Culture medium suitable for cultivating phellinus igniarius on mulberry and its application

Abstract

The application relates to a culture medium suitable for culturing Morus logenicae and Morus logenicae moxa and for making Morus logenicae moxa moxa sticks, and belongs to the technical field of biotechnology and traditional Chinese medicine preparation. The culture medium is suitable for culturing the authentic Morus logenicae fruiting body, and the residual culture medium is also suitable for making Morus logenicae moxa moxa sticks. The Morus logenicae moxa moxa sticks can overcome the defects of the traditional Morus logenicae moxa, such as difficulty in ignition and easy extinguishing, and have the advantages of good moxa application effect, low cost, special fragrance during burning and the like. The application of the culture medium can further improve the added value of the Morus logenicae moxa industry, reduces the environmental pollution caused by the waste of the Morus logenicae moxa culture, and provides a new moxa stick for traditional Chinese medicine acupuncture and moxibustion, in addition to traditional Morus logenicae moxa and moxa sticks.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and traditional Chinese medicine preparation technology, specifically relating to a culture medium suitable for culturing Phellinus linteus from mulberry trees and for making Phellinus linteus moxibustion sticks from mulberry branches, and its application. Background Technology

[0002] Acupuncture and moxibustion, a general term encompassing acupuncture and moxibustion, is a vital medical technique in traditional Chinese medicine and was inscribed on the Representative List of the Intangible Cultural Heritage of Humanity in 2010. Moxibustion involves burning or fumigating specific acupoints on the body with pre-made moxa cones or moxa sticks, using heat stimulation to prevent and treat diseases. In traditional moxibustion, mugwort is the most common ingredient, hence traditional moxibustion is often simply referred to as "moxibustion with mugwort." However, besides moxibustion with mugwort, ancient records also mention other methods such as medicated moxibustion, willow twig moxibustion, and mulberry twig moxibustion.

[0003] Mulberry twig moxibustion is a moxibustion method that involves burning mulberry twigs or mulberry wood to create a burning ember, which is then used to fumigate the affected area. The *Compendium of Materia Medica* records that mulberry twig moxibustion "treats carbuncles and boils that do not rise to the surface, prevents bruising and decay, and treats genital sores, scrofula, phlegm sores, leg ulcers, and stubborn sores; after extinguishing the embers, apply moxibustion twice daily." Chen Shigong's *Orthodox Manual of Surgery* from the Ming Dynasty records that mulberry twig moxibustion "treats various sores and boils, preventing hardness from ulcerating and decay from rotting." Furthermore, *Essential Compendium of Materia Medica*, *Divine Moxibustion Classic*, *Introduction to Medicine*, and *Complete Works of Surgery* all record specific methods of applying mulberry twig moxibustion and consider it to have effects such as "detoxifying and relieving pain, reducing swelling and dispersing blood stasis, assisting yang and promoting tissue regeneration," and "regenerating new flesh after ulceration and decay."

[0004] Despite the numerous benefits of mulberry twig moxibustion mentioned above, and the widespread distribution of mulberry trees making twig moxibustion readily available, it is currently rare. The vast majority of moxibustion methods seen by the public are still moxibustion with twigs, suggesting that the mulberry twig moxibustion method described in ancient texts may have some drawbacks. Based on the patent applicant's numerous attempts, it is speculated that the uncommon nature of mulberry twig moxibustion may be due to its difficulty in igniting, maintaining continuous burning, and tendency to extinguish after burning, making it difficult for traditional Chinese medicine practitioners to adopt throughout history. The Ming Dynasty medical text *Surgical Orthodox Manual* by Chen Shigong records, "Light one end and apply it to the affected area; replace it when the fire goes out, using five or six pieces of wood each time," further confirming the drawback of mulberry twig moxibustion being easily extinguished and requiring repeated replacement.

[0005] Currently, the most widely used moxibustion methods in Traditional Chinese Medicine are acupuncture and moxibustion. Moxibustion sticks made from mugwort have many advantages, such as being easy to burn, having a specific mugwort aroma, and having a suitable burning temperature for moxibustion. However, they also have many cumbersome steps, such as the need for planting, harvesting, and processing mugwort, which makes them more expensive than mulberry branch moxibustion and produces more smoke when burned. Furthermore, moxibustion and mulberry branch moxibustion use different burning materials, resulting in certain differences in the methods of application, the diseases targeted, and the effects of moxibustion. Mulberry branch moxibustion still possesses specific effects that moxibustion does not have, such as "detoxification and pain relief, swelling reduction and blood stasis dispersing, yang-tonifying and tissue-regenerating" and "removing ulcers and necrotic tissue."

[0006] Therefore, to overcome the shortcomings of the ancient method of moxibustion using mulberry branches, while maintaining the efficacy of traditional mulberry branch moxibustion and overcoming the high cost of moxibustion, we need to develop a brand-new moxibustion stick based on traditional medicine and combined with modern technology.

[0007] Sanghuang is a rare medicinal fungus that parasitizes mulberry trees. Its medicinal uses are recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) from the pre-Qin period more than two thousand years ago, and in subsequent medical books throughout history. Modern research shows that Sanghuang has numerous effects, including anti-cancer properties, nodule elimination, antibacterial activity, anti-inflammation, and immune enhancement, earning it the titles of "forest gold" and "king of anti-cancer agents."

[0008] There is only one authentic Sanghuang, namely Sanghuang from the mulberry tree. It was not until 2012 that people truly discovered that the authentic Sanghuang is a previously undescribed species, a new species, namely Sanghuang from the mulberry tree (Sanghuangporus sanghuang) (Wu Shenghua, “The Renaming of the Precious Medicinal Fungus ‘Sanghuang’”, Edible and Medicinal Fungi, 2012(03)). It is a fungus belonging to the family Phytophoraceae and the genus Phytophorus that parasitizes mulberry trees, and it is a specific parasite. The fruiting body has a unique fragrance (Wu Shenghua et al., “Classification and Development Prospects of Sanghuang”, Fungal Research, 2016(4); Fang Rongjun et al., “Research on the Properties of Sanghuang”, Sericultural Science, 2022, 48(2)). The Tang Dynasty’s Zhen Quan’s “Treatise on the Properties of Medicinal Herbs” and many modern studies have shown that Sanghuang from the mulberry tree has no toxic side effects.

[0009] Because genuine wild mulberry Sanghuang is extremely rare, even considered an endangered medicinal herb, its growth requires extremely demanding environmental conditions, making artificial cultivation extremely difficult. Currently, the vast majority of the so-called "Sanghuang" on the market are "imitations" of genuine mulberry Sanghuang. Local cultivation regulations in various provinces also stipulate the use of imitations instead of genuine Sanghuang, such as Poplar Sanghuang (Sanghuangporus vaninii, host is poplar) in Zhejiang and Anhui, and Inonotushispidus (widely used host) in Shandong and Hebei. Currently, the imitations on the market include at least 13 species, such as Poplar Sanghuang, Syringa reticulata Sanghuang, Pine Sanghuang, Lacquer Sanghuang, and Inonotushispidus. While imitations are named after "mulberry," their fruiting bodies in the wild do not exclusively parasitize mulberry trees. Artificial cultivation generally uses other woods (such as oak) instead of mulberry, and some imitations have certain toxic side effects.

[0010] Therefore, if a culture medium can be invented that can cultivate authentic mulberry fruiting bodies, and does not contain miscellaneous trees other than mulberry (such as oak) used in general imitation mulberry or traditional edible fungus substitutes, as well as lime, gypsum, KH2PO4, MgSO4.7H2O and a large amount of cottonseed hulls, bran and other impurities, the substitute material after collecting mulberry fruiting bodies is suitable for making a brand new moxibustion stick - mulberry branch mulberry moxibustion stick, that is, to create a culture medium suitable for cultivating mulberry fruiting bodies and can also be used as mulberry branch mulberry moxibustion stick, then it will have at least the following three benefits: (1) Overcoming the disadvantage that authentic mulberry fruiting bodies are difficult to cultivate; (2) Reducing the environmental pollution of "waste" after mulberry cultivation, turning waste into treasure, and further enhancing the added value of the mulberry fruiting body industry; (3) Innovating the ancient method of mulberry branch moxibustion, providing traditional Chinese medicine with a new type of moxibustion stick other than the ancient method of mulberry branch moxibustion and traditional moxibustion sticks. Summary of the Invention

[0011] Technical problem: The primary technical problem to be solved by this invention is to provide a culture medium suitable for cultivating the fruiting bodies of mulberry mulberry and also for making mulberry twig moxibustion sticks.

[0012] Another technical problem to be solved by the present invention is to provide the application of the above-mentioned culture medium in the cultivation of mulberry fruiting bodies and the preparation of mulberry branch moxibustion sticks.

[0013] Technical Solution: To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0014] The Sanghuang described in this invention uses the mulberry tree Sanghuang strain, which belongs to the Sanghuangporus genus of the family Phytomycetes.

[0015] A culture medium suitable for cultivating Phellinus linteus and making Phellinus linteus moxibustion sticks is composed of: mulberry twig fragments, mulberry leaf powder, sucrose and lemon juice.

[0016] The method for preparing the above-mentioned culture medium includes the following steps:

[0017] (1) Take the mulberry branches that have survived the winter after being pruned in the spring of that year, dry them in the shade, and then crush them into mulberry branch fragments with a diameter of about 0.5cm.

[0018] (2) Pick mature mulberry leaves, remove the petioles, dry them at 40℃ to constant weight, and crush them into powder;

[0019] (3) Take natural lemons, squeeze out the lemon juice, and dilute it with purified water to make lemon water with a concentration of 0.1%;

[0020] (4) Mix the following ingredients in a mass ratio of mulberry twig powder: sucrose: mulberry leaf powder: 0.1% lemon water at 98:1:1:150. Stir well to make a suitable culture medium for cultivating mulberry fungus and making mulberry twig fungus moxibustion sticks.

[0021] (5) Place the above raw materials in a polypropylene culture bag and sterilize them under high temperature and high pressure (121℃, 0.15Mpa) for 2-2.5h. After cooling to room temperature, it can be used as a culture medium suitable for cultivating mulberry mulberry fungus and making mulberry branch mulberry fungus moxibustion sticks.

[0022] The application of the above-mentioned culture medium in the cultivation of Phellinus linteus includes the following steps:

[0023] (1) Inoculate the above culture medium with the mulberry fungus strain, and after inoculation, place it in a culture room at 28℃ for dark and static solid fermentation culture;

[0024] (2) After the mycelium has fully colonized the bag, transfer it to low light (300 lux) and continue to incubate at 28℃;

[0025] (3) After about 7 days, when most of the mycelia in the culture medium turn yellow and the protruding nodular fruiting body primordia begin to appear in the bag, make an opening treatment (make 1-2 two crescent-shaped cuts in the middle of the bag with a blade) and continue to culture at 28°C and weak light (300 lux).

[0026] (4) After about 60 days of opening, when the fruiting bodies of the mulberry tree mature, collect the fruiting bodies of the mulberry tree.

[0027] The fruiting bodies of mulberry trees cultured in the above-mentioned culture medium exhibit the following characteristics:

[0028] (1) The mulberry fruiting bodies cultured in this culture medium have excellent appearance and high yield (an average of 65g of mulberry fruiting bodies can be produced per bag), and high content of medicinal components (the total polyphenol content of the dried fruiting body powder averages 40.63mg / g), making it suitable for high-quality and high-yield cultivation of mulberry fruiting bodies.

[0029] (2) The mycelium of mulberry tree mulberry fungus cultured in this culture medium turns yellow quickly (the mycelium fills the bag and is cultured in weak light for about 7 days, and most of the mycelium will turn yellow). The content of yellow mycelium in the bag is high (the bag is covered with golden yellow mycelium skin). The residue is suitable for making mulberry branch mulberry fungus moxibustion strips.

[0030] The application of the above-mentioned culture medium in the preparation of mulberry twig and Phellinus linteus moxibustion sticks includes the following steps:

[0031] (1) After collecting the fruiting bodies of the mulberry mulberry fungus cultured in the above culture medium, remove the polypropylene plastic bag from the remaining fungal bag, keep the bag opening and the old mycelium around the fungal bag, take out the mulberry branch culture medium residue covered with mulberry mulberry fungus mycelium, and crush it into small pieces with a diameter of about 5cm.

[0032] (2) Place the fragments in an oven, dry them at 40°C until constant weight and then crush them. Sieve them through a sieve with a sieve diameter of 4 mm and keep the powder as the raw material for mulberry branch moxibustion sticks.

[0033] (3) Take the mulberry branch and moxa stick raw material from step 2, use commercially available mulberry bark paper, place it in the moxa stick rolling machine, press the raw material, and roll it into mulberry branch and moxa stick;

[0034] (4) After rolling, use glutinous rice glue to glue the mulberry bark paper joints, and seal both ends of the moxibustion strip with mulberry bark paper. After the glued area dries, the production of mulberry branch and mulberry yellow moxibustion strip is complete.

[0035] The mulberry branch and Phellinus linteus moxibustion strips prepared using the above culture medium have the following characteristics:

[0036] (1) The culture medium contains only mulberry branches (98%) and a very small amount of mulberry leaves (1%), and does not contain miscellaneous trees other than mulberry trees, lime, gypsum, KH2PO4, MgSO4·7H2O and a large amount of cottonseed hulls, bran and other impurities. When the mulberry branch moxibustion strips are burned, the temperature is close to that of the ancient mulberry branch moxibustion method, but it can overcome the defects of the ancient mulberry branch moxibustion method, such as being difficult to ignite, easy to extinguish, unstable burning state and black smoke. Therefore, it is practical.

[0037] (2) Compared with traditional moxibustion sticks, mulberry branch and mulberry fungus moxibustion sticks have similar advantages such as easy ignition, continuous burning, white smoke, suitable burning temperature, and long burning time. When burning, they have the following advantages over traditional moxibustion sticks: less smoke, less choking, and can produce the special aroma of burning mulberry fungus mycelium, which has a certain calming effect.

[0038] (3) The moxibustion effect is good. It has the effects of reducing swelling and dispersing blood stasis, promoting yang and promoting tissue regeneration, which are the traditional methods of mulberry branch moxibustion. It also has the effects of warming the meridians and dispelling cold, promoting qi and unblocking collaterals, which are the traditional methods of mulberry branch moxibustion and moxibustion. It is a new type of moxibustion stick in addition to the traditional methods of mulberry branch moxibustion and moxi

[0039] Beneficial effects:

[0040] This invention is the first to create a culture medium suitable for cultivating *Sanghuang* (a type of fungus) from mulberry trees and for making *Sanghuang* moxibustion sticks from mulberry branches. This culture medium is suitable for high-quality, high-yield cultivation of *Sanghuang* from mulberry trees. Furthermore, the culture medium contains a high mycelium content in the residual fungal bags after *Sanghuang* harvesting, and is free of other trees and debris, making it suitable for producing *Sanghuang* moxibustion sticks from mulberry branches. The produced *Sanghuang* moxibustion sticks from mulberry branches overcome the shortcomings of traditional mulberry branch moxibustion methods, such as difficulty in ignition and easy extinguishing, and are practical. Compared with traditional moxibustion sticks, they have advantages such as less pungent smoke during burning, the emission of a unique fragrance, and lower cost, making them suitable for long-term use by patients with financial difficulties. These moxibustion sticks have good therapeutic effects and represent a new type of moxibustion stick besides traditional mulberry branch moxibustion and moxibustion. The application of this culture medium can further increase the added value of the mulberry *Sanghuang* industry and reduce pollution from *Sanghuang* cultivation waste; the produced *Sanghuang* moxibustion sticks from mulberry branches can provide a new medical material for traditional Chinese medicine acupuncture. Attached Figure Description

[0041] Figure 1 Mulberry fruiting bodies and residual substrate cultured on mulberry branch culture medium. (A) Cultured fruiting bodies; (B) Dried residual substrate from mulberry branch culture medium (arrow indicates the state of the mycelium after drying).

[0042] Figure 2 These are the states of four types of moxibustion sticks during burning: 1. Mulberry twig moxibustion stick, 2. Mulberry twig powder moxibustion stick, 3. Moxa moxibustion stick, and 4. Mulberry twig and Phellinus linteus moxibustion stick. Detailed Implementation

[0043] The present invention will be further described in detail below with reference to embodiments, but the implementation of the present invention is not limited thereto.

[0044] Example 1: Preparation of solid-state fermentation culture medium for mulberry branches and Phellinus linteus

[0045] 1. Cut mulberry branches that have survived the winter after being pruned in the spring of the current year, and let them air dry indoors for about 60 days until they have lost all moisture and are no longer losing weight.

[0046] 2. Use a twig shredder to shred the mulberry twigs into pieces about 0.5cm in diameter, dry them, and store them for later use;

[0047] 3. Pick mature mulberry leaves, remove the petioles, dry at 40℃ to constant weight, crush and put into a Chinese medicine pulverizer, pulverize for 1 minute (25000 rpm), and take the powder for later use.

[0048] 4. Take commercially available fresh lemons, slice them, and use a garlic press to extract the lemon juice. Dilute the juice with purified water to make a 0.1% lemon water solution.

[0049] 4. Mix the following ingredients in a mass ratio of mulberry twig powder: sucrose: mulberry leaf powder: 0.1% lemon water at 98:1:1:150. Stir thoroughly to create a suitable culture medium for cultivating mulberry fungus and making mulberry twig fungus moxibustion sticks (referred to as "mulberry twig fungus solid fermentation culture medium" or "mulberry twig culture medium").

[0050] 5. Prepare culture medium A by mixing mulberry twig fragments, sucrose, and purified water in a mass ratio of 99:1:150. Prepare culture medium B by mixing mulberry twig fragments, sucrose, mulberry leaf powder, and purified water in a mass ratio of 98:1:1:150.

[0051] 6. Place the above three raw materials (mulberry branch and mulberry fungus solid fermentation culture medium raw material, culture medium A raw material, culture medium B raw material) into 250ml tissue culture bottles respectively, and compact them slightly. The culture medium should occupy about 2 / 3 of the culture bottle volume.

[0052] 7. Sterilize in an autoclave at 121℃ for 30 minutes, then cool in a clean bench for later use to prepare three kinds of solid fermentation culture media (mulberry branch and mulberry fungus solid fermentation culture medium, culture medium A, and culture medium B).

[0053] Example 2: Screening of Phellinus linteus strains suitable for fermentation using mulberry branches

[0054] 1. Select various strains of Phellinus linteus preserved in the laboratory on PDA medium, including Phellinus linteus vulgaris, Phellinus pineinatus, Phellinus linteus oakinatus, Phellinus lacquer treeinatus, Phellinus purpurea, Phellinus coarse-haired fibrous fungus, Phellinus poplarinatus, and Phellinus mulberryina (Table 1), and inoculate them into the three solid fermentation media in Example 1, and seal the bottle tightly. Inoculate at least three bottles for each strain.

[0055] 2. Place in complete darkness at 28°C and incubate for 45 days.

[0056] 3. After 45 days of cultivation, open the bottle and observe the growth status of various *Sanghuang* strains in the three culture media mentioned above, and smell the emitted odor. It was found that in the three culture media, *Sanghuang baoma* and *Sanghuang songshu* only produced a small amount of white mycelium, with a large number of mulberry branches remaining uninfected, thus failing to complete the entire solid-state fermentation process; the culture bottles had an off-odor after opening. *Sanghuang guashu*, *Sanghuang rufushu*, *Sanghuang ziyou*, and *Fodium coarsemiflorum* completed the entire fermentation process, but the mycelium turned white and could not form yellow mycelium; the culture bottles had a slight off-odor after opening. *Sanghuang yangshu* completed the entire fermentation process, but only formed a small amount of yellow mycelium; the culture bottles had a slight off-odor after opening. The mycelium of Phellinus linteus can fully grow on all three culture media and complete the entire fermentation process. There is no odor in the culture bottle. However, in culture media A or B, a large amount of yellow mycelium cannot be formed and there is no special fragrance. Only in the solid fermentation culture medium of Phellinus linteus can a large amount of yellow mycelium be formed and there is no odor in the culture bottle, but a special fragrance.

[0057] 4. Remove the above fermentation cultures from the bottles and dry them in a 40°C oven until the moisture is lost and the weight is constant.

[0058] 5. Grind the Chinese herbal medicine using a grinder (25,000 rpm) and sieve it through a sieve with a 4mm aperture. Set aside for later use.

[0059] 6. Ignite the above powders with an igniter and observe the burning state of each powder, as well as the emitted odor. It was found that the powders fermented in the three different culture media produced a lot of black smoke when burned, which was pungent and had an unpleasant odor, making them unsuitable as raw materials for mulberry branch moxibustion sticks. The powders fermented in culture media A or B produced white smoke when burned, which was not pungent but had no aroma; the powders fermented in mulberry branch moxibustion solid fermentation medium produced white smoke when burned, which was not pungent but emitted a unique sandalwood-like aroma similar to that of burning mulberry branch moxibustion mycelium (Table 1). The experimental results show that only the combination of culturing mulberry branch moxibustion strains in mulberry branch moxibustion solid fermentation medium is most suitable for solid fermentation culture using mulberry branches as raw material and is suitable as a raw material for mulberry branch moxibustion sticks. Therefore, only mulberry branch moxibustion strains were selected as the fermentation culture strain for subsequent experiments, and the mulberry branch moxibustion solid fermentation medium was preferred as the culture medium for subsequent experiments.

[0060] Table 1. Comparison of culture of various *Sanghuang* strains on mulberry twig medium and the ignition of the medium after drying.

[0061]

[0062]

[0063] Example 3: Cultivation and Collection of Fruiting Bodies from Mulberry Tree Phellinus linteus

[0064] 1. Prepare the raw materials for solid fermentation culture medium of mulberry branches and Phellinus linteus according to steps 1-4 in Example 1.

[0065] 2. Pack the mulberry branch and Phellinus linteus solid fermentation medium raw material from step 1 into polyethylene bags measuring 17cm x 36cm x 0.0045cm. Insert a 13cm long plastic punch with holes on the top wall into the center and seal the bags with caps. Sterilize at 121℃ under high pressure (0.15MPa) for 2-2.5 hours. When the temperature of the bags drops to around 50℃, transfer them to a sterile room or laminar flow hood to serve as the culture medium for cultivating Phellinus linteus fruiting bodies.

[0066] 3. Once the mushroom bag has cooled to room temperature, remove the plastic punch from the bag and inoculate it with the mulberry fungus strain from Example 2.

[0067] 4. After inoculation, place the bags in a dark, static solid-state fermentation room at 28°C.

[0068] 5. Once the mycelium has fully colonized the bag, transfer it to a 28°C, low-light (300 lux) environment for cultivation.

[0069] 6. Observe the mycelial growth status in the culture medium. It was found that the mycelial growth in the mulberry branch Sanghuang solid fermentation medium was faster, and the mulberry Sanghuang cultured in this medium turned yellow quickly: after only about 7 days of low light culture, most of the mycelia in the culture medium turned yellow, and protruding nodular fruiting body primordia began to appear in the bags.

[0070] 7. Opening treatment: After the protruding nodular fruiting body primordia appear, use a sterilized blade to make 1-2 two crescent-shaped cuts in the middle of the bag. After the opening treatment, the cultivation stage of the Phellinus linteus fruiting bodies and mycelium inside the bag begins.

[0071] 8. Fruiting Body Collection: After the mulberry twig Sanghuang solid fermentation medium (mulberry Sanghuang fruiting body medium) is opened and cultured for about 60 days, the mulberry twig Sanghuang fruiting bodies will turn golden yellow. At this time, the mulberry Sanghuang fruiting bodies are mature and can be collected. Mulberry Sanghuang fruiting bodies cultured in this medium are of excellent quality and high yield (an average of 65g of fruiting bodies can be produced per mulberry twig Sanghuang fruiting body). Figure 1 A).

[0072] 8. Detection of medicinal components: The polyphenol content of *Sanghuangporus sanghuang* fruiting bodies was determined according to the method described in "Optimization of Ultrasonic Extraction Process for Polyphenols from *Sanghuangporus sanghuang* by Orthogonal Experiment" (Feng Ziwang et al., 2012). The results showed that the *Sanghuangporus sanghuang* cultured on this medium had a high content of medicinal components (the average total polyphenol content of the dried fruiting body powder reached 40.63 mg / g). The results indicate that this solid-state fermentation medium for *Sanghuangporus sanghuang* is suitable for high-quality cultivation of authentic *Sanghuangporus sanghuang* fruiting bodies. Furthermore, due to the high content of yellow mycelium inside the bags and the presence of golden-yellow mycelial skin at the bag opening and around the bag, the residue is suitable for making *Sanghuangporus sanghuang* moxibustion sticks.

[0073] Example 4: Preparation of raw materials for moxibustion sticks made from mulberry twigs and Phellinus linteus

[0074] 1. After collecting the fruiting bodies, remove the polypropylene plastic bag outside the mulberry branch and Phellinus linteus solid fermentation medium, leaving the bag opening and the old mycelium around the bag. Take out the mulberry branch medium residue that is covered with golden mulberry mycelium and crush it into small pieces about 5cm in diameter.

[0075] 2. Place the fragments in an oven and dry at 40°C until constant weight. Figure 1 B).

[0076] 3. Grind the Chinese herbal medicine using a grinder (25,000 rpm) for 3-5 minutes, then sieve it through a sieve with a 4mm aperture to collect the powder, which will be used as the raw material for mulberry twig and mulberry fungus moxibustion sticks for later use.

[0077] Example 5: Preparation of Moxibustion Sticks Made from Mulberry Twigs and Phellinus Officinalis

[0078] 1. Take the mulberry twig and mulberry bark moxibustion strip raw material from Example 4, use commercially available mulberry bark paper, place it in an moxibustion strip rolling machine, compress the raw material, and roll it into mulberry twig and mulberry bark moxibustion strips.

[0079] 2. After rolling, use glutinous rice glue to seal the joints of the mulberry bark paper, and seal both ends of the moxa stick with mulberry bark paper.

[0080] 3. Place the moxa stick in a cool place and wait for the pasted area to dry. This completes the preparation of the mulberry twig and mulberry stigma moxa stick.

[0081] Example 6: Burning test of mulberry twig and mulberry wormwood moxibustion sticks

[0082] 1. Take commercially available Qichun moxa sticks (2cm in diameter, 20cm in length, and 26g in weight) and set aside.

[0083] 2. Select the mulberry branch (dried, not crushed, 2cm in diameter) from step 1 of Example 1 as the moxa stick for the ancient method of mulberry branch moxibustion, and set aside.

[0084] 3. Take the mulberry branch fragments from steps 1-2 of Example 1 and grind them into powder according to the method in step 3 of Example 4. Weigh the powder and roll it into mulberry branch powder moxibustion sticks (without mulberry pollen) using mulberry bark paper, following the steps in Example 5. The size and weight of the prepared moxibustion sticks should be the same as those in step 1 (diameter 2cm, length 20cm, weight 26g). Set aside.

[0085] 4. Weigh the raw materials of mulberry twig and mulberry fungus moxibustion strips from Example 4, and make mulberry twig and mulberry fungus moxibustion strips with a diameter of 2cm, a length of 20cm, and a weight of 26g according to the method of Example 5, for later use.

[0086] 5. Ignite the four types of moxa sticks from steps 1-4 above using an igniter, and observe the burning state. Figure 2 The flame temperature of the moxa stick at the burning site was measured using an infrared thermometer. The vertical burning time was calculated under static and windless conditions. The smoke, odor, and ash emitted during the burning process were observed. The results are shown in Table 2.

[0087] Table 2. Comparison of the burning effects of four types of moxibustion sticks

[0088]

[0089]

[0090] 6. Comparison of the results of burning with mulberry twigs and mulberry wormwood sticks:

[0091] 1) The mulberry twig culture medium contains only mulberry twigs (98%) and a very small amount of mulberry leaves (1%). The small amount of sucrose is consumed by the carbon skeleton of mycelial development, and lemon juice is a purely natural substance. Mulberry leaves themselves are a natural substance used for both medicinal and edible purposes. The culture medium does not contain artificial chemicals, nor does it contain any non-mulberry trees (such as oak, quince, etc.) or impurities (such as lime, gypsum, KH2PO4, MgSO4·7H2O, and large amounts of cottonseed hulls, bran, etc.) used in the cultivation of *Sanghuang* or traditional edible fungi. After fermentation by *Sanghuang* mycelium, the culture medium becomes loose and soft, and flammable. The high mycelial content of *Sanghuang* makes it suitable for making mulberry twig *Sanghuang* moxibustion sticks.

[0092] 2) The burning test results show that the burning temperature of the mulberry twig moxibustion stick is similar to that of the ancient mulberry twig moxibustion or moxibustion sticks made of mulberry twig powder. It overcomes the defects of the ancient mulberry twig moxibustion, such as being difficult to ignite, easy to extinguish, unstable burning state, and producing black smoke, and thus has practicality.

[0093] 3) Compared to traditional moxa sticks, mulberry twig and Phellinus linteus moxa sticks have similar advantages such as easy ignition, continuous burning, white smoke, suitable burning temperature, and long burning time. Furthermore, when burning, mulberry twig and Phellinus linteus moxa sticks offer the following advantages over traditional moxa sticks: less smoke that is not pungent, and the unique aroma (sandalwood scent) of burning mulberry and Phellinus linteus mycelium, which has certain calming and soothing effects. Since this moxa stick utilizes waste materials from Phellinus linteus cultivation, it only covers production costs and does not require the complex processes of planting, harvesting, and processing of Artemisia argyi, resulting in low cost. It can serve as a new, low-cost moxa stick alternative to traditional moxa sticks, representing an integrated innovation of medicinal fungal cultivation technology and traditional medicine.

[0094] Example 7: Moxibustion test of mulberry twig and mulberry wormwood moxibustion sticks

[0095] 1. Under the guidance of a traditional Chinese medicine practitioner, moxibustion was applied to the joints, acupoints, or affected areas of volunteers. It was found that the temperature of the mulberry twig and mulberry stigma moxa sticks was suitable when burned, and the patients' skin did not feel a burning sensation, and there was no discomfort in the joints, acupoints, or affected areas. It possesses the effects of traditional mulberry twig moxibustion, such as reducing swelling and dispersing blood stasis, and promoting yang and tissue regeneration. It also has the effects of moxibustion, such as warming the meridians and dispelling cold, and promoting qi circulation and unblocking the collaterals. It has the characteristics of good moxibustion effect.

[0096] 2. Since mulberry bark paper is made from mulberry branches, it is also a traditional Chinese medicine. Traditional Chinese medicine believes it has certain effects in dispelling wind and dampness, and promoting blood circulation and removing blood stasis. Using mulberry bark paper to make mulberry branch moxibustion sticks has even better effects in dispelling dampness and warming the body.

[0097] 3. Compared to traditional moxibustion: Mulberry twig and Phellinus linteus moxibustion sticks do not produce the strong smoke odor of traditional moxibustion. The white smoke is not pungent and instead produces a unique fragrance with calming and soothing effects, without any toxic side effects. As a new type of moxibustion stick, it can serve as a beneficial supplement or alternative to traditional mulberry twig moxibustion or moxa sticks, providing a new medical material for traditional Chinese medicine acupuncture. Since the raw material for mulberry twig and Phellinus linteus moxibustion sticks is the waste residue from the cultivation of Phellinus linteus, it represents a comprehensive utilization of medicinal fungal culture waste. This not only further enhances the added value of the rare medicinal fungus mulberry linteus industry and reduces the environmental pollution caused by Phellinus linteus culture waste, but also has a low cost, making it more suitable for long-term use by patients with financial difficulties.

Claims

1. A method suitable for cultivating Phellinus linteus (a type of medicinal mushroom) Sanghuangporus sanghuang The culture medium, characterized in that, It is composed of mulberry twig fragments, mulberry leaf powder, cane sugar, and lemon juice.

2. The method for cultivating Phellinus linteus (Sanghuang) of mulberry trees as described in claim 1. Sanghuangporus sanghuang The culture medium, characterized in that, The mass ratio of mulberry twig powder, cane sugar, mulberry leaf powder, and lemon juice is 98:1:1:

150.

3. The method for cultivating Phellinus linteus (Sanghuang) of mulberry trees according to claim 1 or 2. Sanghuangporus sanghuang The culture medium, characterized in that, The lemon water is lemon water with a lemon juice content of 0.1% by mass.

4. The method for cultivating Phellinus linteus (Sanghuang) as described in any one of claims 1 to 3. Sanghuangporus sanghuang The method for preparing the culture medium of ) is characterized in that, Includes the following steps: (A1) Take mulberry branches that have survived the winter after being pruned in the spring of that year, dry them in the shade, and then crush them into mulberry branch fragments with a diameter of 0.5cm. (A2) Pick mature mulberry leaves, remove the petioles, dry them at 40℃ to constant weight, and pulverize them into powder; (A3) Take natural lemons, squeeze out the lemon juice, and dilute it with purified water to make lemon water with a mass concentration of 0.1%; (A4) Mix thoroughly according to the mass ratio of mulberry twig powder: sucrose: mulberry leaf powder: 0.1% lemon juice of 98:1:1:150 to make mulberry mushroom extract. Sanghuangporus sanghuang Culture medium raw materials; (A5) After placing the above raw materials in polypropylene bags, sterilize them under high temperature and high pressure, and cool them to room temperature. This is suitable for cultivating mulberry fungus (Sanghuang). Sanghuangporus sanghuang ) culture medium.

5. The method for cultivating mulberry fungus (Sanghuang) according to claim 4. Sanghuangporus sanghuang The method for preparing the culture medium of ) is characterized in that, In step (A5), the sterilization conditions are: temperature 121℃, pressure 0.15Mpa, and time 2-2.5h.

6. The method for cultivating Phellinus linteus (Sanghuang) as described in any one of claims 1 to 3. Sanghuangporus sanghuang The culture medium for culturing mulberry fungus (Sanghuang) Sanghuangporus sanghuang Application in sub-entities.

7. The method for cultivating Phellinus linteus (Sanghuang) as described in any one of claims 1 to 3. Sanghuangporus sanghuang The application of the culture medium of mulberry twig and mulberry fungus in the preparation of moxibustion sticks, wherein the preparation method of the moxibustion sticks includes the following steps: (B1) The suitable mulberry mulberry fungus cultivation method as described in any one of claims 1 to 3 ( Sanghuangporus sanghuang Mulberry fungus (Sanghuang) cultured in culture medium Sanghuangporus sanghuang Sub-entity; (B2) After collecting the fruiting bodies, remove the remaining culture medium residue and crush it into small pieces with a diameter of about 5cm. (B3) Place the fragments in an oven, dry them to a constant weight, crush them, sieve them, and keep the powder as the raw material for mulberry branch and mulberry fungus moxibustion sticks; (B4) Take the mulberry twig and mulberry saffron moxibustion strip raw materials from step (B3) and roll them into mulberry twig and mulberry saffron moxibustion strips using mulberry bark paper; (B5) After rolling, use glutinous rice glue to glue the mulberry bark paper joints together, and seal both ends of the moxibustion strip with mulberry bark paper. After the glued area dries, it is made into a mulberry branch and mulberry yellow moxibustion strip.

8. The application according to claim 7, characterized in that, In step (B3), the sieving is performed using a sieve with a sieve aperture of 4 mm.

9. The application according to claim 7, characterized in that, In step (B3), the drying temperature is 40°C.

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