Hericium erinaceus converted mycelium culture method, chronic gastritis traditional Chinese medicine composition and application

Abstract

The invention provides a hericium erinaceus converted mycelium culture method, a chronic gastritis traditional Chinese medicine composition and application, hericium erinaceus strains are inoculated into a hericium erinaceus culture medium, and the hericium erinaceus culture medium comprises, by weight, 350-490 parts of bran, 100-140 parts of corn bran, 50-70 parts of millet peel and 400-600 parts of water; the biotransformation culture medium comprises the following components in parts by weight: 350-490 parts of bran, 100-140 parts of corn bran, 50-70 parts of millet peel and 400-600 parts of water, sterilizing, inoculating, culturing for 45-55 days at the culture temperature of 20-28 DEG C, taking out mycelia, and drying at the temperature of lower than 65 DEG C to obtain the hericium erinaceus transformed mycelia. The culture medium formed by biotransformation of bran, corn bran and millet bran is adopted, the culture medium and the production process are optimized, the digestion promoting and swelling eliminating effects are good, and the effect of treating chronic non-atrophic gastritis by the hericium erinaceus transformed mycelium and the composition thereof is remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of microbial engineering, and in particular to a method for culturing transformed mycelium of Hericium erinaceus, a traditional Chinese medicine composition for chronic gastritis and an application thereof. Background Art

[0002] Hericium erinaceus is a mushroom (fungus) of the Hericium family. It has been used in traditional Chinese medicine to treat gastrointestinal diseases for more than 2000 years. A series of small molecule compounds, such as erinacines, are believed to have neuroregenerative effects and can cross the blood-brain barrier to repair damaged nerve tissue. Hericium erinaceus also has anti-ulcer, anti-inflammatory, anti-microbial, immunomodulatory, liver function-enhancing, anti-aging, blood sugar and blood lipid lowering, hypoxia resistance, cardiac output increase, and circulation improvement. Medical preparations containing Hericium erinaceus are widely used in China.

[0003] The most important active substances in Hericium erinaceus mycelium are polysaccharides and glycoproteins. The current research on Hericium erinaceus polysaccharides at home and abroad shows that Hericium erinaceus polysaccharides have multiple biological activities and pharmacological effects, can enhance the phagocytic function of macrophages, promote the formation of hemolysin, resist leukocyte decline, reduce blood sugar, anticoagulation, antithrombosis, antimutation and anti-aging, etc. Therefore, Hericium erinaceus polysaccharides have attracted much attention and have become a hot spot for research and development and application in the fields of molecular biology, medicine, food science in recent years. "Chinese Pharmacopoeia" records Hericium erinaceus mycelium (see pages 1614-1615 of the first volume of the 2015 edition), and the Hericium erinaceus mycelium described in the present invention is the Hericium erinaceus mycelium recorded in this standard.

[0004] Biotransformation "mycelium" refers to the biochemical reaction process of decomposing organic matter by utilizing the metabolic function of mycelium (including fungi). With suitable culture medium as nutrition, rich secondary metabolites are produced through the growth metabolism and life activities of mycelium, and different biologically active raw materials can be obtained, which helps to enhance the efficacy or reduce the toxic side effects of drugs. At present, the application of new medicinal mycelium of Hericium erinaceus to gastrointestinal cancer has become the development direction of national key research on new drugs. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a method for culturing transformed mycelium of Hericium erinaceus with good digestion-promoting and bloating-relieving effects, a traditional Chinese medicine composition for chronic gastritis and its application, which significantly improves the effect of transformed mycelium of Hericium erinaceus and its composition in treating chronic non-atrophic gastritis.

[0006] The present invention provides a method for culturing transformed mycelium of Hericium erinaceus, comprising the following steps:

[0007] 1) Cultivation of cultivated species strains: inoculating the Hericium erinaceus strain into a Hericium erinaceus culture medium, controlling the temperature to 20-27° C., the relative humidity to 85%-90%, and culturing for 30-40 days, wherein the Hericium erinaceus culture medium comprises the following components in parts by weight: 350-490 parts of bran, 100-140 parts of corn husks, 50-70 parts of millet husks, and 400-600 parts of water;

[0008] 2) Biotransformation: inoculating the cultivated strain obtained in step 1) into a biotransformation medium, wherein the biotransformation medium comprises the following components in parts by weight: 350-490 parts of bran, 100-140 parts of corn husks, 50-70 parts of millet husks, and 400-600 parts of water, sterilizing, inoculating, controlling the temperature at 20° C.-28° C., culturing for 45-55 days, taking out the mycelium, and drying at a temperature below 65° C. to obtain Hericium erinaceus transformation mycelium.

[0009] Preferably, the bioconversion medium comprises the following components in parts by weight: 420 parts of bran, 120 parts of corn husks, 60 parts of millet husks, and 500 parts of water.

[0010] Preferably, before the culturing of the cultivar strain, the following steps are also included:

[0011] 1) Mother strain culture: inoculate the Hericium erinaceus strain into a mother culture medium and culture at a constant temperature of 20° C.-28° C. for 8-12 days; the mother culture medium comprises 75%-85% potato, 5%-10% agar powder, 4%-8% glucose, 1%-4% yeast extract, 1%-4% peptone, 0%-1% potassium dihydrogen phosphate, 0%-1% dipotassium hydrogen phosphate, 0%-1% magnesium sulfate, 0%-1% vitamin B1, and 0%-1% vitamin B2;

[0012] 2) Cultivation of stock strains: inoculate the strains cultured in step 1) into a stock culture medium and culture at 20° C.-28° C. for 25-35 days; the stock culture medium comprises the following components in parts by weight: 420 parts of cottonseed hulls, 140 parts of wheat bran, 70 parts of corn husks, and 600 parts of water.

[0013] A transformed mycelium of Hericium erinaceus, wherein the transformed mycelium of Hericium erinaceus is obtained according to any of the above-mentioned culture methods.

[0014] A traditional Chinese medicine composition for chronic gastritis comprises 16 parts of the above-mentioned transformed mycelium of Hericium erinaceus, 8 parts of cuttlebone, 4 parts of vinegared Corydalis yanhusuo, 4 parts of wine-cooked white peony root, 4 parts of vinegared Cyperus rotundus and 4 parts of liquorice.

[0015] A method for preparing the above-mentioned traditional Chinese medicine composition for chronic gastritis comprises the following steps: taking 8 parts of transformed mycelium of Hericium erinaceus, decocting twice with water, each time for 2 hours, filtering, combining the filtrate and concentrating under reduced pressure, vacuum drying, and grinding into fine powder; grinding cuttlebone, vinegar Corydalis, wine white peony root, vinegar Cyperus rotundus and liquorice into fine powder, sieving, and sterilizing; taking another 8 parts of transformed mycelium of Hericium erinaceus, grinding into fine powder, mixing with the fine powder obtained in the first two times and an appropriate amount of starch, and putting into capsules to obtain the composition.

[0016] The present invention also provides a use of the transformed mycelium of Hericium erinaceus in preparing a medicament for chronic non-atrophic gastritis.

[0017] The method for culturing transformed mycelium of Hericium erinaceus, the traditional Chinese medicine composition for chronic gastritis and the application of the present invention adopt a culture medium for biological transformation of bran, corn husk and millet husk, optimize the culture medium and optimize the production process, increase the active ingredients of the transformed mycelium of Hericium erinaceus, and the obtained transformed mycelium of Hericium erinaceus and the corresponding traditional Chinese medicine composition have good effects of promoting digestion and relieving swelling, and significantly improve the effect of the transformed mycelium of Hericium erinaceus and the composition thereof in treating chronic non-atrophic gastritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a flow chart of the subject experiment in Example 2. DETAILED DESCRIPTION

[0019] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings.

[0020] Example 1

[0021] A traditional Chinese medicine composition for chronic gastritis comprises 16 parts of transformed mycelium of Hericium erinaceus, 8 parts of cuttlebone, 4 parts of vinegared Corydalis yanhusuo, 4 parts of wineed white peony root, 4 parts of vinegared Cyperus rotundus and 4 parts of liquorice, namely Hericium erinaceus Jianweiling tablets.

[0022] The preparation method of the chronic gastritis Chinese medicine composition is as follows:

[0023] Take 8 parts of transformed mycelium of Hericium erinaceus, add water and boil twice, each time for 2 hours, filter, combine the filtrate and concentrate under reduced pressure to an appropriate amount, vacuum dry, and grind into fine powder; grind the remaining five flavors including cuttlebone, vinegar Corydalis, wine white peony root, vinegar Cyperus rotundus, and licorice powder into fine powder, sieve, and sterilize; take another 8 parts of transformed mycelium of Hericium erinaceus and grind into fine powder, mix with the two fine powders prepared above and an appropriate amount of starch, and put into capsules to obtain.

[0024] The preparation method of the Hericium erinaceus transformed mycelium is as follows:

[0025] First, culture the transformed mycelium of Hericium erinaceus:

[0026] 1) Mother strain culture: The Hericium erinaceus strain is inoculated into a mother culture medium and cultured at a constant temperature of 24° C. for 12 days; the composition of the mother culture medium is 85% potato, 6% agar powder, 8% glucose, 2% yeast extract, 4% peptone, 0.5% potassium dihydrogen phosphate, 0.5% dipotassium hydrogen phosphate, 0.5% magnesium sulfate, 0.5% vitamin B1, and 0.5% vitamin B2.

[0027] 2) Cultivation of stock strains: The strains cultured in step 1) are inoculated into a stock culture medium and cultured at 20° C. for 35 days; the stock culture medium comprises the following components in parts by weight: 420 parts of cottonseed hulls, 140 parts of wheat bran, 70 parts of corn husks, and 600 parts of water.

[0028] 3) Cultivation of cultivar strains: inoculating the Hericium erinaceus strain into a Hericium erinaceus culture medium, controlling the temperature to 27° C. and the relative humidity to 87%, and culturing for 35 days, wherein the Hericium erinaceus culture medium comprises the following components in parts by weight: 420 parts of bran, 120 parts of corn husks, 60 parts of millet husks, and 500 parts of water;

[0029] 4) Biotransformation: inoculating the cultivated strain obtained in step 3) into a biotransformation medium, wherein the biotransformation medium comprises the following components in parts by weight: 420 parts of bran, 120 parts of corn husks, 60 parts of millet husks, and 500 parts of water; inoculating after sterilization, controlling the temperature at 28° C., culturing for 45 days, taking out the mycelium, and drying it at a temperature below 65° C. to obtain Hericium erinaceus transformation mycelium.

[0030] Example 2

[0031] A randomized, double-blind, double-dummy, positive-controlled, non-inferiority clinical trial of the effectiveness and safety of Hericium erinaceus Jianweiling Tablets in Example 1 in treating chronic non-atrophic gastritis.

[0032] 1 Trial Overview

[0033] 1.1 Random

[0034] Block randomization was used. Random numbers were generated using SAS 9.4 software, i.e., all subjects received the corresponding treatment allocation table (trial drugs and control drugs were allocated in a 1:1 ratio). The trial drugs were coded according to the randomization table, and the subjects were enrolled in the order of the trial drug numbers. This clinical study was participated by one research center and was planned to complete 240 cases. The random numbers were reproducible. The block length, seed number, and other parameters were recorded in the blind bottom.

[0035] 1.2 Implementation of the blind method

[0036] This clinical trial adopted the double-blind double-dummy method.

[0037] Biostatisticians used SAS 9.4 software to generate a random number table for drug allocation to ensure that the subjects were selected into the group and the corresponding treatment drugs were allocated according to the random number table. The generation of random numbers, the coding of the study drugs, the use of drugs by the subjects, the recording and evaluation of the study results, the monitoring of the study process, the data management and statistical analysis must remain blind. According to the needs of the blind method, simulants of Hericium erinaceus Jianweiling Tablets and Omeprazole Enteric-coated Tablets were prepared respectively. The shape, size, properties, smell, packaging, labeling and other characteristics of the simulants are consistent with Hericium erinaceus Jianweiling Tablets and Omeprazole Enteric-coated Tablets, but do not contain active ingredients. The test drugs and control drugs are uniformly packaged to ensure that the study drugs and reference drugs are indistinguishable in appearance, and the usage and dosage of the study drugs and control drugs are the same.

[0038] After completing the drug blinding, the blind editor should sign the envelope and submit it to the clinical research unit and the sponsor. After the study is completed, it will be used for statistical analysis and unblinding.

[0039] After data locking and before statistical analysis, the first unblinding is performed to determine whether each drug number belongs to group A or group B. After statistical analysis, the principal investigator writes a summary report based on the statistical report. At the clinical study summary meeting, the sponsor's representative, principal investigator, biostatistician and other experts jointly conduct the second unblinding to determine whether group A and group B are test drugs or control drugs, respectively.

[0040] 1.3 Trial Drugs

[0041] Experimental drugs: Hericium erinaceus Jianweiling tablets, oral, 4 tablets at a time, 3 times a day; Omeprazole enteric-coated tablets simulant, oral, 1 tablet at a time, once a day, swallowed every morning.

[0042] Control drugs: Omeprazole enteric-coated tablets, oral, 1 tablet at a time, once a day, swallowed every morning; Hericium erinaceus simulant, oral, 4 tablets at a time, 3 times a day.

[0043] 1.4 Sample size

[0044] Operational considerations: Taking into account the 20% withdrawal rate in clinical trials and the operability of clinical trials, the sample size of this trial was determined to be 120 cases in each group, for a total of 240 cases.

[0045] 1.5 Visiting Points

[0046] (1) Visit 1: Screening phase (Visit 1, day-10 to 0)

[0047] (2) Visit 2: enrollment and medication (Visit 2, 1st week ± 2 days)

[0048] (3) Visit 3: Medication phase (Visit 3, 3rd week ± 2 days)

[0049] (4) Visit 4: Follow-up period (Visit 4, 5th week ± 5 days)

[0050] 2. Observation of effectiveness and safety

[0051] 2.1 Effectiveness Observation

[0052] 2.1.1 Main efficacy indicators

[0053] (1) Improvement rate of main clinical symptoms and signs

[0054] The main clinical symptoms and signs (stomach pain, gastric bloating) of chronic non-atrophic gastritis of the subjects were scored before administration, 2 weeks after administration, and 4 weeks after administration. The scoring criteria are shown in Table 2-1.

[0055] Table 2-1 Scoring criteria for main symptoms

[0056]

[0057]

[0058] The therapeutic index was calculated as follows: therapeutic index = [(pre-treatment score - post-treatment score) ÷ pre-treatment score] × 100%.

[0059] Judging clinical efficacy:

[0060] Clinical recovery: clinical symptoms basically disappear, efficacy index ≥ 95%;

[0061] Markedly effective: clinical symptoms were significantly improved, 70%≤efficacy index<95%;

[0062] Effective: clinical symptoms improved, 30%≤efficacy index<70%;

[0063] Ineffective: No improvement in clinical symptoms, efficacy index <30%.

[0064] When the efficacy index 4 weeks after administration is ≥30%, the drug is considered to be effective; when the efficacy index is <30%, it is considered ineffective.

[0065] 2.1.2 Secondary efficacy indicators

[0066] (1) VAS score

[0067] The degree of gastric pain of the subjects was scored before administration, 2 weeks after administration, and 4 weeks after administration, using the visual analog scale (VAS) method, which is currently more commonly used in clinical practice internationally, namely the 0-10 Numeric Pain Intensity Scale: a 10 cm line segment was taken, 0 for no pain, and 10 for extreme pain. The patients marked the corresponding positions according to the degree of pain, and the researchers measured the specific scores.

[0068] The VAS score was used to grade the degree of gastric pain: normal: 0cm; mild: 0-3cm; moderate: 4-6cm; severe: 7-10cm.

[0069] (2) Improvement rate of minor clinical symptoms and signs of chronic non-atrophic gastritis.

[0070] The subjects' minor clinical symptoms and signs of chronic non-atrophic gastritis were scored before administration, 2 weeks after administration, and 4 weeks after administration. The scoring criteria for minor symptoms are shown in Table 2-2.

[0071] Table 2-2 Scoring criteria for minor symptoms

[0072]

[0073] (3) Improvement rate of overall clinical symptoms and signs of chronic non-atrophic gastritis

[0074] Total symptom score = major symptom score + minor symptom score.

[0075] (4) Gastric pain disappearance rate / relief rate, relief time / disappearance time

[0076] The subject's gastric pain symptom score dropped to 0 points and remained there until the end of the treatment without recurrence, which was called "disappearance".

[0077] A one-level decrease in the subject's gastric pain symptom score is called "relief."

[0078] (5) Distension disappearance rate / relief rate, relief time / disappearance time

[0079] The subject's bloating symptom score dropped to 0 and remained there until the end of the treatment without recurrence, which was called "disappearance".

[0080] A one-level decrease in the subject's bloating symptom score is called "relief."

[0081] (6) Helicobacter pylori stool antigen test

[0082] Fecal antigen testing was performed 4 weeks after administration to check the negative rate of Helicobacter pylori.

[0083] 2.2 Safety Observation

[0084] 2.2.1 Routine safety observation Routine test items, test frequency and time points

[0085] (1) Physical examination, vital signs (blood pressure, pulse, temperature, respiration)

[0086] (2) Routine blood, urine and stool tests, and urine pregnancy tests for women

[0087] (3) Electrocardiogram

[0088] (4) Liver function (alanine aminotransferase ALT, aspartate aminotransferase AST, total bilirubin TBil, glutamate transpeptidase, alkaline phosphatase) and renal function (urea nitrogen BUN, creatinine Cr)

[0089] The above items should be tested once within 7 days before enrollment and within 2 days after the end of the treatment. Patients who were normal before treatment but abnormal after treatment, or abnormal before treatment but aggravated after treatment, should be rechecked immediately. Those who were confirmed to be abnormal should be followed up and rechecked regularly until they were normal, or rechecked 3 times and confirmed to be unable to return to normal.

[0090] 2.2.2 Adverse events

[0091] All adverse events occurring during the trial were recorded.

[0092] 1. Statistical analysis of population segmentation

[0093] (1) Full analysis set (FAS): refers to all cases that have been randomized and used the study drug at least once and have post-drug evaluation data according to the intention-to-treat (ITT) principle. This data set is composed of all randomized subjects excluded in a minimal and reasonable manner. For example, cases without any follow-up observation data after enrollment (no medication) can be excluded. Missing data related to the efficacy in the FAS will be estimated using the last observation carried forward (LOCF) method. FAS is the main population for efficacy evaluation.

[0094] (2) Per protocol set (PPS): All cases that meet the inclusion and exclusion criteria of the protocol, have good compliance (80% to 120%), have completed all planned visits and completed the main research indicators specified in the CRF, have not used or received drugs or treatments that may affect the efficacy evaluation during the trial, and have no major violations of the trial protocol. The PPS will be finalized during the blind review of the data and is the secondary population for efficacy evaluation.

[0095] (3) Safety set (SS): All patients who have been randomized and used the study drug at least once and have safety evaluation data after medication constitute the safety population of this study. SS is the main population for safety evaluation in this study.

[0096] 2. Statistical analysis methods

[0097] 5.1 Basic Considerations

[0098] The statistical analysis software was SAS 9.4.

[0099] The main efficacy index was estimated by non-inferiority test method, with one-sided test level α = 0.025 and non-inferiority margin of 0.15. When the non-inferiority test P value should be <0.025, the test drug is non-inferior to the control drug. The statistical tests of other indicators were all bilaterally tested, with test level α = 0.05, and the difference was considered statistically significant when P < 0.05.

[0100] Statistical description: Descriptive statistical indicators of quantitative data include number of cases, mean, standard deviation, median, quartile, maximum value, minimum value, etc. The minimum and maximum values ​​retain the same number of decimal places as the original data. Descriptive statistical indicators of qualitative data include number of cases and frequency. Unless otherwise specified, percentages are calculated based on non-missing data; percentages retain one decimal place.

[0101] 5.2 Hypothesis Testing

[0102] This study conducted non-inferiority tests on the main evaluation indicators.

[0103] (1) When A is the control group and B is the experimental group, the non-inferiority hypothesis test is as follows:

[0104] H0:μ A -μ B ≥Δ

[0105] H1:μ A -μ B <Δ

[0106] (2) When A is the experimental group and B is the control group, the non-inferiority hypothesis test is as follows:

[0107] H0:μ A -μ B ≤-Δ

[0108] H1:μ A -μ B >-Δ

[0109] The non-inferiority test is a one-sided test with one-sided α=0.025. When the P value ≤0.025, it indicates that H0 is rejected and the test drug is non-inferior to the control drug. For the first case, when A is the control group and B is the test group, the upper limit of the 95% confidence interval must be less than Δ (non-inferiority margin) to consider that the test drug is non-inferior to the control drug. For the second case, when A is the test group and B is the control group, the lower limit of the 95% confidence interval must be greater than -Δ to consider that the test drug is non-inferior to the control drug. The non-inferiority margin Δ in this test is 0.15.

[0110] The comparison of other secondary efficacy indicators in this study was a difference test, H0: μ A =μ B ; H1: μ A ≠μ B The two-sided test level α = 0.05, when the P value ≤ 0.05, it indicates that H0 is rejected, which indicates that the difference between group A and group B is statistically significant.

[0111] 5.3 Distribution of subjects

[0112] Descriptive statistics were mainly used to describe the distribution of subjects in the form of flow charts, summarizing the number and percentage of cases enrolled and completed, and the number of cases in each analysis set.

[0113] List the subjects who dropped out, including drug number, enrollment time, dropout / exclusion time, and reason;

[0114] List the status of each subject in the analysis data set and indicate the reasons for not including the data set.

[0115] 5.4 Demographic and baseline data

[0116] The analysis was performed based on FAS and PPS.

[0117] Group and count demographic data such as gender, age, height, weight, BMI, ethnicity, etc.

[0118] Baseline data included current medical history, past medical history, past medication history, and baseline examinations.

[0119] Quantitative data were compared using t-test or rank-sum test, and qualitative data were compared between groups using chi-square or Fisher's exact probability method to determine whether the baseline between groups was balanced.

[0120] 5.5 Compliance and concomitant medication

[0121] The analysis was performed based on FAS.

[0122] Medication compliance: The compliance of each group was calculated by classifying them into good compliance and poor compliance.2 Test or Fisher's exact probability analysis. Good compliance means 80% ≤ actual medication dosage / prescribed medication dosage ≤ 120%, and poor compliance means actual medication dosage / prescribed medication dosage < 80% or > 120%.

[0123] Combined medication: Combined medication in group statistical trials is measured using X 2 The inter-group comparison was performed using the P value test or Fisher's exact probability method, and all concomitant medications were listed, including drug number, group, drug name, reason for use, start time, end time, etc.

[0124] 5.6 Security Analysis

[0125] Analysis based on SS.

[0126] 5.7 Effectiveness Evaluation

[0127] The analysis was performed based on FAS and PPS.

[0128] 5.7.1 Symptom score

[0129] Descriptive statistics were performed on the single symptom scores, main symptom scores, minor symptom scores and total scores of Visits 1 to 4 by group. Group t-test or rank-sum test was used for inter-group comparison. Paired t-test or paired rank-sum test was used to compare the scores of Visits 3 and 4 before and after administration.

[0130] 5.7.2 Therapeutic index

[0131] The differences in single symptom scores, major symptom scores, minor symptom scores and total scores before and after administration were calculated, and the improvement of symptoms and signs was evaluated.

[0132] Clinical recovery: clinical symptoms basically disappear, efficacy index ≥ 95%;

[0133] Markedly effective: clinical symptoms were significantly improved, 70%≤efficacy index<95%;

[0134] Effective: clinical symptoms improved, 30%≤efficacy index<70%;

[0135] Ineffective: No improvement in clinical symptoms, efficacy index <30%

[0136] Efficacy index = [(pre-treatment score - post-treatment score) ÷ pre-treatment score] × 100%

[0137] Using X 2 The effectiveness between groups was compared by using the Fisher exact probability method, and the 95% confidence interval of the difference between the groups was calculated by the WALD method.

[0138] 5.7.3 Non-inferiority test of clinical efficacy

[0139] The effective rate of clinical efficacy was statistically analyzed in groups. An efficacy index ≥ 30% was considered effective, and an efficacy index < 30% was considered ineffective. Non-inferiority test was used for inter-group comparison, and the difference in clinical effective rate between the two groups and 95% confidence interval were calculated. The 95% confidence interval of the difference in the effective rate of the main symptom score and the non-inferiority margin were used to determine whether the test drug was non-inferior to the control drug.

[0140] 5.7.4 VAS scoring

[0141] Descriptive statistics were performed on the VAS scores of visits 1 to 4 by group, and group t-test or rank-sum test was used for inter-group comparison. Paired t-test or paired rank-sum test was used to compare the scores of visits 3 and 4 before and after administration.

[0142] 5.7.5 Relief / disappearance time of stomach pain / bloating symptoms

[0143] Descriptive statistics were performed on the results of the relief and disappearance rates of gastric pain / bloating symptoms in each group, and X 2 The inter-group comparisons were performed using Fisher's exact probability method.

[0144] Descriptive statistics were performed on the results of the relief and disappearance time of gastric pain / bloating symptoms in each group, and t-test or rank-sum test was used for comparison between groups.

[0145] 5.7.6 Helicobacter pylori negative rate

[0146] Negative rate: Fecal antigen test was performed 4 weeks after administration, and the negative rate of Helicobacter pylori in each group was statistically analyzed and X 2 The inter-group comparisons were performed using Fisher's exact probability method.

[0147] Negative conversion rate: Calculate the ratio of positive before treatment and negative after treatment in the two groups, and use X 2 The inter-group comparisons were performed using Fisher's exact probability method.

[0148] 5.7.6 Sensitivity analysis

[0149] For subjects with controversial inclusion in the FAS or PPS dataset, a sensitivity analysis of the final results was performed by including the results in the FAS or PPS dataset, and the differences between the two analysis results were compared. If the sensitivity analysis results showed that the results of the two comparisons were the same, the analysis results of the controversial subjects were the final results. If the results were different, the reasons should be analyzed.

[0150] 6. Statistical analysis results

[0151] 6.1 Distribution of Subjects

[0152] 6.1.1 Completion of the test

[0153] This trial planned to enroll 240 subjects, including 120 in group A and 120 in group B. A total of 201 subjects completed the trial, including 101 in group A and 100 in group B.

[0154] 6.1.2 Dataset Division

[0155] A total of 205 cases were included in FAS, including 102 cases in group A and 103 cases in group B; a total of 201 cases were included in PPS, including 101 cases in group A and 100 cases in group B; a total of 206 cases were included in SS, including 102 cases in group A and 104 cases in group B.

[0156] 6.1.3 Subjects who dropped out

[0157] In this trial, 39 subjects did not complete the trial, of which 30 subjects did not complete the trial due to "drop off" and 9 subjects did not complete the trial due to "eliminated from trial".

[0158] 19 subjects in Group A did not complete the trial, including 12 subjects who were “dropped off” and 7 subjects who were “eliminated from trial”.

[0159] Twenty subjects in Group A did not complete the trial, including 18 subjects who were “dropped off” and 2 subjects who were “eliminated from trial”.

[0160] 6.2 Demographic and baseline data

[0161] 6.2.1 Demographic data

[0162] The FAS results are as follows:

[0163] The average age of the subjects in group A was 37.3±12.06 years old, with 56 (54.9%) males and 46 (45.1%) females; 77 (75.5%) married and 25 (24.5%) unmarried; 51 (50.0%) manual laborers and 51 (50.0%) non-manual laborers; the average height was 158.77±10.068cm, the average weight was 74.34±17.140kg, and the average BMI was 29.40±6.854kg / m 2 .

[0164] The average age of the subjects in group B was 37.5±12.60 years old, with 45 (43.7%) males and 58 (56.3%) females; 76 (73.8%) married and 27 (26.2%) unmarried; 35 (34.0%) manual laborers and 68 (66.0%) non-manual laborers; the average height was 158.62±10.012cm, the average weight was 70.36±15.961kg, and the average BMI was 27.80±6.055kg / m 2 .

[0165] The PPS results are as follows:

[0166] The average age of the subjects in group A was 37.3±12.12 years old, with 55 (54.5%) males and 46 (45.5%) females; 76 (75.2%) married and 25 (24.8%) unmarried; 50 (49.5%) manual laborers and 51 (50.5%) non-manual laborers; the average height was 158.87±10.070cm, the average weight was 74.22±17.178kg, and the average BMI was 29.31±6.818kg / m 2 .

[0167] The average age of the subjects in group B was 37.4 (12.63 years old), with 43 (43.0%) males and 57 (57.0%) females; 74 (74.0%) married and 26 (26.0%) unmarried; 34 (34.0%) manual laborers and 66 (66.0%) non-manual laborers; the average height was 158.86±9.799cm, the average weight was 71.04±15.691kg, and the average BMI was 27.99±6.014kg / m 2 .

[0168] The analysis results of the FAS set and the PPS set showed that there was no statistically significant difference between the two groups in terms of age, gender, ethnicity, marital status, height, weight, BMI, etc. (P>0.05), but there was a statistically significant difference in the nature of work (P<0.05), with a larger proportion of non-manual laborers in Group B.

[0169] 6.2.2 Gastroscopy and pathological examination results

[0170] FAS

[0171] During the screening period, the subjects were subjected to gastroscopy and pathological examination, and the results showed that all subjects were diagnosed with non-atrophic gastritis. The results of Hp examination showed that 87 (86.1%) subjects in group A were positive and 14 (13.9%) were negative, and 81 (78.6%) subjects in group B were positive and 22 (21.4%) were negative. The chi-square test was used for inter-group comparison, and there was no statistically significant difference (P = 0.1602).

[0172] Table 6-1 Gastroscopy and pathological biopsy [FAS]

[0173]

[0174] PPS

[0175] During the screening period, the subjects were subjected to gastroscopy and pathological examination, and the results showed that all subjects were diagnosed with non-atrophic gastritis. The results of Hp examination showed that 86 (86.0%) subjects in group A were positive and 14 (14.0%) were negative, and 78 (78.0%) subjects in group B were positive and 22 (22.0%) were negative. The chi-square test was used for inter-group comparison, and the difference was not statistically significant (P = 0.1409).

[0176] Table 6-2 Gastroscopy and pathological biopsy [PPS]

[0177] variable Group A Group B P-value Gastroscopy and biopsy Non-atrophic gastritis 100(100.0％) 100(100.0％) total 100(100.0％) 100(100.0％) HP Check Negative 14(14.0％) 22(22.0％) 0.1409 Positive 86(86.0％) 78(78.0％) total 100(100.0％) 100(100.0％)

[0178] 6.2.3 Past medical history / medication status

[0179] FAS

[0180] The disease course of subjects in Group A was 20.8±6.73 days, 101 (99.0%) subjects had been diagnosed with gastritis, 1 (1.0%) subject had received drug treatment within 2 weeks before the trial, 16 (15.7%) subjects had other diseases, and 16 (15.7%) subjects had received drug treatment due to concurrent diseases.

[0181] The disease course of subjects in Group B was 21.6±6.17 days, 103 (100.0%) subjects had been diagnosed with gastritis, no subject had received drug treatment within 2 weeks before the trial, 15 (14.6%) subjects had other diseases, and 15 (14.6%) subjects had received drug treatment due to concurrent diseases.

[0182] There was no statistically significant difference in the medical history / medication status between the two groups.

[0183] Table 6-3 Medical history / medication history [FAS]

[0184]

[0185]

[0186] PPS

[0187] The disease course of subjects in Group A was 20.8±6.77 days, 100 (99.0%) subjects had been diagnosed with gastritis, 1 (1.0%) subject had received drug treatment within 2 weeks before the trial, 16 (15.8%) subjects had other diseases, and 16 (15.8%) subjects had received drug treatment due to concurrent diseases.

[0188] The disease course of subjects in Group B was 21.6±6.25 days, 100 (100.0%) subjects had been diagnosed with gastritis, no subject had received drug treatment within 2 weeks before the trial, 15 (15.0%) subjects had other diseases, and 15 (15.0%) subjects had received drug treatment due to concurrent diseases.

[0189] There was no statistically significant difference in the medical history / medication status between the two groups.

[0190] Table 6-4 Medical history / medication history [FAS]

[0191]

[0192]

[0193] 6.2.4 Baseline vital signs

[0194] During the screening period, the respiratory rate was 19.2±2.07 (times / min) in group A and 19.4±2.06 (times / min) in group B;

[0195] The pulse rate of Group A was 81.8±9.73, and that of Group B was 82.4±8.75 (times / min); the body temperature of Group A was 37.00±0.165 (℃), and that of Group B was 36.71±2.982 (℃); the systolic blood pressure of Group A was 124.8±11.16 (mmHg), and that of Group B was 123.3±9.93 (mmHg); the diastolic blood pressure of Group A was 80.2±8.81, and that of Group B was 80.0±7.23 (mmHg).

[0196] 6.2.5 Baseline symptom score

[0197] FAS

[0198] During the screening period, the total symptom score was 27.1±7.00 in group A and 29.0±7.82 in group B. There was no statistically significant difference in the total symptom score between the two groups.

[0199] The total score of the main symptom score was 15.8±5.80 in group A and 17.7±6.63 in group B. The difference in the main symptom scores between the two groups was statistically significant.

[0200] The total score of the minor symptom score was 11.2±.86 in group A and 11.2±2.65 in group B. There was no statistically significant difference in the minor symptom scores between the two groups.

[0201] PPS

[0202] During the screening period, the total symptom score was 27.1±7.03 in group A and 29.1±7.90 in group B. There was no statistically significant difference in the total symptom score between the two groups.

[0203] The total score of the main symptom score was 15.9±5.81 in group A and 17.8±6.70 in group B. The difference in the main symptom scores between the two groups was statistically significant.

[0204] The total score of the minor symptom score was 11.2±2.88 in group A and 11.2±2.67 in group B. There was no statistically significant difference in the minor symptom scores between the two groups.

[0205] 6.2.6 Baseline VAS score

[0206] FAS

[0207] During the screening period, the VAS score was 3.7±1.34 in group A and 4.1±1.52 in group B. There was no statistically significant difference in VAS scores between the two groups.

[0208] PPS

[0209] During the screening period, the VAS score was 3.7±1.34 in group A and 4.1±1.53 in group B. There was no statistically significant difference in VAS scores between the two groups.

[0210] 6.3 Compliance and concomitant medication

[0211] 6.3.1 Concomitant medication

[0212] FAS

[0213] During the trial, 16 subjects (15.7%) in group A and 15 (14.6%) in group B took concomitant medications. The chi-square test was used to compare the two groups, and the difference was not statistically significant (P=0.8224).

[0214] PPS

[0215] During the trial, 16 subjects (15.8%) in group A and 15 (15.0%) in group B took concomitant medications. The chi-square test was used to compare the two groups, and the difference was not statistically significant (P=0.8688).

[0216] Table 6-5 Summary of combined medication

[0217]

[0218]

[0219] 6.3.2 Medication compliance

[0220] The results based on PPS showed that the medication compliance of the subjects in Group A and Group B was between 80% and 120%, and the medication compliance was good.

[0221] 6.4 Effectiveness Evaluation

[0222] The analysis was performed based on FAS and PPS.

[0223] 6.4.1 Symptom score

[0224] (1) Symptom scores at each visit point after medication

[0225] FAS

[0226] The main symptom scores of Group A at Visit 3 were 9.3±4.70 points and 4.0±5.24 points at Visit 4; the main symptom scores of Group B at Visit 3 were 10.9±5.46 points and 5.5±6.04 points at Visit 4. There was a significant difference in the main symptom scores of Visit 3 between the two groups (P=0.0104), and the main symptom scores of Group A were lower than those of Group B. There was no significant difference in the main symptom scores of Visit 4 between the two groups (P=0.0776).

[0227] The scores of minor symptoms were 7.1±2.53 points at visit 3 and 4.8±2.15 points at visit 4 in group A; 7.4±2.40 points at visit 3 and 5.3±2.47 points at visit 4 in group B. There was no significant difference in the scores of major symptoms at visit 3 between the two groups (P=0.3884). There was no significant difference in the scores of major symptoms at visit 4 between the two groups (P=0.1847).

[0228] Total symptom score: visit 3 was 16.3±6.23 points in group A, and visit 4 was 8.9±6.64 points; visit 3 was 18.3±6.53 points in group B, and visit 4 was 10.8±7.66 points. There was a significant difference in the main symptom score of visit 3 between the two groups (P=0.0190), and the main symptom score of group A was lower than that of group B. There was no significant difference in the main symptom score of visit 4 between the two groups (P=0.0746).

[0229] Table 6-6 Symptom score [FAS]

[0230]

[0231] PPS

[0232] The main symptom scores of Group A at Visit 3 were 9.2±4.71 points and 3.9±5.21 points at Visit 4; the main symptom scores of Group B at Visit 3 were 10.8±5.51 points and 5.3±5.96 points at Visit 4. There was a significant difference in the main symptom scores of Visit 3 between the two groups (P=0.0148), and the main symptom scores of Group A were lower than those of Group B. There was no significant difference in the main symptom scores of Visit 4 between the two groups (P=0.1157).

[0233] The scores of minor symptoms were 7.1±2.52 points at visit 3 and 4.8±2.07 points at visit 4 in group A; 7.3±2.38 points at visit 3 and 5.2±2.36 points at visit 4 in group B. There was no significant difference in the scores of major symptoms at visit 3 between the two groups (P=0.4666). There was no significant difference in the scores of major symptoms at visit 4 between the two groups (P=0.2599).

[0234] Total symptom score: visit 3 was 16.3±6.23 points in group A, and visit 4 was 8.7±6.52 points; visit 3 was 18.1±6.55 points in group B, and visit 4 was 10.5±7.43 points. There was a significant difference in the main symptom score of visit 3 between the two groups (P=0.0254), and the main symptom score of group A was lower than that of group B. There was no significant difference in the main symptom score of visit 4 between the two groups (P=0.1106).

[0235] Table 6-7 Symptom score [PPS]

[0236]

[0237] (2) Changes in symptom scores at each visit point after medication

[0238] The changes in symptom scores before and after treatment were calculated (score change = before treatment - after treatment) and compared among the groups.

[0239] FAS

[0240] The changes in the main symptom scores were 6.6±4.46 points at visit 3 and 11.8±6.42 points at visit 4 in group A; 6.9±4.93 points at visit 4 in group B and 12.3±6.48 points at visit 4. There was no significant difference in the changes in the main symptom scores at visits 3 and 4 between the two groups.

[0241] The change in the secondary symptom score was 4.1±2.71 points at visit 3 and 6.4±2.84 points at visit 4 in group A; 3.9±2.54 points at visit 4 in group B and 5.9±3.14 points at visit 4. There was no significant difference in the change in the secondary symptom score at visits 3 and 4 between the two groups.

[0242] The changes in total symptom scores were 10.8±6.11 points at visit 3 and 18.2±7.62 points at visit 4 in group A; 10.7±6.22 points at visit 4 in group B and 18.1±8.17 points at visit 4. There was no significant difference in the changes in minor symptom scores at visits 3 and 4 between the two groups.

[0243] Table 6-8 Comparison of symptom scores between groups [FAS]

[0244]

[0245]

[0246] PPS

[0247] The changes in the main symptom scores were 6.6±4.44 points at visit 3 and 12.0±6.34 points at visit 4 in group A; 7.0±4.90 points at visit 4 in group B and 12.6±6.31 points at visit 4. There was no significant difference in the changes in the main symptom scores at visits 3 and 4 between the two groups.

[0248] The change in the secondary symptom score was 4.2±2.69 points at visit 3 and 6.5±2.78 points at visit 4 in group A; 4.0±2.52 points at visit 4 and 6.0±3.06 points at visit 4 in group B. There was no significant difference in the change in the secondary symptom score at visits 3 and 4 between the two groups.

[0249] The changes in total symptom scores were 10.9±6.05 points at visit 3 and 18.4±7.44 points at visit 4 in group A, and 11.0±6.12 points at visit 4 in group B, and 18.6±7.82 points at visit 4. There was no significant difference in the changes in minor symptom scores at visits 3 and 4 between the two groups.

[0250] Table 6-9 Comparison of symptom scores between groups [PPS]

[0251]

[0252]

[0253] 6.4.2 Comparison of symptom score efficacy index

[0254] The efficacy indexes of the main symptom scores, minor symptom scores and total symptom scores of group A and group B were calculated by group, and the differences in efficacy indexes between group A and group B were compared using non-parametric rank sum test.

[0255] FAS

[0256] The total symptom scores of group A: 3 (2.9%) cases were clinically cured, 61 (59.8%) cases were clinically effective, 31 (30.4%) cases were effective, and 7 (6.9%) cases were ineffective; the total symptom scores of group B: 3 (2.9%) cases were clinically cured, 46 (44.7%) cases were clinically effective, 42 (40.8%) cases were effective, and 12 (11.7%) cases were ineffective. The results showed that the difference in the efficacy index of the total symptom score was statistically significant (P=0.0336), and the efficacy index of group A was higher than that of group B.

[0257] PPS

[0258] The total symptom scores of group A were as follows: 3 (3.0%) cases were clinically cured, 61 (60.4%) cases were clinically effective, 31 (30.7%) cases were effective, and 6 (5.9%) cases were ineffective; the total symptom scores of group B were as follows: 3 (3.0%) cases were clinically cured, 46 (46.0%) cases were clinically effective, 41 (41.0%) cases were effective, and 10 (10.0%) cases were ineffective. The results showed that the difference in the efficacy index of the total symptom score was statistically significant (P=0.0468), and the efficacy index of group A was higher than that of group B.

[0259] Table 6-10 Comparison of clinical significance of effective indicators of symptom scoring

[0260]

[0261]

[0262] 6.4.3 Non-inferiority comparison of clinical efficacy of main symptom scores

[0263] The results of FAS and PPS are consistent, as follows:

[0264] The clinical effective rate of FAS group A was 92 (90.2%), and the effective rate of group B was 92 (89.3%). The chi-square test was used for comparison, and there was no statistically significant difference between the two groups (P = 0.8362). The 95% confidence interval of the difference in effective rate between group A and group B was calculated to be -0.0742 to 0.0918. If group A is the experimental group and group B is the control group, the lower limit of the 95% confidence interval of the difference in effective rate between group A and group B is -0.0742, which is greater than -0.15, and the experimental group can be considered non-inferior to the control group. If group A is the control group and group B is the experimental group, the upper limit of the 95% confidence interval of the difference in effective rate between group A and group B is 0.0918, which is less than 0.15, and the experimental group can be considered non-inferior to the control group.

[0265] The effective rate of PPS group A was 92 (91.1%), and the effective rate of group B was 91 (91.0%). The chi-square test was used for comparison, and the difference between the two groups was not statistically significant (P = 0.9824). The 95% confidence limit of the difference in effective rate between group A and group B was calculated to be -0.0781 to 0.0798. If group A is the experimental group and group B is the control group, the lower limit of the 95% confidence interval of the difference in effective rate between group A and group B is -0.0781, which is greater than -0.15, and the experimental group can be considered non-inferior to the control group. If group A is the control group and group B is the experimental group, the upper limit of the 95% confidence interval of the difference in effective rate between group A and group B is 0.0798, which is less than 0.15, and the experimental group can be considered non-inferior to the control group.

[0266] Table 6-11 Comparison of effective indicators of symptom scores between group A and group B

[0267]

[0268]

[0269] *Note 1: Non-inferiority trial 1 of AB refers to the non-inferiority trial of the difference between group A and group B, with a cutoff value of 0.15;

[0270] *Note 2: Non-inferiority trial 2 of BA refers to the non-inferiority trial of the difference between group B and group A, with a cutoff value of 0.15;

[0271] *Note 3: The AB 95% confidence interval refers to the 95% confidence interval of the difference between Group A and Group B;

[0272] * Note 4: When A is the control group, see the results of non-inferiority trial 1. If the upper limit of the 95% confidence interval is less than 0.15, the test group is non-inferior to the control group;

[0273] * Note 5: When B is the control group, see the results of non-inferiority trial 2. If the lower limit of the 95% confidence interval is greater than -0.15, the experimental group is non-inferior to the control group.

[0274] 6.4.4 Non-inferiority comparison of clinical efficacy of minor symptom scores

[0275] The results of FAS and PPS are consistent, as follows:

[0276] FAS

[0277] The clinical effective rate of group A was 93 (91.2%), and the effective rate of group B was 87 (84.5%). The chi-square test was used to compare the difference between the two groups, and there was no statistically significant difference (P = 0.1421). The 95% confidence limit of the difference in effective rate between group A and group B was -0.0219 to 0.1561.

[0278] PPS

[0279] The effective rate of group A was 93 (92.1%), and the effective rate of group B was 87 (87.0%). The chi-square test was used to compare the difference between the two groups, and there was no statistically significant difference (P = 0.2392). The 95% confidence limit of the difference in effective rate between group A and group B was -0.0336 to 0.1352.

[0280] 6.4.5 Non-inferiority comparison of clinical efficacy of total symptom score

[0281] The results of FAS and PPS are consistent, as follows:

[0282] FAS

[0283] The clinical effective rate of group A was 95 (93.1%), and the effective rate of group B was 91 (88.3%). The chi-square test was used to compare the difference between the two groups, and there was no statistically significant difference (P = 0.2372). The 95% confidence limit of the difference in effective rate between group A and group B was -0.0312 to 0.1269.

[0284] PPS

[0285] The effective rate of group A was 95 (94.1%), and the effective rate of group B was 90 (90.0%). The chi-square test was used to compare the difference between the two groups, and there was no statistically significant difference (P=0.2877). The 95% confidence limit of the difference in effective rate between group A and group B was -0.0341 to 0.1153.

[0286] 6.4.6 VAS scoring

[0287] (1) VAS scores at each visit point after medication

[0288] FAS

[0289] VAS score: visit 3 was 1.4±1.35 points in group A, visit 4 was 0.9±1.24 points; visit 3 was 1.6±1.47 points in group B, visit 4 was 1.2±1.40 points. There was no significant difference in the main symptom score of visit 3 between the two groups (P=0.3026), and the main symptom score of group A was lower than that of group B. There was no significant difference in the main symptom score of visit 4 between the two groups (P=0.0834).

[0290] PPS

[0291] VAS score: visit 3 was 1.3±1.35 points in group A, visit 4 was 0.9±1.25 points; visit 3 was 1.5±1.40 points in group B, visit 4 was 1.2±1.31 points. There was a significant difference in the main symptom score of visit 3 between the two groups (P=0.4540), and the main symptom score of group A was lower than that of group B. There was no significant difference in the main symptom score of visit 4 between the two groups (P=0.1438).

[0292] Table 6-12 Comparison of VAS scores between group A and group B

[0293]

[0294]

[0295] (2) Changes in VAS scores at each visit point after medication

[0296] The changes in VAS scores before and after treatment were calculated (score change = before treatment - after treatment) and compared among the groups.

[0297] FAS

[0298] The changes in VAS scores were 2.1±1.39 points at visit 3 and 2.5±1.60 points at visit 4 in group A; 2.0±1.49 points at visit 4 in group B and 2.4±1.54 points at visit 4. There was no significant difference in the changes in the main symptom scores at visits 3 and 4 between the two groups.

[0299] PPS

[0300] The changes in VAS scores were 2.1±1.38 points at visit 3 and 2.5±1.58 points at visit 4 in group A; 2.1±1.47 points at visit 4 in group B and 2.4±1.51 points at visit 4. There was no significant difference in the changes in the main symptom scores at visits 3 and 4 between the two groups.

[0301] Table 6-13 Comparison of VAS scores among groups

[0302]

[0303] 6.4.7 Comparison of VAS score efficacy index

[0304] The efficacy index of VAS score of group A and group B was calculated by group, and the difference of efficacy index between group A and group B was compared by non-parametric rank sum test. The results of FAS and PPS showed no statistically significant difference.

[0305] Table 6-14 Comparison of clinical significance of VAS score efficacy index

[0306]

[0307]

[0308] 6.4.8 Non-inferiority comparison of clinical efficacy based on VAS score

[0309] The results of FAS and PPS are consistent, as follows:

[0310] FAS

[0311] The clinical effective rate of group A was 87 (86.1%), and the effective rate of group B was 81 (78.6%). The chi-square test was used for comparison, and there was no statistically significant difference between the two groups (P = 0.1602). The 95% confidence limit of the difference in effective rate between group A and group B was calculated to be -0.0290 to 0.1789. If group A is the experimental group and group B is the control group, the lower limit of the 95% confidence interval of the difference in effective rate between group A and group B is -0.0290, which is greater than -0.15, and the experimental group can be considered non-inferior to the control group. If group A is the control group and group B is the experimental group, the upper limit of the 95% confidence interval of the difference in effective rate between group A and group B is 0.1789, which is greater than 0.15, and the experimental group cannot be considered non-inferior to the control group.

[0312] PPS

[0313] The effective rate of group A was 87 (87.0%), and the effective rate of group B was 81 (81.0%). The chi-square test was used for comparison, and the difference between the two groups was not statistically significant (P = 0.2472). The 95% confidence limit of the difference in effective rate between group A and group B was calculated to be -0.0413 to 0.1613. If group A is the experimental group and group B is the control group, the lower limit of the 95% confidence interval of the difference in effective rate between group A and group B is -0.0413, which is greater than -0.15, and the experimental group can be considered non-inferior to the control group. If group A is the control group and group B is the experimental group, the upper limit of the 95% confidence interval of the difference in effective rate between group A and group B is 0.1613, which is greater than 0.15, and the experimental group cannot be considered non-inferior to the control group.

[0314] Table 6-15 Comparison of VAS score effectiveness index between group A and group B

[0315]

[0316] *Note 1: Non-inferiority trial 1 of AB refers to the non-inferiority trial of the difference between group A and group B, with a cutoff value of 0.15;

[0317] *Note 2: Non-inferiority trial 2 of BA refers to the non-inferiority trial of the difference between group B and group A, with a cutoff value of 0.15;

[0318] *Note 3: The AB 95% confidence interval refers to the 95% confidence interval of the difference between Group A and Group B;

[0319] * Note 4: When A is the control group, see the results of non-inferiority trial 1. If the upper limit of the 95% confidence interval is less than 0.15, the test group is non-inferior to the control group;

[0320] *Note 5: When B is the control group, see the results of non-inferiority trial 2. If the lower limit of the 95% confidence interval is greater than -0.15, the experimental group is non-inferior to the control group.

[0321] 6.4.9 Time for gastric pain symptoms to be relieved / disappeared

[0322] FAS

[0323] The gastric pain symptom relief rate in group A was 84 (83.2%), and in group B was 86 (86.0%), with no significant difference between the two groups (P=0.5784).

[0324] The disappearance rate of gastric pain symptoms in group A was 63 (62.4%), and that in group B was 52 (52.0%). There was no significant difference between the two groups (P=0.1371).

[0325] The time for gastric pain relief in group A was 13.5±5.15, and that in group B was 11.9±4.69. The difference between the two groups was statistically significant (P=0.0340). The time for gastric pain relief in group B was shorter than that in group A.

[0326] The time for gastric pain symptoms to disappear was 17.7±3.33 in group A and 16.6±4.15 in group B. There was no significant difference between the two groups (P=0.1860).

[0327] Table 6-16 Gastric pain symptoms relieved / disappeared [FAS]

[0328]

[0329]

[0330] PPS

[0331] The gastric pain symptom relief rate in group A was 84 (83.2%), and in group B was 86 (86.0%), with no significant difference between the two groups (P=0.5784).

[0332] The disappearance rate of gastric pain symptoms in group A was 63 (62.4%), and that in group B was 52 (52.0%). There was no significant difference between the two groups (P=0.1371).

[0333] The time for gastric pain relief in group A was 13.5±5.15, and that in group B was 11.9±4.69. The difference between the two groups was statistically significant (P=0.0430). The time for gastric pain relief in group B was shorter than that in group A.

[0334] The time for gastric pain symptoms to disappear was 17.7±3.33 in group A and 16.6±4.15 in group B. There was no significant difference between the two groups (P=0.1860).

[0335] Table 6-17 Gastric pain symptoms relieved / disappeared [PPS]

[0336] variable Group A Group B P-value Whether stomach pain symptoms are relieved no 17(16.8％) 14(14.0％) 0.5784 yes 84(83.2％) 86(86.0％) total 101(100.0％) 100(100.0％) Whether the stomach pain symptoms disappear no 38(37.6％) 48(48.0％) 0.1371 yes 63(62.4％) 52(52.0％) total 101(100.0％) 100(100.0％) Stomach pain relief time Number of cases 84 86 0.0340 Mean(SD) 13.5(5.15) 11.9(4.69) median 12.0 11.0 Quartiles 10.0,18.0 9.0,16.0 Minimum to maximum value 3～26 3～23 Time for stomach pain symptoms to disappear Number of cases 63 52 0.1860 Mean(SD) 17.7(3.33) 16.6(4.15) median 19.0 17.0 Quartiles 16.0,20.0 14.0,19.5 Minimum to maximum value 6～25 7～23

[0337] 6.4.10 Time for gastric bloating to ease / disappear

[0338] FAS

[0339] The relief rate of gastric distension symptoms in group A was 75 (74.3%), and that in group B was 78 (78.0%). There was no significant difference between the two groups (P = 0.5338).

[0340] The disappearance rate of gastric distension symptoms in group A was 65 (64.4%), and in group B was 59 (59.0%), with no significant difference between the two groups (P=0.4348).

[0341] The time for gastric distension symptom relief in group A was 11.2±4.39, and in group B was 12.6±4.99. There was no significant difference between the two groups (P=0.0923).

[0342] The disappearance time of gastric distension symptoms in group A was 13.8±4.59, and that in group B was 16.1±4.64. The difference between the two groups was statistically significant (P=0.0116). The disappearance time of gastric distension symptoms in group A was shorter than that in group B.

[0343] Table 6-18 Relief / disappearance of gastric bloating [FAS]

[0344] variable Group A Group B P-value Is stomach bloating relieved? no 26(25.7％) 22(22.0％) 0.5338 yes 75(74.3％) 78(78.0％) total 101(100.0％) 100(100.0％) Does the bloating disappear? no 36(35.6％) 41(41.0％) 0.4348 yes 65(64.4％) 59(59.0％) total 101(100.0％) 100(100.0％) Stomach bloating relief time Number of cases 75 78 0.0923 Mean(SD) 11.2(4.39) 12.6(4.99) median 11.0 11.5 Quartiles 9.0,12.0 9.0,17.0 Minimum to maximum value 3～23 2～23 Time for stomach bloating to disappear Number of cases 65 59 0.0116 Mean(SD) 13.8(4.59) 16.1(4.64) median 13.0 17.0 Quartiles 10.0,17.0 12.0,19.0 Minimum to maximum value 5～23 6～26

[0345] PPS

[0346] The relief rate of gastric distension symptoms in group A was 75 (74.3%), and that in group B was 78 (78.0%). There was no significant difference between the two groups (P = 0.5338).

[0347] The disappearance rate of gastric distension symptoms in group A was 65 (64.4%), and that in group B was 59 (59.0%). There was no significant difference between the two groups (P=0.4348).

[0348] The time for relief of gastric distension symptoms in group A was 11.2±4.39, and that in group B was 12.6±4.99. There was no significant difference between the two groups (P=0.0923).

[0349] The disappearance time of gastric distension symptoms in group A was 13.8±4.59, and that in group B was 16.1±4.64. The difference between the two groups was statistically significant (P=0.0116). The disappearance time of gastric distension symptoms in group A was shorter than that in group B.

[0350] Table 6-19 Relief / disappearance of gastric bloating [PPS]

[0351]

[0352]

[0353] 6.4.11 Helicobacter pylori negative rate / negative conversion rate

[0354] The Helicobacter pylori negative rates after administration are as follows, and the results of FAS and PPS are consistent:

[0355] Fecal antigen detection was performed 4 weeks after administration. The negative rate of Helicobacter pylori was 71 (70.3%) in group A and 76 (76.0%) in group B. The chi-square test was used for inter-group comparison. There was no significant difference in the negative rate of Helicobacter pylori between the two groups (P=0.3618).

[0356] Table 6-20 Helicobacter pylori stool antigen detection

[0357]

[0358] The negative conversion rate of Helicobacter pylori was calculated, that is, the proportion of positive before medication that turned negative after medication. The results of FAS and PPS were consistent:

[0359] The Helicobacter pylori negative conversion rate was 61 (70.9%) in group A and 56 (71.8%) in group B. The chi-square test was used for inter-group comparison, and there was no significant difference in the Helicobacter pylori negative rate between the two groups (P=0.9027).

[0360] Table 6-21 Helicobacter pylori negative rate

[0361]

[0362] 6.5 Security Analysis

[0363] 6.5.1 Adverse events

[0364] In this trial, 1 subject experienced 1 adverse event, with an adverse event incidence of 0.5% (1 / 206), including an adverse reaction incidence of 0.0% (0 / 206).

[0365] The incidence of adverse events in group A was 0.0% (0 / 102), and the incidence of adverse reactions was 0.0% (0 / 102). In group B, 1 subject had 1 adverse event, and the incidence of adverse events was 1.0% (1 / 104), of which the incidence of adverse reactions was 0.0% (0 / 104).

[0366] 6.5.2 Laboratory tests

[0367] Complete blood test

[0368] Group A: Hemoglobin test results of 2 subjects changed from normal before medication to abnormal after medication, which had no clinical significance; Red blood cells test results of 8 subjects changed from normal before medication to abnormal after medication, which had no clinical significance; White blood cells test results of 8 subjects changed from normal before medication to abnormal after medication, which had no clinical significance; Platelets test results of 7 subjects changed from normal before medication to abnormal after medication, which had no clinical significance.

[0369] Group B: Hemoglobin test results of 8 subjects changed from normal before medication to abnormal after medication, which had no clinical significance; Red blood cells test results of 6 subjects changed from normal before medication to abnormal after medication, which had no clinical significance; White blood cells test results of 5 subjects changed from normal before medication to abnormal after medication, which had no clinical significance; Platelets test results of 6 subjects changed from normal before medication to abnormal after medication, which had no clinical significance.

[0370] Blood biochemistry

[0371] Group A: AlanineAminotransferase test results of 9 subjects changed from normal before administration to abnormal after administration, which had no clinical significance; AspartateAminotransferase test results of 10 subjects changed from normal before administration to abnormal after administration, which had no clinical significance; GamaGlutamylTransferase test results of 3 subjects changed from normal before administration to abnormal after administration, which had no clinical significance; Alkalinephosphatase test results of 6 subjects changed from normal before administration to abnormal after administration, which had no clinical significance; Totalbilirubin test results of 3 subjects changed from normal before administration to abnormal after administration, which had no clinical significance; BloodUreanitrogen test results of 0 subjects changed from normal before administration to abnormal after administration, which had no clinical significance; Creatinine test results of 8 subjects changed from normal before administration to abnormal after administration, which had no clinical significance.

[0372] Group B: AlanineAminotransferase test results of 8 subjects changed from normal before administration to abnormal after administration without clinical significance; AspartateAminotransferase test results of 9 subjects changed from normal before administration to abnormal after administration without clinical significance, and AspartateAminotransferase test result of 1 subject changed from abnormal before administration without clinical significance to abnormal after administration with clinical significance; GamaGlutamylTransferase test results of 5 subjects changed from normal before administration to abnormal after administration without clinical significance, and GamaGlutamylTransferase test results of 3 subjects changed from abnormal before administration without clinical significance to abnormal after administration with clinical significance; Alkalinephosphatase test results of 12 subjects changed from normal before administration to abnormal after administration without clinical significance; Totalbilirubin test results of 2 subjects changed from normal before administration to abnormal after administration without clinical significance; BloodUrea test results of 0 subjects The nitrogen test results changed from normal before medication to abnormal after medication, which had no clinical significance. The Creatinine test results of 14 subjects changed from normal before medication to abnormal after medication, which had no clinical significance.

[0373] Urinalysis

[0374] Group A: A total of 13 subjects' Protein test results changed from normal before medication to abnormal after medication, which had no clinical significance; a total of 1 subject's Glucose test result changed from normal before medication to abnormal after medication, which had no clinical significance; a total of 12 subjects' White blood cell test results changed from normal before medication to abnormal after medication, which had no clinical significance; a total of 18 subjects' Red blood cell test results changed from normal before medication to abnormal after medication, which had no clinical significance.

[0375] Group B: A total of 10 subjects' Protein test results changed from normal before medication to abnormal after medication, which had no clinical significance; a total of 3 subjects' Glucose test results changed from normal before medication to abnormal after medication, which had no clinical significance; a total of 9 subjects' White blood cell test results changed from normal before medication to abnormal after medication, which had no clinical significance; a total of 16 subjects' Red blood cell test results changed from normal before medication to abnormal after medication, which had no clinical significance.

[0376] 6.5.3 Vital signs

[0377] There were no significant differences in respiration, body temperature, pulse, systolic blood pressure and diastolic blood pressure between Group A and Group B before and after administration.

[0378] 6.5.4 Electrocardiogram

[0379] In Group A and Group B, 0 subjects were normal before administration and abnormal after administration, and 0 subjects who were abnormal before administration were still abnormal after administration.

[0380] 6.5.5 Physical examination

[0381] In Group A and Group B, 0 subjects were normal before administration and abnormal after administration, and 0 subjects who were abnormal before administration were still abnormal after administration.

[0382] 7. Conclusions of statistical analysis

[0383] 7.1 Effectiveness Evaluation

[0384] FAS: The clinical effective rate of group A was 92 (90.2%), and the effective rate of group B was 92 (89.3%). The chi-square test was used for comparison, and there was no statistically significant difference between the two groups (P = 0.8362). The 95% confidence interval of the difference in effective rate between group A and group B was calculated to be -0.0742 to 0.0918. If group A is the experimental group and group B is the control group, the lower limit of the 95% confidence interval of the difference in effective rate between group A and group B is -0.0742, which is greater than -0.15, and the experimental group can be considered non-inferior to the control group. If group A is the control group and group B is the experimental group, the upper limit of the 95% confidence interval of the difference in effective rate between group A and group B is 0.0918, which is less than 0.15, and the experimental group can be considered non-inferior to the control group.

[0385] PPS: The effective rate of group A was 92 (91.1%), and the effective rate of group B was 91 (91.0%). The chi-square test was used for comparison, and the difference between the two groups was not statistically significant (P = 0.9824). The 95% confidence limit of the difference in effective rate between group A and group B was calculated to be -0.0781 to 0.0798. If group A is the experimental group and group B is the control group, the lower limit of the 95% confidence interval of the difference in effective rate between group A and group B is -0.0781, which is greater than -0.15, and the experimental group can be considered non-inferior to the control group. If group A is the control group and group B is the experimental group, the upper limit of the 95% confidence interval of the difference in effective rate between group A and group B is 0.0798, which is less than 0.15, and the experimental group can be considered non-inferior to the control group.

[0386] Conclusion: The test drug was non-inferior to the control drug.

[0387] 7.2 Safety evaluation

[0388] In this trial, no adverse events or adverse reactions occurred in Group A; in Group B, 1 subject experienced 1 adverse event, with an adverse event incidence of 1.0% (1 / 104), of which the adverse reaction incidence was 0.0% (0 / 104).

[0389] In this trial, 1 subject experienced 1 adverse event, with an adverse event incidence of 0.5% (1 / 206), including an adverse reaction incidence of 0.0% (0 / 206).

[0390] Conclusion: No serious adverse events occurred in this trial, and the safety was good.

[0391] A person skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of protection of the present application is limited to these examples. In line with the concept of the present application, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of different aspects of one or more embodiments of the present application as described above, which are not provided in detail for the sake of simplicity.

[0392] One or more embodiments of the present application are intended to cover all such substitutions, modifications and variations that fall within the broad scope of the present application. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of one or more embodiments of the present application should be included in the protection scope of the present application.

Claims

1. A method for culturing transformed mycelium of Hericium erinaceus, characterized in that: The steps include: 1) Cultivation of cultivated species strains: inoculating the Hericium erinaceus strain into a Hericium erinaceus culture medium, controlling the temperature to 20-27° C., the relative humidity to 85%-90%, and culturing for 30-40 days, wherein the Hericium erinaceus culture medium comprises the following components in parts by weight: 350-490 parts of bran, 100-140 parts of corn husks, 50-70 parts of millet husks, and 400-600 parts of water; 2) Biotransformation: The cultivated strain obtained in step 1) is inoculated into a biotransformation medium, wherein the biotransformation medium comprises the following components in parts by weight: 350-490 parts of bran, 100-140 parts of corn husks, 50-70 parts of millet husks, and 400-600 parts of water. After sterilization, the inoculation is carried out, the temperature is controlled at 20° C.-28° C., and the culture is carried out for 45-55 days. The mycelium is taken out and dried at a temperature below 65° C. to obtain the transformed mycelium of Hericium erinaceus.

2. The method for culturing transformed mycelium of Hericium erinaceus according to claim 1, characterized in that: The bioconversion culture medium comprises the following components in parts by weight: 420 parts of bran, 120 parts of corn husks, 60 parts of millet husks, and 500 parts of water.

3. The method for culturing transformed mycelium of Hericium erinaceus according to claim 1, characterized in that: Before the cultivation of the cultivar strain, the following steps are also included: 1) Mother strain culture: inoculate the Hericium erinaceus strain into a mother culture medium and culture at a constant temperature of 20° C.-28° C. for 8-12 days; the mother culture medium comprises 75%-85% potato, 5%-10% agar powder, 4%-8% glucose, 1%-4% yeast extract, 1%-4% peptone, 0%-1% potassium dihydrogen phosphate, 0%-1% dipotassium hydrogen phosphate, 0%-1% magnesium sulfate, 0%-1% vitamin B1, and 0%-1% vitamin B2; 2) Cultivation of stock strains: inoculate the strains cultured in step 1) into a stock culture medium and culture at 20° C.-28° C. for 25-35 days; the stock culture medium comprises the following components in parts by weight: 420 parts of cottonseed hulls, 140 parts of wheat bran, 70 parts of corn husks, and 600 parts of water.

4. A transformed mycelium of Hericium erinaceus, characterized in that: The transformed mycelium of Hericium erinaceus is obtained by the culturing method according to any one of claims 1 to 3.

5. A Chinese medicine composition for chronic gastritis, characterized in that: The method comprises 16 parts of the transformed mycelium of Hericium erinaceus as claimed in claim 4, and further comprises 8 parts of cuttlebone, 4 parts of vinegar-cooked Corydalis yanhusuo, 4 parts of wine-cooked white peony root, 4 parts of vinegar-cooked Cyperus rotundus and 4 parts of liquorice.

6. A method for preparing the Chinese medicine composition for chronic gastritis as claimed in claim 5, characterized in that: Take 8 parts of transformed mycelium of Hericium erinaceus, add water and boil twice, each time for 2 hours, filter, combine the filtrate and concentrate under reduced pressure, vacuum dry, and grind into fine powder; grind cuttlebone, vinegar Corydalis, wine white peony root, vinegar Cyperus rotundus and liquorice into fine powder, sieve and sterilize; take another 8 parts of transformed mycelium of Hericium erinaceus and grind into fine powder, mix with the fine powder obtained from the first two times and an appropriate amount of starch, and put into capsules to obtain.

7. Use of the transformed mycelium of Hericium erinaceus according to claim 4 in preparing a drug for chronic non-atrophic gastritis.

Images