine content of Example 3 and Example 4 groups is also higher than that of Example 1 group. This result shows that the effects of test drug 3 and test drug 4 in increasing the 5-hydroxytryptamine level are better than those of test drug 1, and the effects of test drug 3 and test drug 4 in increasing the dopamine content are better than those of test drug 1.

[0067] 3.3 Melatonin content in serum Figure 4 is a graph showing the content of melatonin in serum of the normal group, the model group, the Example 1 group, the Example 2 group, the Example 3 group and the Example 4 group. Figure 4It can be seen that compared with the normal group, the melatonin content in the serum of the model group was significantly reduced (P < 0.05); compared with the model group, the melatonin content in the serum of Example 1, Example 2, Example 3 and Example 4 groups were significantly increased (P < 0.05). This result shows that the test drug can effectively improve the level of melatonin in the serum of model rats, thereby having a certain improvement effect on the depression of insomnia rats. Figure 4 It can also be seen that the melatonin content of Example 3 and Example 4 groups is higher than that of Example 1 group. This result shows that the effects of test drug 3 and test drug 4 on upregulating melatonin levels are better than test drug 1.

[0068] In summary, the present invention provides a Chinese medicine composition with sleep-improving and anti-depressant effects, and also provides a method for preparing a reagent using the Chinese medicine composition with sleep-improving and anti-depressant effects as a raw material. The effects of the Chinese medicine composition on the sleep-onset time of insomnia model rats, the 5-hydroxytryptamine content and dopamine content in rat brain tissue, and the melatonin content in serum are studied. The results show that the Chinese medicine composition can shorten the sleep-onset time of insomnia rats, increase the 5-hydroxytryptamine content and dopamine content in rat brain tissue, increase the melatonin content in rat serum, and ultimately improve the insomnia symptoms and depression of insomnia rats.

[0069] The above description is only an exemplary embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent structural changes made by using the contents of the present invention specification and drawings under the technical concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A Chinese medicine composition having sleep-improving and anti-depressant effects, characterized in that: The invention comprises the following raw materials in parts by weight: 5-100 parts of sour jujube seeds, 5-100 parts of peach kernels, 5-100 parts of gardenias, 5-100 parts of light black beans, 10-200 parts of oysters, 1-20 parts of longan meat, and 1-10 parts of gamma-aminobutyric acid.

2. The Chinese medicine composition having sleep-improving and anti-depressant effects according to claim 1, characterized in that: The invention also comprises the following raw materials in parts by weight: 5-100 parts of bupleurum root, 5-100 parts of julibrissin bark, and 5-100 parts of curcuma.

3. The Chinese medicine composition having sleep-improving and anti-depressant effects as claimed in claim 2, characterized in that: The invention also comprises the following raw materials in parts by weight: 5-100 parts of Poria cocos and 5-100 parts of Rhizoma Anemarrhenae.

4. The Chinese medicine composition having sleep-improving and anti-depressant effects as claimed in claim 3, characterized in that: The invention comprises the following raw materials in parts by weight: 50-100 parts of spiny jujube seeds, 30-60 parts of peach kernels, 40-100 parts of gardenias, 40-100 parts of light fermented black beans, 100-200 parts of oysters, 10-20 parts of longan pulp, 50-100 parts of bupleurum, 50-100 parts of julibrissin barks, 50-100 parts of turmeric, 50-100 parts of poria, 50-100 parts of anemarrhena, and 2-4 parts of gamma-aminobutyric acid.

5. Use of the Chinese medicinal composition having sleep-improving and anti-depressant effects as claimed in any one of claims 1 to 4 in the preparation of a preparation for improving sleep disorders and anti-depressant effects.

6. A method for preparing a preparation, characterized in that: The following steps are involved: S1. Obtaining a Chinese medicine composition according to the raw material ratio described in any one of claims 1 to 4; S2, crushing the Chinese jujube seeds, peach kernels and gardenia in the traditional Chinese medicine composition respectively to obtain crushed Chinese jujube seeds, crushed peach kernels and crushed gardenia; S3, mixing the crushed material of spiny jujube seeds, crushed material of peach kernels, crushed material of gardenia, and each raw material in the Chinese medicine composition except γ-aminobutyric acid, spiny jujube seeds, peach kernels, and gardenia to obtain a mixture, soaking the mixture in water, heating and boiling, filtering the mixture to obtain a filtrate, and concentrating the filtrate under reduced pressure for the first time to obtain a first concentrated solution; S4, mixing the first concentrated solution with an ethanol solution to obtain a mixed solution, and concentrating the mixed solution under reduced pressure for a second time to obtain a second concentrated solution; S5. Mixing the second concentrated solution and γ-aminobutyric acid to obtain the preparation.

7. The preparation method according to claim 6, characterized in that: In step S3: The mass of the water is 5 to 15 times that of the mixture; and / or, The soaking time is 20 to 90 minutes; and / or, The heating and boiling temperature is 80-120°C; and / or, The heating and boiling time is 40 to 120 min; and / or, The temperature of the first reduced pressure concentration is 60-90° C.; and / or, The pressure of the first reduced pressure concentration is -0.05 ~ -0.1 MPa.

8. The preparation method according to claim 6, characterized in that: In step S4: The ethanol solution is an ethanol aqueous solution with an ethanol volume fraction of 95%; and / or, The volume fraction of ethanol in the mixed solution is 20-60%; and / or, The temperature of the second reduced pressure concentration is 40-70° C.; and / or, The pressure of the second reduced pressure concentration is -0.05 ~ -0.1 MPa.

9. The preparation method according to claim 6, characterized in that: The step S4 comprises: The first concentrated solution is mixed with the ethanol solution to obtain a mixed solution, the mixed solution is allowed to stand for 12 to 24 hours and then filtered, the filtrate is collected, and the filtrate is concentrated under reduced pressure for a second time to obtain a second concentrated solution.

10. A preparation, characterized in that The preparation is prepared according to the preparation method according to any one of claims 6 to 9.

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