BIOLOGICAL MICROBIAL DRUG FOR TREATING SKIN DISEASES, AND PREPARATION METHOD AND APPLICATION THEREOF

Abstract

A biological microbial drug for treating skin diseases as well as a preparation method and application thereof. The biological microbial drug includes Pleurotus ostreatus cultivated in a medium added with a Traditional Chinese Medicine (TCM) combination, and the TCM combination includes Sangusis draconis, Astragalus membranaceus, Schisandra chinensis, Herba epimedii, Eucommia ulmoides, Chinese wolfberry and Cortex dictamni. The dry biological microbial drug can be brewed like tea or boiled with water, and is non-bitter, non-toxic, tasteless, free of side effects, green, natural, and suitable for the treatment of psoriasis, eczema, urticaria and neurodermatitis.

Claims

1. A biological microbial drug for treating skin diseases, comprising: Pleurotus ostreatus cultivated in a Pleurotus ostreatus medium added with a traditional Chinese medicine (TCM) combination, wherein, the TCM combination comprises Sangusis draconis, Astragalus membranaceus, Schisandra chinensis, Herba epimedii, Eucommia ulmoides, Chinese wolfberry and Cortex dictamni.

2. The biological microbial drug for treating the skin diseases according to claim 1, wherein a weight ratio of the TCM combination to the Pleurotus ostreatus medium is (89.5 -110.5):(162-324), and the Pleurotus ostreatus medium added with the TCM combination has a water content of 55%-65% and a pH value of 6.5-8.

3. The bacteriological microbial drug for treating the skin diseases according to claim 1, wherein the TCM combination comprises, by weight, 3.5%-6.5% of the Sangusis draconis, 18.5%-21.5% of the Astragalus membranaceus, 13.5%-16.5% of the Schisandra chinensis, 8.5%-11.5% of the Herba epimedii, 3.5%-6.5% of the Eucommia ulmoides, 28.5%-31.5% of the Chinese wolfberry and 13.5%-16.5% of the Cortex dictamni.

4. The bacteriological microbial drug for treating the skin diseases according to claim 1, wherein the Pleurotus ostreatus medium comprises a cottonseed husk/broadleaf tree sawdust/corn core/bean straw powder/peanut shell/bamboo, a soybean cake powder, a maize meal, a bran, a lime powder, a gypsum powder and a brown sugar/white sugar, with a weight ratio of (100-200):(10-20):(20-40):(20-40):(6-12):(4-8):(2-4).

5. A method for preparing a biological microbial drug for treating the skin diseases, comprising the following steps: 1) preparing and sterilizing a Pleurotus ostreatus culture medium added with a TCM combination having a water content of 55%-65% and a pH value of 6.5-8, wherein the TCM combination comprises Sangusis draconis, Astragalus membranaceus, Schisandra chinensis, Herba epimedii, Eucommia ulmoides, Chinese wolfberry and Cortex dictamni, and the weight ratio of the TCM combination to the Pleurotus ostreatus medium is (89.5-110.5):(162-324); 2) under sterile conditions, inoculating a strain of Pleurotus ostreatus by a 4%-9% v/v inoculum size, with an incubation temperature below 30 C. and a humidity of 60%-70%; 3) keeping a humidity of 80%-95% when mushrooms appear; and 4) when growing to a harvesting standard, the Pleurotus ostreatus is harvested and dried in time to obtain the biological microbial drug.

6. The method for preparing the biological microbial drug for treating the skin diseases according to claim 5, wherein the Pleurotus ostreatus medium added with the TCM combination comprises 3.5-6.5 parts of the Sangusis draconis, 18.5-21.5 parts of the Astragalus membranaceus, 13.5-16.5 parts of the Schisandra chinensis, 8.5-11.5 parts of the Herba epimedii, 3.5-6.5 parts of the Eucommia ulmoides, 28.5-31.5 parts of the Chinese wolfberry, 13.5-16.5 parts of the Cortex dictamni, 100-200 parts of a cottonseed husk/broadleaf tree sawdust/corn core/bean straw powder/peanut shell/bamboo crumbs, 10-20 parts of a soybean cake powder, 20-40 parts of a maize meal, 20-40 parts of a bran, 6-12 parts of a lime powder, 4-8 parts of a gypsum powder and 2-4 parts of a brown sugar/white sugar.

7. A method for using the biological microbial drug for treating the skin diseases according to claim 1, comprising the following step: applying the biological microbial drug for treating the skin diseases.

8. The method according to claim 7, wherein the skin diseases comprise psoriasis, eczema, urticaria and neurodermatitis.

9. The method to claim 7, wherein the Pleurotus ostreatus is brewed or boiled with water and is taken once a day.

10. The method according to claim 7, wherein a weight ratio of the TCM combination to the Pleurotus ostreatus medium is (89.5-110.5):(162-324), and the Pleurotus ostreatus medium added with the TCM combination has a water content of 55%-65% and a pH value of 6.5-8.

11. The method according to claim 7, wherein the TCM combination comprises, by weight, 3.5%-6.5% of the Sangusis draconis, 18.5%-21.5% of the Astragalus membranaceus, 13.5%-16.5% of the Schisandra chinensis, 8.5%-11.5% of the Herba epimedii, 3.5%-6.5% of the Eucommia ulmoides, 28.5%-31.5% of the Chinese wolfberry and 13.5%-16.5% of the Cortex dictamni.

12. The method according to claim 7, wherein the Pleurotus ostreatus medium comprises a cottonseed husk/broadleaf tree sawdust/corn core/bean straw powder/peanut shell/bamboo, a soybean cake powder, a maize meal, a bran, a lime powder, a gypsum powder and a brown sugar/white sugar, with a weight ratio of (100-200):(10-20):(20-40):(20-40):(6-12):(2-4).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIG. 1 is the comparative photos of No. 1 patient before and after treatment with the biological microbial drug of the invention, (a) before treatment, and (b) after treatment.

[0033] FIG. 2 is the comparative photos of No. 2 patient before and after treatment with the biological microbial drug of the invention, (a) before treatment, and (b) after treatment.

[0034] FIG. 3 is the comparative photos of No. 3 patient before and after treatment with the biological microbial drug of the invention, (a) before treatment, and (b) after treatment.

[0035] FIG. 4 is the comparative photos of No. 4 patient before and after treatment with the biological microbial drug of the invention, (a) before treatment, and (b) after treatment.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0036] The technical solutions of the invention are clearly and completely described below with reference to embodiments. Clearly, the embodiments in the following description are only illustrative ones of the invention. Based on these illustrative embodiments, all other embodiments obtained by those ordinarily skilled in the field without creative efforts are all within the protection scope of the invention.

Embodiments

[0037] (I) Inoculation and Culture of Pleurotus ostreatus Strain, Original Strain and Production Strain

[0038] 1. (Grade I) Inoculation and Culture of Parent Strain

[0039] 1) Strain: Pleurotus ostreatus c Gm CC 5.729, purchased from the Institute of Microbiology, Chinese Academy of Sciences. Other strains of Pleurotus ostreatus may also be used in the invention, and the invention has no limitation in this aspect.

[0040] 2) Medium Formula:

[0041] 200 g of wheat grains, 20 g of glucose, 3 g of potassium dihydrogen phosphate, 1.5 g of magnesium sulfate, 10 g of peptone, 4 tablets of vitamin B, 20 g of agar, and 1000mL of water.

[0042] 3) Inoculation

[0043] a) First the wheat grainswith soft fire for 20 minutes, and then filter.

[0044] b) Add water and heat, and then boil the agar, glucose, magnesium sulfate, monopotassium phosphate, peptone, and vitamin B until complete dissolution. The mixed solution is measured with measuring glass to check if it is 1000 mL; if it is not sufficient, make up with boiling water.

[0045] c) Dispense the medium into tubes with 10-12 mL of liquid in each tube. Pay attention not to allow the medium to get on the test tube when dispensing, which may otherwise contaminate the cotton plug with contaminated bacteria. Then put on the cotton plug, tighten moderately, tie up the test tubes by 10/bundle, wrap the cotton plug and test tube opening with kraft paper, put it into the pressure cooker and sterilize for 40 minutes at the pressure of 1.5 kg, turn off the fire, and wait until the pressure gauge is zero before exhaustion and open the cover, and take out the test tubes and place on the table inclined, wherein the length of the slope is that of the tubes.

[0046] d) Start inoculation when the temperature of the test tubes drop below 30 degrees, and carry out inoculation under strict aseptic conditions (e.g., ultra-clean work bench and aseptic chamber).

[0047] 4) Culture

[0048] After being inoculated into the test tube, the strain placed in a 25 C. incubator and is cultured for 7-12 days. After test tube is full of the mycelium and checked without any other bacteria, the second-class original strain is inoculated.

[0049] 2. (Grade II) Inoculation and Cultivation of Original Strain p 1) Formula of Culture Materials:

[0050] 20 kg of broadleaf tree sawdust, 20 kg of corn cob, 5 kg of soybean cake powder, 5 kg of maize meal, 500 g of sugar, 250 g of magnesium sulfate, 250 g of monopotassium phosphate, 500 g of gypsum powder, and 500 g of lime powder.

[0051] 2) Inoculation and Culture

[0052] Add 65% of water to the culture materials (i.e., the water content of the medium is 65%), mix the water and the culture materials, dispense the culture materials into strain bottles or the strain bags, and sterilize the culture materials thoroughly with conventional method. When the temperature drops to 30 C., transfer the culture materials to the inoculation room or inoculation chamber (all sterile) and inoculate the parent strain. After inoculation, culture at 25-30 C. for 25-30 days, and the strain can be used after being tested as bacteria free.

[0053] 3. (Grade III) Inoculation and Cultivation of Production Strain

[0054] In the same way as the original strain produced, the times of production transference can only be increased to obtain sufficient amount of strain for production.

[0055] (II) Biological Microbial DrugInoculation of Pleurotus ostreatus

[0056] 1. Preparation of the Medium:

[0057] 5 parts of Sangusis draconis (powder), 20 parts of Astragalus membranaceus (granules), 15 parts of Schisandra chinensis (twig powder/trunk sawdust), 10 parts of Herba epimedii (powder), 5 parts of Eucommia ulmoides (granules/powder), 30 parts of Chinese wolfberry (trunk sawdust) and 15 parts of Cortex dictamni trunk (powder); there are 100 parts of TCM materials in total.

[0058] In addition, 100 parts of cottonseed husk/broadleaf tree sawdust, 10 parts of soybean cake powder, 20 parts of maize meal, 20 parts of bran, 6 parts of lime powder, 4 parts of gypsum powder and 2 parts of brown sugar.

[0059] The water content is 65% and the pH is 6.5-8.

[0060] The formula is the optimal one of the invention. The content of each component in the TCM combination, the composition and content in the Pleurotus ostreatus medium, and the added amount of the TCM combination in the Pleurotus ostreatus medium can be adjusted by those skilled in this field within a proper range according to practical requirements, but the invention is not limited thereto.

[0061] The optimal formula of the TCM combination is finally determined by the applicant of the invention after numerous times of experiment. If the active ingredients released by various TCM materials are too high, the emergency of mycelium of Pleurotus ostreatus will be inhibited, and even the culture materials will undergo acid deterioration, and contamination by other bacteria, and the active ingredients become low, and the grown Pleurotus ostreatus has no TCM effect or a slight TCM effect.

[0062] It is noteworthy that in the TCM materials selected in this application, Schisandra chinensis and Chinese wolfberry used are in the form of twig powder and trunk sawdust respectively, instead fruits. This is because the applicant found during the test that the organic acid content in their fruits is too high, causing complete failure of the culture, while after smashing of Chinese wolfberry and Schisandra chinensis trees into scraps (sawdust), the nutrition provided by these offals can not only achieve a corresponding curative effect of the cultured Pleurotus ostreatus, without bringing negative impact to the culture.

[0063] 2. Sterilization for Bag Package

[0064] 1) Stack all raw materials together on the concrete floor, rack over for 3 times for drying, stir while adding tap water, so that the raw materials can fully absorb the water. After the raw materials are mixed, grip tightly with the hand until 3-5 drops come out of the culture medium, measure if the pH value is qualified: if the pH value is low, add a small amount of lime powder for adjustment;

[0065] 2) After the completion of step 1), stack the culture materials in a trapezoidal shape, punch numerous holes from the top, so that the culture materials are ventilated up and down, and then cover with a plastic film, and begin to fill in bags 24 hours later;

[0066] 3) Use 20*42 cm plastic bags, wherein each bag can hold 1.8 kg of dry substance. After filling the bag, sterilize immediately in autoclave (100 kPa, 121 C.) for 3 hours or at atmospheric pressure (100 C.) for 16 hours. In both sterilization at normal pressure or high pressure, there should be designated person for observation until finally achieving the effect of completely killing all bacteria.

[0067] 3. Inoculation

[0068] Biological microbial drugthe Pleurotus ostreatus bag is relatively large, so it should not be inoculated in the sterile chamber, but generally inoculated in the inoculation room or simple inoculation curtain.

[0069] 1) First clean the inoculation room or curtain, and transfer the sterilized bags, strain, tools, and alcohol lamp into the inoculation room or curtain, and fumigate and sterilize by 4 g of smoke disinfectant per cubic meter, and keep the door and window sealed for 24 hours.

[0070] 2) The next morning, the staff quickly enters the inoculation room or curtain to completely disinfect all the tools and hands with 75% alcohol, and then open the bag and inoculate by the inoculation amount of 5-8% (generally 500 g of bacterial strain for 20 bags). The inoculation amount should not be too large or too small: if a too large inoculation amount may easily cause early mushrooming, thus inhibiting the growth of mycelium; a too small inoculation amount may cause slow growth rate, affecting the time of mushrooming, and easily cause contamination of other bacteria.

[0071] 3) About 2 days after inoculation, the mycelium begins to germinate; after 30 days, the whole bag is full of hyphae, white and thick, with good elasticity. During the period of mushrooming, the bacteria bag should be controlled below 30 C. (optically 20-28 C., relative humidity of 60-70%), and the contaminated bags should be taken out in time for destruction.

[0072] 4. Mushrooming Management

[0073] Biological microbial drugAfter the mushroom buds appear on the Pleurotus ostreatus bag, pay attention to the ventilation and humidification. Both ends of the bacteria bags should be opened to facilitate ventilation, and the relative humidity in the air should be increased by spraying water to the ground, walls and spaces and maintained at about 80-95%. It is forbidden to spray water directly into the mushroom buds. When the mushroom buds form the size of the soybean granules, increase the ventilation volume, and provide scattered light. As the mushrooms grow, increase the humidity in the mushroom room. Perform light spray, frequent spray, and fine spray to strengthen ventilation and air exchange, keep the air fresh and moist to facilitate normal growth and development of the sporocarp. At this time, control the light intensity at the standard of about 1000-1500 lux, and the carbon dioxide within 0.5%.

[0074] 5. Harvesting Standard

[0075] Biological microbial drugthe harvesting standard for Pleurotus ostreatus is that the mushroom cap is fully spread, but with inwardly curled edge, and the color gradually becomes paler, and the white hair starts to appear in the concave part. Early harvesting affects the production, and late harvesting makes the edge of mushroom cap become thin, the flesh become loose, and the stalk become aged, thick and hard, thus leads to degraded quality, poor taste, and reduced weight, which affects the growth of the next generation of mushrooms or affects the mushrooming.

[0076] 6. Processing

[0077] When the mushroom cap is fully spread and ready to mature, collect the whole Pleurotus ostreatus, and tear the whole mushroom into a single piece or strips for drying (control the temperature at 40 C. for the first 2 hours, then increase to 60 C. until dry). Do not dry under the sun, because the ultraviolet rays will destroy the efficacy and value of Pleurotus ostreatus.

[0078] After being dried, put in a plastic bag and seal the opening to prevent moisturizing again.

APPLICATION EXAMPLES

[0079] The applicant provided the biological microbial drug of the invention to 60 patients with psoriasis, and the patients with psoriasis were randomized evenly into 2 groups which correspondingly take the biological microbial drug prepared in Embodiment 1 once a day; the efficacy includes effective, ineffective, and recovered. During the medication, other stimulant drugs should not be taken, and blood routine and urine routine examinations should be performed regularly every month.

[0080] Ineffective: the symptoms of psoriasis are significantly improved;

[0081] Ineffective: no obvious effect;

[0082] Recovered: the psoriasis symptoms disappear without recurrence.

[0083] The numbers of effective and recovered cases in clinical trial are counted in 3 months and 6 months. The results are shown in Tables 1-3, respectively.

TABLE-US-00001 TABLE 1 Statistics on the number of effective and recovered cases after taking the biological microbial drug of the invention for 3 months in the 2 groups of psoriasis patients Group Cases Effective Ineffective Recovered Group 1 30 30 0 23 Group 2 30 30 0 24

TABLE-US-00002 TABLE 2 Statistics on the number of effective and recovered cases after taking the biological microbial drug of the invention for 6 months in the 2 groups of psoriasis patients Group Cases Effective Ineffective Recovered Group 1 30 30 0 28 Group 2 30 30 0 30

TABLE-US-00003 TABLE 3 Statistics on the number of effective and recovered cases after taking the biological microbial drug of the invention for 3 and 6 months respectively in the 2 groups of psoriasis patients Effective Recovered Time Effective Ineffective Recovered rate rate 3 months 60 0 47 100% 78% 6 months 60 0 58 100% 97%

[0084] It can be seen from Table 1-3 that the biological microbial drug for treating psoriasis provided by the invention can achieve a recovered rate higher than 78% and an effective rate of 100% in 3 months, which means that the biological microbial drug has a very good efficacy; it is observed in the trial that the skin of the recovered patients is restored to the original state within 30 days, without recurrence observed. The liver and kidney functions of the patient are normal during the drug administration, and no adverse injuries have been caused after taking the medicine.

[0085] In order to explain the authenticity of the effect of the invention, the treatment records of the patients taking the biological microbial drug of the invention are taken as the examples, and the consents have been obtained from the patients for the disclosure of the patient information and the informed consent forms have been signed.

[0086] Clinical Case 1

[0087] No. 1 patient, Liu * Mei, (151****7326, Building No. 3, Zhanqian Resident, Yichun District, Yichun City, Heilongjiang Province), suffered from psoriasis for 32 years, and was recovered after taking the biological microbial drug for 6 months, and completely rehabilitated after consolidation therapy for 1 month. The skin conditions before and after treatment are shown in FIG. 1.

[0088] Clinical Case 2

[0089] No. 2 patient, Cao*shun, (159****0780, Xiaoyun Village, Xiaoyun Town, Jinxiang County, Shandong Province), suffered from psoriasis for 6 years, and smooth all over the body after taking the biological microbial drug for 4 months, and was completely recovered after taking it for another 2 months. The skin conditions before and after treatment are shown in FIG. 2.

[0090] Clinical Case 3

[0091] No. 3 patient, Zhen * shuai, (152 **** 5927, No. 7, Fangzhuang, Peixian Development Zone, Jiangsu Province), suffered from joint-type psoriasis for 2 and a half years, with purple tinea areas all over the body, especially, the joints of the extremities were prominent, and the conditions were serious. The skin turned back normal after taking the biological microbial drug for 3 months, and complete recovery was achieved after consolidation for a month. The skin conditions before and after treatment are shown in FIG. 3.

[0092] Clinical Case 4

[0093] No. 4 patient, Ding * liang, (158****6569, Dinglou Village, Zhaozhuang Town, Feng County, Jiangsu Province), suffered from psoriasis vulgaris on the head, chest, back, buttocks and thighs and legs for 3 years, and complete recovery was achieved after taking the microbial drug for 4 months, and consolidation therapy for another month. The skin conditions before and after treatment are shown in FIG. 4.

[0094] Clinical Case 5

[0095] No. 5 patient, daughter of Zhang * yan (134****2164, Zhangzhuang Village Xuzhai Town, Shan County, Shandong Province), suffered from pustular psoriasis for 15 years, with scald head up to 0.5 cm thick all around, without any healthy area all over the body. The psoriasis is thick and sticky, brown in color, with pus and bleeding. The patient was up to 39-41 C. in severe cases. Over the years, the patient have been seeking medical advices all around, such as Beijing 301 Hospital, Shanghai Changhai Dermatology Hospital as well as Wuhan, Shijiazhuang and Shandong Skin Hospitals, and Jining Medical College Affiliated Hospital, etc., but the patient lost confidence in recovery, even the most basic control cannot be achieved. Out of desperation, the patient found our biological microbial drug. After 4 months, the skin turned back shiny as it was, and complete recovery was achieved in 6 months.

[0096] From the first patient taking biological microbial drug in 2010 until Jul. 09, 2019, the total number of psoriasis patients taking biological microbial drug is about 400, and generally significant effective was achieved after about 15 days of taking the drug, and the effect was prominent in 30 days. The effective rate is 100%, and the recovered rate is over 90%.

[0097] Clearly, compared with other existing drugs, the drug for treating psoriasis provided by this invention has the advantages of quick and effective treatment of psoriasis, without toxic or side effects, less relapse, convenient medication, and low cost, etc.