WO2020211571A1 - Soil preparation capable of preventing and treating neuroendocrine disorder-related diseases, preparation method therefor and application thereof - Google Patents

Abstract

A soil preparation, which is prepared from sterilized soil and oral probiotics or auxiliary materials, and an application of the soil preparation in preparing a drug for preventing and treating neuroendocrine disorder-related diseases.

Description

Soil preparation capable of preventing and treating neuroendocrine disorders related diseases, and preparation method and application thereof Technical field

The invention relates to a soil preparation, in particular to a soil preparation that can be used to prevent and treat neuroendocrine disorders related diseases and a preparation method thereof.

Background technique

Interstitial cystitis refers to a disease with urinary discomfort, manifested by varying degrees of frequency, urgency, and often accompanied by varying degrees of pain in the bladder area, upper or lower abdomen, and other symptoms. The medical history exceeds 6 weeks, and infection and other diseases must be ruled out before the diagnosis of interstitial cystitis can be made. Interstitial cystitis is often associated with lower urinary tract symptoms. The two main symptoms of the disease are frequent urination and pain, which seriously affect the quality of life of patients. More than 80% of patients have different degrees of sleep disturbance, depression and anxiety, and the prevalence is increasing year by year.

There have been a large number of studies on the causes of interstitial cystitis, but there is still no uniform etiological standard. Neurobiological pathogenesis believes that when sensory nerves are activated, they release signal molecules such as substance P, neurokinin A and calcitonin, which can easily cause neuroinflammatory reactions, further causing bladder mucosal damage and increased permeability. Mast cells in ulcerative interstitial cystitis increase significantly. Mast cells contain a large number of inflammatory mediators, leading to symptoms such as pain, frequent urination, and edema. However, the number of mast cells in nonulcerative interstitial cystitis is normal in most cases.

The treatment of interstitial cystitis currently mainly uses tricyclic antidepressants to antagonize the effects of some neurotransmitters, and has certain therapeutic effects in clinical applications; antihistamines are used to reduce mast cell activation and infiltration , And then achieve the effect of relieving pain; Sodium pentamethylene sulfide can form a protective film on the surface of the bladder to reduce the damage of toxins to the bladder nerve. Patients with more severe symptoms are often treated with bladder perfusion, cystectomy, and urinary diversion.

Trigeminal neuralgia is a common clinical refractory disease. The pain is unbearable at the onset of onset. It is mainly manifested as paroxysmal rapid electric shock-like, knife-cut-like and tear-like pain, which often stops suddenly, and the course of the disease is longer and serious. It affects the patient's normal life and work, reduces the patient's quality of life, and is known as "the first pain in the world".

The disease mostly occurs in the elderly over the age of 50, and generally has a unilateral attack. There are more women than men, and the incidence is about 1/15,000. There are mainly two types of trigeminal neuralgia, primary and secondary. Secondary trigeminal neuralgia refers to the facial pain of patients innervated by the trigeminal nerve due to the stimulation and compression of tumors and vascular diseases.

The pathogenesis of primary trigeminal neuralgia is not yet clear. Most people think that trigeminal neuralgia is caused by multiple factors. Among them, central disease and peripheral disease, as well as changes in immune factors and neurotransmitters are important causes of trigeminal neuralgia.

At present, the treatment methods for trigeminal neuralgia include Western medicine treatment, combined Chinese and Western medicine treatment, and surgical treatment (such as microvascular decompression, radiofrequency thermocoagulation, stereotactic radiotherapy, etc.). Western medicine treatment, as the patient's drug resistance increases, the factors of more drug response will lead to the further development of the disease. The repetition of TCM treatment is poor, and surgical treatment is more commonly used clinically, but the side effects of surgical treatment are often more serious. Therefore, the current treatment of this disease is more difficult.

Through experimental screening, it is found that the present invention can regulate the structure and composition of intestinal microbes, and further regulate neuroendocrine activities, thereby achieving the treatment and prevention of the occurrence of the aforementioned diseases.

Summary of the invention

The main technical problem solved by the present invention is to overcome the shortcomings of the prior art, provide a good safety, low side effects, and have better prevention and treatment of qualitative cystitis, primary trigeminal neuralgia, depression, sleep disorders, youth A soil preparation for acne or constipation.

Technical solutions. To solve the above technical problems, the present invention adopts the following technical solutions:

A soil preparation is provided, which is prepared from sterilized soil and oral probiotics or auxiliary materials.

In a preferred embodiment of the present invention, the soil is uncontaminated soil, including red soil, loess, black soil and cinnamon soil.

In a preferred embodiment of the present invention, the probiotics include Bifidobacterium, Lactobacillus, Streptococcus, Propionibacterium, Lactococcus lactis, or Leuconostoc mesenteroides.

In a preferred embodiment of the present invention, the auxiliary materials are edible materials, including protein, starch or cellulose food materials.

In a preferred embodiment of the present invention, the soil preparation is used to prepare the prevention and/or treatment of human neuroendocrine disorders, including primary interstitial cystitis, interstitial pneumonia, primary trigeminal neuralgia, Drugs for multiple sclerosis, alopecia areata, psoriasis, depression, sleep disorders, acne, or constipation.

A preparation method for preparing the above soil preparation includes the following steps:

(1) Collect pollution-free soil and do trace element analysis to screen the soil with qualified heavy metal content; (2) Autoclave, autoclave at 121℃ for 60 minutes;

(3) After cooling, mix it evenly with probiotics or auxiliary materials in a predetermined ratio;

(4) Prepare into capsules or granules.

In a preferred embodiment of the present invention, the soil contains the following elements in mass percentages: Na 0.5-1%, Mg 1-3%, Al 5-10%, Si 20-35%, K 1-5%, Ca 0.5-10%, Ti 0-5%, Fe 2-10%, O 40-50%.

In a preferred embodiment of the present invention, the toxic trace elements include Hg, Pb, As, Cd and Cu; wherein Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤ 2.0mg/kg, Cu≤20.0mg/kg.

In step (1), no toxic organic matter can be detected in the conventional soil method.

In a preferred embodiment of the present invention, the soil preparation is made into a solid or fluid form that can be taken orally.

The beneficial effects of the present invention are:

1) The soil preparation of the present invention can further regulate neuroendocrine function by regulating the intestinal microecology: a number of studies have shown that intestinal microbes and their metabolites can stimulate the intestinal epithelial cells to produce serotonin (more than 95% of the human body The serotonin comes from the intestine). Serotonin is an important neurotransmitter in neuroendocrine, which is closely related to the occurrence of depression and many other diseases. Therefore, the present invention can adjust the structure and composition of human intestinal microbes, thereby regulating human neuroendocrine activities, and further achieve the prevention and treatment of interstitial cystitis, primary trigeminal neuralgia, multiple sclerosis, depression, sleep The effect of diseases such as obstacles.

2) Since the active ingredients in the preparation are mainly inorganic and organic matter in the soil, the ingredients in the soil are relatively stable under dry conditions, so the storage, transportation and production costs are relatively low.

detailed description

The preferred embodiments of the present invention are described in detail below, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, so as to make a clearer and clearer definition of the protection scope of the present invention.

Example 1 Black clay preparation for preventing and treating interstitial cystitis.

(1) Preparation of preparation: select black loam soil, sample sample element average content (mass percentage): Na 0.95%, Mg 2.03%, Al 8.28%, Si 27.19%, K 2.56%, Ca 6.45%, Ti 0.58%, Fe 5.57 %, O 46.39%, of which heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 1.7% of the dry weight of the soil. The high-pressure fire fungus is reserved for use. The auxiliary materials are high-quality dry-fried peanut powder and sucrose. The soil after the high-pressure fire fungus is mixed with dry-fried peanut powder and sucrose. The mixing ratio is soil: peanut powder: sucrose. 1:1:0.5 (mass ratio) to obtain a black clay preparation.

(2) Interstitial cystitis research subjects: 40 patients were selected as the experimental subjects, the age range was 20-85 years old, and the average age was 50.1±14.1 years, including 15 male patients and 25 female patients. The patient's course of illness was 10 to 70 months, with a course interval of 30.1±11.7 months. Among them, 10 cases were first diagnosed and 30 cases were revisited.

The patient met the 2016 diagnostic criteria of the Canadian Urological Association (CUA). Inclusion criteria: 1) Medical history of more than 6 weeks, no urinary tract infection, ultrasound kidney, ureter, bladder no abnormal findings; 2) Frequent urination, lower abdominal discomfort or pain when holding back urine, lower abdominal discomfort or pain relieve symptoms after urination; 3 ) After anesthesia, under the pressure of 80cmH2O (1cmH2O=0.098kPa), the bladder continued to expand for 5 minutes. Cystoscopy found Hunner ulcer or diffuse bladder mucosal hemorrhage, that is: 10 or more bleeding points per field of view, and more than 3 fields of view .

Exclusion criteria: 1) The patient's maximum bladder function capacity in the awake state> 350ml; 2) No urgency sensation when filling the bladder with 150ml of fluid; 3) Symptom duration <6 weeks; 4) Symptoms relieved after antibiotics or antispasmodics; 5) The frequency of urination during the day is less than 8 times, and nocturia is less than 2 times. 6) Complicated with urethral or vaginal infections, bacterial cystitis, tuberculous cystitis, radiation cystitis, chemical cystitis, pelvic inflammatory disease, endometriosis, sexually transmitted diseases, urinary calculi, bladder tumors Wait.

(3) Research content: ①Age, gender, course of disease, main symptoms of treatment, etc.; ②Urinary Diary; ③O'Leary Sant score sheet, including two parts: Symptom Index (ICSI) and Problem Index (ICPI); ④Pelvic cavity Pain and urinary frequency/urgency symptom scale (PUF); ⑤Visual Analogue Scale/Score (VAS); ⑥Athens Insomnia Scale (AIS); ⑦Anxiety Self-Rating Scale (Self- rating Anxiety scale, SAS), Depression Self-rating Scale (Se] f-rating Depression scale, SDS). The initial condition of all IC/BPS patients before treatment and the follow-up conditions at 3 and 6 months after treatment were recorded. All questionnaires (except for the urination diary) were filled out and retrieved by the investigator, and the urination diary was retrieved after 2 to 3 days.

(3) Treatment method: The black clay preparation of step (1) in the treatment group was taken orally, twice a day, with a dosage of 5g/time, and it could be mixed with food. The treatment effect was evaluated after 3 months of medication. The curative effect was evaluated after taking it for three months.

(4) Evaluation of treatment effect: The collected data is statistically analyzed using SPSS 18.0 software. It is effective if the IC symptom score drops> 5 points, or> 25%.

(5) Results: According to statistical analysis, there were significant differences before and after treatment in the treatment group (P<0.001). No adverse reactions. As shown in Table 1 and Table 2.

(6) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 3), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 1. Comparison of 24h urination frequency, ICSI, ICPI, PUF, VAS scores of patients before and after treatment

time	N	24h urination frequency	ICSI	ICPI	PUF	VAS
before therapy	40	26.60±12.35	14.28±4.78	9.56±2.43	14.66±4.19	6.11±2.22

Table 2. Anxiety and depression scores of IC patients

To	N	SAS	SDS	AIS
Before treatment for IC patients	40	45.67±6.77	52.46±11.31	5.68±3.32
After IC patient treatment	40	32.23±5.69	45.32±8.12	2.14±1.02
Chinese norm	1628/1158	29.87±10.07	41.88±10.57	To

Note: Compared before and after treatment, P＜0.001

Table 3 Random forest analysis of types of significantly increased intestinal bacteria content in the treatment group

Example 2 A preparation of Bifidobacterium loessum used for the prevention and treatment of primary trigeminal neuralgia.

(1) Preparation of preparation: select yellow loam soil, sample sample element average content: Na 0.36%, Mg 1.49%, Al 9.26%, Si 28.3%, K 2.29%, Ca 4.22%, Ti 0.73%, Fe 7.95%, O 45.4 %, where the heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. , And conventional methods cannot detect toxic organic matter. Organic matter components accounted for 1.2% of the dry weight of the soil, autoclaved for later use, while adding cellulose as an auxiliary material, and Bifidobacterium longum subsp.infantis ATCC 15697 as a probiotic. After autoclaving, the soil, cellulose and Bifidobacterium longum dry powder are mixed in a ratio of soil: cellulose: Bifidobacterium longum dry powder 1:1:1 (mass ratio) to prepare a loess preparation.

(2) Selection of subjects for primary trigeminal neuralgia: 40 patients were randomly divided into treatment group and control group. There were 20 patients in the treatment group, 8 males and 12 females, aged 15-80 years old, with an average of (51.8±18.2) years old. The course of illness was 5 days to 16 years, with an average of (3.5±1.6) years; the control group had 20 cases, 9 males and 11 females, aged 16 to 78 years, with an average of (55.3±14.8) years; the course of illness was 5 days to 20 years, with an average (4.04±1.8) years. There was no statistical difference in gender, age, and course of disease between the two groups. The selected cases all met the diagnostic criteria for primary trigeminal neuralgia.

The diagnostic criteria refer to the diagnostic criteria of primary trigeminal neuralgia in the 2004 International Classification of Headaches (Second Edition) of the International Headache Society Classification Committee. 1) Sudden onset pain on the face, generally less than 2 minutes in duration, invading one or several branches of the trigeminal nerve distribution area; 2) The pain has at least one of the following symptoms: severe, acupuncture-like, burning-like or even knife Cut sample; can be triggered by trigger point or related trigger factors and cause sudden attack; 3) clinically no positive signs of nervous system; 4) cannot be attributed to other diseases and dysfunction.

Inclusion criteria: 1) Meet the relevant diagnostic criteria of Chinese and Western medicine; 2) Men or women aged 15 years or older but less than 80 years old, the two groups of observation subjects in the course of disease, age and various indicators before and after treatment, after statistical processing There is no obvious difference, and it is comparable; 3) A week before the observation, it is necessary to stop taking various Chinese and Western medicines and stop other treatment methods; 4) The condition is stable and the consciousness is clear; 5) The frequency of attacks is greater than or equal to 1 time per day. 6) The patient understands the trial process and participates voluntarily, and signs an informed consent form;

Exclusion criteria: 1) Those who do not meet the above diagnostic criteria; 2) Patients with secondary trigeminal neuralgia; 3) Women who are breast-feeding or pregnant; 4) Those with other primary diseases such as severe cardio-cerebrovascular and coagulopathy; 5) Those who are participating in other medical trials; 6) Those who have severe nerve damage and cognitive impairment that affect pain assessment.

Grouping: Randomly divided into treatment group of 20 cases and conventional treatment group of 20 cases. There was no statistical difference in gender, age, and course of disease between the two groups.

(3) Treatment method: The treatment group takes the loess preparation of step (1), 3 times a day, the dosage is 3-6g/time, and can be mixed with food. The control group was a placebo made of cellulose: Bifidobacterium longum dry powder with a mass ratio of 1:1, and the dose was the same as the treatment group. The treatment effect was evaluated after 2 months of medication.

(4) Evaluation of treatment effect: After 2 months of treatment, observe the treatment effect according to the "Clinical Guidelines for the Treatment of Trigeminal Neuralgia with New Traditional Chinese Medicine" promulgated in 2002, and evaluate the effect in accordance with clinical treatment experience.

① Follow-up recovery: the pain symptoms disappeared completely within 12 months from the date of treatment;

②The follow-up is effective: most of the pain disappeared within 12 months from the treatment date;

③ Effective follow-up: pain relief within 12 months from the treatment date;

④ Follow-up is invalid: The pain has not been significantly relieved within 12 months from the date of treatment.

Effective rate = (number of cures + number of significant effects + effective number) X 100% / total number

(5) Results: According to statistical analysis, there are significant differences in the clinical efficacy of the treatment groups (P<0.001). No adverse reactions. See Table 4 and Table 5.

(6) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 6), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 4 Pain scores before and after treatment in the two groups

Group

N

before therapy

After treatment

Difference

Control group	20	19.11±4.03	13.42±4.16	5.69±3.68
therapy group	20	19.14±4.48	2.98±1.99	14.66±5.39

Note: The difference between the two groups of patients before and after treatment is in accordance with the normal distribution, after the paired sample t test, the treatment group t=14.66, P＜0.001

Table 5 Comparison of curative effect between the two groups before and after treatment

Curative effect	N	get well	Markedly effective	effective	invalid	Efficient
therapy group	20	12	3	2	3	85%
Control group	20	3	3	1	13	35%

Note: After chi-square test: df=3, X ² =1.98, P=0.007.

Table 6 Random forest analysis of types of significantly increased intestinal bacteria content in the treatment group

Example 3 A red clay preparation for the prevention and treatment of multiple sclerosis.

(1) Preparation of preparation: select red loam, and the average element content of the sample sample: Na 0.48%, Mg 1.37%, Al 8.93%, Si 30.69%, K 2.87%, Ca 1.68%, Ti 0.04%, Fe 6.18%, O 47.76 %, where the heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 2.1% of the dry soil weight. Autoclave for later use. The auxiliary material is starch. The soil after autoclaving is mixed with starch, and the mixing ratio is soil: starch 1:1 (mass ratio) to prepare a red clay preparation.

(2) Participants selection: Among the 20 experimental groups, 8 were male patients and 12 were female patients; the youngest was 19 years old and the oldest was 70 years old, with an average of 44.23±15.14 years old; the average course of disease was 2.1±4.4 months. Among the 20 control groups, 9 were male patients and 11 were female patients; the youngest was 18 and the oldest was 68 years old, with an average of 43.35±14.66 years old; the average course of disease was 2.4±5.1 months. The clinical manifestations of the two groups were not statistically different (P>0.05).

(3) Treatment method: The treatment group took the red clay preparation of step (1) twice a day, 5g each time, which can be mixed with food. The control group took starch as a placebo, and the dosage was the same as the treatment group. The treatment effect was evaluated after 3 months of medication.

(4) Comparison of EDSS scores between the two groups after treatment

After 3 months of treatment, the EDSS scores of the experimental group and the control group had statistically significant differences before and after treatment (P<0.05). After treatment, the EDSS score of the experimental group decreased significantly more than that of the control group, which was statistically significant (P=0.000). The specific values are shown in Table 7.

(5) Comparison of EDSS scores and recurrence times between the two groups one year after treatment

One year after treatment, compared with the control group, the experimental group had a significant difference in the changes of EDSS score before and after treatment (P＜0.05). The decrease in EDSS score of the experimental group was significantly different than that of the control group (P＜0.05). 0.05). In addition, the number of relapses after 1 year between the two groups of patients was 1 in the experimental group and 2 in the control group. There was no statistical difference (P>0.05). The specific values are shown in Table 8.

(6) Comparison of treatment effects

According to statistical analysis, there is a significant difference in clinical efficacy between the experimental group and the control group (P<0.001). No adverse reactions. See Table 9 for details.

(7) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 10), The type and number of bacteria in Actinomycetes increased significantly (P＜0.01)

Table 7 Comparison of EDSS scores of two groups of patients with multiple sclerosis after 3 months of treatment

Group	before therapy	After treatment	t value	P value
Experimental group n=20	5.13±1.09	2.12±1.36	7.723	＜0.001
Control group n=20	5.07±2.01	4.44±1.67	1.078	0.288
t value	To	-4.817	To	To
P value	To	＜0.001	To	To

Table 8 Comparison of EDSS scores and recurrence times of two groups of patients one year after treatment

Group	before therapy	1 year after treatment	t value	P value
Experimental group n=20	5.13±1.09	2.19±1.19	8.142	＜0.001
Control group n=20	5.07±2.01	4.96±1.96	0.175	0.862
t value	To	-5.402	To	To
P value	To	＜0.001	To	To

Table 9 Comparison of the treatment effect of two groups of multiple sclerosis patients

Group	Number of cases	Markedly effective	effective	invalid	Total effective rate
Control group	20	1	5	14	30%
test group	20	12	6	2	90%

Note: After chi-square test: df=1, X ² ＝15.000, P＜0.001

Table 10 Random forest analysis of types of significantly increased intestinal bacteria content in the treatment group

Example 4 Study on curative effect of loess preparation on acne

(1) Preparation of preparation: select yellow loam soil, sample sample element average content (mass percentage): Na 0.84%, Mg 1.42%, Al 9.17%, Si 30%, K 2.37%, Ca 1.48%, Ti 0.67%, Fe 6.94 %, O 47.11%, where the heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. , And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 2.1% of the dry weight of the soil, autoclaved for later use. The auxiliary materials are high-quality dry-fried peanut powder and starch. The soil after autoclaving and dry-fried peanut powder and starch are mixed in a ratio of soil: peanut powder: starch 1:1:1 (mass ratio) to obtain a loess preparation.

(2) Selection of acne patients: 80 patients were randomly divided into treatment group and control group. There were 40 cases in the treatment group, 20 males and 20 females, aged 10-25 years old, with an average of (15.7±5.1) years old. The course of disease was 1 to 7 years; there were 40 cases in the control group, 20 males and 20 females, aged 11 to 26 years, with an average of (15.3±6.3) years old; the course of disease was 1 to 7 years. There was no statistical difference in gender, age, and course of disease between the two groups. The selected case was diagnosed as acne by an experienced dermatologist.

(3) Treatment method: The treatment group takes orally every day. Step (1) Loess preparation, 3 times a day, 5 grams each time. The control group took peanut powder with a mass ratio of 1:1: starch as a placebo, and the dosage was the same as that in the treatment group. The treatment effect was evaluated after 4 weeks of medication.

(4) Evaluation of treatment effect: According to the "Diagnosis and Curative Effect Criteria for Diseases of Traditional Chinese Medicine" combined with clinical development Cure: Skin damage subsided, and symptoms disappeared and improved; Improvement: Subjective symptoms were significantly reduced, and skin lesions subsided by more than 30%; Unhealed: Skin lesions and symptoms did not change or resolved less than 30%. The effective rate is calculated based on the total number of cured and improved cases.

(5) Results: The results show the effective rate of treatment, see Table 11. According to statistical analysis, there is a significant difference in clinical efficacy between the treatment group and the control group (P<0.001). No adverse reactions.

(6) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 12), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 11 Comparison of curative effect between the two groups of patients after 4 weeks of treatment (n,%)

Group	N	cure	Get better	invalid	Effective number of cases	Efficient
therapy group	40	twenty one	14	5	35	87.5%
Control group	40	5	5	30	10	25%

Note: After chi-square test: df=1, X ² ＝31.746, P＜0.001.

Table 12. Random forest analysis of the types of significantly increased intestinal bacteria content in the treatment group

Example 5 Study on the function of red clay preparation in auxiliary treatment of depression

(1) Preparation of preparation: select red loam soil, sample sample element average content (mass percentage): Na 0.69%, Mg 2.32%, Al 7.34%, Si 28.04%, K 2.54%, Ca 9.17%, Ti 1.51%, Fe 5.78 %, O 42.61%, of which heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. , And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 1.5% of the dry weight of the soil, autoclaved for later use. The auxiliary materials are high-quality bran and corn flour. The soil after autoclaving is mixed with bran corn flour, and the mixing ratio is soil: bran: corn flour 1 :1:1 (mass ratio), prepared into red clay preparation.

(2) Selection of subjects for depression: All 60 volunteers met the diagnostic criteria for depression ("Chinese Mental Disorder Classification and Diagnostic Criteria", 3rd edition). The subjects were randomly divided into a treatment group, a total of 30 cases, 15 male patients, 15 female patients, aged 13-65 years old, average (38.5±9.9) years old, average course of disease (17.8±11.7) months; control group 30 cases There were 15 male patients and 15 female patients, aged from 14 to 65 years old, with an average age of (39.5±10.8) years and an average course of (16.4±10.1) months. There was no statistical difference between the two groups of patients in indicators such as age, gender, and disease course.

(3) Treatment method: Before administration, the treatment group and the control group were set to wash out for 2 weeks to avoid the influence of other depression drugs on this study. The observation group took the red clay preparation of step (1) every day, 3 times a day, 4 grams each time. The control group took the same proportion of auxiliary materials (quality ratio 1:1 bran and corn flour) as a placebo, and the dosage was the same as that of the treatment group. The treatment effect was evaluated after 6 months of medication.

(4) Evaluation of the treatment effect: The Hamilton Depression Scale (HAMD) score and comparative analysis were used for different treatment periods. Before treatment, the HAMD scores of the two groups were basically the same, and there was no statistically significant difference between the groups . At 6 weeks and 12 weeks after treatment, the HAMD score of the observation group was significantly improved compared with the control group, and the difference between the groups was statistically significant (P<0.05), as shown in Table 13.

Table 13 HAMD index scores of the two groups of patients

Group	Number of participants	before therapy	6 weeks of treatment	Treatment for 12 weeks
Observation group	28	28.69±7.97	10.45±4.36	11.22±4.63
Control group	28	28.54±6.85	14.22±5.74	15.44±6.36
t	To	0.076	-2.768	-2.838
P	To	0.940	0.008	0.006

(5) Comparison of adverse reactions between the two groups of patients: During the treatment period, both groups of patients experienced adverse reactions such as dry mouth, sweating, nausea, vomiting, weight gain, food blur, etc. However, the incidence of adverse reactions in the observation group was both Significantly lower than those in the control group. Among them, the incidence of adverse reactions such as sweating, dry mouth, nausea and vomiting, weight gain, blurred vision, and headaches were statistically different between the groups, as shown in Table 14.

(6) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 15), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 14 Comparison of adverse reactions during treatment between the two groups of patients

Group	Sweating	Dry mouth	feel sick and vomit	Weight gain	Blurred vision	dizziness	Constipation/diarrhea	headache
Observation group	2	3	5	1	2	0	2	1
Control group	7	12	11	8	4	4	3	7
P	0.069	0.007	0.076	0.011	0.388	0.120	1.000	0.056

Table 15. Random forest analysis of types of significantly increased intestinal bacteria content in the treatment group

Example 6 Study on the treatment of functional constipation with black soil preparation

(1) Preparation of preparation: select black loam soil, sample sample element average content (mass percentage): Na 1.13%, Mg 0.87%, Al 7.24%, Si 32.7%, K 1.99%, Ca 1.39%, Ti 0.63%, Fe 6.22 %, O 47.83%, of which heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 2.4% of the dry weight of the soil, autoclaved for later use. The auxiliary materials are wheat flour and corn flour. The soil after autoclaving is mixed with wheat flour and corn flour, and the mixing ratio is soil: wheat flour: corn flour 1:1:1 (Mass ratio) to obtain a black soil preparation.

(2) Selection of subjects with functional constipation: 70 patients were randomly divided into treatment group and control group. There were 35 patients in the treatment group, including 17 males and 18 females, aged 30-66 years old, with an average of (41.9±7.5) years old. The course of illness was 1-8 years, with an average of (3.5±2.3) years; the control group had 35 cases, 18 males and 17 females, aged from 31 to 67 years, with an average of (42.1±8.5) years; the course of illness was 1-8 years, with an average of (4.1 ±3.2) years. There was no statistical difference in gender, age, and course of disease between the two groups. All selected cases met the diagnostic criteria for functional constipation (Rome II criteria).

(3) Treatment method: The treatment group takes orally every day Step (1) Black soil preparation, 3 times a day, 5 grams each time. The control group took wheat flour: corn flour with the same mass ratio of 1:1 as placebo, and the dosage was the same as that of the treatment group. The treatment effect was evaluated 14 days after the medication.

(4) Evaluation of treatment effect: It is formulated with reference to the therapeutic effect standard of adult constipation in the "Standards for Diagnosis and Curative Effect of TCM Diseases". (1) Cure: Defecation once within 1 day, normal stool quality, unobstructed defecation, no recurrence within 1 month; (2) Significant effect: 1 defecation within 2 days, stool quality tends to be normal, unobstructed defecation, none within 1 month Relapse; (3) Effective: Defecation once within 3 days, the quality of the stool is turned, and the defecation is not smooth; (4) Ineffective: No significant improvement in clinical symptoms. The effective rate is calculated based on the total number of cured and markedly effective cases.

(5) Results: The results show the effective rate of treatment, see Table 16. According to statistical analysis, there is a significant difference in clinical efficacy between the treatment group and the control group (P<0.001). No adverse reactions.

(6) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 17), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 16 Comparison of curative effect between two groups of patients after 14 days of treatment (n,%)

Group	N	cure	Markedly effective	effective	invalid	Effective number of cases	Efficient
therapy group	35	20	8	3	4	31	88.5%
Control group	35	3	2	8	twenty two	13	37.1%

Note: After Chi-square test: df=1, X ² ＝19.82, P＜0.001

Table 17 Random forest analysis of types of significantly increased intestinal bacteria content in the treatment group

Example 7 Study on treatment of alopecia areata with black soil preparation

(1) Preparation of preparation: select black loam soil, sample average element content (mass white content ratio): Na 0.58%, Mg 1.31%, Al 8.71%, Si 29.07%, K 2.22%, Ca 2.69%, Ti 0.54%, Fe 7.56%, O 47.32%, of which heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 2.5% of the dry weight of the soil, autoclaved for later use. The auxiliary materials are red bean flour and corn flour. The soil after autoclaving is mixed with red bean flour and corn flour. The mixing ratio is soil: red bean flour: corn flour is 1 :1:1 (mass ratio) to obtain a black clay preparation.

(2) Selection of patients with alopecia areata: 70 patients were randomly divided into treatment group and control group. There were 35 patients in the treatment group, including 18 males and 17 females, aged from 16 to 55 years old, with an average of (28.5±10.4) years old. The course of illness was 3-10 weeks, with an average of (7.5±4.5) weeks; the control group had 35 cases, 18 males and 17 females, aged 17-57 years, with an average of (26.5±9.8) years old; the course of illness was 4-10 weeks, with an average of (8.1 ±4.7) weeks. There was no statistical difference between the two groups in gender, age, and course of disease.

(3) The diagnostic criteria for alopecia areata is drawn up with reference to "Clinical Dermatology" and "Handbook of Integrated Traditional Chinese and Western Medicine Skin and Venereology":

①The disease can occur at any age, but it is more common in young adults.

②The patient's scalp suddenly appeared round or oval, 1-10cm diameter, unequal number, and clearly defined hair loss areas on the scalp. Single or multiple hairs. In severe cases, the entire scalp hair will fall off completely, which means "full baldness", or hair on other parts of the body, such as eyebrows, armpit hair, and pubic hair will fall off, which means "general alopecia".

③In the advanced stage of the disease, the hair on the edge of the hair loss area becomes loose, and the hair is easily pulled out. In the hair pulling experiment (+), the hair that fell off the affected area was observed under a microscope. It can be seen that the proximal end of the hair shaft was atrophy, which appeared to be thick and thin at the top. Exclamation mark" phenomenon.

④The skin at the lesion is smooth, without inflammation, no atrophy or scar.

(4) Treatment method: The treatment group was taken orally every day. Step (1) Black soil preparation, 2 times a day, 12 grams each time. The control group took red bean flour: corn flour with the same mass ratio of 1:1 as placebo, and the dosage was the same as that of the treatment group. The treatment effect was evaluated after 180 days of medication.

(5) Observation index: Observe the patient's hair loss area and range before and after treatment, as well as the scalp hair regrowth, hair pulling experiment and other results, and record these results. Then, according to the skin lesion scoring standard, the scores of the above observation items before and after the treatment of the subject's skin lesion were calculated. According to the observational indicators of hair loss in the third series of "Guiding Principles for Clinical Research of New Chinese Medicines", the specific scoring standards are as follows:

A. Hair loss area and location: 0 points: no obvious hair loss areas; 1 point: limited to flaky hair loss on the head epidermis, and the area of hair loss is less than or equal to 10% of the total scalp area; 2 points: only on the head epidermis Patchy hair loss occurs, the area of hair loss is greater than 10% of the total scalp area, but less than 20%; 3 points: the area of hair loss is greater than or equal to 20%

B: Hair pull test: 0 points: negative; 1 point: positive.

C: Hair growth: (according to four-level scoring method) ① Hair regeneration in the lesion area: 3 points: <30% or still falling; 2 points: >30% and <70%, no shedding; 1 point: >70 % And <95%, no off; 0 points: 100%, no off. ②New-born hair density: 3 points: the healthy hair area is sparse; 2 points: the healthy hair area is sparse; 1 point: the healthy hair area is sparse; 0 points: no difference from the healthy hair area. ③ New-born hair thickness: 3 points: obviously softer than healthy hair area; 2 points: softer hair area; 1 point: slightly softer hair area; 0 point: no difference from healthy hair area. ④ Newborn hair color: 3 points: white color; 2 points: gray or yellow color; 1 point: brown color; 0 point: black color.

(6) Evaluation of treatment effect: The evaluation standard of curative effect refers to "Guiding Principles for Clinical Research of New Chinese Medicines." Clinical recovery: The patient's head hair loss ceases, and the shed hair on the scalp completely resumes growth. The density and color of the newly grown hair is the same as that of the normal scalp hair; the therapeutic index is ≥95%. Significant effect: the patient's head hair stops falling off, and the newly grown scalp hair is ≥70%, and its density and color are close to normal scalp hair; 60%≤curative index <95%. Effective: the patient's head hair stops falling off, 30%≤newly grown scalp hair≤70%, sparse hair and white growth can be seen in the skin lesion area; 30%≤curative index <60%. Invalid: newly grown scalp hair <30% or continue to fall out; curative effect index <30%.

Total effective rate = (number of cured cases + number of markedly effective cases + number of effective cases) / total number of cases in this group × 100%

(7) Results: The results show the effective rate of treatment, see Tables 18 and 19. According to statistical analysis, there is a significant difference in clinical efficacy between the treatment group and the control group (P<0.001). No adverse reactions.

(8) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 20), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 18 Comparison of the total scores of the two groups before and after treatment

Group	Number of cases	Total score before treatment	Total score after treatment
therapy group	35	10.00±4.36	2.31±1.43
Control group	35	10.12±3.65	9.56±5.07
P value	To	P>0.05	P＜0.001

Table 19 Comparison of curative effect between two groups of patients after 180 days of treatment (n,%)

Group	N	cure	Markedly effective	effective	invalid	Total effective rate
therapy group	35	15	8	8	4	88.6%
Control group	35	4	2	8	twenty one	40%

Note: After chi-square test: df=1, x2=17.98, p＜0.001

Table 20. Random forest analysis of the types of significantly increased intestinal bacteria content in the treatment group

Example 8 Study on the treatment of psoriasis vulgaris with red clay preparation

(1) Preparation of preparation: select black loam soil, sample sample element average content (mass percentage): Na 0.56%, Mg 1.34%, Al 8.01%, Si 28.03%, K 2.21%, Ca 4.11%, Ti 0.34%, Fe 7.85 %, O 47.55%, of which heavy metal content Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg/kg, As≤2.0mg/kg, Cu≤20.0mg/kg. And conventional methods cannot detect toxic organic matter. The organic matter component accounts for 2.7% of the dry weight of the soil. Autoclave sterilizes for later use. The auxiliary materials are peanut flour and wheat flour. The soil after autoclaving is mixed with wheat flour and corn flour. The mixing ratio is soil: peanut flour: wheat flour 1:1:1 (Mass ratio) to obtain red clay preparation.

(2) Psoriasis vulgaris participant selection: 80 patients were randomly divided into treatment group and control group. There were 40 patients in the treatment group, 25 males and 15 females, aged 40-69 years old, with an average of (54.5±7.8) years old. The course of disease was 2-16 years, with an average of (8.5±3.6) years; the control group had 40 cases, 28 males and 12 females, aged 41-69 years, with an average of (57.5±7.9) years; the course of disease was 2-17 years, with an average of (8.1 ±4.2) years. There was no statistical difference in gender, age, and course of disease between the two groups.

(3) Western medicine diagnostic criteria for psoriasis vulgaris: The Western medicine diagnostic criteria for psoriasis vulgaris refer to "Chinese Clinical Dermatology" (2010 Edition Jiangsu Science and Technology Press): the skin damage of this disease is red inflammatory papules and spots Papules and patches of varying sizes are the main ones, covered with multiple layers of silvery white scales. A bright film can be seen when the scales are scraped, and there may be a bit of bleeding under the film. The rash can occur all over the surface of the body. In the case of the scalp, the hair is bundled; in the case of the nail plate (finger, toe), it may have a pit like a thimble or the nail plate may be uneven, yellow and thick. It can be accompanied by varying degrees of itching. The course of the disease is slow and easy to relapse.

(4) Treatment method: The treatment group takes orally every day Step (1) Black soil preparation, 3 times a day, 10 grams each time. The control group took peanut flour: wheat flour with the same mass ratio of 1:1 as the placebo, and the dosage was the same as that of the treatment group. The treatment effect was evaluated after 180 days of medication.

(5) Curative effect scoring standard

①The lesion area and severity index of patients with psoriasis refer to the PASI score: the lesion area and severity index of patients with psoriasis refer to the Global PASI (Psoriasis area and severity index, PASI) scoring standard. Before treatment for the two groups of patients Record the PASI points after treatment.

②Scoring of the degree of pruritus: Score the pruritus of the three groups of patients before and after treatment, observe the improvement of the degree of pruritus before and after treatment, using a four-level scoring method of 0, 3, 6, and 9 points. 0 points: no itching; 3 points: slight itching, but does not affect sleep and work; 6 points: paroxysmal itching, which affects sleep and work, requires medication; 9 points: severe itching, which seriously affects sleep and work.

(6) Evaluation of therapeutic effect: The evaluation standard of therapeutic effect shall refer to "Guiding Principles for Clinical Research of New Chinese Medicines"

Clinical recovery: almost all skin lesions disappear, curative effect index> 95%

Markedly effective: most of the skin lesions disappeared, 95%＞curative effect index ≤70%

Effective: the skin lesions partly disappeared, 70%＞curative effect index ≤55%

Invalid: the skin lesions are not resolved obviously, the curative effect index is less than 55%

Efficacy index=(integral before treatment-integral after treatment)/integral before treatment×100%.

The total effective rate includes the number of cured cases, the number of markedly effective cases, and the total number of effective cases as a percentage of all cases in this group. That is, the total effective rate=[(number of cured cases+number of markedly effective cases+number of effective cases)/total number of cases in this group]×100%.

(7) Results: The results show the effective rate of treatment, see Tables 21 to 23. According to statistical analysis, there is a significant difference in clinical efficacy between the treatment group and the control group (P<0.001). No adverse reactions.

(8) Changes in intestinal microbes: 16s rRNA was sequenced and analyzed by a high-throughput sequencer. The results showed that the diversity of intestinal microbes in the treatment group increased significantly (P＜0.01). Random Forests analysis showed (Table 24), The species and number of bacteria in Actinomycetes increased significantly (P<0.01).

Table 21 Comparison of PASI scores before and after treatment between the two groups

Group	Number of cases	Pre-treatment score	Score after treatment
therapy group	40	15.17±3.72	2.96±1.88
Control group	40	15.81±3.96	14.86±4.73
P value	To	P>0.05	P＜0.001

Table 22 Comparison of pruritus scores before and after treatment in the two groups

Group	Number of cases	Pre-treatment score	Score after treatment
therapy group	40	8.29±1.07	1.34±1.46
Control group	40	8.45±1.69	8.11±2.25
P value	To	P>0.05	P＜0.001

Table 23 Comparison of curative effect between the two groups of patients after 180 days of treatment (n,%)

Group	N	cure	Markedly effective	effective	invalid	Efficient
therapy group	40	10	18	6	6	85%
Control group	40	2	2	8	28	30%

Note: After chi-square test: df=1, X ² ＝18.66, P＜0.001

Table 24 Random forest analysis of types of significantly increased intestinal bacteria content in the treatment group

The above are only the embodiments of the present invention, and do not limit the scope of the present invention. Any equivalent structure or equivalent process transformation made by using the content of the present invention, or directly or indirectly applied to other related technical fields, are all The same reason is included in the scope of patent protection of the present invention.

Claims (9)

1. A soil preparation is characterized in that it is prepared from sterilized soil and oral probiotics or auxiliary materials.
2. The soil formulation according to claim 1, wherein the soil includes red soil, loess, black soil and cinnamon soil.
3. The soil preparation according to claim 1, wherein the probiotic bacteria include Bifidobacterium, Lactobacillus, Streptococcus, Propionibacterium, Lactococcus lactis, or Leuconostoc mesenteroides.
4. The soil preparation according to claim 1, wherein the auxiliary material comprises protein, fat, sugar, starch or cellulose food material.
5. The method for preparing a soil preparation according to any one of claims 1 to 4, characterized in that it comprises the following steps:

   (1) Collect pollution-free soil and do trace element analysis to screen out pollution-free soil with qualified heavy metal content;

   (2) Then autoclave, sterilize under water steam, high temperature and high pressure for 60 to 120 minutes;

   (3) After cooling, mix it evenly with probiotics or auxiliary materials in a predetermined ratio;

   (4) Prepare into oral preparation. Capsules or granules.
6. The method for preparing a soil preparation according to claim 5, characterized in that the soil contains the following elements in mass percentage: Na 0.5-1%, Mg 1-3%, Al 5-10%, Si 20-35% , K 1～5%, Ca 0.5～10%, Ti 0～5%, Fe 2～10%, O 40～50%.
7. The method for preparing a soil preparation according to claim 5, wherein the heavy metals include Hg, Pb, As, Cd and Cu; wherein Hg≤0.2mg/kg, Pb≤5.0mg/kg, Cd≤0.3mg /kg, As≤2.0mg/kg, Cu≤20.0mg/kg.
8. The use of the soil preparation according to any one of claims 1 to 4 in the preparation of drugs for preventing and treating neuroendocrine disorders related diseases.
9. The soil preparation according to any one of claims 1 to 4 is used for preparing and preventing primary interstitial cystitis, interstitial pneumonia, primary trigeminal neuralgia, multiple sclerosis, alopecia areata, psoriasis, depression , Sleep disorders, acne or constipation drugs.